Taurocholic acid improves hepatocyte lipid deposition by regulating macrophage polarization through TGR5

Qin MA; Lincheng JIANG; Yuehua YOU; Hongbing NI; Xiaojing LIN; Xiaoqiu XIAO; Jibin LI

doi:10.16016/j.2097-0927.202304057

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Taurocholic acid improves hepatocyte lipid deposition by regulating macrophage polarization through TGR5

Monographic Reports: Nutritional intervention for chronic diseases | 更新时间：2026-01-09

- Taurocholic acid improves hepatocyte lipid deposition by regulating macrophage polarization through TGR5
- Journal of Army Medical University Vol. 45, Issue 15, Pages: 1612-1620(2023)
- 作者机构：
  
  1.400016 重庆，重庆医科大学: 公共卫生学院营养与食品卫生学教研室
  2.400016 重庆，重庆医科大学: 附属第一医院重大代谢性疾病转化医学重点实验室
- 作者简介：
- 基金信息：
- DOI：10.16016/j.2097-0927.202304057
  CLC： R575.5;R967;R977.9
- Received：13 April 2023，
  
  Revised：2023-05-22，
  
  Published：15 August 2023
- 稿件说明：
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Qin MA, Lincheng JIANG, Yuehua YOU, et al. Taurocholic acid improves hepatocyte lipid deposition by regulating macrophage polarization through TGR5[J]. Journal of Army Medical University, 2023, 45(15): 1612-1620.
DOI：

Qin MA, Lincheng JIANG, Yuehua YOU, et al. Taurocholic acid improves hepatocyte lipid deposition by regulating macrophage polarization through TGR5[J]. Journal of Army Medical University, 2023, 45(15): 1612-1620. DOI： 10.16016/j.2097-0927.202304057.

摘要

目的

观察牛磺石胆酸(taurocholic acid

TLCA)调节棕榈酸(palmitic acid

PA)诱导的巨噬细胞极化，探讨其对肝细胞脂质代谢的影响。

方法

分别采用TLCA、PA处理小鼠单核巨噬细胞(RAW264.7)，实验分为对照(CON)组、TLCA组、PA组和PA+TLCA组。24 h后收集各组条件培养基(conditioned medium

CM)，分别处理C57 BL/6J小鼠肝脏原代细胞和AML-12肝细胞；使用氨苯蝶啶(triamterene

Tr)抑制巨噬细胞武田G蛋白偶联受体5 (Takeda G-protein receptor 5，TGR5)，以证实其在调节巨噬细胞极化以及降低肝细胞脂质沉积中的作用。采用BODIPY荧光染色检测肝细胞内脂质积聚；RT-qPCR检测炎症因子及脂质代谢相关基因mRNA水平；蛋白质免疫印迹法检测巨噬细胞TGR5和肝细胞固醇调节元件结合蛋白-1c(sterol regulatory element-binding proteins，SREBP-1c)表达水平；细胞免疫荧光染色观察细胞内iNOS、CD206蛋白表达。

结果

BODIPY荧光染色显示，PA组较CON组肝细胞脂质沉积显著增多，PA+TLCA组脂肪沉积得到缓解(

＜0.05)；与PA组比较，PA+TLCA组原代肝细胞和AML-12细胞脂质合成基因ACC1、FASN、SREBP-1c的mRNA水平以及SREBP-1c蛋白表达水平显著降低(

＜0.05)，ACOX1和CPT1A表达水平在各组间无显著变化；RT-qPCR结果显示，PA诱导巨噬细胞向M1型极化，促炎因子(iNOS、IL-6、IL-1β、TNF-α)水平显著升高，而联合TLCA处理后可降低炎症因子水平且增加RAW264.7细胞M2型极化相关基因(Arg1、IL-4、IL-10)mRNA表达水平(

＜0.05)；与CON组比较，PA组M1型巨噬细胞标志物iNOS显著增加，M2型巨噬细胞标志物CD206明显降低；TLCA可促进TGR5蛋白表达(

=0.071 4)，氨苯蝶啶抑制TGR5后，显著抑制TLCA调节巨噬细胞极化和减少肝细胞脂质沉积的作用。

结论

TLCA通过巨噬细胞TGR5抑制PA诱导的M1型极化且促进M2型极化，促炎症因子减少，进而降低肝细胞脂质合成，改善肝细胞脂质沉积。

Abstract

Objective

To investigate the effect of taurocholic acid (TLCA) regulating palmitic acid (PA)-induced macrophage polarization on hepatocyte lipid metabolism.

Methods

Mouse mononuclear macrophage (RAW264.7) cells

treated with TLCA and PA

were divided into control (CON)

TLCA

PA and PA+TLCA groups. The conditioned medium (CM) was collected for culturing primary hepatocytes from C57 BL/6J and AML-12 liver cells. Triamterene was used to inhibit macrophage Takeda G-protein receptor 5(TGR5)

which could investigate the role of TGR5 in regulating macrophage polarization and reducing lipid deposition in hepatocytes. BODIPY fluorescence staining was performed to detect lipid accumulation in hepatocytes; The mRNA levels of genes related to inflammatory factors and lipid metabolism were detected by RT-qPCR; Western blotting was performed to detect the level of protein expression of macrophage TGR5 and hepatic cell SREBP-1c. The expression of iNOS and CD206 were observed by immunofluorescence staining.

Results

BODIPY fluorescence staining showed that PA group displayed significantly increased lipid accumulation

compared with CON group

while TLCA treatment could reduce the PA-induced lipid accumulation(

0.05); Compared with the PA group

the PA+TLCA group had decreased mRNA levels of ACC1

FASN and SREBP-1c and protein expression of SREBP-1c (

0.05). However

there were no significant changes of expression of ACOX1 and CPT1A among these groups mentioned above. The outcome of RT-qPCR showed that PA stimulated RAW264.7 into M1-type polarization and significantly increased the levels of proinflammatory factors (iNOS

IL-6

IL-1β

TNF-α). PA combined with TLCA could reduce the levels of inflammatory factors and increased the mRNA expression levels of M2-type polarization-related genes

Arg1

IL-4 and IL-10 (

0.05). Compared with CON group

the PA group showed increased M1-type marker iNOS and decreased M2-type marker CD206

while TLCA could promote the protein expression of TGR5(

=0.0714). TGR5 inhibited by triamterene could suppress macrophage polarization regulated by TLCA and reduce hepatocyte lipid accumulation.

Conclusion

TLCA inhibits PA-induced M1-type polarization and promotes M2-type polarization

via

TGR5

which reduces proinflammatory factors

reduces hepatic lipogenesis and promotes hepatocyte lipid accumulation.

关键词

Keywords

references

European Association for the Study of the Liver (EASL), European Association for the Study of Diabetes (EASD), European Association for the Study of Obesity (EASO). EASL-EASD-EASO Clinical Practice Guidelines for the management of non-alcoholic fatty liver disease[J]. J Hepatol, 2016, 64(6): 1388-1402. DOI: 10.1016/j.jhep.2015.11.004.

LE M H, YEO Y H, LI X H, et al . 2019 global NAFLD prevalence: a systematic review and meta-analysis[J ] . Clin Gastroenterol Hepatol, 2022, 20(12): 2809-2817.e28. DOI: 10.1016/j.cgh.2021.12.002.

ZHOU J H, ZHOU F, WANG W X, et al . Epidemiological features of NAFLD from 1999 to 2018 in China[J ] . Hepatology, 2020, 71(5): 1851-1864. DOI: 10.1002/hep.31150.

SUN Y Y, LI X F, MENG X M, et al . Macrophage phenotype in liver injury and repair[J ] . Scand J Immunol, 2017, 85(3): 166-174. DOI: 10.1111/sji.12468.

MURRAY P J. Macrophage polarization[J]. Annu Rev Physiol, 2017, 79: 541-566. DOI: 10.1146/annurev-physiol-022516-034339.

CAI Y L, LI H G, LIU M Y, et al . Disruption of adenosine 2A receptor exacerbates NAFLD through increasing inflammatory responses and SREBP1c activity[J ] . Hepatology, 2018, 68(1): 48-61. DOI: 10.1002/hep.29777.

WANG D Q H, CAREY M C. Therapeutic uses of animal biles in traditional Chinese medicine: an ethnopharmacological, biophysical chemical and medicinal review[J]. World J Gastroenterol, 2014, 20(29): 9952-9975. DOI: 10.3748/wjg.v20.i29.9952.

KAWAMATA Y, FUJII R, HOSOYA M, et al . A G protein-coupled receptor responsive to bile acids[J ] . J Biol Chem, 2003, 278(11): 9435-9440. DOI: 10.1074/jbc.M209706200.

KEITEL V, DONNER M, WINANDY S, et al . Expression and function of the bile acid receptor TGR5 in Kupffer cells[J ] . Biochem Biophys Res Commun, 2008, 372(1): 78-84. DOI: 10.1016/j.bbrc.2008.04.171.

SHI Y, SU W T, ZHANG L, et al . TGR5 regulates macrophage inflammation in nonalcoholic steatohepatitis by modulating NLRP3 inflammasome activation[J ] . Front Immunol, 2021, 11: 609060. DOI: 10.3389/fimmu.2020.609060.

HOLTER M M, CHIRIKJIAN M K, BRIERE D A, et al . Compound 18 improves glucose tolerance in a hepatocyte TGR5-dependent manner in mice[J ] . Nutrients, 2020, 12(7): 2124. DOI: 10.3390/nu12072124.

王妙然, 李月, 张芸绮, 等. 基于网络药理学牛磺石胆酸抗炎作用及机制[J]. 中国药理学通报, 2021, 37(11): 1614-1619. DOI: 10.3969/j.issn.1001-1978.2021.11.023.

WANG M R, LI Y, ZHANG Y Q, et al . Anti-inflammatory effect and mechanism of taurocholic acid based on network pharmacology[J ] . Chin Pharmacol Bull, 2021, 37(11): 1614-1619. DOI: 10.3969/j.issn.1001-1978.2021.11.023.

MA L Q, LI H G, HU J B, et al . Indole alleviates diet-induced hepatic steatosis and inflammation in a manner involving myeloid cell 6-phosphofructo-2-kinase/fructose-2, 6-biphosphatase 3[J ] . Hepatology, 2020, 72(4): 1191-1203. DOI: 10.1002/hep.31115.

WANG X X, RAO H Y, ZHAO J M, et al . STING expression in monocyte-derived macrophages is associated with the progression of liver inflammation and fibrosis in patients with nonalcoholic fatty liver disease[J ] . Lab Investig, 2020, 100(4): 542-552. DOI: 10.1038/s41374-019-0342-6.

HU Q, LYON C J, FLETCHER J K, et al . Extracellular vesicle activities regulating macrophage- and tissue-mediated injury and repair responses[J ] . Acta Pharm Sin B, 2021, 11(6): 1493-1512. DOI: 10.1016/j.apsb.2020.12.014.

LIU C, RAJAPAKSE A G, RIEDO E, et al . Targeting arginase-Ⅱ protects mice from high-fat-diet-induced hepatic steatosis through suppression of macrophage inflammation[J ] . Sci Rep, 2016, 6(1): 1-13. DOI: 10.1038/srep20405.

QUEK J, CHAN K E, WONG Z Y, et al . Global prevalence of non-alcoholic fatty liver disease and non-alcoholic steatohepatitis in the overweight and obese population: a systematic review and meta-analysis[J ] . Lancet Gastroenterol Hepatol, 2023, 8(1): 20-30. DOI: 10.1016/S2468-1253(22)00317-X.

KANG K, REILLY S M, KARABACAK V, et al . Adipocyte-derived Th2 cytokines and myeloid PPARδ regulate macrophage polarization and insulin sensitivity[J ] . Cell Metab, 2008, 7(6): 485-495. DOI: 10.1016/j.cmet.2008.04.002.

MENGHINI R, MENINI S, AMORUSO R, et al . Tissue inhibitor of metalloproteinase 3 deficiency causes hepatic steatosis and adipose tissue inflammation in mice[J ] . Gastroenterology, 2009, 136(2): 663-672.e4. DOI: 10.1053/j.gastro.2008.10.079.

IPSEN D H, LYKKESFELDT J, TVEDEN-NYBORG P. Molecular mechanisms of hepatic lipid accumulation in non-alcoholic fatty liver disease[J]. Cell Mol Life Sci, 2018, 75(18): 3313-3327. DOI: 10.1007/s00018-018-2860-6.

张翔, 伍若楠, 刘家静, 等. M1型巨噬细胞促进非酒精性脂肪性肝炎[J]. 中华消化杂志, 2022, 42(2): 73-82. DOI: 10.3760/cma.j.cn311367-20210914-00497.

ZHANG X, WU R N, LIU J J, et al . M1 macrophages promote non-alcoholic steatohepatitis[J ] . Chin J Dig, 2022, 42(2): 73-82. DOI: 10.3760/cma.j.cn311367-20210914-00497.

RAJAGOPAL S, PONNUSAMY M. Metabotropic GPCRs: TGR5 and P2Y receptors in health and diseases[M ] . Singapore: Springer Nature Singapore Pte Ltd, 2008: 1-85. https://link.springer.com/book/10.1007/978-981-13-1571-8 https://link.springer.com/book/10.1007/978-981-13-1571-8 .

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Related Author

JIANG Lincheng

YOU Yuehua

NI Hongbing

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XIAO Xiaoqiu

LI Jibin

MA Qin

ZHAO Wenxia

Related Institution

Department of Nutrition and Food Hygiene， School of Public Health， Chongqing Medical University

The Chongqing Key Laboratory of Translational Medicine in Major Metabolic Diseases，Laboratory of Translational Medicine for Major Metabolic Diseases， the First Hospital of Chongqing Medical University

Department of Gastroenterology， the First Affiliated Hospital of Henan University of Chinese Medicine

Department of Infectious Diseases, the First Affiliated Hospital of Chongqing Medical University

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