Contributions of White and Brown Adipose Tissues to the Circadian Regulation of Energy Metabolism

✅ 全文

白色和棕色脂肪组织对能量代谢昼夜节律调控的贡献

作者 Isabel Heyde; Kimberly Begemann; Henrik Oster 期刊 Endocrinology 发表日期 2021 ISSN 0013-7227 DOI 10.1210/endocr/bqab009 类型 原创研究 (Original Research)

📄 英文摘要 English Abstract

EN

The term energy metabolism comprises the entirety of chemical processes associated with uptake, conversion, storage, and breakdown of nutrients. All these must be tightly regulated in time and space to ensure metabolic homeostasis in an environment characterized by cycles such as the succession of day and night. Most organisms evolved endogenous circadian clocks to achieve this goal. In mammals, a ubiquitous network of cellular clocks is coordinated by a pacemaker residing in the hypothalamic suprachiasmatic nucleus. Adipocytes harbor their own circadian clocks, and large aspects of adipose physiology are regulated in a circadian manner through transcriptional regulation of clock-controlled genes. White adipose tissue (WAT) stores energy in the form of triglycerides at times of high energy levels that then serve as fuel in times of need. It also functions as an endocrine organ, releasing factors in a circadian manner to regulate food intake and energy turnover in other tissues. Brown adipose tissue (BAT) produces heat through nonshivering thermogenesis, a process also controlled by the circadian clock. We here review how WAT and BAT contribute to the circadian regulation of energy metabolism. We describe how adipose rhythms are regulated by the interplay of systemic signals and local clocks and summarize how adipose-originating circadian factors feed-back on metabolic homeostasis. The role of adipose tissue in the circadian control of metabolism becomes increasingly clear as circadian disruption leads to alterations in adipose tissue regulation, promoting obesity and its sequelae. Stabilizing adipose tissue rhythms, in turn, may help to combat disrupted energy homeostasis and obesity.

📄 中文摘要 Chinese Abstract

中文
能量代谢涵盖营养物质的摄取、转化、储存与分解的化学过程,这些过程必须在时间和空间上受到严格调控,以维持昼夜节律等周期性环境中的代谢稳态。大多数生物已进化出内源性昼夜节律钟来实现这一目标。在哺乳动物中,一个遍布全身的细胞时钟网络由下丘脑视交叉上核(SCN)中的起搏器协调。脂肪细胞拥有自己的昼夜节律钟,脂肪生理学的诸多方面通过时钟控制基因的转录调控以昼夜节律方式受到调节。白色脂肪组织(WAT)以甘油三酯形式储存能量,并作为内分泌器官释放因子以调节食物摄入和能量周转。棕色脂肪组织(BAT)通过非颤抖性产热产生热量,同样受昼夜节律钟控制。分子钟由*Bmal1*、*Clock*、*Cry1/2*和*Per1-3*等时钟基因在转录-翻译反馈环路(TTFL)中相互作用构成。SCN中的主起搏器通过光输入使内部节律与外界光暗周期同步,而外周组织则通过行为、神经和体液信号实现同步。

📋 英文结构化总结 English Structured Summary

全文整理

EN

Header:

Background Energy metabolism encompasses the chemical processes of nutrient uptake, conversion, storage, and breakdown, which must be tightly regulated in time and space to maintain metabolic homeostasis in an environment with cycles such as day and night. Most organisms have evolved endogenous circadian clocks to achieve this. In mammals, a ubiquitous network of cellular clocks is coordinated by a pacemaker in the hypothalamic suprachiasmatic nucleus (SCN). Adipocytes harbor their own circadian clocks, and large aspects of adipose physiology are regulated in a circadian manner through transcriptional regulation of clock-controlled genes. White adipose tissue (WAT) stores energy as triglycerides and functions as an endocrine organ, releasing factors to regulate food intake and energy turnover. Brown adipose tissue (BAT) produces heat via nonshivering thermogenesis, also controlled by the circadian clock. The molecular clock consists of clock genes such as *Bmal1*, *Clock*, *Cry1/2*, and *Per1-3* interacting in a transcriptional-translational feedback loop (TTFL). A master pacemaker in the SCN aligns internal rhythms with the external light-dark cycle via light input, while peripheral tissues are synchronized by behavioral, neuronal, and humoral signals.

Header:

Methods N/A – Review article

Header:

Results The provided excerpt does not contain detailed experimental results; it reviews the contributions of WAT and BAT to circadian regulation of energy metabolism. Key findings described include: WAT stores energy and releases adipokines in a circadian manner; BAT produces heat via nonshivering thermogenesis under clock control; circadian disruption leads to alterations in adipose tissue regulation, promoting obesity and its sequelae.

Header:

Data Summary No quantitative data or statistics are provided in the excerpt. The text describes the molecular organization of the circadian clock (TTFL with *Bmal1*, *Clock*, *Per*, *Cry*, *ROR*, *REV-ERB*, and D-box loops) and the hierarchical network with the SCN as master pacemaker, but no numerical results are presented.

Header:

Conclusions Stabilizing adipose tissue rhythms may help to combat disrupted energy homeostasis and obesity. The role of adipose tissue in the circadian control of metabolism is becoming increasingly clear, as circadian disruption alters adipose tissue regulation and promotes obesity.

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Practical Significance Stabilizing adipose tissue rhythms may help to combat disrupted energy homeostasis and obesity, highlighting the potential for therapies targeting circadian regulation of adipose tissue to mitigate metabolic disorders.

📋 中文结构化总结 Chinese Structured Summary

中文

背景:

能量代谢涵盖营养物质的摄取、转化、储存与分解的化学过程,这些过程必须在时间和空间上受到严格调控,以维持昼夜节律等周期性环境中的代谢稳态。大多数生物已进化出内源性昼夜节律钟来实现这一目标。在哺乳动物中,一个遍布全身的细胞时钟网络由下丘脑视交叉上核(SCN)中的起搏器协调。脂肪细胞拥有自己的昼夜节律钟,脂肪生理学的诸多方面通过时钟控制基因的转录调控以昼夜节律方式受到调节。白色脂肪组织(WAT)以甘油三酯形式储存能量,并作为内分泌器官释放因子以调节食物摄入和能量周转。棕色脂肪组织(BAT)通过非颤抖性产热产生热量,同样受昼夜节律钟控制。分子钟由*Bmal1*、*Clock*、*Cry1/2*和*Per1-3*等时钟基因在转录-翻译反馈环路(TTFL)中相互作用构成。SCN中的主起搏器通过光输入使内部节律与外界光暗周期同步,而外周组织则通过行为、神经和体液信号实现同步。

方法:

不适用——综述文章

结果:

所提供的摘录不包含详细的实验结果;其综述了白色脂肪组织(WAT)和棕色脂肪组织(BAT)对能量代谢昼夜节律调控的贡献。所述关键发现包括:WAT以昼夜节律方式储存能量并释放脂肪因子;BAT在节律钟控制下通过非颤抖性产热产生热量;昼夜节律紊乱导致脂肪组织调控的改变,促进肥胖及其后续病变。

数据摘要:

摘录中未提供定量数据或统计数据。文本描述了昼夜节律钟的分子组织结构(包含*Bmal1*、*Clock*、*Per*、*Cry*、*ROR*、*REV-ERB*及D-box环路的TTFL)以及以SCN为主起搏器的层级网络,但未呈现数值结果。

结论:

稳定脂肪组织节律可能有助于对抗能量稳态紊乱和肥胖。脂肪组织在代谢昼夜节律控制中的作用日益明确,因为昼夜节律紊乱会改变脂肪组织的调控并促进肥胖。

实际意义:

稳定脂肪组织节律可能有助于对抗能量稳态紊乱和肥胖,这凸显了针对脂肪组织昼夜节律调控的疗法在缓解代谢紊乱方面的潜力。

📖 英文全文 English Full Text

EN

https://academic.oup.com/endo      1 Endocrinology, 2021, Vol. 162, No. 3, 1–14 doi:10.1210/endocr/bqab009

Mini-Review ISSN Online 1945-7170 This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial- NoDerivs licence (http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial reproduction and distribution of the work, in any medium, provided the original work is not altered or transformed in any way, and that the work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

© The Author(s) 2021. Published by Oxford University Press on behalf of the Endocrine Society.

Mini-Review Contributions of White and Brown Adipose

Tissues to the Circadian Regulation of Energy Metabolism

Isabel Heyde, Kimberly Begemann, and Henrik Oster Institute of Neurobiology, University of Lübeck, Lübeck 23562, Germany

ORCiD number: 0000-0002-1414-7068 (Henrik Oster).

Abbreviations: AgRP, agouti-related peptide; AMPK, adenosine monophosphate–activated protein kinase C; Atgl, adipose triglyceride lipase; Bmal1, brain and muscle aryl hydrocarbon receptor nuclear translocator-like protein 1; BAT, brown adipose tissue; CD36, cluster of differentiation 36; Clock, circadian locomotor output cycles kaput; Cry1/2, cryptochrome 1 and 2; DIT, diet-induced thermogenesis; FAs, fatty acids; FGF21, fibroblast growth factor  21; GCs, glucocorticoids;

GLUT4, glucose transporter 4; IL, interleukin; LPL, lipoprotein lipase; mRNA, messenger RNA; Nampt, nicotinamide phosphoriboysltransferase; Npas2, neuronal PAS domain protein 2; Per1-3, period 1-3; PGC-1, PPAR gamma coactivator 1;

POMC, proopiomelanocortin; PUFAs, polyunsaturated fatty acids; REV-ERBα/β, reverse erythroblastoma; RORα-γ, retinoic acid-related orphan receptors; ROREs, retinoic acid-related orphan receptor response elements; SCN, suprachiasmatic nucleus; SNS, sympathetic nervous system; SREBP-1c, stimulatory factor-1/2/regulatory element-binding protein-1c; TGs, triglycerides; TNF-α, tumor necrosis factor α; TTFL, transcriptional-translational feedback loop; UCP1, uncoupling protein

1; WAT, white adipose tissue 20211623114 Received: 23 November 2020; Editorial Decision: 12 January 2021; First Published Online: 16 January 2021; Corrected and

Typeset: 5 February 2021. 2021 Abstract The term energy metabolism comprises the entirety of chemical processes associated with uptake, conversion, storage, and breakdown of nutrients. All these must be tightly regulated in time and space to ensure metabolic homeostasis in an environment char- acterized by cycles such as the succession of day and night. Most organisms evolved endogenous circadian clocks to achieve this goal. In mammals, a ubiquitous network of cellular clocks is coordinated by a pacemaker residing in the hypothalamic supra- chiasmatic nucleus. Adipocytes harbor their own circadian clocks, and large aspects of adipose physiology are regulated in a circadian manner through transcriptional regu- lation of clock-controlled genes. White adipose tissue (WAT) stores energy in the form of triglycerides at times of high energy levels that then serve as fuel in times of need. It also functions as an endocrine organ, releasing factors in a circadian manner to regulate food intake and energy turnover in other tissues. Brown adipose tissue (BAT) produces heat through nonshivering thermogenesis, a process also controlled by the circadian clock. We here review how WAT and BAT contribute to the circadian regulation of energy metabolism. We describe how adipose rhythms are regulated by the interplay of sys- temic signals and local clocks and summarize how adipose-originating circadian factors feed-back on metabolic homeostasis. The role of adipose tissue in the circadian control

2  Endocrinology, 2021, Vol. 162, No. 3 of metabolism becomes increasingly clear as circadian disruption leads to alterations in adipose tissue regulation, promoting obesity and its sequelae. Stabilizing adipose tissue rhythms, in turn, may help to combat disrupted energy homeostasis and obesity.

Key Words: circadian clocks, energy metabolism, adipose tissue, BAT, WAT, circadian rhythm, hormones, adipokines, cytokines, thermogenesis, obesity

Molecular and Cellular Circadian Networks Many aspects of the environment show regular rhythms.

For many organisms the most prominent of these rhythms is the 24-hour solar cycle resulting in changes in, for ex- ample, illumination, temperature, or food availability. Species throughout all phyla have adapted to these predictable vari- ations by evolving internal timekeepers, so-called circadian clocks (from the Latin “circa diem,” meaning “around a day”).

At the molecular level, the circadian clock of mam- mals consists of clock genes such as brain and muscle aryl hydrocarbon receptor nuclear translocator-like protein

1 (Bmal1, aka Arntl), circadian locomotor output cycles kaput (Clock) and its analogue, neuronal PAS domain pro- tein 2 (Npas2), cryptochrome 1 and 2 (Cry1/2), and period

1-3 (Per1-3) (Fig. 1A) (1). They interact in a transcriptional- translational feedback loop (TTFL), which comprises the heart of the clock machinery (2). Briefly, in the morning

BMAL1 and CLOCK heterodimerize and bind to E-box enhancers in the promotors of Per and Cry genes (3). Over the course of the day PER and CRY proteins translocate into the nucleus, where they inhibit CLOCK:BMAL1–me- diated transcription, including their own (4). Gradual deg- radation of PER and CRY during the night is controlled by the casein kinases and F-box ubiquitin transferases, thus resulting in disinhibition of CLOCK:BMAL1 toward the next morning (1). This core loop is stabilized by a second

TTFL in which retinoic acid-related orphan receptors (RORα-γ) and reverse erythroblastoma (REV-ERBα/β, aka

NR1D1/2) proteins compete for binding to retinoic acid- related orphan receptor response elements (ROREs) in the promotor of Bmal1 (5). Thereby, RORs activate whereas

REV-ERBs inhibit Bmal1 gene expression. A  third feed- back loop consists of D-site albumin promotor binding protein and nuclear factor interleukin 3 regulated (NFIL3, aka E4BP4), which modulate gene expression by binding to D-boxes in the promotors of several clock genes, for ex- ample, Per1-3, Rev-Erbα/β, Rorα/β, and Cry1 (5-10). The role of D-boxes as additional feedback loops is still a cur- rent research topic. So far it is assumed that D-box regu- lation is involved in circadian signaling. However, mice deficient for E4bp4 do not show differences in circadian oscillation (6). These interlocked TTFLs drive circadian gene expression of thousands of tissue-specific clock- controlled genes throughout the day (11, 12).

Molecular clocks are found in all cells and tissues.

This circadian clock network is organized in a hierarch- ical manner with a master pacemaker located in the supra- chiasmatic nucleus (SCN) of the hypothalamus (13). The most prominent, the zeitgeber—an external time signal entraining the endogenous circadian clock—is light. Light reaches melanopsin (OPN4) expressing intrinsically photo- sensitive retinal ganglion cells that directly project to the

SCN through the retinohypothalamic tract (14, 15). In this way, the SCN aligns its internal rhythm with the external light-dark cycle. In the absence of regular light input, the endogenous circadian rhythm is maintained, showing a

“free-running” species-specific period close to 24 hours (2).

From the SCN, behavioral, neuronal, and humoral signals transmit internal time to peripheral tissues (Fig. 1B) (16).

Apart from light, other zeitgebers can influence the circa- dian clock network. The timing of food intake, for example, is a potent zeitgeber of peripheral tissue clocks while only marginally affecting the master pacemaker (17, 18).

Adipose Functions in Metabolism Most adipose tissue depots in mammals are classified as

“white” (19) with spherical cells of variable size (25-200 µm) and a single large lipid droplet. Brown adipocytes, on the other hand, are smaller (15-60 µm) and contain multiple small lipid droplets as well as much more mitochondria (20). One main function of white adipose tissue (WAT) is the storage of energy.

Stimulated by insulin, glucose and lipoprotein-derived fatty acids (FAs) are taken up by the adipocyte where they are con- verted into triglycerides (TGs) (21, 22). If energy levels drop, stored TGs are broken down during lipolysis and released as glycerol and FAs that can then both be used as energy sources by other organs (23, 24). WAT is also an important endocrine organ. White adipocytes release adipocytokines such as leptin and adiponectin to regulate metabolic functions in other per- ipheral tissues and the brain (24).

Brown adipose tissue (BAT) differs morphologically and functionally from WAT. Brown adipocytes convert en- ergy into heat by nonshivering thermogenesis (20). Cold exposure as well as noradrenergic stimulation lead to β-adrenergic excitation, which then induces lipolysis in brown adipocytes (22, 25). The resulting FAs activate un- coupling protein 1 (UCP1), which forms a proton leak in

Endocrinology, 2021, Vol. 162, No. 3 3 the inner mitochondrial membrane to produce heat during oxidative phosphorylation (25, 26).

Oscillating Signals Regulate White Adipose Tissue Metabolism

Metabolic homeostasis is regulated by the interaction of different organs, including liver, pancreas, adrenal, and adipose tissue, in a circadian manner. Involved tissues re- lease factors or stimulate the autonomic nervous system to mediate the metabolic state to other participating or- gans. WAT is a target tissue that receives and integrates numerous signals. WAT metabolism is strongly controlled by the interplay of 2 pancreatic hormones, β-cell–derived insulin and α-cell–derived glucagon, both of which are regulated by energy intake while also showing underlying circadian rhythms in plasma levels controlled by the SCN (27). Besides regulation by the SCN, cell-autonomous clocks in α and β cells control the circadian release of lo- cally produced hormones (28-31). α- and β-Cell clocks show distinct phases in vivo and in vitro, with the α-cell clock being phase delayed by 1 to 2 hours compared to β cells. Cell type–specific clocks regulate the transcription of key genes involved in glucose sensing and hormone re- lease (28). Together, the findings show that insulin—but also glucagon—release is regulated by the intrinsic clock in the distinct cell types and is affected by metabolic state.

Food intake increases circulating FAs, amino acids, and glucose, which triggers the release of insulin from pan- creatic β cells, resulting in a peak of blood insulin levels in the middle of the active phase (27, 28). In adipocytes, insulin inhibits lipolysis and activates lipogenesis (Fig. 2) (32-34). Insulin-mediated repression of lipolysis is regu- lated via the mammalian target of rapamycin complex 1 (mTORC1)-early growth response element 1 pathway, which inhibits adipose triglyceride lipase (Atgl) promotor activity, encoding a key enzyme of lipolysis (32), and other key proteins of the lipolytic pathway including hormone- sensitive lipase (HSL) and perilipin (35-39). On the other hand, insulin increases glucose and FA uptake as well as FA synthesis and TG storage by glucose transporter 4 (GLUT4, aka SLC2A4) translocation and activating upstream stimu- latory factor-1/2/regulatory element-binding protein-1c (SREBP-1c)- and carbohydrate-response element binding protein-α/β (ChREBP-α/β)-mediated target gene expres- sion, respectively (40). Insulin sensitivity is gated by the clock (41-43). In vitro studies suggest that insulin action, that is, activation of the protein kinase B–pathway, depends on adipose clock function at least in subcutaneous adipose tissue (41). Glucagon, released from pancreatic α cells as a signal of low energy, is a strong counterregulatory signal of insulin and displays high levels in the transition of rest to activity phase. Although it is still not clear whether adipo- cytes actually express the glucagon receptor, the hormone induces adipose lipolysis. Glucagon’s effects may be me- diated by stimulation of the sympathetic nervous system (SNS), which induces lipolysis via β-adrenergic pathways (44-47).

Fibroblast growth factor 21 (FGF21) is regulated by

NFIL3 (aka E4BP4) and peaks during the fasting period (48). It is predominantly synthesized in the liver and regu- lates carbohydrate and lipid metabolism (48-50). In lean

Figure 1.  A, The core transcriptional-translational feedback loop (TTFL) comprises the transcription factors brain and muscle aryl hydrocarbon re- ceptor nuclear translocator-like protein 1 (BMAL1) and circadian locomotor output cycles kaput (CLOCK), inducing expression of period 1-3 (PERs) and cryptochrome 1 and 2 (CRYs), which in turn repress BMAL1:CLOCK activity. The circadian clock is modulated and stabilized by auxiliary loops.

Nuclear factor interleukin 3 regulated (NFIL3) and D-site albumin promotor binding protein (DBP) inhibit and activate expression of several clock genes, respectively. Retinoic acid-related orphan receptors (RORs) and reverse erythroblastoma (REV-ERBs) are controlled by BMAL1:CLOCK and function as activators or inhibitors of Bmal1 expression, respectively. B, The zeitgeber light aligns the suprachiasmatic nucleus (SCN) with the ex- ternal light-dark cycle. Peripheral clocks, for example, white adipose tissue (WAT), gut, liver, pancreas, and brown adipose tissue (BAT), are synchron- ized by the SCN via the autonomic nervous system (ANS), hormones, and behavior. The circadian clock network drives rhythms in lipogenesis and lipolysis as well rhythmic release of adipokines from WAT. BAT produces heat via nonshivering thermogenesis.

4  Endocrinology, 2021, Vol. 162, No. 3 mice, FGF21 increases adipose lipid uptake through cluster of differentiation 36 (CD36) and lipoprotein lipase (LPL).

Conversely, in insulin-resistant obese mice, FGF21 in- creases catabolism of TGs in BAT while inducing WAT browning by an increase in PGC-1α protein levels (51-53).

FGF21 potently lowers blood glucose levels by increasing glucose uptake into the liver, WAT, and BAT (54, 55) via adiponectin (56, 57). Moreover, adiponectin seems to me- diate FGF21-induced energy expenditure (57). Ghrelin, a stomach-derived hormone, shows a diurnal oscillation with increased levels during fasting and low levels during feeding phases (58-60), and its messenger RNA (mRNA) expression and release are disrupted in Bmal1 knockouts (60). Ghrelin promotes lipogenic gene expression in WAT via autonomic stimulation (61, 62) or directly through ac- tivation of its receptor, growth hormone secretagogue re- ceptor, which is expressed in adipocytes of old, but hardly detectable in young, mice (63).

Glucocorticoids (GCs) are secreted from the adrenal in a circadian manner and in response to stress. The adrenal clock gates the sensitivity to incoming signals and regu- lates GC rhythms (64). The local clock is important for rhythmic production of steroidogenic genes but seems to be dispensable for rhythmic GC output (65, 66). Global clock disruption deletes rhythmic GC output (64, 67). GCs promote adipocyte differentiation (68-70). Dampened

GC rhythms result in increased lipid accumulation and body weight gain due to upregulation of the FA trans- porter Cd36 (71). Hypercortisolemia increases lipolysis while at the same time promoting visceral adiposity, prob- ably by enhanced preadipocyte differentiation (69, 72).

GCs stimulate the expression of Hsl, Atgl, and Lpl, key enzymes of the lipolytic pathway, in a dose-dependent manner (73-76). At the same time, GCs promote adi- pose insulin resistance by downregulating the expression of insulin receptor substrate-1 and inhibiting GLUT4 plasma membrane translocation (77-79). GC effects are dependent on 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1), which might be a promising target to coun- teract GC-mediated adiposity and insulin resistance (74).

In subcutaneous, but not in visceral adipose tissue, GC decreases lipolysis by reducing the expression of Hsl (80).

This strongly suggests that the effects of GCs on adipose tissue metabolism are depot dependent.

Together, WAT physiology is regulated by numerous in- coming oscillating signals in a circadian manner. Insulin and ghrelin, signaling the feeding:fasting state, activate lipogenic gene expression. Insulin and FGF21 increase lipid and glucose uptake at different times of day to maintain en- ergy homeostasis. Glucagon and GCs, both peaking around activity onset, stimulate lipolysis to fuel the body in the transition from rest to the active state.

White Adipose Tissue Clocks Drive Rhythmic Gene Expression

Adipose physiology is regulated by adipocyte clocks.

Together with the circadian-regulated incoming signals,

Figure 2.  White adipose tissue (WAT) function is regulated by numerous rhythmic signals originating from other peripheral or central tissues. The circadian clocks in white adipocytes are essential for proper WAT function. WAT releases adipokines and cytokines in a circadian manner, regulating food intake, insulin sensitivity, and inflammation. ANS indicates autonomic nervous system; IL, interleukin; TNF, tumor necrosis factor.

Endocrinology, 2021, Vol. 162, No. 3 5 adipose tissue adapts to daily variations of energy avail- ability and needs to maintain energy homeostasis.

Adipocyte clocks regulate the local transcriptome in a cir- cadian manner including key enzymes of lipogenesis and lipolysis, for example, Atgl, Hsl, caveolin 2, acyl-CoA synthetase (Acsl1), phosphatidate phosphatase (Lpin1),

Lpl, peroxisome proliferator-activated receptor α (Pparα),

PPAR gamma coactivator 1-β (Pgc1β), Srepb1-α, and Pparγ2 (81-86). BMAL1:CLOCK activate gene expression of Atgl and Hsl via binding to E-box promotor elements (81). Most of those diurnal rhythms are lost in obese in- dividuals with type 2 diabetes (83, 87). The physiological significance of adipose tissue clocks has been demonstrated in numerous studies. Bmal1-deficient embryonic fibro- blasts show impaired differentiation into adipocytes (82).

BMAL1:CLOCK dimers regulate adipogenesis via the Wnt signaling pathway (88, 89). Genetic disruption of the cir- cadian clock by mutations in Clock leads to increased adi- posity on regular chow in a genetic background–dependent manner. In both strains, Clock mutants show blunted or lost rhythms in serum TGs, FAs, and glycerol, indicating impairments in fat absorption and lipolysis (81, 90). The latter was attributed to decreased transcription of Atgl and Hsl (81). Adipocyte-specific knockout of Bmal1 leads to obesity under high-fat diet conditions, probably due to changes in circulating polyunsaturated FAs that centrally affect food intake rhythms. This idea is supported by the aberrant expression of hypothalamic neuropeptides in- volved in appetite regulation in those mice (91). Together, the existence of a circadian adipocyte-hypothalamus axis emphasizes the importance of a functional adipose tissue clock for the circadian regulation of energy homeostasis.

Overexpression of Bmal1 increases the expression of lipogenesis-related genes in WAT (82). BMAL1:CLOCK in- duce Ppar expression via E-box binding (92, 93). PPARγ in particular is crucial for adipocyte differentiation and adipogenesis (94-96). PPARs also feed-back on Clock gene expression. PPARγ induces Rev-Erbα expression in adi- pose tissue whereas PPARα activates Bmal1 transcription in the liver (97, 98). PER2, being part of the negative limb of the core TTFL, directly inhibits expression of PPARγ target gene expression by suppressing PPARγ binding to

PPAR response elements (99). The clock modulator–dif- ferentiated embryo chronodrocyte protein 1 (DEC1) pre- vents PPARγ-mediated target gene expression, thereby promoting circadian oscillations in these genes. In line with this, Dec1 deficiency leads to a pronounced increase in gene expression related to FA biosynthesis, lipid storage, and lipolysis in the dark phase as well as a loss of circadian variation in serum FAs (100). Expression of the nuclear re- ceptors Rev-Erbα/β and RORα/β peaks at the end of the light phase in mice. Srebp-1c, Pparγ, as well as adiponectin and leptin also exhibit diurnal mRNA rhythms peaking at night (101). Genetic disruption of Rev-Erbα is associ- ated with decreased SREBP1 and SREBP2 activity (102).

REV-ERBα regulates SREBPs activity and bile acid metab- olism in the liver (102). In muscle, REV-ERBβ is recruited to the Srebp-1c promotor, inducing gene expression (103).

Thus, REV-ERB agonists may be promising candidates to treat obesity by increasing energy expenditure and, thereby, improving dyslipidemia and hyperglycemia through al- terations in circadian gene expression of metabolic genes (104).

In summary, the circadian clock is important for adipo- cyte differentiation and the rhythmic expression of lipogenic and lipolytic genes. Clock proteins regulate the activity of proteins involved in WAT metabolism that, in turn, control rhythmic FA release. FAs signal the metabolic state to the brain, adjusting food intake. Circadian disruptions in WAT metabolism lead to alterations in adipogenesis, lipid mobil- ization, and food intake, promoting obesity.

Rhythmic Output from White Adipose Tissue Regulates Metabolism

Metabolic homeostasis is regulated by factors released by, inter alia, WAT in a circadian manner. Such factors modu- late the physiology of other tissues and are integrated at a central level to control food intake. One of the main func- tions of WAT is the storage of lipids and release of lipo- lytic products. The latter—as FAs and glycerol—exhibits a prominent circadian rhythm in mice and humans that is only partly driven by food intake (81, 90, 100, 105).

A loss of polyunsaturated FA (PUFA) rhythms induces al- terations in the expression of appetite-regulating neuro- peptides and increases food intake. Decreased PUFA levels are accompanied by reduced expression of long-chain fatty acid elongase 5/6 (Elovl5/6) and stearoyl-CoA desaturase

1 (Scd1), key enzymes in PUFA biosynthesis (91, 106).

Restoration of PUFA content in the hypothalamus rescues food intake rhythms, body weight development, appetite- regulated neuropeptide expression, and energy homeostasis (91). This clearly shows the crucial role of oscillating lipo- lytic output for whole-body metabolic homeostasis.

WAT is also an endocrine organ releasing adipokines in a circadian manner that contribute to the regulation of me- tabolism throughout the body (Fig. 2). Secretion of leptin, one of the best studied adipokines, is stimulated by insulin but is also regulated by adipose tissue clocks (107, 108). The diurnal leptin pattern is maintained under regular feeding of

6 meals a day but is abolished by lesioning the SCN, which suggests that leptin secretion is controlled by the circadian clock (109). In vitro studies reveal that the adipocyte clock regulates leptin secretion. Although mRNA expression of

6  Endocrinology, 2021, Vol. 162, No. 3 leptin is not rhythmic in adipocytes, leptin release changes throughout the day (107). Leptin levels correlate with body fat content and decrease during fasting (110-112). It crosses the blood-brain barrier and acts as a satiety hormone, regulating energy expenditure and food intake (113-116).

Leptin binds to leptin receptors throughout the central nervous system, for example, in the arcuate nucleus. The arcuate nucleus contains neuropeptide Y (NPY)-/agouti- related peptide (AgRP)-positive and proopiomelanocortin (POMC)-/ cocaine- and amphetamine-related transcript (CART)-positive neurons, which regulate food intake and energy expenditure (117). Leptin suppresses food intake by inhibiting the expression of orexigenic neuropeptides Agrp and Npy and stimulating the expression of anorexigenic

Pomc (118-121). Chronic jet lag promotes obesity probably by central leptin resistance and downregulation of leptin transcription (122). Leptin’s effects on energy expenditure may in part be mediated by BAT activation. A recent dis- covery suggests that the thermogenic effect of leptin may be regulated by PGC-1β expressing POMC-neurons (123).

Furthermore, leptin stimulates β-adrenergic receptors and, thereby, increases expression of Ucp1 in BAT (124).

Despite the effects on BAT, leptin activates β-oxidation in peripheral tissues, for example, muscle and liver, via adeno- sine monophosphate–activated protein kinase C (AMPK) signaling (125). Thereby, leptin prevents lipid accumulation in such tissues. Furthermore, leptin directly suppresses the release of GCs, which play an important role in glucose and lipid homeostasis (70, 126, 127). Adiponectin is expressed in a circadian manner in adipose tissue (128) as a target gene of PPARγ and PGC1β, which are regulated by the cir- cadian clock (129). As mentioned earlier, it plays a crucial role in the regulation of energy expenditure and insulin sensitivity (56, 57). Adiponectin increases β-oxidation and glucose uptake and, thus, decreases body weight (130-132).

It also induces browning of WAT via a sirtuin 1 (SIRT1)- AMPK–mediated upregulation of Ucp1 expression to af- fect energy expenditure (133, 134). However, in BAT itself, adiponectin seems to inhibit Ucp1 expression via inhibition of β-adrenergic receptor expression (135). Expression of adiponectin receptors (Adipor1 and 2) exhibits a circadian oscillation in adipose tissue and in the mediobasal hypo- thalamus (128, 136). The adipokine signals the peripheral metabolic state to the brain, which in combination with blood glucose levels results in adjustment of food intake (137, 138). Its action is mediated by AMPK signaling (139). Adiponectin also induces Bmal1 expression in the mediobasal hypothalamus that then locally regulates the expression of orexigenic neuropeptides (136). These find- ings suggest a mechanism by which peripheral circadian clock disruption my alter food intake rhythms, promoting the development of metabolic disorders.

Serum visfatin levels, encoded by nicotinamide phosphoriboysltransferase (Nampt), exhibit a diurnal rhythm that is inversely related to leptin (140-143). Nampt expression rhythms are shifted by sleep deprivation, negatively affecting glucose homeostasis (142). Visfatin/

NAMPT also catalyzes the rate-limiting step in the nico- tinamide adenine dinucleotide (NAD+) salvage pathway.

Nampt expression is regulated by the circadian clock and modulates the core TTFL by regulating the activity of the histone deacetylase SIRT1 (143). Its insulin-mimetic func- tion is controversial and still under investigation (144- 146). However, it has become more evident that visfatin has a proinflammatory function and most studies agree on a positive correlation between fat mass and visfatin levels (147-150). Infiltrated immune cells might be a major source of visfatin expression (149, 151). Circulating visfatin levels are positively correlated with proinflammatory cytokines such as interleukin-6 (IL-6) and C-reactive protein, and visfatin expression is strongly correlated with expression of tumor necrosis factor α (Tnf-α) and Il-6 (148, 150,

152). Thus, increased visfatin may have deleterious effects on energy homeostasis. In fact, visfatin induces insulin re- sistance in the liver partly via induction of inflammatory pathways and induces FA-mediated neuroinflammation (152, 153).

Resistin reduces insulin sensitivity and shows a circa- dian rhythm trailing that of insulin and suggesting a nega- tive feedback on insulin action (154). Circadian rhythms in resistin expression are stimulated by rhythmic input of insulin. In obesity, concomitant with insulin resistance, rhythmic resistin expression is blunted or abolished (155,

156). In humans, resistin is mainly expressed by macro- phages whereas its main source in rodents are adipocytes.

Gene and protein structures differ between humans and rodents, accounting for their different functional roles (157). Human resistin activates circadian expression of proinflammatory cytokines, such as TNFα, IL-6, and IL-12, which contribute to development of insulin resistance and inflammation (158, 159). In turn, such cytokines enhance resistin expression (160). Neutrophils are the first immune cells to infiltrate adipose tissues after a dietary challenge (161). Neutrophils, in turn, attract further immune cells including macrophages, which then reduce insulin sensi- tivity and induce chronic inflammation (162, 163). TNFα, predominantly released by macrophages, promotes lip- olysis (164) and inhibits the expression of perilipin, a pro- tein associated with fat storage (164, 165). Furthermore, it decreases GLUT4 and LPL expression (166). As such, high

TNFα levels inhibit insulin-mediated glucose uptake (167,

168) and promote the development of insulin resistance, for example, in obese individuals (167, 169). GC treat- ment inhibits TNFα-mediated insulin resistance but also

Endocrinology, 2021, Vol. 162, No. 3 7 decreases its lipolytic effects, which contribute to fat accu- mulation (170).

Taken together, through rhythmic release of FAs and adipokine hormones, WAT plays a pivotal role in the cir- cadian modulation of energy homeostasis. It regulates food intake rhythms, energy expenditure, insulin sensitivity, and metabolic inflammation. In laboratory rodents, WAT rhythm disruption promotes overeating and obesity.

Circadian Aspects of Brown Adipose Tissue Metabolism Crosstalk

The main function of BAT is the conversion of energy into heat by nonshivering thermogenesis (20). Heat production in BAT is achieved by β-oxidation or through uncoupling of mitochondrial proton transport from energy production by UCP1 (171). BAT heat generation allows mammals to keep their body temperature more constant and cope with cold temperature environments. On a cold stimulus, auto- nomic activation leads to a release of noradrenaline at BAT terminals. Activation of β 3-adrenergic receptors in brown adipocytes results in G protein–controlled activation of the protein kinase A pathway and further gene expression of metabolic genes, including Ucp1 (171). Additionally, this pathway activates ATGL to yield FAs that then further stimulate UCP1 (26, 171).

Heat production in BAT is also induced by a carbohydrate-rich meal (diet-induced thermogenesis or

DIT; Fig. 3) through autonomic adrenergic activation (172,

173). Hormones originating from the gut as well as bile acids are also able to stimulate DIT (174). Gut-derived cholecystokinin (CCK) and secretin activate BAT thermo- genesis via vagal afferents and sympathetic efferents and

UCP1 activation (174). Ghrelin rhythms affect Ucp1 ex- pression, and its secretion is reduced after food intake (175,

176). By using DIT, our body is able to partly reduce ex- cessive energy uptake from food and thereby avoid energy storage in the form of fat (171). The induction of BAT post- prandial thermogenesis leading to glucose and FA uptake might be stimulated by insulin (173). Interestingly, UCP1 seems to be essential because mice with ablated UCP1 no longer show diet-induced thermogenesis but gain weight in- stead (177). Endocrine circadian factors such as the pineal hormone melatonin (156, 157) modulate the capacity of

BAT for nonshivering thermogenesis (171). In rodents, GCs downregulate UCP1 and, thereby, BAT thermogenesis. In humans, they have the opposite effect (178).

Like WAT, BAT is also involved in the circadian regula- tion of energy metabolism. However, because BAT function is not primarily endocrine, the focus of metabolic regu- lation is connected to the clearance of metabolic factors from the bloodstream as well as affecting the capacity of thermogenesis in brown adipocytes. The circadian clock and metabolic regulation of BAT are tightly connected.

Chronic rhythm disruption by repeated shifting of the light-dark cycle leads not only to whitening of the BAT but also reduces UCP1 expression in rats (179).

Glucose as well as TG/FA uptake in BAT is rhythmic, with a maximum at the end of the inactive or in the begin- ning of the active phase, respectively (180-183). Enzymes involved in TG breakdown such as LPL also show their maximum activity and expression in the beginning of the active phase (180, 183). This indicates a role of BAT in circadian gating of FA, TG, as well as glucose clearance (180, 183, 184). In line with this, thermogenesis is higher in the active phase (182). Because high BAT activity in hu- mans is associated with reduced glycemia, these data sug- gest that sufficient BAT could stabilize glucose fluctuations throughout the day and thereby maintain glucose homeo- stasis (184). Interestingly, glucose uptake in human BAT is increased in the morning, correlating with high Ucp1 and

Glut4 gene expression (184, 185). FGF21 seems to be im- portant for BAT glucose clearance. It inhibits temperature decreases in BAT and ameliorates normal glucose clearance by controlling stable BAT temperature and, thereby, BAT thermogenesis (186). Interestingly, BAT thermogenic ac- tivity also increases the local release of FGF21, indicating the existence of a paracrine feedback (187).

Circadian thermogenic plasticity is controlled by REV- ERBα as shown in mice that cope better with cold tem- peratures at times of low REV-ERBα expression (189).

REV-ERBα represses UCP1. In turn, cold temperatures downregulate Rev-Erbα, leading to an induction of UCP1 to increase thermogenesis. REV-ERBα depletion leads to the complete loss of body temperature rhythms and BAT activity with overall higher body temperature (189). This emphasizes the importance of the circadian regulation of

BAT metabolism. Interestingly, the SCN itself is involved in plasma TG variations by regulating lipid uptake into

BAT through the control of REV-ERBα (182, 190). This fuels the assumption that disturbed circadian rhythms con- tribute to hyperlipidemia (190).

In summary, circadian regulation of metabolism by BAT mainly involves regulating the uptake and clearance of glu- cose and lipids from the circulation. In addition, the cap- acity of thermogenesis is regulated by the circadian clock within BAT, but also by circadian hormones such as mela- tonin and GCs (see Fig. 3).

Conclusion The role of WAT and BAT depots in the circadian regulation of metabolism becomes increasingly clear. Both fat depots harbor intrinsic clocks that regulate the tissue-specific

8  Endocrinology, 2021, Vol. 162, No. 3 transcription of many key genes involved in lipogenesis and lipolysis in WAT and thermogenesis in BAT. WAT regulates feeding behavior, carbohydrate metabolism, and energy ex- penditure by releasing adipokines and FAs in a circadian manner. In this way, metabolism is modulated as a func- tion of the peripheral energy state through central integra- tion of feedback signals to respond to the organism’s needs.

BAT, in turn, contributes to the circadian regulation of en- ergy substrates in the circulation. In recent years, research has mostly focused on characterizing the effects of (tissue) clock disruption on energy metabolism. Circadian studies have helped us understand how incoming signals and

WAT physiology are integrated at the tissue level to gen- erate coherent output that, in turn, regulates food intake and modulates the physiology of other organs involved in metabolic homeostasis. Thus, adipose tissues not only pos- sess a lipid storage and thermogenesis function, but are im- portant regulators of energy homeostasis. From a clinical point of view, it will be important to dissect the regulatory mechanisms controlling circadian rhythms in adipose tis- sues. Together, recent findings have changed our view on adipose tissue as storage tissue and emphasize its role as a possible therapeutic target. Such knowledge may help us devise adipose depot-specific chronopharmacological approaches to counteract the misbalanced energy homeo- stasis that has become so prevalent in modern societies.

Clock modulators like nobiletin, a RORα/β agonist, may be putative therapeutic agents. Nobiletin promotes adipocyte differentiation, stimulates lipolysis by induction of, for ex- ample, Pparγ expression, improves insulin resistance, and decreases adipocytokine expression (191-194). Another possible therapeutic approach may be the use of light. Very recent findings show that light increases lipolysis and mito- chondrial activity via Opsin3 signaling in adipose tissues (195, 196). Chronotargeted approaches might be useful to modulate adipose tissue activity, and future studies are needed to evaluate their beneficial effects in humans.

Acknowledgments Financial Support: This work was supported by the German

Research Foundation (DFG; grant Nos. GRK1957, OS353-7/1,

OS353-10/1, and CRC296).

Additional Information Correspondence: Henrik Oster, PhD, Institute of Neurobiology,

University of Lübeck, Ratzeburger Allee 160, 23562 Lübeck, Ger- many. Email: henrik.oster@uni-luebeck.de.

Disclosures: The authors have nothing to disclose.

Data Availability: This review manuscript does not contain any original data. Data sharing is not applicable to this article because no data sets were generated or analyzed during the present study.

Figure 3.  Brown adipose tissue (BAT) function is modulated by numerous rhythmic signals originating from other peripheral or central tissues as well as circulating metabolites such as glucose and fatty acids (FAs). The circadian clock in brown adipocytes is essential for proper BAT function.

BAT produces heat via nonshivering thermogenesis, affecting body temperature. ANS indicates autonomic nervous system; CCK, cholecystokinin;

SNS, sympathetic nervous system.

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以下是对该学术英文段落的中文翻译,严格保留专业术语的准确性:

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**白色与棕色脂肪组织对能量代谢昼夜节律调控的贡献**

伊莎贝尔·海德、金伯利·贝格曼、亨里克·奥斯特 德国吕贝克大学神经生物学研究所,吕贝克 23562 ORCiD编号:0000-0002-1414-7068(亨里克·奥斯特)

**缩写词**:AgRP,刺鼠相关肽;AMPK,腺苷一磷酸激活蛋白激酶C;Atgl,脂肪甘油三酯脂肪酶;Bmal1,脑和肌肉芳香烃受体核转位样蛋白1;BAT,棕色脂肪组织;CD36,分化簇36;Clock,昼夜节律运动输出周期蛋白;Cry1/2,隐花色素1和2;DIT,饮食诱导产热;FAs,脂肪酸;FGF21,成纤维细胞生长因子21;GCs,糖皮质激素;GLUT4,葡萄糖转运蛋白4;IL,白细胞介素;LPL,脂蛋白脂肪酶;mRNA,信使RNA;Nampt,烟酰胺磷酸核糖转移酶;Npas2,神经元PAS结构域蛋白2;Per1-3,周期蛋白1-3;PGC-1,PPARγ共激活因子1;POMC,阿黑皮素原;PUFAs,多不饱和脂肪酸;REV-ERBα/β,反向成红细胞瘤;RORα-γ,维甲酸相关孤儿受体;ROREs,维甲酸相关孤儿受体反应元件;SCN,视交叉上核;SNS,交感神经系统;SREBP-1c,刺激因子1/2/调节元件结合蛋白-1c;TGs,甘油三酯;TNF-α,肿瘤坏死因子α;TTFL,转录-翻译反馈环;UCP1,解偶联蛋白1;WAT,白色脂肪组织

**摘要** “能量代谢”一词涵盖与营养物质的摄取、转化、储存和分解相关的全部化学过程。这些过程必须在时间和空间上受到严格调控,以确保在具有昼夜节律等周期性变化的环境中维持代谢稳态。大多数生物进化出内源性昼夜节律钟(即生物钟)来实现这一目标。在哺乳动物中,一个遍布全身的细胞生物钟网络由位于下丘脑视交叉上核(SCN)的主起搏器协调。脂肪细胞自身拥有生物钟,且脂肪组织的许多生理功能通过时钟控制基因的转录调控以昼夜节律方式运行。白色脂肪组织(WAT)在能量充足时以甘油三酯形式储存能量,在需要时作为燃料供能;同时,它还作为内分泌器官,以昼夜节律方式释放因子,调控其他组织的食物摄入和能量转换。棕色脂肪组织(BAT)通过非颤抖性产热产生热量,这一过程同样受生物钟调控。本文综述了WAT和BAT如何参与能量代谢的昼夜节律调控,描述脂肪节律如何受系统信号与局部生物钟相互作用的调节,并总结源自脂肪组织的昼夜节律因子如何反馈调节代谢稳态。随着研究深入,脂肪组织在代谢昼夜调控中的角色日益清晰:昼夜节律紊乱会导致脂肪组织功能异常,促进肥胖及其并发症;而稳定脂肪组织节律则有助于对抗能量稳态失衡和肥胖。

**关键词**:生物钟;能量代谢;脂肪组织;BAT;WAT;昼夜节律;激素;脂肪因子;细胞因子;产热;肥胖

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**分子与细胞昼夜节律网络** 环境中许多方面呈现规律性节律变化,其中最显著的是由太阳周期引起的24小时光照、温度或食物可用性等变化。各门类的生物均已进化出内部计时器——即“生物钟”(源自拉丁语“circa diem”,意为“约一天”)以适应这些可预测的变化。

在分子层面,哺乳动物生物钟由以下时钟基因组成:脑和肌肉芳香烃受体核转位样蛋白1(Bmal1,又称Arntl)、昼夜节律运动输出周期蛋白(Clock)及其类似物神经元PAS结构域蛋白2(Npas2)、隐花色素1和2(Cry1/2)以及周期蛋白1-3(Per1-3)(图1A)(1)。它们通过一个称为转录-翻译反馈环(TTFL)的机制相互作用,构成生物钟的核心机制(2)。简而言之,在早晨,BMAL1与CLOCK形成异源二聚体,结合于Per和Cry基因启动子区的E-box增强子元件(3)。随着白天推进,PER和CRY蛋白转入细胞核,抑制CLOCK:BMAL1介导的转录(包括自身基因的转录)(4)。夜间PER和CRY蛋白逐渐降解,该过程由酪蛋白激素和F-box泛素转移酶调控,从而使CLOCK:BMAL1在次日早晨解除抑制(1)。这一核心环路由第二个TTFL稳定:维甲酸相关孤儿受体(RORα-γ)与反向成红细胞瘤蛋白(REV-ERBα/β,又称NR1D1/2)竞争结合Bmal1启动子区的维甲酸相关孤儿受体反应元件(ROREs)(5)。其中,RORs激活Bmal1表达,而REV-ERBs则抑制其表达。第三个反馈环包含D位点白蛋白启动子结合蛋白和核因子白细胞介素3调控因子(NFIL3,又称E4BP4),它们通过结合多个时钟基因(如Per1-3、Rev-Erbα/β、Rorα/β和Cry1)启动子中的D-box元件来调控基因表达(5–10)。目前关于D-box作为额外反馈环的作用仍是研究热点。现有证据表明D-box参与昼夜节律信号传导,但E4bp4基因敲除小鼠并未表现出昼夜振荡差异(6)。这些相互嵌套的TTFL驱动数千个组织特异性时钟控制基因在一天中呈现节律性表达(11, 12)。

分子生物钟存在于所有细胞和组织中。该昼夜节律网络呈层级结构,主起搏器位于下丘脑视交叉上核(SCN)(13)。最重要的授时因子(zeitgeber)——即同步内源性生物钟的外部时间信号——是光。光被表达黑视蛋白(OPN4)的内在光敏视网膜神经节细胞感知,这些细胞通过视网膜下丘脑束直接投射至SCN(14, 15)。由此,SCN将其内部节律与外界光暗周期同步。在缺乏规律光输入的情况下,内源性昼夜节律仍可维持,表现为接近24小时的“自由运行”物种特异性周期(2)。从SCN出发,行为、神经和体液信号将内部时间信息传递至外周组织(图1B)(16)。除光外,其他授时因子也可影响生物钟网络。例如,进食时间是外周组织生物钟的强效授时因子,但对主起搏器影响较小(17, 18)。

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**脂肪组织在代谢中的功能** 哺乳动物大多数脂肪库被归类为“白色”(19),其细胞呈球形,大小不一(25–200 µm),含单个大脂滴。棕色脂肪细胞较小(15–60 µm),含多个小脂滴及大量线粒体(20)。白色脂肪组织(WAT)的主要功能是能量储存。在胰岛素刺激下,葡萄糖和脂蛋白来源的脂肪酸(FAs)被脂肪细胞摄取并转化为甘油三酯(TGs)(21, 22)。当能量水平下降时,储存的TGs在脂解过程中分解为甘油和FAs,供其他器官利用(23, 24)。WAT也是重要的内分泌器官,白色脂肪细胞释放瘦素和脂联素等脂肪因子,调节外周组织和大脑的代谢功能(24)。

棕色脂肪组织(BAT)在形态和功能上均不同于WAT。棕色脂肪细胞通过非颤抖性产热将能量转化为热量(20)。寒冷暴露或去甲肾上腺素能刺激引发β-肾上腺素能兴奋,进而诱导棕色脂肪细胞脂解(22, 25)。产生的FAs激活解偶联蛋白1(UCP1),后者在线粒体内膜形成质子漏,在氧化磷酸化过程中产热(25, 26)。

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**振荡信号调控白色脂肪组织代谢** 代谢稳态由肝脏、胰腺、肾上腺和脂肪组织等多个器官以昼夜节律方式相互作用调控。相关组织释放因子或刺激自主神经系统,向其他参与器官传递代谢状态。WAT作为靶组织,接收并整合多种信号。WAT代谢受两种胰腺激素的强烈调控:β细胞分泌的胰岛素和α细胞分泌的胰高血糖素。两者均受能量摄入调节,同时其血浆水平也表现出由SCN控制的昼夜节律(27)。除SCN调控外,α和β细胞自身的细胞自主生物钟也控制局部激素的节律性释放(28–31)。体内和体外研究表明,α细胞生物钟较β细胞延迟1–2小时。细胞类型特异性生物钟调控葡萄糖感应和激素释放相关关键基因的转录(28)。综上,胰岛素和胰高血糖素的释放均受各自细胞内源性生物钟调控,并受代谢状态影响。

进食增加循环中的FAs、氨基酸和葡萄糖,触发胰腺β细胞释放胰岛素,使血液胰岛素水平在活动中期达到峰值(27, 28)。在脂肪细胞中,胰岛素抑制脂解并促进脂质生成(图2)(32–34)。胰岛素通过哺乳动物雷帕霉素靶蛋白复合物1(mTORC1)-早期生长反应元件1通路抑制脂肪甘油三酯脂肪酶(Atgl)启动子活性(Atgl是脂解关键酶),并抑制脂解通路中其他关键蛋白如激素敏感脂肪酶(HSL)和脂滴包被蛋白(perilipin)的表达(35–39)。另一方面,胰岛素通过促进葡萄糖转运蛋白4(GLUT4,又称SLC2A4)的膜转位,以及激活上游刺激因子1/2/调节元件结合蛋白-1c(SREBP-1c)和碳水化合物反应元件结合蛋白-α/β(ChREBP-α/β)介导的靶基因表达,分别增强葡萄糖和FAs摄取、FAs合成及TGs储存(40)。胰岛素敏感性受生物钟门控(41–43)。体外研究表明,胰岛素作用(即蛋白激酶B通路激活)至少皮下脂肪组织中依赖于脂肪生物钟功能(41)。

胰高血糖素由胰腺α细胞在低能量状态下释放,是胰岛素的强效拮抗信号,在静息向活动过渡阶段水平较高。尽管尚不清楚脂肪细胞是否表达胰高血糖素受体,但该激素可诱导脂肪脂解,其效应可能通过刺激交感神经系统(SNS)经β-肾上腺素能通路介导(44–47)。

成纤维细胞生长因子21(FGF21)受NFIL3(又称E4BP4)调控,在禁食期达峰(48)。它主要在肝脏合成,调节糖脂代谢(48–50)。在瘦小鼠中,FGF21通过分化簇36(CD36)和脂蛋白脂肪酶(LPL)增加脂肪脂质摄取。相反,在胰岛素抵抗肥胖小鼠中,FGF21促进BAT中TGs分解,并通过上调PGC-1α蛋白水平诱导WAT褐变(51–53)。FGF21通过脂联素(56, 57)显著降低血糖,增强肝脏、WAT和BAT的葡萄糖摄取(54, 55)。此外,脂联素可能介导FGF21诱导的能量消耗(57)。

胃源性激素胃饥饿素呈昼夜振荡,禁食期升高,进食期降低(58–60),其mRNA表达和释放在Bmal1敲除小鼠中被破坏(60)。胃饥饿素通过自主神经刺激(61, 62)或直接激活其受体(生长激素促分泌素受体,该受体在老年小鼠脂肪细胞中表达,但在幼年小鼠中几乎检测不到)(63)促进WAT中脂质生成基因表达。

糖皮质激素(GCs)由肾上腺以昼夜节律方式分泌,并响应应激。肾上腺生物钟门控对传入信号的敏感性并调节GC节律(64)。局部生物钟对类固醇生成基因的节律性产生重要,但对GC输出的节律性似乎非必需(65, 66)。全局生物钟破坏会消除GC输出的节律性(64, 67)。GCs促进脂肪细胞分化(68–70)。GC节律减弱会导致脂质积累增加和体重上升,原因可能是脂肪酸转运蛋白Cd36上调(71)。高皮质醇血症促进脂解,同时可能通过增强前脂肪细胞分化促进内脏肥胖(69, 72)。GCs以剂量依赖性方式刺激脂解通路关键酶Hsl、Atgl和Lpl的表达(73–76)。同时,GCs通过下调胰岛素受体底物-1表达和抑制GLUT4膜转位,促进脂肪胰岛素抵抗(77–79)。GC效应依赖于11β-羟基类固醇脱氢酶1型(11β-HSD1),该酶可能是对抗GC介导的肥胖和胰岛素抵抗的潜在靶点(74)。在皮下(而非内脏)脂肪组织中,GC通过降低Hsl表达抑制脂解(80),强烈提示GC对脂肪组织代谢的作用具有库位依赖性。

综上,WAT生理功能受多种以昼夜节律方式传入的振荡信号调控。胰岛素和胃饥饿素(信号指示进食/禁食状态)激活脂质生成基因表达;胰岛素和FGF21在不同时段增加脂质和葡萄糖摄取以维持能量稳态;胰高血糖素和GCs(均在活动起始时达峰)刺激脂解,为身体从静息向活动状态过渡提供燃料。

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**白色脂肪组织生物钟驱动节律性基因表达** 脂肪生理功能受脂肪细胞生物钟调控。结合昼夜节律性传入信号,脂肪组织适应每日能量可用性的变化,以维持能量稳态。脂肪细胞生物钟以昼夜节律方式调控局部转录组,包括脂解和脂质生成的关键酶,如Atgl、Hsl、小窝蛋白2、酰基辅酶A合成酶(Acsl1)、磷脂酸磷酸酶(Lpin1)、LPL、过氧化物酶体增殖物激活受体α(Pparα)、PPARγ共激活因子1-β(Pgc1β)、Srebp1-α和Pparγ2(81–86)。BMAL1:CLOCK通过结合E-box启动子元件激活Atgl和Hsl基因表达(81)。在2型糖尿病肥胖个体中,这些昼夜节律大多消失(83, 87)。

脂肪组织生物钟的生理意义已在多项研究中证实。Bmal1缺陷的胚胎干细胞向脂肪细胞分化受损(82)。BMAL1:CLOCK二聚体通过Wht信号通路调控脂肪生成(88, 89)。Clock基因突变导致的生物钟遗传破坏,在常规饮食背景下以遗传背景依赖性方式增加肥胖。两品系Clock突变体均表现出血清TGs、FAs和甘油节律减弱或消失,提示脂肪吸收和脂解受损(81, 90)。后者归因于Atgl和Hsl转录减少(81)。脂肪细胞特异性Bmal1敲除在高脂饮食条件下导致肥胖,可能与循环多不饱和FAs变化影响中枢食物摄入节律有关。这一观点得到下丘脑食欲调控神经肽异常表达的支持(91)。综上,存在“脂肪细胞-下丘脑”昼夜节律轴,强调功能性脂肪组织对能量稳态昼夜调控的重要性。

Bmal1过表达可增加WAT中脂质生成相关基因表达(82)。BMAL1:CLOCK通过E-box结合诱导Ppar表达(92, 93)。PPARγ尤其对脂肪细胞分化和脂肪生成至关重要(94–96)。PPARs也反馈调控Clock基因表达:PPARγ诱导脂肪组织中Rev-Erbα表达,而PPARα激活肝脏中Bmal1转录(97, 98)。PER2作为核心TTFL负性臂成员,通过抑制PPARγ与PPAR反应元件结合,直接抑制PPARγ靶基因表达(99)。生物钟调节因子——分化胚胎软骨细胞蛋白1(DEC1)阻止PPARγ介导的靶基因表达,从而促进这些基因的昼夜振荡。与此一致,Dec1缺失导致暗期中脂肪酸合成、脂质储存和脂解相关基因表达显著增加,且血清FAs昼夜节律消失(100)。核受体Rev-Erbα/β和RORα/β表达在小鼠光照期结束时达峰。Srebp-1c、Pparγ以及脂联素和瘦素的mRNA也呈现夜间达峰的昼夜节律(101)。Rev-Erbα遗传破坏与SREBP1和SREBP2活性降低相关(102)。REV-ERBα调控肝脏中SREPBs活性和胆汁酸代谢(102)。在肌肉中,REV-ERBβ被招募至Srebp-1c启动子,诱导基因表达(103)。因此,REV-ERB激动剂可能通过改变代谢基因的昼夜节律表达,增加能量消耗,从而改善血脂异常和高血糖,成为治疗肥胖的有前景候选药物(104)。

总之,生物钟对脂肪细胞分化及脂质生成/脂解基因的节律性表达至关重要。时钟蛋白调控WAT代谢相关蛋白活性,后者进而控制FAs的节律性释放。FAs将代谢状态信号传递至大脑,调节食物摄入。WAT代谢的昼夜节律紊乱会导致脂肪生成、脂质动员和食物摄入异常,促进肥胖。

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**白色脂肪组织的节律性输出调控代谢** 代谢稳态受WAT等组织以昼夜节律方式释放的因子调控。这些因子调节其他组织生理功能,并在中枢水平整合以控制食物摄入。WAT主要功能之一是储存脂质并释放脂解产物。后者(FAs和甘油)在小鼠和人类中呈现显著的昼夜节律,该节律仅部分由进食驱动(81, 90, 100, 105)。多不饱和脂肪酸(PUFA)节律丧失会改变食欲调控神经肽表达并增加食物摄入。PUFA水平降低伴随长链脂肪酸延长酶5/6(Elovl5/6)和硬脂酰-CoA去饱和酶1(Scd1)(PUFA合成关键酶)表达减少(91, 106)。恢复下丘脑中PUFA含量可挽救食物摄入节律、体重发育、食欲调控神经肽表达及能量稳态(91)。这清楚表明,振荡性脂解输出对全身代谢稳态至关重要。

WAT也是内分泌器官,以昼夜节律方式释放脂肪因子,参与全身代谢调控(图2)。瘦素是研究最深入的脂肪因子之一,其分泌受胰岛素刺激,也受脂肪组织生物钟调控(107, 108)。每日6次规律进食下瘦素节律仍可维持,但SCN损毁后该节律消失,提示瘦素分泌受生物钟控制(109)。体外研究表明,脂肪细胞生物钟调控瘦素分泌。尽管脂肪细胞中瘦素mRNA表达无节律性,但瘦素释放全天变化(107)。瘦素水平与体脂含量相关,禁食时下降(110–112)。它穿过血脑屏障,作为饱腹激素调节能量消耗和食物摄入(113–116)。瘦素与中枢神经系统(如弓状核)中瘦素受体结合。弓状核包含神经肽Y(NPY)/刺鼠相关肽(AgRP)阳性神经元和阿黑皮素原(POMC)/可卡因-苯丙胺调节转录肽(CART)阳性神经元,调控食物摄入和能量消耗(117)。瘦素通过抑制促食欲神经肽Agrp和Npy表达,刺激抑食欲神经肽Pomc表达,从而抑制食物摄入(118–121)。慢性时差可能通过中枢瘦素抵抗和瘦素转录下调促进肥胖(122)。瘦素对能量消耗的作用可能部分由BAT激活介导。近期发现提示,瘦素的产热效应可能受表达PGC-1β的POMC神经元调控(123)。此外,瘦素刺激β-肾上腺素能受体,从而增加BAT中Ucp1表达(124)。尽管对BAT有影响,瘦素还通过腺苷一磷酸激活蛋白激酶C(AMPK)信号通路激活外周组织(如肌肉和肝脏)中的β-氧化(125),从而防止脂质在这些组织中积累。此外,瘦素直接抑制GCs释放,而GCs在糖脂稳态中起重要作用(70, 126, 127)。

脂联素在脂肪组织中呈昼夜节律表达(128),是受生物钟调控的PPARγ和PGC1β的靶基因(129)。如前所述,它在调节能量消耗和胰岛素敏感性中起关键作用(56, 57)。脂联素增加β-氧化和葡萄糖摄取,从而降低体重(130–132)。它还通过沉默信息调节因子1(SIRT1)-AMPK介导的Ucp1表达上调,诱导WAT褐变,影响能量消耗(133, 134)。然而,在BAT本身,脂联素似乎通过抑制β-肾上腺素能受体表达来抑制Ucp1表达(135)。脂联素受体(Adipor1和2)在脂肪组织和下丘脑内侧基底部呈昼夜振荡表达(128, 136)。该脂肪因子将外周代谢状态信号传递至大脑,结合血糖水平调节食物摄入(137, 138)。其作用由AMPK信号通路介导(139)。脂联素还诱导下丘脑内侧基底部Bmal1表达,进而局部调控促食欲神经肽表达(136)。这些发现提示一种机制:外周生物钟紊乱可能改变食物摄入节律,促进代谢性疾病发展。

由烟酰胺磷酸核糖转移酶(Nampt)编码的血清内脂素水平呈昼夜节律,与瘦素呈负相关(140–143)。睡眠剥夺会改变Nampt表达节律,对糖稳态产生负面影响(142)。内脂素/NAMPT还催化烟酰胺腺嘌呤二核苷酸(NAD+)补救合成通路中的限速步骤。Nampt表达受生物钟调控,并通过调节组蛋白去乙酰化酶SIRT1活性影响核心TTFL(143)。其拟胰岛素功能尚存争议,仍在研究中(144–146)。然而,越来越多的证据表明内脂素具有促炎功能,多数研究一致认为脂肪量与内脂素水平呈正相关(147–150)。浸润的免疫细胞可能是内脂素表达的主要来源(149, 151)。循环内脂素水平与促炎细胞因子如白细胞介素-6(IL-6)和C反应蛋白呈正相关,且内脂素表达与肿瘤坏死因子α(Tnf-α)和Il-6表达高度相关(148, 150, 152)。因此,内脂素升高可能对能量稳态产生有害影响。事实上,内脂素通过诱导炎症通路部分导致肝脏胰岛素抵抗,并诱导FAs介导的神经炎症(152, 153)。

抵抗素降低胰岛素敏感性,其昼夜节律滞后于胰岛素,提示其对胰岛素作用有负反馈调节(154)。抵抗素表达的节律性受胰岛素节律性输入刺激。在肥胖伴随胰岛素抵抗状态下,抵抗素表达的节律性减弱或消失(155, 156)。在人类中,抵抗素主要由巨噬细胞表达,而在啮齿类动物中主要来源为脂肪细胞。人与啮齿类动物在基因和蛋白结构上存在差异,导致功能角色不同(157)。人抵抗素激活促炎细胞因子(如TNFα、IL-6和IL-12)的昼夜表达,促进胰岛素抵抗和炎症发展(158, 159)。反过来,这些细胞因子增强抵抗素表达(160)。中性粒细胞是饮食挑战后最先浸润脂肪组织的免疫细胞(161)。中性粒细胞进一步吸引巨噬细胞等免疫细胞,后者降低胰岛素敏感性并诱导慢性炎症(162, 163)。主要由巨噬细胞释放的TNFα促进脂解(164),并抑制脂滴包被蛋白(与脂肪储存相关)表达(164, 165)。此外,它降低GLUT4和LPL表达(166)。因此,高TNFα水平抑制胰岛素介导的葡萄糖摄取(167, 168),促进胰岛素抵抗发展(如肥胖个体中)(167, 169)。GC治疗可抑制TNFα介导的胰岛素抵抗,但也减弱其脂解作用,促进脂肪积累(170)。

综上,通过FAs和脂肪激素的节律性释放,WAT在能量稳态的昼夜调节中发挥核心作用。它调控食物摄入节律、能量消耗、胰岛素敏感性和代谢性炎症。在实验啮齿动物中,WAT节律紊乱促进过度进食和肥胖。

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**棕色脂肪组织代谢的昼夜节律交互作用** BAT的主要功能是通过非颤抖性产热将能量转化为热量(20)。BAT产热通过β-氧化实现,或通过UCP1将线粒体质子转运与能量产生解偶联(171)。BAT产热使哺乳动物体温更稳定,应对寒冷环境。寒冷刺激下,自主神经激活导致BAT末梢释放去甲肾上腺素。棕色脂肪细胞中β3-肾上腺素能受体激活引发G蛋白控制的蛋白激酶A通路激活,进而诱导代谢基因(包括Ucp1)表达(171)。此外,该通路激活ATGL产生FAs,进一步刺激UCP1(26, 171)。

BAT产热也可由富含碳水化合物的餐食(饮食诱导产热,DIT;图3)通过自主肾上腺素能激活诱导(172, 173)。肠道来源的激素和胆汁酸也能刺激DIT(174)。肠道源性胆囊收缩素(CCK)和促胰液素通过迷走神经传入和交感神经传出激活BAT产热,并激活UCP1(174)。胃饥饿素节律影响Ucp1表达,且其分泌在进食后减少(175, 176)。通过DIT,机体可部分减少食物中过量能量摄取,避免以脂肪形式储存能量(171)。BAT餐后产热诱导(导致葡萄糖和FAs摄取)可能受胰岛素刺激(173)。有趣的是,UCP1似乎是必需的,因为UCP1消融小鼠不再表现饮食诱导产热,反而体重增加(177)。内分泌性昼夜节律因子(如松果体激素褪黑素)调节BAT非颤抖性产热能力(171)。在啮齿类动物中,GCs下调UCP1从而抑制BAT产热;在人类中则作用相反(178)。

与WAT类似,BAT也参与能量代谢的昼夜节律调控。然而,由于BAT功能主要非内分泌性,其代谢调控重点在于清除血液循环中的代谢因子,以及影响棕色脂肪细胞的产热能力。BAT的生物钟与代谢调控紧密相连。反复光暗周期转换导致的慢性节律紊乱不仅引起BAT“白化”,还降低大鼠UCP1表达(179)。

BAT中葡萄糖及TGs/FAs摄取呈节律性,分别在非活动期末或活动期初达峰(180–183)。参与TG分解的酶(如LPL)活性和表达也在活动期初最高(180, 183)。这表明BAT在FAs、TGs和葡萄糖清除的昼夜门控中发挥作用(180, 183, 184)。与此一致,产热在活动期更高(182)。由于人类高BAT活性与低血糖相关,这些数据提示充足BAT可稳定全天血糖波动,维持葡萄糖稳态(184)。有趣的是,人BAT葡萄糖摄取在早晨增加,与Ucp1和Glut4基因高表达相关(184, 185)。FGF21似乎对BAT葡萄糖清除重要。它通过稳定BAT温度进而调控BAT产热,抑制BAT温度下降并改善正常葡萄糖清除(186)。有趣的是,BAT产热活性也增加局部FGF21释放,提示存在旁分泌反馈(187)。

昼夜节律性产热可塑性受REV-ERBα调控。研究表明,在REV-ERBα表达较低时段,小鼠更能耐受低温(189)。REV-ERBα抑制UCP1。反过来,低温下调Rev-Erbα,诱导UCP1以增加产热。REV-ERBα缺失导致体温和BAT活动节律完全丧失,整体体温升高(189)。这强调了BAT代谢昼夜调控的重要性。有趣的是,SCN本身通过控制REV-ERBα参与血浆TGs变化,调控脂质向BAT摄取(182, 190)。这支持了昼夜节律紊乱可能导致高脂血症的假设(190)。

总之,BAT对代谢的昼夜调控主要涉及调节循环中葡萄糖和脂质的摄取与清除。此外,产热能力受BAT内部生物钟调控,也受褪黑素和GCs等昼夜激素调节(见图3)。

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**结论** WAT和BAT在代谢昼夜节律调控中的作用日益清晰。两种脂肪库均具有内源性生物钟,调控组织特异性转录——包括WAT中脂质生成/脂解关键基因和BAT中产热相关基因。WAT通过以昼夜节律方式释放脂肪因子和FAs,调节进食行为、碳水化合物代谢和能量消耗。由此,代谢通过外周能量状态的反馈信号在中枢整合,响应机体需求进行调节。BAT则参与循环能量底物的昼夜调控。近年来,研究多聚焦于(组织)生物钟破坏对能量代谢的影响。昼夜节律研究帮助我们理解传入信号与WAT生理如何在组织水平整合,产生协调输出,进而调控食物摄入并调节其他代谢稳态相关器官的生理功能。因此,脂肪组织不仅是脂质储存和产热器官,更是能量稳态的重要调节器。从临床角度看,解析调控脂肪组织昼夜节律的机制至关重要。近期研究改变了我们仅将脂肪组织视为储存组织的观点,强调其作为潜在治疗靶点的价值。此类知识有助于开发脂肪库特异性时辰药理学方法,对抗现代社会日益普遍的能