目的 探讨维生素D通过法尼醇X受体（farnesoid X receptor,FXR）途径对油酸（oleic acid,OA）诱导的肝细胞脂质蓄积改善作用。方法 将HepG2细胞分为三组,对照组不作任何处理,模型组采用OA（0.5 mmol/L）诱导构建HepG2细胞脂质蓄积模型,干预组在给予OA（0.5 mmol/L）的同时给予1,25(OH)2D3（0.05 mmol/L）进行干预;油红O染色观察细胞脂质蓄积;对FXR进行免疫荧光染色并用DAPI荧光染料标记细胞核,观察FXR在细胞核的表达水平;蛋白质免疫印迹法检测FXR、SREBP-1c、SCD-1、FAS、PPARα和CPT-1A蛋白表达水平。结果 与对照组相比,模型组细胞内出现大量脂滴蓄积;予以1,25(OH)2D3干预后脂质蓄积显著改善;进一步的分子机制研究结果显示, 1,25(OH)2D3干预使细胞核中FXR表达显著增加,同时脂肪酸合成关键调控信号分子SREBP-1c、SCD-1和FAS蛋白表达显著降低,而脂肪酸β氧化分解关键调控信号分子PPARα和CPT-1A蛋白表达显著增加。结论 1,25(OH)2D3有可能通过FXR途径改善OA诱导的HepG2细胞脂质蓄积。
Abstract
Objective To explore the effect of vitamin D on oleic acid (OA)-induced lipid accumulation in hepatocytes through the farnesoid X receptor (FXR) pathway. Methods HepG2 cells were divided into three groups. The control group was not treated. The OA group was treated with OA (0.5mmol/L) to induce lipid accumulation in HepG2 cells, and the OA+VD group was treated with OA (0.5mmol/L) and 1,25(OH)2D3 (0.05mmol/L). The lipid accumulation in the cells was observed by Oil Red O staining. FXR expression in the nucleus was observed with the merged images, in which the expression of FXR in the cells was detected with immunofluorescence staining and that in the cell nuclei were labeled with DAPI. The protein expressions of FXR, SREBP-1c, SCD-1, FAS, PPARα, and CPT-1A were assayed by Western blot. Results Compared with the control group, OA treatment resulted in significant intracellular lipid droplet accumulation, and was significantly ameliorated by the simultaneous treatment of 1,25(OH)2D3. The results of mechanistic investigations revealed that 1,25(OH)2D3 treatment significantly enhanced the protein expression of FXR in the nuclei compared to the OA treatment. Meanwhile, 1,25(OH)2D3 treatment also significantly down-regulated the expression of key proteins involved in fatty acid synthesis (SREBP-1c, SCD-1, and FAS) and up-regulated the expression of key proteins involved in β-oxidation of fatty acid (PPARα and CPT-1A). Conclusion 1,25(OH)2D3 has the potential to ameliorate OA-induced lipid accumulation in HepG2 cells via the FXR pathway.
关键词
维生素D /
HepG2细胞 /
油酸 /
脂质蓄积 /
FXR
Key words
vitamin D3 /
HepG2 cells /
oleic acid /
lipid accumulation /
FXR
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参考文献
[1] 中华医学会肝病学分会.代谢相关(非酒精性)脂肪性肝病防治指南(2024年版)[J].实用肝脏病杂志,2024,27:494–510.
[2] Rinella ME, Neuschwander-Tetri BA, Siddiqui MS, et al. AASLD practice guidance on the clinical assessment and management of nonalcoholic fatty liver disease[J]. Hepatology, 2023,77:1797-1835.
[3] Teng ML, Ng CH, Huang DQ, et al. Global incidence and prevlence of nonalcoholic fatty liver disease[J]. Clin Mol Hepatol, 2023,29(Suppl):S32–S42.
[4] Le MH, Yeo YH, Zou B, et al. Forecasted 2040 global prevalence of nonalcoholic fatty liver disease using hierarchical Bayesian approach[J]. Clin Mol Hepatol, 2022,28:841–850.
[5] Zhou J, Zhou F, Wang W, et al. Epidemiological Features of MAFLD From 1999 to 2018 in China[J]. Hepatology, 2020,71:1851–1864.
[6] Yuan S, Larsson SC. Inverse association between serum 25-Hydroxyvitamin D and nonalcoholic fatty liver disease[J]. Clin Gastroenterol Hepatol, 2023,21:398–405.e4.
[7] Mohamed AA, Halim AA, Mohamed S, et al. The effect of high oral loading dose of cholecalciferol in non-alcoholic fatty liver disease patients: a randomized placebo controlled trial[J]. Front Pharmacol, 2023,14:1149967.
[8] Zhang XL, Chen L, Yang J, et al. Vitamin D alleviates non-alcoholic fatty liver disease via restoring gut microbiota and metabolism[J]. Front Microbiol, 2023,14:1117644.
[9] Xiang J, Zhang Z, Xie H, et al. Effect of different bile acids on the intestine through enterohepatic circulation based on FXR[J]. Gut Microbes, 2021,13:1949095.
[10] Chiang JYL, Ferrell JM.Bile acid receptors FXR and TGR5 signaling in fatty liver diseases and therapy[J]. Am J Physiol Gastrointest Liver Physiol, 2020,318:G554–G573.
[11] Yang Z, Danzeng A, Liu Q, et al. The role of nuclear receptors in the pathogenesis and treatment of non-alcoholic fatty liver disease[J]. Int J Biol Sci, 2024,20:113–126.
[12] Zhou S, You H, Qiu S, et al. A new perspective on MAFLD: Focusing on the crosstalk between peroxisome proliferator-activated receptor alpha (PPARα) and farnesoid X receptor (FXR)[J]. Biomed Pharmacother, 2022,54:113577.
[13] Alshawsh MA, Alsalahi A, Alshehade SA, et al. A comparison of the gene expression profiles of non-alcoholic fatty liver disease between animal models of a high-fat diet and methionine-choline-deficient diet[J]. Molecules, 2022,27:858.
[14] Zhang M, Bai X, Du Q, et al. the different mechanisms of lipid accumulation in hepatocytes induced by oleic acid/palmitic acid and high-fat diet[J]. Molecules, 2023,28:6714.
[15] Chen Q, Zhao L, Mei L, et al. Vitamin C and vitamin D3 alleviate metabolic-associated fatty liver disease by regulating the gut microbiota and bile acid metabolism via the gut-liver axis[J]. Front Pharmacol, 2023,14:1163694.
[16] Lee SB, Jin MH, Yoon JH.The contribution of vitamin D insufficiency to the onset of steatotic liver disease among individuals with metabolic dysfunction[J]. Sci Rep, 2024,14:6714.
[17] Chang E.Vitamin D Mitigates hepatic fat accumulation and inflammation and increases SIRT1/AMPK expression in AML-12 hepatocytes[J]. Molecules, 2024,29:1401.
[18] Stofan M, Guo GL.Bile acids and FXR: novel targets for liver diseases[J]. Front Med, 2020,7:544.
[19] Song Z, Xiaoli AM, Yang F.Regulation and metabolic significance of de novo lipogenesis in adipose tissues[J]. Nutrients, 2018,10:1383.
[20] Hu P, Li K, Peng X, et al. Nuclear receptor PPARα as a therapeutic target in diseases associated with lipid metabolism disorders[J]. Nutrients, 2023,15:4772.
[21] Donde H, Ghare S, Joshi-Barve S, et al. Tributyrin inhibits ethanol-induced epigenetic repression of CPT-1A and attenuates hepatic steatosis and injury[J]. Cell Mol Gastroenterol Hepatol, 2020,9:569–585.
[22] Tahri-Joutey M, Andreoletti P, Surapureddi S, et al. Mechanisms mediating the regulation of peroxisomal fatty acid beta-oxidation by PPARα[J]. Int J Mol Sci, 2021,22:8969.
[23] Cominguez DC, Park YJ, Kang YM, et al. Clitorin ameliorates western diet-induced hepatic steatosis by regulating lipogenesis and fatty acid oxidation in vivo and in vitro[J]. Sci Rep, 2022,12:4154.
[24] He J, Yang Y, Zhang F, et al. Effects of Poria cocos extract on metabolic dysfunction-associated fatty liver disease via the FXR/PPARα-SREBPs pathway[J]. Front Pharmacol, 2022,13:1007274.
[25] Meng Q, Duan XP, Wang CY, et al. Alisol B 23-acetate protects against non-alcoholic steatohepatitis in mice via farnesoid X receptor activation[J]. Acta Pharmacol Sin, 2017,38:69–79.
[26] Ke Z, Fan C, Li J, et al. Nobiletin intake attenuates hepatic lipid profiling and oxidative stress in hfd-induced nonalcoholic-fatty-liver-disease mice[J]. Molecules, 2023,28:2570.
[27] Du T, Xiang L, Zhang J, et al. Vitamin D improves hepatic steatosis in MAFLD via regulation of fatty acid uptake and β-oxidation[J]. Front Endocrinol, 2023,14:1138078.
基金
国家重点研发计划项目（No.2024YFF1106000）; 山西省应用基础研究(No.202303021211124); 山西省高等教育“百亿工程”科技引导专项
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