Involvement of TRP Channels in Adipocyte Thermogenesis: An Update

✅ 全文

TRP通道在脂肪细胞产热中的作用:最新进展

作者 Wuping Sun; Yixuan Luo; Fei Zhang; Shuo Tang; Tao Zhu 期刊 Frontiers in Cell and Developmental Biology 发表日期 2021 ISSN 2296-634X DOI 10.3389/fcell.2021.686173 类型 原创研究 (Original Research)

📄 英文摘要 English Abstract

EN

Obesity prevalence became a severe global health problem and it is caused by an imbalance between energy intake and expenditure. Brown adipose tissue (BAT) is a major site of mammalian non-shivering thermogenesis or energy dissipation. Thus, modulation of BAT thermogenesis might be a promising application for body weight control and obesity prevention. TRP channels are non-selective calcium-permeable cation channels mainly located on the plasma membrane. As a research focus, TRP channels have been reported to be involved in the thermogenesis of adipose tissue, energy metabolism and body weight regulation. In this review, we will summarize and update the recent progress of the pathological/physiological involvement of TRP channels in adipocyte thermogenesis. Moreover, we will discuss the potential of TRP channels as future therapeutic targets for preventing and combating human obesity and related-metabolic disorders.

📄 中文摘要 Chinese Abstract

中文
肥胖患病率已成为严重的全球健康问题,其成因是能量摄入与消耗之间的失衡。棕色脂肪组织(BAT)是哺乳动物非颤抖性产热或能量耗散的主要场所。因此,调节棕色脂肪组织的产热功能可能成为体重控制和肥胖预防的有前景的应用方向。TRP通道是非选择性钙通透性阳离子通道,主要位于细胞膜上。作为研究热点,TRP通道已被报道参与脂肪组织产热、能量代谢和体重调节。肥胖是一个严重的公共卫生问题,可引发多种疾病,包括糖尿病、高血压、冠心病和癌症,已受到广泛关注。根据基于1975年至2014年200个国家数据的预测,到2025年,全球肥胖患病率男性将达到18%,女性将达到21%。此外,肥胖不仅在发达国家日益流行,在发展中国家也日趋普遍。因此,迫切需要制定策略来预防和逆转肥胖及相关代谢性疾病。

📋 英文结构化总结 English Structured Summary

全文整理

EN

Background:

Obesity prevalence became a severe global health problem and it is caused by an imbalance between energy intake and expenditure. Brown adipose tissue (BAT) is a major site of mammalian non-shivering thermogenesis or energy dissipation. Thus, modulation of BAT thermogenesis might be a promising application for body weight control and obesity prevention. TRP channels are non-selective calcium-permeable cation channels mainly located on the plasma membrane. As a research focus, TRP channels have been reported to be involved in the thermogenesis of adipose tissue, energy metabolism and body weight regulation. Obesity is a severe public health problem causing various diseases including diabetes, hypertension, coronary heart diseases and cancer, which has received considerable attention as a major public health concern. According to a prediction based on the data from 1975 to 2014 in 200 countries, the prevalence of global obesity will reach to 18% for men and 21% for women by 2025. In addition, obesity is becoming prevalent not only in the developed countries, but also in the developing countries. Therefore, urgent strategies are required for the prevention and reversal of obesity and related metabolic diseases.

Methods:

N/A - Review article

Results:

It has assessed that BAT thermogenesis was decreased in obese mice by oxygen consumption measurement. UCP1 expression level in BAT was decreased in almost all obese animals whereas increased in lean animals. UCP1 knockout (UCP1KO) mice exhibited obesity phenotypes with increased body fat after six months high fat diet (HFD) feeding. On the other hand, cold stimulation and/or β3-adrenergic receptor agonist treatment decreased body fat amount by enhancing BAT activity. Cold exposure also increased BAT volume and activity, thus increasing energy consumption and promoting weight loss of obese people. Several studies have reported that there was a negative correlation between BAT activity/amount and body mass index (BMI) in humans. Imaging data have revealed that patients with higher BMI have lower BAT activity. Moreover, a single nucleotide substitution at –3826A to G of UCP1 gene polymorphism has been found in human, which decreased the mRNA expression of Ucp1 and enhanced the age-related obesity and BAT degradation. UCP1 is expressed in the mitochondria inner membranes of brown adipocytes, which uncouples ATP synthesis from oxidative phosphorylation, thereby dissipating energy as heat.

Data Summary:

According to a prediction based on the data from 1975 to 2014 in 200 countries, the prevalence of global obesity will reach to 18% for men and 21% for women by 2025. UCP1 knockout (UCP1KO) mice exhibited obesity phenotypes with increased body fat after six months high fat diet (HFD) feeding.

Conclusions:

Therefore, BAT might play critical role in the regulation of body weight and energy homeostasis. Modulation of BAT thermogenesis might be a promising application for body weight control and obesity prevention. TRP channels have been reported to be involved in the thermogenesis of adipose tissue, energy metabolism and body weight regulation, and we will discuss the potential of TRP channels as future therapeutic targets for preventing and combating human obesity and related-metabolic disorders.

Practical Significance:

Modulation of BAT thermogenesis might be a promising application for body weight control and obesity prevention. TRP channels are discussed as future therapeutic targets for preventing and combating human obesity and related-metabolic disorders. Urgent strategies are required for the prevention and reversal of obesity and related metabolic diseases.

📋 中文结构化总结 Chinese Structured Summary

中文

背景:

肥胖患病率已成为严重的全球健康问题,其成因是能量摄入与消耗之间的失衡。棕色脂肪组织(BAT)是哺乳动物非颤抖性产热或能量耗散的主要场所。因此,调节棕色脂肪组织的产热功能可能成为体重控制和肥胖预防的有前景的应用方向。TRP通道是非选择性钙通透性阳离子通道,主要位于细胞膜上。作为研究热点,TRP通道已被报道参与脂肪组织产热、能量代谢和体重调节。肥胖是一个严重的公共卫生问题,可引发多种疾病,包括糖尿病、高血压、冠心病和癌症,已受到广泛关注。根据基于1975年至2014年200个国家数据的预测,到2025年,全球肥胖患病率男性将达到18%,女性将达到21%。此外,肥胖不仅在发达国家日益流行,在发展中国家也日趋普遍。因此,迫切需要制定策略来预防和逆转肥胖及相关代谢性疾病。

方法:

不适用——综述类文章

结果:

通过耗氧量测定评估发现,肥胖小鼠的棕色脂肪组织产热功能下降。几乎所有肥胖动物棕色脂肪组织中UCP1表达水平均降低,而在瘦弱动物中则升高。UCP1基因敲除(UCP1KO)小鼠在高脂饮食(HFD)喂养六个月后表现出肥胖表型,体脂增加。另一方面,冷刺激和/或β3-肾上腺素能受体激动剂处理可通过增强棕色脂肪组织活性来减少体脂量。冷暴露还可增加棕色脂肪组织的体积和活性,从而增加能量消耗并促进肥胖人群的体重减轻。多项研究报道,人体中棕色脂肪组织活性/数量与体重指数(BMI)呈负相关。影像学数据显示,BMI较高的患者棕色脂肪组织活性较低。此外,在人体中发现UCP1基因多态性存在-3826A到G的单核苷酸替换,该突变降低了Ucp1的mRNA表达,并加剧了年龄相关性肥胖和棕色脂肪组织退化。UCP1在棕色脂肪细胞线粒体内膜上表达,使ATP合成与氧化磷酸化解偶联,从而以热能形式耗散能量。

数据摘要:

根据基于1975年至2014年200个国家数据的预测,到2025年,全球肥胖患病率男性将达到18%,女性将达到21%。UCP1基因敲除(UCP1KO)小鼠在高脂饮食(HFD)喂养六个月后表现出肥胖表型,体脂增加。

结论:

因此,棕色脂肪组织可能在体重调节和能量稳态中发挥关键作用。调节棕色脂肪组织产热功能可能成为体重控制和肥胖预防的有前景的应用方向。TRP通道已被报道参与脂肪组织产热、能量代谢和体重调节,我们将讨论TRP通道作为未来预防和治疗人类肥胖及相关代谢性疾病治疗靶点的潜力。

实际意义:

调节棕色脂肪组织产热功能可能成为体重控制和肥胖预防的有前景的应用方向。TRP通道被讨论作为未来预防和治疗人类肥胖及相关代谢性疾病的有前景的治疗靶点。迫切需要制定策略来预防和逆转肥胖及相关代谢性疾病。

📖 英文全文 English Full Text

EN

REVIEW published: 24 June 2021 doi: 10.3389/fcell.2021.686173

Involvement of TRP Channels in Adipocyte Thermogenesis: An Update Wuping Sun 1† , Yixuan Luo 2† , Fei Zhang 2 , Shuo Tang 3* and Tao Zhu 4* 1

Department of Pain Medicine and Shenzhen Municipal Key Laboratory for Pain Medicine, Shenzhen Nanshan People’s Hospital and The 6th Affiliated Hospital of Shenzhen University Health Science Center, Shenzhen, China, 2 Department of Cardiovascular Surgery, Shenzhen Nanshan People’s Hospital and The 6th Affiliated Hospital of Shenzhen University Health Science Center, Shenzhen, China, 3 Department of Orthopaedics, The Eighth Affiliated Hospital, Sun Yat-sen University, Shenzhen, China, 4 Department of Respiratory Medicine, Second Affiliated Hospital of Chongqing Medical University, Chongqing, China

Edited by: Maria Elena De Lima, Grupo Santa Casa BH, Brazil Reviewed by: Karla S. Fernandes, Instituto de Ensino e Pesquisa Santa Casa BH, Brazil Célio José Castro Junior, Grupo Santa Casa BH, Brazil *Correspondence: Tao Zhu zhutao063020@163.com Shuo Tang tangshuo1205@163.com

Obesity prevalence became a severe global health problem and it is caused by an imbalance between energy intake and expenditure. Brown adipose tissue (BAT) is a major site of mammalian non-shivering thermogenesis or energy dissipation. Thus, modulation of BAT thermogenesis might be a promising application for body weight control and obesity prevention. TRP channels are non-selective calcium-permeable cation channels mainly located on the plasma membrane. As a research focus, TRP channels have been reported to be involved in the thermogenesis of adipose tissue, energy metabolism and body weight regulation. In this review, we will summarize and update the recent progress of the pathological/physiological involvement of TRP channels in adipocyte thermogenesis. Moreover, we will discuss the potential of TRP channels as future therapeutic targets for preventing and combating human obesity and related-metabolic disorders. Keywords: TRP channels, calcium, thermogenesis, energy metabolism, brown adipocytes, beige adipocytes, obesity

equally to this work Specialty section: This article was submitted to Cellular Biochemistry, a section of the journal Frontiers in Cell and Developmental Biology Received: 26 March 2021 Accepted: 02 June 2021 Published: 24 June 2021 Citation: Sun W, Luo Y, Zhang F, Tang S and Zhu T (2021) Involvement of TRP Channels in Adipocyte Thermogenesis: An Update. Front. Cell Dev. Biol. 9:686173. doi: 10.3389/fcell.2021.686173

ADIPOSE TISSUES AND OBESITY Obesity is a severe public health problem causing various diseases including diabetes, hypertension, coronary heart diseases and cancer, which has received considerable attention as a major public health concern (Nguyen and El-Serag, 2010; Blüher, 2019). According to a prediction based on the data from 1975 to 2014 in 200 countries, the prevalence of global obesity will reach to 18% for men and 21% for women by 2025 (NCD Risk Factor Collaboration, 2016). In addition, obesity is becoming prevalent not only in the developed countries, but also in the developing countries (Maharani and Tampubolon, 2016). Therefore, urgent strategies are required for the prevention and reversal of obesity and related metabolic diseases. Abbreviations: BMI, body mass index; BAT, brown adipose tissue; CT, computed tomography; FDG, fluorodeoxyglucose; HFD, high fat diet; iBAT, interscapular BAT; WAT, white adipose tissue; UCP1, uncoupling protein-1; PET, positron emission tomography; PGC1α, peroxisome proliferator-activated receptor gamma coactivator 1-alpha; ROS, reactive oxygen species; TM, trans-membrane; TRP channel, Transient receptor potential channel; TRPV, TRP Vanilloid; TRPV2KO, TRPV2knockout; TRPC, TRP Canonical; TRPM, TRP Melastatin; TRPML, TRP Mucolipin; TRPN, TRP NomPC; TRPP, TRP Polycystin; TRPA, TRP Ankyrin; WT, wild-type; [Ca2+ ]i , intracellular Ca2+ levels; 4α-PDD, 4α-phorbol-12, 13-didecanoate.

Frontiers in Cell and Developmental Biology | www.frontiersin.org 1 June 2021 | Volume 9 | Article 686173 Sun et al. TRP Channels in Adipocyte Thermogenesis

Obesity is accompanied by the imbalance of caloric intake and consumption (Hall and Guo, 2017). There is evidence that adipose tissue is involved in the long-term regulation of energy metabolism and fat quality. Adipose tissue is a highly specialized tissue and plays a key role in energy mobilization regulation (Reilly and Saltiel, 2017; Zhai et al., 2020). Two types of adipose tissue have been found in mammals so far, called white adipose tissue (WAT) and brown adipose tissue (BAT) (Cannon and Nedergaard, 2004; Wu et al., 2020). WAT is generally thought as an organ stores excess energy which maintains energy in the form of triglyceride in lipid droplets. However, a new type of brown-like adipocyte was termed beige/brite adipocyte or inducible brown adipocyte has recently been found in human WAT (Sharp et al., 2012; Cypess et al., 2013; Lidell et al., 2013). BAT, which consumes energy and produce heat rapidly, was first discovered in mammalian hibernation research (Ricquier and Kader, 1976). This thermogenic function is mainly mediated by uncoupling protein-1 (UCP1), a polypeptide that exists in the mitochondrial inner membrane of brown adipocytes (Kajimura et al., 2015; Bertholet et al., 2017; Cannon et al., 2020). It has assessed that BAT thermogenesis was decreased in obese mice by oxygen consumption measurement (Martinez-Botas et al., 2000; Ussher et al., 2010). UCP1 expression level in BAT was decreased in almost all obese animals whereas increased in lean animals (Shirkhani et al., 2018). UCP1 knockout (UCP1KO) mice exhibited obesity phenotypes with increased body fat after six months high fat diet (HFD) feeding (Kontani et al., 2005). On the other hand, cold stimulation and/or β3-adrenergic receptor agonist treatment decreased body fat amount by enhancing BAT activity (Lowell and Spiegelman, 2000; Cannon and Nedergaard, 2004). Cold exposure also increased BAT volume and activity, thus increasing energy consumption and promoting weight loss of obese people (Hanssen et al., 2015a,b; Leiria et al., 2019). Several studies have reported that there was a negative correlation between BAT activity/amount and body mass index (BMI) in humans. Imaging data have revealed that patients with higher BMI have lower BAT activity (Cypess et al., 2009; Pfannenberg et al., 2010; Ouellet et al., 2011). Moreover, a single nucleotide substitution at –3826A to G of UCP1 gene polymorphism has been found in human, which decreased the mRNA expression of Ucp1 and enhanced the age-related obesity and BAT degradation (Nagai et al., 2007; Yoneshiro et al., 2013). Therefore, BAT might play critical role in the regulation of body weight and energy homeostasis.

Classical brown fat is primarily distributed around interscapular BAT (iBAT), axillary, paravertebral, and perirenal sites (Park et al., 2014). Mitochondria and multilocular lipid droplets were enriched in brown adipocytes, which makes it have remarkable capacity to dissipate energy in the form of heat (Song et al., 2020). UCP1 is expressed in the mitochondria inner membranes of brown adipocytes, which uncouples ATP synthesis from oxidative phosphorylation, thereby dissipating energy as heat. It is well known that BAT non-shivering thermogenesis is controlled directly by sympathetic nervous system innervation and activation. BAT thermogenesis is induced and regulated by the release of norepinephrine from sympathetic nerve terminals and its subsequent binding by β3-adrenergic receptors (Nedergaard et al., 2005; Feldmann et al., 2009). Several studies have shown that how UCP1 is activated, and long chain fatty acid is essential for H+ transport (Fedorenko et al., 2012). In addition, another proposed mechanism is that mitochondrial reactive oxygen species (ROS) production regulates UCP1 sulfenylation and thermogenesis (Chouchani et al., 2016). However, signaling pathways for thermogenesis in the downstream of β3-adrenergic receptor activation still have not been well clarified. Beige adipocyte (UCP1-positive adipocyte) is known to be surrounded by numerous UCP1-negative adipocytes in human WAT (Wu et al., 2012). Beige adipocytes could be recruited after a short-term cold challenge or treatment with β3-adrenergic receptor agonists (Saito et al., 2020). They are very similar to brown adipocytes with high UCP1 expression and thermogenesis (Ye et al., 2013; Li et al., 2014). There are two groups that are a BAT-positive group (subjects have detectable FDG uptake upon cold stimulation) and a BAT-negative group (subjects have undetectable FDG uptake) in humans. Energy metabolism was higher in the BAT-positive group than the BAT-negative group after an acute cold exposure (Orava et al., 2011; Yoneshiro et al., 2011). These studies clearly revealed a critical function for brown and beige adipocytes in cold-induced thermogenesis in humans. Therefore, approaches to modulate brown or beige adipocyte activities might be potential way to prevent and treat human obesity and related metabolic diseases.

TRP CHANNELS Transient receptor potential (TRP) ion channels are a major class of calcium-permeable channels, most of which are non-selective cation channels (Montell and Rubin, 1989). TRP channels contain six trans-membrane (TM) domains (TM1–TM6) with a pore loop between TM5 and TM6 (Cao et al., 2013b; Liao et al., 2013; Paulsen et al., 2015; Huynh et al., 2016; Zubcevic et al., 2016). TRP channel superfamily is now subdivided into seven subfamilies and contains 27 channels: TRPV (Vanilloid), TRPC (Canonical), TRPM (Melastatin), TRPML (Mucolipin), TRPN (NomPC), TRPP (Polycystin), and TRPA (Ankyrin) based on their primary amino acid sequences (Ramsey et al., 2006; Wu et al., 2010; Gees et al., 2012). The main signaling pathways in which TRP channels triggered are based on calcium influx through the channels, leading to increases in intracellular Ca2+ levels ([Ca2+ ]i ). Numerous studies have shown that some TRP channels are expressed in adipocytes and are involved in energy

THERMOGENESIS IN BROWN AND BEIGE ADIPOCYTES BAT was thought to be restricted only in infants (Lean, 1989; Enerback, 2010). However, previous works have reported that BAT was also found in adult humans by using fluorodeoxyglucose (FDG)-positron emission tomography (PET) in combination with computed tomography (CT) techniques (Cypess et al., 2009; van Marken Lichtenbelt et al., 2009). This novel finding highlights the critical role for BAT in the regulation of energy metabolism and fat deposition (Nedergaard and Cannon, 2010; Nedergaard et al., 2011). Frontiers in Cell and Developmental Biology | www.frontiersin.org

prevalence of human obesity in eastern Asian countries was decreased by increasing the consumption of hot foods containing capsaicin (Wahlqvist and Wattanapenpaiboon, 2001). It has also been reported that capsaicin injection induced adrenaline secretion, this effect was significantly reduced in TRPV1KO mice (Uchida et al., 2017). Capsaicin directly binds to TRPV1 in gastrointestinal tract, produce afferent signal, subsequently transmit to ventromedial hypothalamic nucleus of central nervous system, and finally send signal to WAT. This could promote the expression of β2-adrenoceptor and the production of PRDM16 protein, thus promoting the generation of beige adipocytes, resulting in increased systemic energy expenditure (Ohyama et al., 2016; Saito et al., 2020). Catechins in green tea may activate and recruit BAT by acting on TRPV1/TRPA1 of gastrointestinal sensory neurons in the same way as capsaicin (Mako et al., 2015). Besides, topical application of capsaicin cream in mice resulted in weight loss and adipose tissue weight (Lee et al., 2013). However, whether the regulatory effect of topical capsaicin on obesity is through TRPV1 to activate central nervous system remains to be further studied. These studies clearly demonstrated that targeting TRPV1 and modulation its activity with capsaicin and analogs could be effective approaches for human obesity treatment and prevention, although the anti-obesity effect of TRPV1 activation may be involved not only in adipose tissue, but also in nervous system.

metabolism and inflammation of adipose tissues, suggesting the potential role of TRP channels in human obesity treatment and prevention (Bishnoi et al., 2018; Uchida et al., 2018; Gao et al., 2019; Zhai et al., 2020). In the present review, we will provide a systematic and brief summary of TRP channels in the regulation of adipocyte thermogenesis and update the recent progress.

TRPV1 TRPV1 is well-known as a receptor of capsaicin, the pungent ingredient in “hot” chili peppers (Caterina et al., 1997). TRPV1 is activated by a variety of stimuli, including heat (Cao et al., 2013a), protons and capsaicin (Dhaka et al., 2009). In addition, TRPV1 is activated by some compounds in garlic, onion (Salazar et al., 2008), black pepper (Okumura et al., 2010), and other foods, such as gingerol (Iwasaki et al., 2006). TRPV1 has been reported to be expressed in both WAT and BAT (Bishnoi et al., 2013; Kida et al., 2016). TRPV1 expression level is increased in the differentiated HB2 brown adipocytes than in pre-adipocytes (Kida et al., 2016). Moreover, activation of TRPV1 up-regulates the expression of thermogenic genes and induced “browning” in 3T3-L1 adipocytes (Figure 1; Baboota et al., 2014). TRPV1 is expressed in 3T3-L1 pre-adipocytes, adipose tissue of mice and fat tissue of obese humans (Zhang et al., 2007). TRPV1 is activated by dietary capsaicin, a process that induces calcium influx and prevents adipogenesis in 3T3-L1 cells (Zhang et al., 2007) and probably occurs through a calcineurin pathway (Cioffi, 2007). Besides, dietary capsaicin treatment prevented HFDinduced obesity in wild-type (WT) mice in vivo, but not in TRPV1KO mice (Zhang et al., 2007; Chen J. et al., 2015; Chen N. et al., 2015). Moreover, TRPV1 was involved in the regulation of energy intake and glucose homeostasis in WAT during HFDinduced obesity (Lee et al., 2015). Absence of TRPV1 exacerbated obese and insulin resistance associated with HFD and aging (Lee et al., 2015). It has also been reported that monoacylglycerol up-regulated UCP1 expression level in brown adipocytes and suppressed accumulation of visceral fat in mice fed with high fat and sucrose through activation of TRPV1 (Iwasaki et al., 2011). Fish oil intake induced UCP1 up-regulation in both brown and white adipose tissues in a TRPV1 dependent manner (Kim et al., 2015; Lund et al., 2018). Oleoylethanolamide, a newly reported TRPV1 ligand, is also involved in the regulation of energy intake and consumption, feeding behavior and weight control (Laleh et al., 2019). Human studies have showed that capsaicin ingestion enhanced fat oxidation and energy metabolism during aerobic exercise (Shin and Moritani, 2007). A continuous consumption of chili increased the energy metabolism in the middle-aged subjects (Ahuja et al., 2006). Capsinoids, a non-pungent capsaicin analogs, for continuous 1–3 months also increased energy expenditure and fat oxidation with a reduction in abdominal adiposity in overweight and obese subjects (Inoue et al., 2007; Snitker et al., 2009). Moreover, capsaicin and capsinoids as food ingredients enhanced BAT thermogenesis, subsequently decreased fat mass in humans (Yoneshiro et al., 2012; Saito and Yoneshiro, 2013). An epidemiological study suggested that the energy metabolism was enhanced and the Frontiers in Cell and Developmental Biology | www.frontiersin.org

TRPV2 TRPV2 was initially reported to be activated by noxious heat with an activation temperature threshold of higher than 52◦ C (Caterina et al., 1999) and found to be activated by several chemicals, e.g., 2-aminoethoxydiphenyl borate (2APB) and lysophosphatidylcholine (LPC) (Juvin et al., 2007; Monet et al., 2009). TRPV2 was also reported to be activated by mechanical stimulation and/or cell swelling (Muraki et al., 2003; Iwata et al., 2009). TRPV2 is expressed in both WAT and BAT (Sun et al., 2017a). TRPV2 is highly expressed in mouse brown adipocytes compared with TRPV1, TRPV3, TRPV4 and TRPM8 (Sun et al., 2016a,b). The expression of TRPV2 was up-regulated at mRNA, protein and functional levels in the differentiated brown adipocytes (Sun et al., 2016b, 2017b). Primary TRPV2-deficient (TRPV2KO) adipocytes show decreased mRNA levels of multiple genes involved in mitochondrial oxidative metabolism, such as Ucp1 and peroxisome proliferator-activated receptor gamma coactivator 1-alpha (Pgc1α). Besides, TRPV2KO adipocytes showed decreased responses to a β-adrenergic receptor agonist, isoproterenol, which might be due to the lack of TRPV2mediated calcium influx. These results suggested that TRPV2mediated calcium influx is involved in thermogenic gene induction upon β-adrenergic receptor activation (Figure 1). TRPV2KO mice showed cold intolerance and significantly smaller increases in Ucp1 mRNA and protein upon cold stimulation at 4◦ C without changes in their activities. On the other hand, sympathetic nerve activity was not changed in TRPV2KO mice. TRPV2KO mice showed impaired iBAT adaptive thermogenesis upon administration of a β3-adrenergic 3

FIGURE 1 | TRP channel-mediated adipocyte thermogenesis. A schematic figure of how TRPV1, TRPV2, TRPV4, TRPM8, and TRPC1-mediated calcium influx regulates thermogenic gene expression in adipocytes, which causes enhanced thermogenesis. Moreover, the increase in sympathetic nerve activity causes norepinephrine release from the sympathetic nerves and activation of β3-adrenergic receptor (β3ADR) in brown adipocytes, TRPV2 synergistically collaborated with β3ADR to involve in the regulation of peroxisome proliferator-activated receptor gamma coactivator-1 a (PGC1a) and uncoupling protein 1 (UCP1), subsequently enhances thermogenesis. On the other hand, TRPV4-mediated calcium influx negatively regulates thermogenic gene expression in adipocytes and subsequently inhibits thermogenesis.

an anti-adipogenic role in vivo (Cheung et al., 2015). Besides, berberine alleviates olanzapine-induced obesity by targeting TRPV1/TRPV3 in hypothalamus of mice (Singh et al., 2020). These studies suggested that targeting TRPV3 could be an intriguing approach for the treatment and prevention of obesity. However, the expression of TRPV3 and its role in human obesity needs further exploration.

receptor agonist, BRL37344. Importantly, TRPV2KO mice had significant increases in body weight and adipose tissues upon a HFD treatment (Sun et al., 2016a). Up-regulation of TRPV2 was also observed in obese and diabetic (db/db) mice (Sun et al., 2016a, 2017a). It has also been reported that tart cherry may attenuate adipogenesis by acting directly on the adipose tissue and down-regulating the HFD-induced mRNA expression of TRPV1 and TRPV2 (Cocci et al., 2021). These findings suggested that TRPV2 might be contributed to adipocyte thermogenesis. However, it is necessary to further examine the expression and function of TRPV2 in human BAT and develop specific ligands of TRPV2.

TRPV4 TRPV4 was reported to be activated by osmolarity changes or mechanical stimuli (Liedtke et al., 2000; Strotmann et al., 2000; Watanabe et al., 2002a). TRPV4 is also activated by diverse chemical compounds, including a synthetic phorbol ester, 4α-phorbol-12, 13-didecanoate (4α-PDD) and GSK1016790A (Watanabe et al., 2002b; Willette et al., 2008) as well as moderate warmth (temperature threshold higher than 27◦ C) (Guler et al., 2002; Watanabe et al., 2002b). TRPV4 is expressed in BAT and WAT as well (Sun et al., 2017a, 2020; Uchida et al., 2018). It has been reported that insulin regulates TRPV4-mediated metabolic homeostasis in human white adipocytes (Sanchez et al., 2016). TRPV4 is involved in the modulation of thermogenic and inflammatory pathways in adipose tissue. Knockdown of TRPV4 enhanced the basal and norepinephrine-induced induction of the expression of Pgc1a and Ucp1 (Ye et al., 2012). ERK1/2 were reported to be activated by TRPV4-mediated calcium signaling (Thodeti et al., 2009), and TRPV4 activation-induced calcium influx caused a rapid phosphorylation of ERK1/2 and JNK1/2, which further suppressed the expression of thermogenic genes in 3T3-F442A adipocytes (Figure 1; Ye et al., 2012). Knockdown of TRPV4 also reduced adipose tissue inflammation by inhibiting a number of pro-inflammatory genes (Ye et al., 2012). The expression of TRPV4 in WAT was higher than that in BAT (Sun et al., 2017a). The significant up-regulation of thermogenic gene expression upon TRPV4 inhibition led to the

TRPV3 TRPV3 is a member of the TRPV subfamily which is different from TRPV1 and TRPV2. TRPV3 is well-known to be activated by innocuous temperature around body temperature but initially activated by a high noxious threshold which is over 50◦ C (Liu and Qin, 2017). The chemical agonists of TRPV3 include camphor, carvacrol, (-)-epicatechin, 2APB, and endogenous ligand farnesyl pyrophosphate (Cheung et al., 2015; Broad et al., 2016). TRPV3 could form heteromeric channels with TRPV1 (Cheng et al., 2012), which also involves in the regulation of adipogenesis and HFD-induced obesity (Cheung et al., 2015). TRPV3 has been reported to be expresses in BAT and WAT (Bishnoi et al., 2013). The expression of TRPV3 was dramatically down-regulated in visceral adipose tissue of obesity mice, including HFD-induced obesity mice, ob/ob and db/db mice (Cheung et al., 2015; Sun et al., 2017a). HFD feeding up-regulated TRPV3 in the medial nucleus tractus solitaries and hypoglossal nucleus, which is accompanied by a reduced expression of proopiomelanocortin and resulted in increased food intake and a gain of bodyweight (Hu et al., 2011). Activation of TRPV3 by (-)-epicatechin prevented adipogenesis in 3T3-L1 preadipocytes and played Frontiers in Cell and Developmental Biology | www.frontiersin.org

by its ligands, such as menthol and icilin, mimics adipocyte thermogenesis and might constitute a promising approach to prevent overweight and obesity. However, randomized clinical trials of topical menthol in obese patients are necessary.

occurrence of WAT “browning” (Ye et al., 2012). TRPV4KO mice exhibited increased muscle energy oxidation and resistance to HFD-induced obese in mice (Kusudo et al., 2012). It has also been reported that treadmill running and rutin ameliorate HFDinduced obesity in mice by suppressing the expression of TRPV4 in adipocytes (Chen N. et al., 2015). Besides, dietary intervention in obese dams protects male offspring from WAT induction of TRPV4, adiposity, and hyperinsulinemia (Janoschek et al., 2016). A human subject-based study has revealed that polymorphisms of TRPV4 gene affects BMI and body fat mass in subjects in Taiwan (Duan et al., 2015). These results revealed an opposite role of TRPV4 in the modulation of adipocyte thermogenesis without knowing the potential mechanisms. Examine the expression and function of TRPV4 in human obesity and developing TRPV4 specific antagonist and in vivo examination of the new compounds is warranted.

TRPA1 TRPA1 was initially reported as a noxious cold-activated channel with a temperature threshold around 17◦ C (Story et al., 2003), However, later studies have initiated a heated debate over the role of TRPA1 as a cold sensor. But its cold sensitivity has been disputed later, and the contribution of TRPA1 to cold sensing is currently a matter of strong debate (Bautista et al., 2006; Talavera et al., 2020). TRPA1 is potentially activated by several food components, like allyl isothiocyanate, icilin, menthol, cinnamaldehyde and capsinoids (Laursen et al., 2015). TRPA1 is involved in adipocyte thermogenesis and energy metabolism (Watanabe and Terada, 2015). In HFD-induced obesity mice, oral administration of allyl isothiocyanate reduces body weight, accumulation of lipid droplets in the liver, and white adipocyte size (Lo et al., 2018). It has been reported that cinnamaldehyde reduces visceral fat deposition in HFDtreated mice by stimulating BAT between scapulae (Tamura et al., 2012). Cinnamaldehyde activates TRPA1 in mouse gastric epithelial cells and up-regulates fatty acid oxidation-related genes in adipose tissue (Camacho et al., 2015). Oleuropein aglycone, as an agonist of TRPA1 and TRPV1, enhances the expression of UCP1 in BAT and promote fat thermogenesis by promoting the secretion of norepinephrine (Oi-Kano et al., 2016). It has been hypothesized that menthol-induced thermogenesis in adipocyte probably involved a TRPA1 mechanism as well (Sakellariou et al., 2016). Moreover, TRPA1 activation induces adrenaline secretion and prevent fat accumulation and obese in rodents (Watanabe and Terada, 2015). Intravenous injection of AITC induces adrenaline secretion, and adrenaline promotes the thermogenesis of BAT by activating β3-adrenergic receptor (Saito et al., 2020). These studies suggested that TRPA1 regulates heat production of BAT through central nervous system (Zsombok and Derbenev, 2016). Therefore, activation of TRPA1 by its ligands might be a promising approach for human obesity treatment and prevention. However, the anti-obesity mechanism which TRPA1 and its ligands involved need further exploration. Randomized clinical trials of TRPA1 activation in obese patients are warranted as well.

TRPM8 The TRPM subfamily consists of eight different subunits, TRPM1 to TRPM8 (Boesmans et al., 2011). TRPM8 is wellknown as a menthol receptor which has been reported in the year of 2002 (McKemy et al., 2002). In a human adipocyte cell line, menthol-induced TRPM8 activation increased UCP1 expression, mitochondrial activation and heat production (Figure 1; Rossato et al., 2014). The mRNA and protein expression levels of TRPM8 are significantly increased in the differentiated adipocytes, suggesting the importance of TRPM8 for adipocyte thermogenesis (Rossato et al., 2014). In cultured adipocytes, menthol induced an up-regulation of UCP1 expression which may through a protein kinase A pathway, which subsequently increases BAT thermogenesis and WAT “browning” (Ma et al., 2012; Jiang et al., 2017; Sanders et al., 2020). Besides, it has been reported that cold-sensing TRPM8 channel participates in the regulation of clock and clock-controlled genes in BAT thermogenesis (Moraes et al., 2017). Bioavailable menthol induces energy expending phenotype in differentiating adipocytes (Khare et al., 2019). In vivo studies have revealed that dietary menthol supplementation dramatically increased the core body temperatures and locomotor activity in WT mice, but not in TRPM8KO and UCP1KO mice. Menthol supplementation in diet alleviated HFD-induced obesity and insulin resistance as well (Ma et al., 2012; Jiang et al., 2017). And the preventive effect of menthol against HFD-induced obesity and related complications probably involve a glucagon mechanism (Khare et al., 2018). These results suggested that activation of TRPM8 could enhance BAT thermogenesis, which paves a new approach for the treatment and prevention of obesity. TRPM8-dependent increase in core body temperature upon a menthol treatment or cold exposure, which may be mediated by a UCP1 up-regulation (Tajino et al., 2011). Intragastric administration of menthol also enhanced BAT thermogenesis in vivo (Tajino et al., 2007; Masamoto et al., 2009). In addition, TRPM8 polymorphism has been reported to be closely correlated with metabolic syndrome in Turkish population (Tabur et al., 2015). Topical menthol appears to increase core body temperature and metabolic rate in adults (Valente et al., 2015). In summary, activation of TRPM8 Frontiers in Cell and Developmental Biology | www.frontiersin.org

TRPC1 AND TRPC5 TRPC subfamily includes seven members (TRPC1–7). TRPC channels are usually formed by homo- or heteromeric TRPC proteins (Huang et al., 2011). There is no evidence demonstrate TRPC channels have thermosensitive property so far. TRPC1 is highly expressed in adipocyte depots including BAT and that TRPC1-deficient mice are prone to weight gain and manifest reduced metabolic control (Wolfrum et al., 2018). TRPC1 regulates brown adipocyte activity in a PPARγ-dependent manner, suggesting that TRPC1 is a downstream component 5

demonstrated to play critical roles in the regulation of energy metabolism for the treatment and prevention of human obesity. In the present review, we summarized and updated the recent progress of the involvement of several TRP channels in adipocyte thermogenesis. It’s worth noting that several concerns still need to be further explored. First of all, the underlying mechanisms which TRP channel-mediated in the thermogenesis process of adipocytes are still controversial, which need to be clearly addressed. Secondly, novel specific ligands of TRP channels are warranted to be developed since there is no specific ligands for TRP channels so far. Thirdly, how do TRP channels exert tissuespecific effects in adipose tissues? These issues are warranted to be addressed by further animal and clinical studies in the future. In conclusion, targeting TRP channels could be promising strategies for clinical treatment and prevention of human obesity and related-metabolic diseases.

of a mechanism that translates metabolic or environmental stimuli into output in the form of BAT activity (Figure 1; Wolfrum et al., 2018). However, an opposite observation has been reported that fat mass and fasting glucose concentrations were lower in TRPC1KO mice that were fed a HFD (45% fat) (Krout et al., 2017). Besides, a mechanically activated TRPC1like current in white adipocytes was observed (El Hachmane and Olofsson, 2018). It has been reported that either knockdown of TRPC1/TRPC5 in vitro or conditional knockout of TRPC5 in vivo has increased adiponectin generation in mouse (Sukumar et al., 2012). In addition, both exogenous and endogenous pituitary adenylate cyclase activating polypeptides stimulate proopiomelanocortin neurons and increase energy consumption by activating TRPC1 and TRPC5 channels, which suggests that it is possible to promote BAT thermogenesis by activating TRPC1/TRPC5 in central nervous system (Chang et al., 2020). These studies demonstrated the involvement of TRPC1/TRPC5 in the regulation of energy homeostasis. Further examination of the expression of TRPC1/TRPC5 in human adipose tissues and developing TRPC1/TRPC5 specific agonist are needed.

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# 瞬时受体电位通道在脂肪细胞产热中的作用:最新研究进展

**摘要**

肥胖已成为严重的全球健康问题,其根本原因在于能量摄入与消耗之间的失衡。棕色脂肪组织(BAT)是哺乳动物非颤抖性产热或能量耗散的主要场所。因此,调控BAT产热可能成为控制体重和预防肥胖的有效策略。瞬时受体潜在通道(TRP通道)是一类主要位于细胞膜上的非选择性钙离子通透性阳离子通道。作为研究热点,TRP通道已被报道参与脂肪组织产热、能量代谢及体重调节。本综述将总结并更新TRP通道在脂肪细胞产热中病理/生理作用的最新研究进展。此外,我们还将探讨TRP通道作为未来预防和治疗人类肥胖及相关代谢性疾病潜在靶点的可能性。

**关键词:** TRP通道、钙离子、产热、能量代谢、棕色脂肪细胞、米色脂肪细胞、肥胖

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## 脂肪组织与肥胖

肥胖是一个严重的公共卫生问题,可引发多种疾病,包括糖尿病、高血压、冠心病和癌症,已引起广泛关注(Nguyen and El-Serag, 2010; Blüher, 2019)。根据基于1975年至2014年200个国家数据的预测,到2025年,全球肥胖患病率男性将达到18%,女性将达到21%(NCD Risk Factor Collaboration, 2016)。此外,肥胖不仅在发达国家日益流行,在发展中国家也日趋普遍(Maharani and Tampubolon, 2016)。因此,迫切需要制定预防肥胖及相关代谢性疾病并使其逆转的策略。

肥胖伴随着热量摄入与消耗之间的失衡(Hall and Guo, 2017)。有证据表明,脂肪组织参与能量代谢和脂肪质量的长期调节。脂肪组织是一种高度特化的组织,在能量动员调控中发挥关键作用(Reilly and Saltiel, 2017; Zhai et al., 2020)。迄今为止,在哺乳动物中已发现两种类型的脂肪组织,即白色脂肪组织(WAT)和棕色脂肪组织(BAT)(Cannon and Nedergaard, 2004; Wu et al., 2020)。WAT通常被认为是储存多余能量的器官,以脂滴中甘油三酯的形式储存能量。然而,最近在人WAT中发现了一种新型的棕色样脂肪细胞,称为米色/棕褐色脂肪细胞或可诱导棕色脂肪细胞(Sharp et al., 2012; Cypess et al., 2013; Lidell et al., 2013)。BAT能够快速消耗能量并产生热量,最初是在哺乳动物冬眠研究中发现的(Ricquier and Kader, 1976)。这一产热功能主要由解偶联蛋白-1(UCP1)介导,UCP1是一种存在于棕色脂肪细胞线粒体内膜中的多肽(Kajimura et al., 2015; Bertholet et al., 2017; Cannon et al., 2020)。通过耗氧量测定发现,肥胖小鼠的BAT产热降低(Martinez-Botas et al., 2000; Ussher et al., 2010)。几乎所有肥胖动物BAT中UCP1表达水平均下降,而在瘦动物中则升高(Shirkhani et al., 2018)。UCP1基因敲除(UCP1KO)小鼠在高脂饮食(HFD)喂养6个月后表现出肥胖表型,体脂增加(Kontani et al., 2005)。另一方面,寒冷刺激和/或β3-肾上腺素能受体激动剂处理可通过增强BAT活性来减少体脂量(Lowell and Spiegelman, 2000; Cannon and Nedergaard, 2004)。寒冷暴露还可增加BAT体积和活性,从而增加能量消耗并促进肥胖者体重减轻(Hanssen et al., 2015a,b; Leiria et al., 2019)。多项研究表明,人体中BAT活性/数量与体重指数(BMI)呈负相关。影像学数据显示,BMI较高的患者BAT活性较低(Cypess et al., 2009; Pfannenberg et al., 2010; Ouellet et al., 2011)。此外,在人类中发现了UCP1基因多态性中-3826A到G的单核苷酸替换,其降低了Ucp1的mRNA表达并加剧了年龄相关性肥胖和BAT退化(Nagai et al., 2007; Yoneshiro et al., 2013)。因此,BAT可能在体重和能量稳态调节中发挥关键作用。

经典棕色脂肪主要分布于肩胛间BAT(iBAT)、腋窝、椎旁和肾周部位(Park et al., 2014)。棕色脂肪细胞富含线粒体和多层脂滴,使其具有以热量形式耗散能量的卓越能力(Song et al., 2020)。UCP1在棕色脂肪细胞线粒体内膜上表达,使ATP合成与氧化磷酸化解偶联,从而以热量形式耗散能量。众所周知,BAT非颤抖性产热直接受交感神经系统神经支配和激活的控制。BAT产热由交感神经末梢释放的去甲肾上腺素及其随后与β3-肾上腺素能受体的结合所诱导和调节(Nedergaard et al., 2005; Feldmann et al., 2009)。多项研究揭示了UCP1的激活机制,长链脂肪酸对H+转运至关重要(Fedorenko et al., 2012)。此外,另一种提出的机制是线粒体活性氧(ROS)的产生调控UCP1的亚磺酰化和产热(Chouchani et al., 2016)。然而,β3-肾上腺素能受体激活下游的产热信号通路仍未完全阐明。

米色脂肪细胞(UCP1阳性脂肪细胞)已知在人类WAT中被大量UCP1阴性脂肪细胞包围(Wu et al., 2012)。米色脂肪细胞可在短期寒冷刺激或β3-肾上腺素能受体激动剂处理后募集(Saito et al., 2020)。它们与棕色脂肪细胞非常相似,具有高UCP1表达和产热能力(Ye et al., 2013; Li et al., 2014)。在人类中,存在BAT阳性组(受试者在寒冷刺激下可检测到FDG摄取)和BAT阴性组(受试者FDG摄取不可检测)。急性寒冷暴露后,BAT阳性组的能量代谢高于BAT阴性组(Orava et al., 2011; Yoneshiro et al., 2011)。这些研究清楚地揭示了棕色和米色脂肪细胞在人类寒冷诱导产热中的关键作用。因此,调控棕色或米色脂肪细胞活性的方法可能是预防和治疗人类肥胖及相关代谢性疾病的潜在途径。

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## TRP通道

瞬时受体电位(TRP)离子通道是一大类钙离子通透性通道,其中大多数为非选择性阳离子通道(Montell and Rubin, 1989)。TRP通道包含六个跨膜结构域(TM1–TM6),TM5和TM6之间有一个孔环(Cao et al., 2013b; Liao et al., 2013; Paulsen et al., 2015; Huynh et al., 2016; Zubcevic et al., 2016)。根据其一级氨基酸序列,TRP通道超家族目前分为七个亚家族,包含27个通道:TRPV(香草素受体)、TRPC(经典型)、TRPM(黑色素抑制素)、TRPML(黏脂蛋白)、TRPN(NomPC)、TRPP(多囊蛋白)和TRPA(锚蛋白)(Ramsey et al., 2006; Wu et al., 2010; Gees et al., 2012)。TRP通道触发的主要信号通路基于通过通道的钙离子内流,导致细胞内Ca2+浓度([Ca2+]i)升高。大量研究表明,一些TRP通道在脂肪细胞中表达,并参与脂肪组织的能量代谢和炎症,提示TRP通道在人类肥胖治疗和预防中的潜在作用(Bishnoi et al., 2018; Uchida et al., 2018; Gao et al., 2019; Zhai et al., 2020)。在本综述中,我们将系统而简要地总结TRP通道在脂肪细胞产热调控中的作用,并更新最新研究进展。

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## TRPV1

TRPV1作为辣椒素("辣"辣椒中的辛辣成分)的受体而广为人知(Caterina et al., 1997)。TRPV1可被多种刺激激活,包括热(Cao et al., 2013a)、质子和辣椒素(Dhaka et al., 2009)。此外,TRPV1还可被大蒜、洋葱(Salazar et al., 2008)、黑胡椒(Okumura et al., 2010)中的某些化合物以及姜辣素(Iwasaki et al., 2006)等其他食物成分激活。据报道,TRPV1在WAT和BAT中均有表达(Bishnoi et al., 2013; Kida et al., 2016)。分化后的HB2棕色脂肪细胞中TRPV1表达水平高于前脂肪细胞(Kida et al., 2016)。此外,TRPV1的激活上调产热基因的表达,并在3T3-L1脂肪细胞中诱导"褐变"(图1;Baboota et al., 2014)。TRPV1在3T3-L1前脂肪细胞、小鼠脂肪组织和肥胖人脂肪组织中均有表达(Zhang et al., 2007)。膳食辣椒素可激活TRPV1,这一过程诱导钙离子内流并阻止3T3-L1细胞中的脂肪生成(Zhang et al., 2007),可能通过钙调磷酸酶途径发挥作用(Cioffi, 2007)。此外,膳食辣椒素处理可预防野生型(WT)小鼠体内HFD诱导的肥胖,但在TRPV1KO小鼠中无此效果(Zhang et al., 2007; Chen J. et al., 2015; Chen N. et al., 2015)。此外,TRPV1参与HFD诱导肥胖期间WAT中能量摄入和葡萄糖稳态的调节(Lee et al., 2015)。TRPV1缺失加剧了HFD和衰老相关的肥胖和胰岛素抵抗(Lee et al., 2015)。还有报道称,单酰甘油通过激活TRPV1上调棕色脂肪细胞中UCP1的表达水平,并抑制高脂高糖饮食小鼠内脏脂肪的积累(Iwasaki et al., 2011)。鱼油摄入以TRPV1依赖性方式诱导棕色和白色脂肪组织中UCP1的上调(Kim et al., 2015; Lund et al., 2018)。油酰乙醇胺作为一种新发现的TRPV1配体,也参与能量摄入和消耗、摄食行为和体重控制的调节(Laleh et al., 2019)。

人体研究表明,辣椒素摄入增强了有氧运动期间的脂肪氧化和能量代谢(Shin and Moritani, 2007)。持续食用辣椒可增加中年受试者的能量代谢(Ahuja et al., 2006)。辣椒素类物质(一种非辛辣的辣椒素类似物)持续服用1-3个月也可增加超重和肥胖受试者的能量消耗和脂肪氧化,并减少腹部脂肪(Inoue et al., 2007; Snitker et al., 2009)。此外,辣椒素和辣椒素类物质作为食品成分可增强BAT产热,从而减少人体脂肪量(Yoneshiro et al., 2012; Saito and Yoneshiro, 2013)。一项流行病学研究表明,东亚国家通过增加含辣椒素的热食消费降低了人类肥胖的患病率(Wahlqvist and Wattanapenpaiboon, 2001)。还有报道称,辣椒素注射可诱导肾上腺素分泌,而TRPV1KO小鼠中此效应显著减弱(Uchida et al., 2017)。辣椒素直接与胃肠道中的TRPV1结合,产生传入信号,随后传递至中枢神经系统的腹内侧下丘脑核,最终向WAT发送信号。这可促进β2-肾上腺素能受体和PRDM16蛋白的表达,从而促进米色脂肪细胞的生成,导致全身能量消耗增加(Ohyama et al., 2016; Saito et al., 2020)。绿茶中的儿茶素可能通过以与辣椒素相同的方式作用于胃肠道感觉神经元的TRPV1/TRPA1来激活和募集BAT(Mako et al., 2015)。此外,在小鼠中外用辣椒素乳膏可导致体重和脂肪组织重量减轻(Lee et al., 2013)。然而,外用辣椒素对肥胖的调节作用是否通过TRPV1激活中枢神经系统仍有待进一步研究。这些研究清楚地表明,靶向TRPV1并利用辣椒素及其类似物调节其活性可能是人类肥胖治疗和预防的有效方法,尽管TRPV1激活的抗肥胖作用可能不仅涉及脂肪组织,还涉及神经系统。

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## TRPV2

TRPV2最初被报道可被有害热激活,激活温度阈值高于52°C(Caterina et al., 1999),并发现可被多种化学物质激活,如2-氨基乙氧基二苯硼酸盐(2APB)和溶血磷脂酰胆碱(LPC)(Juvin et al., 2007; Monet et al., 2009)。TRPV2还被报道可被机械刺激和/或细胞肿胀激活(Muraki et al., 2003; Iwata et al., 2009)。TRPV2在WAT和BAT中均有表达(Sun et al., 2017a)。与TRPV1、TRPV3、TRPV4和TRPM8相比,TRPV2在小鼠棕色脂肪细胞中高表达(Sun et al., 2016a,b)。TRPV2在分化后的棕色脂肪细胞中在mRNA、蛋白和功能水平上表达上调(Sun et al., 2016b, 2017b)。原代TRPV2缺陷型(TRPV2KO)脂肪细胞显示参与线粒体氧化代谢的多个基因(如Ucp1和过氧化物酶体增殖物激活受体γ共激活因子1-α(Pgc1α))的mRNA水平降低。此外,TRPV2KO脂肪细胞对β-肾上腺素能受体激动剂异丙肾上腺素的反应降低,这可能是由于缺乏TRPV2介导的钙离子内流。这些结果表明,TRPV2介导的钙离子内流参与β-肾上腺素能受体激活时产热基因的诱导(图1)。TRPV2KO小鼠表现出冷不耐受,在4°C寒冷刺激下Ucp1 mRNA和蛋白的增加显著减小,但其活动量未发生变化。另一方面,TRPV2KO小鼠的交感神经活动未改变。TRPV2KO小鼠在给予β3-肾上腺素能受体激动剂BRL37344后,iBAT适应性产热受损。重要的是,TRPV2KO小鼠在HFD处理后体重和脂肪组织显著增加(Sun et al., 2016a)。在肥胖和糖尿病(db/db)小鼠中也观察到TRPV2的上调(Sun et al., 2016a, 2017a)。还有报道称,酸樱桃可能通过直接作用于脂肪组织并下调HFD诱导的TRPV1和TRPV2的mRNA表达来减轻脂肪生成(Cocci et al., 2021)。这些发现表明TRPV2可能有助于脂肪细胞产热。然而,有必要进一步检测TRPV2在人BAT中的表达和功能,并开发TRPV2的特异性配体。

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## TRPV3

TRPV3是TRPV亚家族的一个成员,与TRPV1和TRPV2不同。TRPV3以被约体温的无害温度激活而广为人知,但最初被超过50°C的高有害阈值激活(Liu and Qin, 2017)。TRPV3的化学激动剂包括樟脑、香芹酚、(-)-表儿茶素、2APB和内源性配体法尼基焦磷酸(Cheung et al., 2015; Broad et al., 2016)。TRPV3可与TRPV1形成异源多聚体通道(Cheng et al., 2012),也参与脂肪生成和HFD诱导肥胖的调节(Cheung et al., 2015)。据报道,TRPV3在BAT和WAT中表达(Bishnoi et al., 2013)。肥胖小鼠(包括HFD诱导肥胖小鼠、ob/ob和db/db小鼠)内脏脂肪组织中TRPV3的表达显著下调(Cheung et al., 2015; Sun et al., 2017a)。HFD喂养上调了孤束核和舌下神经核中TRPV3的表达,伴随阿黑皮素原表达降低,导致食物摄入增加和体重增加(Hu et al., 2011)。(-)-表儿茶素激活TRPV3可阻止3T3-L1前脂肪细胞中的脂肪生成,并发挥抗脂肪生成作用(Cheung et al., 2015)。此外,小檗碱通过靶向小鼠下丘脑中的TRPV1/TRPV3来减轻奥氮平诱导的肥胖(Singh et al., 2020)。这些研究表明,靶向TRPV3可能是治疗和预防肥胖的一种有前景的方法。然而,TRPV3的表达及其在人类肥胖中的作用有待进一步探索。

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## TRPV4

TRPV4被报道可被渗透压变化或机械刺激激活(Liedtke et al., 2000; Strotmann et al., 2000; Watanabe et al., 2002a)。TRPV4还可被多种化学化合物激活,包括合成佛波酯4α-佛波醇-12,13-二癸酸酯(4α-PDD)和GSK1016790A(Watanabe et al., 2002b; Willette et al, 2008),以及中等温暖(温度阈值高于27°C)(Guler et al., 2002; Watanabe et al., 2002b)。TRPV4在BAT和WAT中均有表达(Sun et al., 2017a, 2020; Uchida et al., 2018)。据报道,胰岛素调控人白色脂肪细胞中TRPV4介导的代谢稳态(Sanchez et al., 2016)。TRPV4参与脂肪组织中产热和炎症通路的调节。TRPV4的敲低增强了Pgc1a和Ucp1基础表达和去甲肾上腺素诱导的表达(Ye et al., 2012)。据报道,ERK1/2被TRPV4介导的钙信号激活(Thodeti et al., 2009),TRPV4激活诱导的钙离子内流引起ERK1/2和JNK1/2的快速磷酸化,进一步抑制3T3-F442A脂肪细胞中产热基因的表达(图1;Ye et al., 2012)。TRPV4的敲低还通过抑制多种促炎基因来减少脂肪组织炎症(Ye et al., 2012)。WAT中TRPV4的表达高于BAT(Sun et al., 2017a)。TRPV4抑制后产热基因表达显著上调,导致WAT"褐变"的发生(Ye et al., 2012)。TRPV4KO小鼠表现出肌肉能量氧化增加和对HFD诱导肥胖的抗性(Kusudo et al., 2012)。还有报道称,跑步机和芦丁通过抑制脂肪细胞中TRPV4的表达来改善小鼠HFD诱导的肥胖(Chen N. et al., 2015)。此外,肥胖母体的饮食干预可保护雄性后代免受WAT中TRPV4诱导、肥胖和高胰岛素血症的影响(Janoschek et al., 2016)。一项基于人体的研究揭示了TRPV4基因多态性影响台湾受试者的BMI和体脂量(Duan et al., 2015)。这些结果表明,TRPV4在脂肪细胞产热调节中发挥相反的作用,但其潜在机制尚不清楚。检测TRPV4在人类肥胖中的表达和功能,并开发TRPV4特异性拮抗剂以及新化合物的体内研究是必要的。

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## TRPA1

TRPA1最初被报道为一种有害冷激活通道,温度阈值约为17°C(Story et al., 2003)。然而,后续研究围绕TRPA1作为冷感受器的角色展开了激烈争论。其冷敏感性后来受到质疑,TRPA1对冷感知的贡献目前仍存在激烈争议(Bautista et al., 2006; Talavera et al., 2020)。TRPA1可被多种食物成分潜在激活,如异硫氰酸烯丙酯、icilin、薄荷醇、肉桂醛和辣椒素类物质(Laursen et al., 2015)。TRPA1参与脂肪细胞产热和能量代谢(Watanabe and Terada, 2015)。在HFD诱导肥胖小鼠中,口服异硫氰酸烯丙酯可减少体重、肝脏中脂滴积累和白色脂肪细胞大小(Lo et al., 2018)。据报道,肉桂醛通过刺激肩胛间BAT减少HFD处理小鼠的内脏脂肪沉积(Tamura et al., 2012)。肉桂醛激活小鼠胃上皮细胞中的TRPA1,并上调脂肪组织中脂肪酸氧化相关基因(Camacho et al., 2015)。橄榄苦苷作为TRPA1和TRPV1的激动剂,通过促进去甲肾上腺素分泌来增强BAT中UCP1的表达并促进脂肪产热(Oi-Kano et al., 2016)。据推测,薄荷醇诱导的脂肪细胞产热可能也涉及TRPA1机制(Sakellariou et al., 2016)。此外,TRPA1激活诱导肾上腺素分泌,并防止啮齿动物脂肪积累和肥胖(Watanabe and Terada, 2015)。静脉注射AITC可诱导肾上腺素分泌,肾上腺素通过激活β3-肾上腺素能受体促进BAT产热(Saito et al., 2020)。这些研究表明,TRPA1通过中枢神经系统调节BAT的产热(Zsombok and Derbenev, 2016)。因此,通过其配体激活TRPA1可能是人类肥胖治疗和预防的有前景的方法。然而,TRPA1及其配体参与的抗肥胖机制有待进一步探索。在肥胖患者中进行TRPA1激活的随机临床试验也是必要的。

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## TRPM8

TRPM亚家族包含八个不同的亚基,TRPM1至TRPM8(Boesmans et al., 2011)。TRPM8作为薄荷醇受体而广为人知,于2002年被报道(McKemy et al., 2002)。在人类脂肪细胞系中,薄荷醇诱导的TRPM8激活增加了UCP1表达、线粒体激活和产热(图1;Rossato et al., 2014)。分化后的脂肪细胞中TRPM8的mRNA和蛋白表达水平显著增加,提示TRPM8对脂肪细胞产热的重要性(Rossato et al., 2014)。在培养的脂肪细胞中,薄荷醇诱导UCP1表达上调,可能通过蛋白激酶A途径,随后增加BAT产热和WAT"褐变"(Ma et al., 2012; Jiang et al., 2017; Sanders et al., 2020)。此外,据报道,冷感受器TRPM8通道参与BAT产热中时钟基因和时钟控制基因的调节(Moraes et al., 2017)。生物可利用的薄荷醇在分化中的脂肪细胞中诱导能量消耗表型(Khare et al., 2019)。体内研究表明,膳食补充薄荷醇可显著增加WT小鼠的核心体温和运动活动,但在TRPM8KO和UCP1KO小鼠中无此效果。膳食中的薄荷醇补充还可减轻HFD诱导的肥胖和胰岛素抵抗(Ma et al., 2012; Jiang et al., 2017)。薄荷醇对HFD诱导肥胖及相关并发症的预防作用可能涉及胰高血糖素机制(Khare et al., 2018)。这些结果表明,激活TRPM8可增强BAT产热,为肥胖的治疗和预防开辟了新途径。薄荷醇处理或寒冷暴露后TRPM8依赖性的核心体温升高可能通过UCP1上调介导(Tajino et al., 2011)。胃内给予薄荷醇也可在体内增强BAT产热(Tajino et al., 2007; Masamoto et al., 2009)。此外,据报道TRPM8多态性与土耳其人群中的代谢综合征密切相关(Tabur et al., 2015)。外用薄荷醇似乎可增加成人的核心体温和代谢率(Valente et al., 2015)。总之,通过其配体(如薄荷醇和icilin)激活TRPM8可模拟脂肪细胞产热,可能构成预防超重和肥胖的有前景的方法。然而,在肥胖患者中进行外用薄荷醇的随机临床试验是必要的。

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## TRPC1和TRPC5

TRPC亚家族包含七个成员(TRPC1-7)。TRPC通道通常由同源或异源TRPC蛋白形成(Huang et al., 2011)。迄今为止,没有证据表明TRPC通道具有温度敏感特性。TRPC1在脂肪库(包括BAT)中高表达,TRPC1缺陷小鼠易出现体重增加和代谢控制降低(Wolfrum et al., 2018)。TRPC1以PPARγ依赖性方式调节棕色脂肪细胞活性,提示TRPC1是将代谢或环境刺激转化为BAT活动输出的机制的一个下游组分(图1;Wolfrum et al., 2018)。然而,有相反的报道指出,喂食HFD(45%脂肪)的TRPC1KO小鼠的脂肪量和空腹葡萄糖浓度较低(Krout et al., 2017)。此外,在白色脂肪细胞中观察到了机械激活的TRPC1样电流(El Hachmane and Olofsson, 2018)。据报道,体外敲低TRPC1/TRPC5或体内条件性敲除TRPC5均可增加小鼠的脂联素生成(Sukumar et al., 2012)。此外,外源性和内源性垂体腺苷酸环化酶激活多肽通过激活TRPC1和TRPC5通道来刺激阿黑皮素原神经元并增加能量消耗,这表明可能通过在中枢神经系统中激活TRPC1/TRPC5来促进BAT产热(Chang et al., 2020)。这些研究表明TRPC1/TRPC5参与能量稳态的调节。需要进一步检测TRPC1/TRPC5在人脂肪组织中的表达,并开发TRPC1/TRPC5特异性激动剂。

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## 总结与展望

TRP通道已被证明在能量代谢的调节中发挥关键作用,对人类肥胖的治疗和预防具有重要意义。在本综述中,我们总结并更新了TRP通道参与脂肪细胞产热的最新研究进展。值得注意的是,仍有几个问题需要进一步探索。首先,TRP通道介导的脂肪细胞产热过程的潜在机制仍存在争议,需要明确阐明。其次,由于目前尚无TRP通道的特异性配体,有必要开发新型特异性配体。第三,TRP通道如何在脂肪组织中发挥组织特异性作用?这些问题有待未来通过进一步的动物和临床研究来解决。总之,靶向TRP通道可能是临床治疗和预防人类肥胖及相关代谢性疾病的有前景的策略。