The role of organic anion transport protein 1a4 in drug delivery and diseases: a review

✅ 全文

有机阴离子转运蛋白1a4在药物递送和疾病中的作用:综述

作者 Daopeng Tan; Jianmei Wang; Qianru ZHANG; Lin Qin; Yuhe Wang; Yuqi He 期刊 Food Science and Technology 发表日期 2023 ISSN 0101-2061 DOI 10.1590/fst.114122 类型 原创研究 (Original Research)

📄 英文摘要 English Abstract

EN

OATP1A4 is an important member of the family of organic anion transporting polypeptides (OATPs), which is generally thought to mediate cellular uptake of endogenous and exogenous substances, such as bile acids, drugs and environmental toxins. Recent studies have found that Oatp1a4 plays an important role in drug passage through the blood-brain barrier and is expected to be an important target for drugs used to treat central diseases. Oatp1a4 has also been associated with various diseases such as cholestasis. differences in Oatp1a4 across age and sex have also become an area of concern for precision drug administration. Therefore, this paper presents a systematic review of Oatp1a4 expression in drug transport and various physiopathological states.

📄 中文摘要 Chinese Abstract

中文
有机阴离子转运多肽(OATPs)是一类属于溶质载体(SLC)家族的膜转运蛋白,介导包括胆汁酸、激素和多种药物在内的大量底物的摄取。OATPs在肝脏、肾脏、脑和肠道等多个人体器官中均有表达。在临床相关性方面,OATPs蛋白常作为药物穿越血脑屏障的关键转运载体。有机阴离子转运蛋白1a4(Oatp1a4)是Oatp家族的原型成员,该家族为高度同源的转运蛋白,在肝脏、大脑和骨骼肌纤维中高表达。Oatp1a4在药物穿越血脑屏障过程中的重要作用已引起越来越多的关注。此外,Oatp1a4还与多种疾病相关。因此,本文系统综述了Oatp1a4在给药、胆汁淤积及其他病理状态中的表达情况,以及其在性别和年龄方面的差异。

📋 英文结构化总结 English Structured Summary

全文整理

EN

Background:

Organic anion transporting polypeptides (OATPs) are a class of membrane transporters belonging to the solute carrier (SLC) family that mediate the uptake of a large number of substrates including bile acids, hormones and many drugs. OATPs have been found to be expressed in several human organs including liver, kidney, brain and intestine. In terms of clinical relevance, OATPs proteins often serve as key transport vehicles for drugs across the blood-brain barrier. Organic anion transport protein 1a4 (Oatp1a4) is the prototypical member of the Oatp family of highly homologous transport proteins that are highly expressed in the liver, cerebrum, and skeletal myofibres. The important role of Oatp1a4 in the passage of drugs through the blood-brain barrier has attracted increasing attention. In addition to this, Oatp1a4 has been associated with a variety of diseases. Therefore, this paper systematically reviewed the expression of Oatp1a4 in drug administration, cholestasis and other pathological states and its differences across gender and age.

Methods:

N/A - Review article

Results:

Oatp1a4 is female-predominant in mouse livers; hepatic Oatp1a4 mRNA levels were decreased by both androgens and male-pattern growth hormone administration. Oatp1a4 mRNA expression is age-dependent: it was hardly detectable in fetal rat livers, low at birth, rapidly increased after weaning (21 d), reached the peak at 60 d, remained stable between 60-180 d, and decreased at elderly (540 and/or 800 d). In the hyperthyroid state, the expression of Oatp1a4 is downregulated. High sucrose diet could reduce gene expressions of Oatp1a4. Calorie restriction could increase the female-predominantly expressed Oatp1a4. Maternal consumption of a high-fat diet during pregnancy and lactation increases the mRNA expression of Oatp1a4. Inosine induces a decrease in Oatp1a4 expression levels through activation of xanthine oxidase induced oxidative stress. It has been demonstrated that the transport activity of Oatp1a4 in the BBB is directly regulated by TGF-β/ALK1 signaling. Statins, as transport substrates for Oatp1a4, improve functional neurological outcomes in patients. Pitavastatin, rosuvastatin, pravastatin, taurolite, digoxin, ochratoxin A and [d-penicillamine (2,5)]-enkephalin are currently identified as substrates of Oatp1a4.

Data Summary:

Oatp1a4 mRNA expression in rat livers peaked at 60 days of age, remained stable from 60 to 180 days, and decreased at 540 and/or 800 days. In mouse livers, Oatp1a4 is female-predominant. Hepatic Oatp1a4 mRNA levels were decreased by both androgens and male-pattern growth hormone administration. High sucrose diet reduced gene expressions of Oatp1a4, while calorie restriction increased the female-predominantly expressed Oatp1a4. In the hyperthyroid state, Oatp1a4 expression is downregulated.

Conclusions:

OATP1A4 plays an important role in drug passage through the blood-brain barrier and is expected to be an important target for drugs used to treat central diseases. Oatp1a4 has also been associated with various diseases such as cholestasis. Differences in Oatp1a4 across age and sex have become an area of concern for precision drug administration. This paper systematically reviewed the expression of Oatp1a4 in drug administration, cholestasis and other pathological states and its differences across gender and age.

Practical Significance:

Important for precise clinical use of drugs. The aim is to bring more attention to the role of Oatp1a4 in the precise clinical administration of medications.

📋 中文结构化总结 Chinese Structured Summary

中文

背景:

有机阴离子转运多肽(OATPs)是一类属于溶质载体(SLC)家族的膜转运蛋白,介导包括胆汁酸、激素和多种药物在内的大量底物的摄取。OATPs在肝脏、肾脏、脑和肠道等多个人体器官中均有表达。在临床相关性方面,OATPs蛋白常作为药物穿越血脑屏障的关键转运载体。有机阴离子转运蛋白1a4(Oatp1a4)是Oatp家族的原型成员,该家族为高度同源的转运蛋白,在肝脏、大脑和骨骼肌纤维中高表达。Oatp1a4在药物穿越血脑屏障过程中的重要作用已引起越来越多的关注。此外,Oatp1a4还与多种疾病相关。因此,本文系统综述了Oatp1a4在给药、胆汁淤积及其他病理状态中的表达情况,以及其在性别和年龄方面的差异。

方法:

不适用——综述类文章

结果:

Oatp1a4在小鼠肝脏中呈雌性优势表达;雄激素和雄性模式生长激素给药均可降低肝脏Oatp1a4 mRNA水平。Oatp1a4 mRNA表达具有年龄依赖性:在胎鼠肝脏中几乎检测不到,出生时水平较低,断奶后(21天)迅速增加,60天时达到峰值,60至180天保持稳定,老年期(540天和/或800天)则下降。在甲状腺功能亢进状态下,Oatp1a4表达下调。高蔗糖饮食可降低Oatp1a4的基因表达。热量限制可增加雌性优势表达的Oatp1a4。妊娠和哺乳期间母体摄入高脂饮食可增加Oatp1a4的mRNA表达。肌苷通过激活黄嘌呤氧化酶诱导的氧化应激导致Oatp1a4表达水平下降。研究表明,血脑屏障中Oatp1a4的转运活性直接受TGF-β/ALK1信号通路调控。他汀类药物作为Oatp1a4的转运底物,可改善患者的功能性神经学结局。匹伐他汀、瑞舒伐他汀、普伐他汀、牛磺胆酸、地高辛、赭曲霉毒素A和[d-青霉胺(2,5)]-脑啡肽目前已被鉴定为Oatp1a4的底物。

数据总结:

大鼠肝脏中Oatp1a4 mRNA表达在60日龄时达到峰值,60至180天保持稳定,540天和/或800天时下降。在小鼠肝脏中,Oatp1a4呈雌性优势表达。雄激素和雄性模式生长激素给药均可降低肝脏Oatp1a4 mRNA水平。高蔗糖饮食降低Oatp1a4的基因表达,而热量限制则增加雌性优势表达的Oatp1a4。在甲状腺功能亢进状态下,Oatp1a4表达下调。

结论:

OATP1A4在药物穿越血脑屏障过程中发挥重要作用,有望成为治疗中枢疾病药物的重要靶点。Oatp1a4还与胆汁淤积等多种疾病相关。Oatp1a4在年龄和性别方面的差异已成为精准给药领域关注的焦点。本文系统综述了Oatp1a4在给药、胆汁淤积及其他病理状态中的表达情况,以及其在性别和年龄方面的差异。

实际意义:

对临床精准用药具有重要意义。旨在引起更多对Oatp1a4在临床精准给药中作用的关注。

📖 英文全文 English Full Text

EN

a Review Article ISSN 0101-2061 (Print) ISSN 1678-457X (Online) Food Science and Technology DOI: https://doi.org/10.1590/fst.114122

The role of organic anion transport protein 1a4 in drug delivery and diseases: a review Daopeng TAN1 , Jianmei WANG1, Qianru ZHANG1, Lin QIN1, Yuhe WANG2, Yuqi HE1* Abstract OATP1A4 is an important member of the family of organic anion transporting polypeptides (OATPs), which is generally thought to mediate cellular uptake of endogenous and exogenous substances, such as bile acids, drugs and environmental toxins. Recent studies have found that Oatp1a4 plays an important role in drug passage through the blood-brain barrier and is expected to be an important target for drugs used to treat central diseases. Oatp1a4 has also been associated with various diseases such as cholestasis. differences in Oatp1a4 across age and sex have also become an area of concern for precision drug administration. Therefore, this paper presents a systematic review of Oatp1a4 expression in drug transport and various physiopathological states. Keywords: organic anion transporting polypeptides; OATP1A4; cholestasis; drug delivery. Practical Application: Important for precise clinical use of drugs.

Currently, foods have gained more attention in terms of their health support and disease-controlling ability(Kesika et al., 2022). Moreover, the study of organic anion transporters is becoming more and more important (Ji et al., 2022; Tan et al., 2022; Tao et al., 2023). Membrane transporters play an important role in the absorption, distribution and elimination of endogenous and xenobiotics, such as bile acids and drugs. Organic anion transporting polypeptides (OATPs) are a class of membrane transporters belonging to the solute carrier (SLC) family that mediate the uptake of a large number of substrates including bile acids, hormones and many drugs (Hagenbuch & Stieger, 2013). OATPs have been found to be expressed in several human organs including liver, kidney, brain and intestine (Gong et al., 2011; Hagenbuch & Stieger, 2013). In terms of clinical relevance, OATPs proteins often serve as key transport vehicles for drugs across the blood-brain barrier. On the other hand, the expression level of OATPs varies significantly across multiple disease states (Kim, 2003; Shitara et al., 2003). In recent years, much progress has been made in the identification of endogenous substrates of OATPs and their role in drug transport (Hagenbuch & Meier, 2004). Organic anion transport protein 1a4 (Oatp1a4) (Figure 1) is the prototypical member of the Oatp family of highly homologous transport proteins that are highly expressed in the liver, cerebrum (Imai et al., 2013), and skeletal myofibres (Sakamoto et al., 2008). The important role of Oatp1a4 in the passage of drugs through the blood-brain barrier has attracted increasing attention. In addition to this, Oatp1a4 has been associated with a variety of diseases. Therefore, this paper systematically reviewed the expression of Oatp1a4 in drug administration, cholestasis and other pathological states and its differences across gender and age. The aim is to bring more attention to the role of Oatp1a4 in the precise clinical administration of medications.

Organic anion transporting polypeptides (Oatps) play an important role in transporting endogenous substances and xenobiotics to the liver and have been implicated in drug-drug interactions. Many factors, such as gender, age and diet, may influence their expression, leading to altered drug disposition, efficacy and toxicity. Organic anion transport protein 1a4 (Oatp1a4) is the prototypical member of the Oatp family of highly homologous transport proteins that are highly expressed in the liver, cerebrum (Imai et al., 2013), and skeletal myofibres (Sakamoto et al., 2008), while its placental expression is low (St-Pierre et al., 2004). In mouse livers, Oatp1a4 is female-predominant. Hepatic Oatp1a4 mRNA levels were decreased by both androgens and male-pattern growth hormone administration (Cheng et al., 2006). This sex-specific difference is also reflected in the functional expression of blood-brain barrier Oatp1a4 (Brzica et al., 2018). And, Oatp1a4 mRNA expression is age-dependent. It was hardly detectable in fetal rat livers, low at birth, Subsequently, rapidly increased after weaning (21 d), and reached the peak at 60 d, and then remained stable during the age between 60-180 d, at last decreased at elderly (540 and/or 800 d) (Hou et al., 2014). In clinical practice thyroid dysfunction varies with age and asymptomatic manifestations of hyperthyroidism are frequently observed in the elderly. In the hyperthyroid state, the expression of Oatp1a4 is downregulated (Engels et al., 2015). In terms of diet, high sucrose diet could reduce gene expressions of Oatp1a4 (Zagorova et al., 2015). Calorie restriction (CR) is one of the most effective anti-aging interventions in mammals. CR could increase the female-predominantly expressed Oatp1a4 (Fu & Klaassen, 2014). Maternal consumption of a high-fat diet during pregnancy and lactation also increases the mRNA expression of Oatp1a4 (Tanaka et al., 2018). Inosine induces a decrease in Oatp1a4 expression levels through activation of xanthine oxidase

Received 01 Oct., 2022 Accepted 28 Nov., 2022 1 Guizhou Engineering Research Center of Industrial Key-technology for Dendrobium Nobile, School of Pharmacy, Zunyi Medical University, Zunyi, China 2 Department of Pharmacy, Affiliated Hospital of Zunyi Medical University, Zunyi, China *Corresponding author: yqhe.pharm@foxmail.com

Food Sci. Technol, Campinas, 43, e114122, 2023 1 The role of Oatp1a4 in drug delivery and diseases currently identified as substrates of Oatp1a4, which mediates the brain-blood and blood-brain transport of these drugs across the blood-brain barrier (Ose et al., 2010). There is growing evidence that bumetanide also affects brain disorders, including autism, neonatal seizures, and epilepsy; however, the low brain levels of bumetanide after systemic administration severely limit its clinical use in the treatment of brain disorders. In vivo experiments have shown that restricted passive diffusion and active efflux transport, mediated by Oat3 as well as the organic anion transporting peptide (Oatp) Oatp1a4 and multidrug resistance protein 4, explain the extremely low brain concentrations achieved after systemic administration of bumetanide (Römermann et al., 2017). Pain is a major symptom associated with inflammation, and inhibition of inflammatory pain by the anti-inflammatory drug diclofenac attenuates these changes in Oatp1a4 functional expression, suggesting that peripheral inflammation can modulate BBB transporters (Ronaldson et al., 2011). 3.2 Role of OATP1A4 in the blood retinal barrier and bloodarachnoid barrier Figure 1. The theoretical 3D structures of OATP1A4 from AlphaFold v2.0, a protein structure database (Jumper et al., 2021).

induced oxidative stress, as confirmed by experiments in rats fed inosine (Tsujimoto et al., 2013).

3 OATP1A4 and drug delivery 3.1 Role of OATP1A4 in the blood-brain barrier The physical and biochemical properties of the blood-brain barrier (BBB) make the treatment of central nervous system disorders extremely difficult because drug delivery to the central nervous system (CNS) is greatly limited by BBB. In contrast, drug delivery to CNS can be achieved by targeting drug uptake transporters such as Oatp1a4, which is also thought to be a strategy that could improve drug delivery to the brain. It has been demonstrated that the transport activity of Oatp1a4 in the BBB is directly regulated by TGF-β/ALK1 signaling, and this pathway could be a target for controlling the CNS delivery of OATP substrate drugs. (Abdullahi et al., 2018), and activation of activin receptor-like kinase (ALK)-1 using bone morphogenetic protein (BMP)-9 increased the expression of Oatp1a4 protein in rat brain microvessels in vivo (Abdullahi et al., 2017a). Hence, targeting Oatp1a4 regulation represents an opportunity to control Oatp1a4 functional expression for the purpose of delivering therapeutics to the CNS (Abdullahi et al., 2017b). The human ortholog of Oatp1a4 is OATP1A2. It has been shown that OATP1A2 could be regulated by transforming growth factor-β/activator receptor-like kinase 1 signaling in humans (Ronaldson et al., 2021). Statins (3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors), as transport substrates for Oatp1a4 improve functional neurological outcomes in patients, and Oatp1a4 statin uptake is higher in the cerebral cortex relative to the hippocampus and cerebellum (Betterton et al., 2022). Pitavastatin, rosuvastatin, pravastatin, taurolite, digoxin, ochratoxin A and [d-penicillamine (2,5)]-enkephalin are 2

Oatp1a4 is also expressed on the blood retinal barrier (BRB) and is a major determinant in controlling the entry of anionic drugs into the retina. Immunoblotting and immunohistochemical analysis showed that Oatp1a4 is localized to the apical membrane of the retinal pigment epithelium (Akanuma et al., 2013). Recent studies have shown that Oatp1a4 is involved in the transport of nutrients including riboflavin, L-ornithine, β-alanine and L-histidine in the blood to the retina, and pravastatin in the BRB (Kubo et al., 2018). In addition, there is evidence that Oatp1a4 plays a role in drug clearance at the blood-arachnoid barrier (BAB) and may play an important role in cerebrospinal fluid (CSF) detoxification in vivo by limiting the distribution of organic anions in the brain and spinal cord (Yaguchi et al., 2019). 3.3 Role of OATP1A4 in hepatic metabolism It is well known that the liver is the primary organ of drug metabolism and organic anion transporting peptides (Oatps) are involved in hepatic transport of a variety of organic anion compounds and drugs. The function of Oatp1a4 dominates the region around the central hepatic vein (CV) in the rat liver lobules and can affect the distribution of substrates such as sulfanilamide-101, digoxin, quinine, d-verapamil, 17betaestradiol-d-17beta-glucuronide in the liver (Akanuma et al., 2019). It has been shown that Oatp1a4 plays a major role in the hepatic accumulation of cardiac glycosides (Takano et al., 2018) and berberine (Chen et al., 2015) in mice. Oatp1a4 plays a major role in the hepatic uptake of beta-lactam antibiotics in humans, and probably corresponds functionally to OATP1B3 in rat liver (Nakakariya et al., 2008). Eprosartan is an angiotensin II receptor antagonist used clinically for the treatment of hypertension and heart failure. It is transported by multiple Oatps (at least Oatp1a1 and Oatp1a4)/Mrp2 in rats and at least OATP1B1/MRP2 in humans (Sun et al., 2014). Adverse effects of statins are usually the result of drug-drug interactions, and inhibition of the Oatp1a4 and Oatp1b2 transporters in rats by the bacteriostatic agent fusidic acid (FA) may attenuate hepatic uptake of statins, leading to increased blood and tissue Food Sci. Technol, Campinas, 43, e114122, 2023

concentrations that exhibit musculoskeletal toxicity (Eng et al., 2016). Pregnenolone-16alpha-carbonitrile (PCN) regulates the expression of some transporters, namely, Oatp1a4 and Mrp3 in liver via a PXR-mediated mechanism (Cheng & Klaassen, 2006). 3.4 Role of OATP1A4 in drug metabolism The liver plays an important role in the in vivo metabolism of drugs, and hepatic transporters contribute to the absorption and excretion of drugs by the liver. Dexamethasone (DEX) treatment of hepatocytes increased the expression of CYP3A1/2, Oatp1a4 and Mrp2 and decreased the expression of Ntcp (Turncliff et al., 2004). Hepatotoxicity of methotrexate was increased during human treatment with dexamethasone, and analysis of methotrexate transporter expression in the liver showed upregulation of Mrp2, Oatp1a4, and Oat2 and downregulation of Mrp3, resulting in reduced biliary elimination and leading to increased hepatotoxicity of the drug in combination with DEX (Fuksa et al., 2010). Perfluorooctanoic acid (PFOA) and perfluorodecanoic acid (PFDA) have been detected in wildlife and humans worldwide. Both play a central role in the downregulation of Oatp1a1, 1a4, 1b2 (Cheng & Klaassen, 2008). Perinatal exposure to polybrominated diphenyl ethers (PBDEs) resulted in increased expression of the hepatic efflux transporters Mdr1 (multidrug resistance), Mrp2 (multidrug resistanceassociated protein) and Mrp3, and the influx transporter Oatp1a4 mRNA (Szabo et al., 2009). Patients with nonalcoholic steatohepatitis (NASH) may ingest both Pravastatin and herbs containing Evodiamine, which upregulates the expression of Oatp1a1, Oatp1a4, and Oatp1b2 decreasing systemic exposure to pravastatin (Liang et al., 2022). The regulation of Oatp1a4 by some natural products or traditional drugs has also been reported. For example, monoammonium glycyrrhizinate (MAG) increases the expression of Oatp1a4 and decreases the expression of Mrp2 (Zhou et al., 2016). Silymarin inhibits protein expression of Oatp1b2 and Oatp1a4 in NASH animals (Lynch et al., 2021). Imperatorin, Isoimperatorin, and Angelica dahurica extract can significantly downregulate the expression of Oatp1a1, Oatp1a4, and Oatp1b2 of liver in mice (Wang et al., 2020).

4 OATP1A4 and diseases 4.1 OATP1A4 and cholestasis Cholestasis is caused by interruption of bile flow and is associated with a variety of liver diseases, and organic anion transporting polypeptides are thought to have an important function in bile acid (BA) transport. Uptake of conjugated BA by the liver may be affected by down-regulation of OATP1A1 and up-regulation of OATP1A4 (Slijepcevic et al., 2015). Oatp1a4 is upregulated in the liver and downregulated in the intestine after bile duct ligation (BDL) in rats (Giroux et al., 2022), this seems to indicate that Oatp1a4 is associated with cholestasis. However, in another experiment using 7-day BDL-induced liver dysfunction rats, it was found that the mRNA expression of Oatp1a4 in the liver of BDL rats decreased to 84.8% of that of normal rats (Horiuchi et al., 2009). This difference in results may be caused by the experimental period. Under lipopolysaccharide (LPS)-induced inflammatory conditions, lymphocyte deficiency altered basal and inflammatory IL-6 mRNA expression, and Food Sci. Technol, Campinas, 43, e114122, 2023

B-cell deletion also attenuated Oatp1a4 mRNA in LPS-treated mice, suggesting that IL-6 signaling may play a critical role (Bodeman et al., 2013). Reverse regulation of Mrp2 and 3 is thought to represent an adaptive response to cholestatic injury in hepatocytes, and downregulation of periportal Ntcp and induction of Oatp1a4 and Oatp1b2 in BDL and LPS-treated rat livers may be adaptive mechanisms for reducing cholestatic injury in hepatocytes with profound downregulation of Bsep and Mrp2 (Donner et al., 2007). Mice deficient in ATP11C are characterized by conjugated hyperbilirubinemia and nonconjugated hyperbilirubinemia, and the basal bile salt uptake transporters OATP1B2, OATP1A1, OATP1A4, and Ntcp are almost absent in central hepatocytes of ATP11C-deficient livers with features very similar to those of Rotor syndrome, suggesting that deletion of OATP expression potentially predisposes to Rotor syndrome (de Waart et al., 2016). Transmembrane protein 30A (TMEM30A) is a β-subunit essential for the function of ATP11C, and the expression and membrane localization of ATP11C were significantly reduced in Tmem30a LKO mice, which correlated with impaired expression and localization of BS transporters, such as OATP1A4, OATP1B2, NTCP, BSEP, and MRP2, suggesting that TMEM30A deficiency can lead to intrahepatic cholestasis in mice by impairing the expression and localization of BS transporters and associated nuclear receptors (Liu et al., 2017). Alpha-naphthyl isothiocyanate (ANIT) is a hepatotoxic agent that causes acute intrahepatic cholestasis in rodents. In an experiment on the effect of Dioscin on ANIT-induced cholestasis, it was found that co-treatment of ANIT with Dioscin prevented adaptive downregulation of Oatp1a1, 1b2 and promoted upregulation of Oatp1a4, multidrug resistance-associated protein (Mrp)2 and bile salt export pump (Bsep), suggesting that Dioscin may prevent hepatic transporters by restoring expression to prevent impairment of liver function (A. Zhang et al., 2016). Pyrazinamide (PZA), a first-line drug for the treatment of tuberculosis, causes severe hepatotoxicity, with a 2-fold increase in serum levels of both ALT and AST, a 10-fold increase in total bile acids in serum, and significantly altered mRNA and protein expression of bile acid synthesis and transporters in PZA-treated rats, with FXR, Bsep, Mrp2, Mdr2, Ostα/β Oatp1a1, Oatp1b2, and Cyp8b1 were decreased, whereas Mrp3, Ntcp, Oatp1a4, and Cyp7a1 were increased (Guo et al., 2016). In estrogen-induced cholestasis, ethinylestradiol significantly increased cholestasis markers, decreased bile bile acid excretion, downregulated hepatocyte transporters (Ntcp/Oatp1b2/Oatp1a4/Mrp2), and upregulated Mrp3 (Muchova et al., 2015). Yinchenhao Decoction (YCHD) is a famous traditional Chinese formula used for treating cholestasis. The cholestatic effect of YCHD was shown to be related to its modulating effect on the expression of metabolic enzymes and transporters in cholestatic liver. YCHD significantly increased the expression of UGT1A1, bile salt export pump, MRP2 and OATP1A4 in cholestatic rats, which had a great ameliorating effect on cholestasis (Yi et al., 2018). In an experiment using Oatp1a4-deficient mice to investigate the physiological role of Oatp1a4 in BA homeostasis, female Oatp1a4-deficient mice showed no significant alterations in BA concentrations in serum or liver, whereas male Oatp1a4 null mice showed significant alterations in BA homeostasis, including increased concentrations of deoxycholic acid (DCA) in serum, liver, and 3

intestinal contents. Loss of Oatp1a4 function did not reduce BA accumulation in serum or liver of bile duct ligated mice, suggesting that Oatp1a4 is unlikely to be an uptake transporter of BA, but plays an important role in secondary BA metabolism in male mice (Zhang et al., 2013). 4.2 OATP1A4 and liver diseases The liver plays an important role in the detoxification process of foreign bodies. Hepatobiliary transporters contribute to the process of foreign body uptake and elimination by the liver. The expression of these transporters may be regulated in the presence of liver failure. For example, in the Long-Evans Cinnamon (LEC) rat hepatitis model, decreased hepatic expression of three sinusoidal organic anion transporters, Ntcp, Oatp1a1 and Oatp1a4, was found (Chiba et al., 2007). Progressive loss of mRNA for transporters such as Ntcp, Bsep, Mrp2, Oatp1/Oatp1a1, Oatp2/Oatp1a4 and Oatp4/Oatp1b2 was found at 20 and 32 weeks of hepatocarcinogenesis in rats (Monte et al., 2005). Plasma bilirubin levels are increased in rat models and in patients with alcoholic liver disease (ALD). The constructive androgen receptor (CAR) is a known xenobiotic receptor that induces bilirubin detoxification and transport, and the CAR agonist phenobarbital (PB) downregulates bilirubin levels in the serum of ALD patients, while selectively upregulating the expression levels of OATP1A1, OATP1A4, UGT1A1 and MRP2 (Wang et al., 2017). After hepatic ischemia-reperfusion (IR) injury, mRNA expression of hepatic transporter Oatp1a1, Oatp1a4, Oatp1b2, Ntcp, Mdr2 and Bsep decreased, while Mdr1b expression increased (Tanaka et al., 2006). Chronic high-fat diets are a key factor in obesity, and progressive obesity can subsequently lead to liver damage, kidney damage, and intestinal atrophy. Transporters expressed in the liver, kidney and intestine play an important role in the deposition of nutrients and drugs. In HFD-fed mice, the relative expression of Oat2 was increased 4.08-fold and the protein expression of Oat2 was upregulated at 24 weeks, while the mRNA expression of the uptake transporters Oct1, Oatp1b2 and Oatp1a4 was decreased by 79%, 61% and 19%, respectively (Lu et al., 2019). 4.3 OATP1A4 and other diseases Autosomal dominant polycystic kidney disease (ADPKD), a common form of hereditary polycystic kidney disease (PKD), is a major cause of renal failure, and increased bile acids in the liver of rats with polycystic kidney (PCK) are associated with decreased protein expression of Mrp2 and Oatp1a4 in the liver (Bezençon et al., 2019). Protein expression of Oatp1a4 is significantly decreased in the brains of Alzheimer’s disease (AD) mice (Wen et al., 2021). Cerebral hypoxia/reoxygenation stress (H/R) is a component of multiple diseases, and in rat brain microvessels, Oatp1a4 expression is significantly increased under H/R conditions (Thompson et al., 2014). Elevated systemic levels of cytokines in rheumatoid arthritis (RA) can alter the expression of metabolic enzymes and transporters. mRNA levels of intestinal Cyp3a1 and hepatic Cyp2c6, Cyp2c7, Cyp3a1, Oatp1a1, Oatp1b2, Oatp1a4 and Mrp2 were significantly decreased in rats with collagen-induced arthritis (CIA) (Lin et al., 2017). 4

5 Conclusions Oatp1a4 is an important Organic anion transporting polypeptides which is known to play an important role in the transport of many drugs and endogenous substances, such as bile acids. Oatp1a4 is highly expressed in the liver and brain and has significant age and sex differences, which are important for individualized drug effectiveness and safety. And significantly altered expression of Oatp1a4 has been found in a variety of diseases. However, the regulatory mechanisms of Oatp1a4 are still poorly understood and more studies are needed to elucidate them.

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# 有机阴离子转运蛋白1a4在药物递送和疾病中的作用:综述

**谭道鹏1**,**王健梅1**,**张倩茹1**,**林琴1**,**王宇鹤2**,**何玉琪1\***

**摘要**

OATP1A4是有机阴离子转运多肽(OATP)家族的重要成员,通常被认为介导内源性和外源性物质(如胆汁酸、药物和环境毒素)的细胞摄取。近期研究发现,Oatp1a4在药物通过血脑屏障的过程中发挥重要作用,有望成为治疗中枢神经系统疾病药物的重要靶点。Oatp1a4还与胆汁淤积等多种疾病相关。Oatp1a4在年龄和性别方面的差异也成为精准给药领域关注的重点。因此,本文系统综述了Oatp1a4在药物转运及多种生理病理状态下的表达情况。

**关键词:** 有机阴离子转运多肽;OATP1A4;胆汁淤积;药物递送

**实际应用:** 对临床精准用药具有重要意义。

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目前,食品在健康支持和疾病控制能力方面受到了更多关注(Kesika等,2022)。此外,有机阴离子转运蛋白的研究正变得越来越重要(Ji等,2022;Tan等,2022;Tao等,2023)。膜转运蛋白在内源性和外源性物质(如胆汁酸和药物)的吸收、分布和消除过程中发挥着重要作用。有机阴离子转运多肽(OATPs)是一类属于溶质载体(SLC)家族的膜转运蛋白,可介导包括胆汁酸、激素和多种药物在内的大量底物的摄取(Hagenbuch & Stieger,2013)。OATPs在肝脏、肾脏、大脑和肠道等多个器官中均有表达(Gong等,2011;Hagenbuch & Stieger,2013)。在临床相关性方面,OATP蛋白通常是药物穿越血脑屏障的关键转运载体。另一方面,OATPs的表达水平在多种疾病状态下存在显著差异(Kim,2003;Shitara等,2003)。近年来,OATP内源性底物的鉴定及其在药物转运中的作用研究取得了很大进展(Hagenbuch & Meier,2004)。

有机阴离子转运蛋白1a4(Oatp1a4)(图1)是Oatp家族的原型成员,该家族是一组高度同源的转运蛋白,在肝脏、大脑(Imai等,2013)和骨骼肌纤维(Sakamoto等,2008)中高表达。Oatp1a4在药物穿越血脑屏障过程中的重要作用引起了越来越多的关注。此外,Oatp1a4还与多种疾病相关。因此,本文系统综述了Oatp1a4在给药、胆汁淤积及其他病理状态下的表达情况及其在性别和年龄方面的差异,旨在引起人们对Oatp1a4在临床精准用药中作用的更多关注。

有机阴离子转运多肽(Oatps)在将内源性和外源性物质转运至肝脏的过程中发挥重要作用,并参与药物-药物相互作用。性别、年龄和饮食等多种因素可能影响其表达,从而导致药物分布、疗效和毒性的改变。有机阴离子转运蛋白1a4(Oatp1a4)是Oatp家族的原型成员,在肝脏、大脑(Imai等,2013)和骨骼肌纤维(Sakamoto等,2008)中高表达,而在胎盘中的表达较低(St-Pierre等,2004)。在小鼠肝脏中,Oatp1a4呈雌性优势表达。雄激素和雄性模式生长激素的给予均可降低肝脏Oatp1a4 mRNA水平(Cheng等,2006)。这种性别特异性差异也反映在血脑屏障Oatp1a4的功能表达中(Brzica等,2018)。此外,Oatp1a4 mRNA的表达具有年龄依赖性。在胎鼠肝脏中几乎检测不到,出生时表达水平较低,断奶后(21天)迅速增加,60天时达到峰值,随后在60-180天之间保持稳定,最终在老年期(540天和/或800天)下降(Hou等,2014)。

在临床实践中,甲状腺功能异常随年龄变化,老年人群中甲状腺功能亢进的无症状表现较为常见。在甲状腺功能亢进状态下,Oatp1a4的表达下调(Engels等,2015)。在饮食方面,高蔗糖饮食可降低Oatp1a4的基因表达(Zagorova等,2015)。热量限制(CR)是哺乳动物中最有效的抗衰老干预措施之一。CR可增加雌性优势表达的Oatp1a4(Fu & Klaassen,2014)。妊娠期和哺乳期母体摄入高脂饮食也会增加Oatp1a4的mRNA表达(Tanana等,2018)。肌苷通过激活黄嘌呤氧化酶诱导的氧化应激降低Oatp1a4的表达水平,这一现象已在喂食肌苷的大鼠实验中得到证实(Tsujimoto等,2013)。

## 3 OATP1A4与药物递送

### 3.1 OATP1A4在血脑屏障中的作用

血脑屏障(BBB)的物理和生化特性使中枢神经系统疾病的治疗极为困难,因为药物向中枢神经系统(CNS)的递送受到BBB的极大限制。相反,通过靶向Oatp1a4等药物摄取转运蛋白可以实现向CNS的药物递送,这被认为是一种改善药物向大脑递送的策略。研究表明,BBB中Oatp1a4的转运活性直接受TGF-β/ALK1信号通路的调控,该通路可作为控制OATP底物药物CNS递送的靶点(Abdullahi等,2018)。使用骨形态发生蛋白(BMP)-9激活激活素受体样激酶(ALK)-1可增加大鼠脑微血管中Oatp1a4蛋白的表达(Abdullahi等,2017a)。因此,靶向Oatp1a4调控代表了控制Oatp1a4功能表达以向CNS递送治疗药物的机会(Abdullahi等,2017b)。

Oatp1a4的人类直系同源物是OATP1A2。研究表明,OATP1A2在人类中可受转化生长因子-β/激活素受体样激酶1信号通路的调控(Ronaldson等,2021)。他汀类药物(3-羟基-3-甲基戊二酰辅酶A还原酶抑制剂)作为Oatp1a4的转运底物可改善患者的功能性神经学结局,且Oatp1a4对他汀的摄取在大脑皮层中相对于小脑和脑干更高(Betterton等,2022)。匹伐他汀、瑞舒伐他汀、普伐他汀、牛磺罗辛、地高辛、赭曲霉毒素A和[d-青霉胺(2,5)]-脑啡肽是目前已鉴定的Oatp1a4底物,Oatp1a4介导这些药物穿越血脑屏障的血液-脑和脑-血液转运(Ose等,2010)。越来越多的证据表明,布美他尼也会影响脑部疾病,包括自闭症、新生儿癫痫和癫痫症;然而,全身给药后布美他尼的脑内水平较低,严重限制了其治疗脑部疾病的临床应用。体内实验表明,受限的被动扩散以及由Oat3、有机阴离子转运肽(Oatp)Oatp1a4和多药耐药蛋白4介导的主动外排转运,解释了布美他尼全身给药后脑内浓度极低的原因(Römermann等,2017)。疼痛是与炎症相关的主要症状,抗炎药物双氯芬酸对炎症性疼痛的抑制可减弱Oatp1a4功能表达的这些变化,表明外周炎症可调节BBB转运蛋白(Ronaldson等,2011)。

### 3.2 OATP1A4在血视网膜屏障和血蛛网膜屏障中的作用

Oatp1a4也在血视网膜屏障(BRB)上表达,是控制阴离子药物进入视网膜的主要决定因素。免疫印迹和免疫组化分析表明,Oatp1a4定位于视网膜色素上皮细胞的顶膜(Akanuma等,2013)。近期研究表明,Oatp1a4参与营养物质(包括核黄素、L-鸟氨酸、β-丙氨酸和L-组氨酸)从血液向视网膜的转运,以及普伐他汀在BRB中的转运(Kubo等,2018)。此外,有证据表明Oatp1a4在血蛛网膜屏障(BAB)的药物清除中发挥作用,并可能通过限制有机阴离子在脑和脊髓中的分布,在体内脑脊液(CSF)解毒中发挥重要作用(Yaguchi等,2019)。

### 3.3 OATP1A4在肝脏代谢中的作用

众所周知,肝脏是药物代谢的主要器官,有机阴离子转运肽(Oatps)参与多种有机阴离子化合物和药物的肝脏转运。Oatp1a4的功能在大鼠肝小叶的中央静脉(CV)周围区域占主导地位,可影响磺胺-101、地高辛、奎宁、d-维拉帕米、17β-雌二醇-d-17β-葡萄糖醛酸苷等底物在肝脏中的分布(Akanuma等,2019)。研究表明,Oatp1a4在小鼠心脏糖苷类(Takano等,2018)和黄连素(Chen等,2015)的肝脏蓄积中起主要作用。Oatp1a4在人肝脏β-内酰胺类抗生素的摄取中起主要作用,在功能上可能与大鼠肝脏中的OATP1B3相对应(Nakakariya等,2008)。厄贝沙坦是一种血管紧张素II受体拮抗剂,临床上用于治疗心力衰竭和心力衰竭。在大鼠中,它由多种Oatps(至少Oatp1a1和Oatp1a4)/Mrp2转运;在人类中,至少由OATP1B1/MRP2转运(Sun等,2014)。他汀类药物的不良反应通常是药物-药物相互作用的结果,在大鼠中,抑菌剂夫西地酸(FA)对Oatp1a4和Oatp1b2转运蛋白的抑制可能减弱他汀类药物的肝脏摄取,导致血液和组织中表现出肌肉骨骼毒性的浓度升高(Eng等,2016)。孕烯醇酮-16α-甲腈(PCN)通过PXR介导的机制调节肝脏中某些转运蛋白的表达,即Oatp1a4和Mrp3(Cheng & Klaassen,2006)。

### 3.4 OATP1A4在药物代谢中的作用

肝脏在药物体内代谢中发挥重要作用,肝脏转运蛋白参与肝脏对药物的吸收和排泄。地塞米松(DEX)处理肝细胞可增加CYP3A1/2、Oatp1a4和Mrp2的表达,并降低Ntcp的表达(Turncliff等,2004)。在人类地塞米松治疗期间,甲氨蝶呤的肝毒性增加,对肝脏中甲氨蝶呤转运蛋白表达的分析显示Mrp2、Oatp1a4和Oat2上调,Mrp3下调,导致胆汁排泄减少,从而在与DEX联合用药时增加药物的肝毒性(Fuksa等,2010)。全氟辛酸(PFOA)和全氟癸酸(PFDA)已在全球野生动物和人类中检出。两者在Oatp1a1、1a4、1b2的下调中均发挥核心作用(Cheng & Klaassen,2008)。围产期暴露于多溴联苯醚(PBDEs)导致肝脏外排转运蛋白Mdr1(多药耐药蛋白)、Mrp2(多药耐药相关蛋白)和Mrp3以及内流转运蛋白Oatp1a4 mRNA的表达增加(Szabo等,2009)。非酒精性脂肪性肝炎(NASH)患者可能同时摄入普伐他汀和含有吴茱萸碱的草药,后者上调Oatp1a1、Oatp1a4和Oatp1b2的表达,从而降低普伐他汀的全身暴露量(Liang等,2022)。一些天然产物或传统药物对Oatp1a4的调节也有报道。例如,甘草酸单铵(MAG)增加Oatp1a4的表达并降低Mrp2的表达(Zhou等,2016)。水飞蓟素抑制NASH动物中Oatp1b2和Oatp1a4的蛋白表达(Lynch等,2021)。欧前胡素、异欧前胡素和白芷提取物可显著下调小鼠肝脏中Oatp1a1、Oatp1a4和Oatp1b2的表达(Wang等,2020)。

## 4 OATP1A4与疾病

### 4.1 OATP1A4与胆汁淤积

胆汁淤积由胆汁流动中断引起,与多种肝脏疾病相关,有机阴离子转运多肽被认为在胆汁酸(BA)转运中具有重要功能。OATP1A1的下调和OATP1A4的上调可能影响肝脏对结合型BA的摄取(Slijepcevic等,2015)。在大鼠胆管结扎(BDL)后,Oatp1a4在肝脏中上调,在肠道中下调(Giroux等,2022),这似乎表明Oatp1a4与胆汁淤积相关。然而,在另一项使用7天BDL诱导的肝功能异常大鼠的实验中,发现BDL大鼠肝脏中Oatp1a4的mRNA表达降低至正常大鼠的84.8%(Horiuchi等,2009)。这种结果差异可能由实验周期不同所致。在脂多糖(LPS)诱导的炎症条件下,淋巴细胞缺乏改变了基础和炎症性IL-6 mRNA的表达,B细胞缺失也减弱了LPS处理小鼠中Oatp1a4 mRNA的表达,表明IL-6信号通路可能发挥关键作用(Bodeman等,2013)。Mrp2和3的逆转调节被认为代表了肝细胞对胆汁淤积性损伤的适应性反应,BDL和LPS处理的大鼠肝脏中门静脉周围Ntcp的下调以及Oatp1a4和Oatp1b2的诱导可能是减少Bsep和Mrp2显著下调的肝细胞中胆汁淤积性损伤的适应性机制(Donner等,2007)。ATP11C缺陷小鼠以结合型高胆红素血症和非结合型高胆红素血症为特征,ATP11C缺陷肝脏中央肝细胞中基底胆汁酸盐摄取转运蛋白OATP1B2、OATP1A1、OATP1A4和NTCP几乎缺失,其特征与Rotor综合征非常相似,表明OATP表达的缺失可能易导致Rotor综合征(de Waart等,2016)。跨膜蛋白30A(TMEM30A)是ATP11C功能所必需的β亚基,Tmem30a LKO小鼠中ATP11C的表达和膜定位显著降低,这与BS转运蛋白(如OATP1A4、OATP1B2、NTCP、BSEP和MRP2)的表达和定位受损相关,表明TMEM30A缺乏可通过损害BS转运蛋白及相关核受体的表达和定位导致小鼠肝内胆汁淤积(Liu等,2017)。α-萘基异硫氰酸酯(ANIT)是一种肝毒性物质,可引起啮齿动物急性肝内胆汁淤积。在薯蓣皂苷对ANIT诱导的胆汁淤积影响的实验中,发现ANIT与薯蓣皂苷联合处理可防止Oatp1a1、1b2的适应性下调,并促进Oatp1a4、多药耐药相关蛋白(Mrp2)和胆汁酸盐外排泵(Bsep)的上调,表明薯蓣皂苷可能通过恢复转运蛋白的表达来防止肝功能受损(A. Zhang等,2016)。吡嗪酰胺(PZA)是治疗结核病的一线药物,可引起严重的肝毒性,PZA处理大鼠血清中ALT和AST水平升高2倍,血清总胆汁酸升高10倍,胆汁酸合成和转运蛋白的mRNA和蛋白表达发生显著改变,其中FXR、Bsep、Mrp2、Mdr2、Ostα/β、Oatp1a1、Oatp1b2和Cyp8b1降低,而Mrp3、Ntcp、Oatp1a4和Cyp7a1升高(Guo等,2016)。在雌激素诱导的胆汁淤积中,炔雌醇显著增加胆汁淤积标志物,减少胆汁酸排泄,下调肝细胞转运蛋白(Ntcp/Oatp1b2/Oatp1a4/Mrp2),并上调Mrp3(Muchova等,2015)。茵陈蒿汤(YCHD)是著名的治疗胆汁淤积的传统中药方剂。YCHD的利胆作用与其对胆汁淤积肝脏中代谢酶和转运蛋白表达的调节作用相关。YCHD显著增加胆汁淤积大鼠中UGT1A1、胆汁酸盐外排泵、MRP2和OATP1A4的表达,对胆汁淤积具有显著的改善作用(Yi等,2018)。在一项使用Oatp1a4缺陷小鼠研究Oatp1a4在BA稳态中生理作用的实验中,雌性Oatp1a4缺陷小鼠血清或肝脏中的BA浓度未见显著变化,而雄性Oatp1a4缺陷小鼠的BA稳态发生显著改变,包括血清、肝脏和

# 翻译

肠道内容物。Oatp1a4功能的缺失并未减少胆管结扎小鼠血清或肝脏中胆汁酸(BA)的积累,这表明Oatp1a4不太可能是胆汁酸的摄取转运体,但在雄性小鼠的次级胆汁酸代谢中发挥重要作用(Zhang等,2013)。

## 4.2 OATP1A4与肝脏疾病

肝脏在外源性物质的解毒过程中发挥重要作用。肝胆转运体参与了肝脏对外源性物质的摄取和清除过程。在肝功能衰竭的情况下,这些转运体的表达可能受到调控。例如,在Long-Evans Cinnamon(LEC)大鼠肝炎模型中,发现三种窦状隙有机阴离子转运体Ntcp、Oatp1a1和Oatp1a4的肝脏表达均下降(Chiba等,2007)。在大鼠肝癌发生过程中,20周和32周时Ntcp、Bsep、Mrp2、Oatp1/Oatp1a1、Oatp2/Oatp1a4和Oatp4/Oatp1b2等转运体的mRNA呈进行性丢失(Monte等,2005)。酒精性肝病(ALD)大鼠模型及患者血浆胆红素水平均升高。组成型雄激素受体(CAR)是一种已知的异源物受体,可诱导胆红素解毒和转运,CAR激动剂苯巴比妥(PB)可降低ALD患者血清中胆红素水平,同时选择性上调OATP1A1、OATP1A4、UGT1A1和MRP2的表达水平(Wang等,2017)。肝脏缺血-再灌注(IR)损伤后,肝脏转运体Oatp1a1、Oatp1a4、Oatp1b2、Ntcp、Mdr2和Bsep的mRNA表达下降,而Mdr1b表达升高(Tanaka等,2006)。慢性高脂饮食是肥胖的关键因素,进行性肥胖可继发导致肝脏损伤、肾脏损伤和肠道萎缩。在肝脏、肾脏和肠道中表达的转运体在营养物质和药物的沉积中发挥重要作用。在高脂饮食(HFD)喂养的小鼠中,24周时Oat2的相对表达增加了4.08倍,Oat2蛋白表达上调,而摄取转运体Oct1、Oatp1b2和Oatp1a4的mRNA表达分别下降了79%、61%和19%(Lu等,2019)。

## 4.3 OATP1A4与其他疾病

常染色体显性多囊肾病(ADPKD)是遗传性多囊肾病(PKD)的常见类型,是肾衰竭的主要原因,多囊肾(PCK)大鼠肝脏中胆汁酸升高与肝脏中Mrp2和Oatp1a4蛋白表达下降相关(Bezençon等,2019)。阿尔茨海默病(AD)小鼠大脑中Oatp1a4蛋白表达显著下降(Wen等,2021)。脑缺氧/再氧合应激(H/R)是多种疾病的组成部分,在H/R条件下,大鼠脑微血管中Oatp1a4表达显著升高(Thompson等,2014)。类风湿关节炎(RA)中细胞因子系统水平升高可改变代谢酶和转运体的表达。在胶原诱导性关节炎(CIA)大鼠中,肠道Cyp3a1及肝脏Cyp2c6、Cyp2c7、Cyp3a1、Oatp1a1、Oatp1b2、Oatp1a4和Mrp2的mRNA水平显著下降(Lin等,2017)。

## 5 结论

Oatp1a4是一种重要的有机阴离子转运多肽,已知在多种药物和内源性物质(如胆汁酸)的转运中发挥重要作用。Oatp1a4在肝脏和大脑中高度表达,具有显著的年龄和性别差异,这对个体化用药的有效性和安全性具有重要意义。在多种疾病中均发现Oatp1a4的表达发生显著改变。然而,Oatp1a4的调控机制仍知之甚少,需要更多研究加以阐明。

## 伦理声明

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