人源PCSK9D374Y加重蛋氨酸胆碱缺乏饮食诱导的小鼠非酒精性脂肪性肝炎

doi:10.16352/j.issn.1001-6325.2025.05.0637

基础医学与临床 ›› 2025, Vol. 45 ›› Issue (5): 637-643.doi: 10.16352/j.issn.1001-6325.2025.05.0637

人源PCSK9^D374Y加重蛋氨酸胆碱缺乏饮食诱导的小鼠非酒精性脂肪性肝炎

阿比旦·阿卜杜如苏力¹, 陈小翠², 崔元峰¹, 托罗娜依·米吉提¹, 邓李惠¹, 陈邦党^1,2*

1.新疆医科大学基础医学院新疆乌鲁木齐 830000;
2.新疆医科大学第一附属医院临床医学研究院,新疆乌鲁木齐 830000

收稿日期:2024-11-04 修回日期:2025-03-01 出版日期:2025-05-05 发布日期:2025-04-23
通讯作者: ^* chenbangdang@126.com
基金资助:
新疆维吾尔自治区自然科学基金重点项目(2022D01D16);国家自然科学基金-新疆联合基金本地青年人才培养专项(U1903304)

Human PCSK9^D374Y exacerbates methionine choline deficiency diet-induced nonalcoholic steatohepatitis in mice

Abidan·ABUDURUSULI¹, CHEN Xiaocui², CUI Yuanfeng¹, Tuoluonayi·MIJITI¹, DENG Lihui¹, CHEN Bangdang^1,2*

1. School of Basic Medical Sciences, Xinjiang Medical University, Urumqi 830000;
2. Clinical Medical Research Institute, the First Affiliated Hospital of Xinjiang Medical University, Urumqi 830000, China

Received:2024-11-04 Revised:2025-03-01 Online:2025-05-05 Published:2025-04-23
Contact: ^* chenbangdang@126.com

摘要/Abstract

摘要： 目的探讨人源前蛋白转化酶枯草杆菌素/Kexin 9型(PCSK9)基因功能获得性突变(hPCSK9^D374Y)对蛋氨酸胆碱缺乏饮食(MCD)诱导的小鼠非酒精性脂肪性肝炎(NASH)的影响。方法选用16只C57BL/6J野生型小鼠,随机分为hPCSK9^D374Y组与对照GFP组,MCD喂养6周,检测肝脏三酰甘油含量及血清丙氨酸氨基转移酶(ALT)和天冬氨酸氨基转移酶(AST)水平。通过油红O及天狼星红染色评估肝脏脂质浸润及纤维化程度;免疫组化分析F4/80阳性细胞浸润情况。Western blot检测脂质合成及炎性反应相关蛋白,RT-qPCR分析相关mRNA表达。结果 hPCSK9^D374Y组小鼠肝脏hPCSK9蛋白及mRNA显著上调,LDLR蛋白表达下调,血清ALT与AST水平显著升高(P＜0.05)。肝脏脂肪变性和纤维化程度加重,F4/80阳性细胞显著增多(P＜0.01)。Western blot分析显示hPCSK9^D374Y组FASN和SCD1蛋白显著上调,PPARα下调;TLR4和p-P65表达升高,而IκBα表达降低(P＜0.001)。RT-qPCR结果表明炎性因子TNF-α、IL-1β、IL-6和MCP-1的mRNA水平显著增加,纤维化相关mRNA(collagen Ⅰα和collagen Ⅲα)显著上调(P＜0.001)。结论人源PCSK9基因功能获得性突变(hPCSK9^D374Y)加重了MCD诱导的小鼠肝脏脂肪变性、炎性反应和纤维化。

关键词: 非酒精性脂肪性肝炎, 前蛋白转化酶枯草溶菌素9(PCSK9), 蛋氨酸胆碱缺乏饮食(MCD), 炎性因子

Abstract: Objective To investigate the effect of mutation human proprotein convertase subtilism/kexin type 9(hPCSK9^D374Y) in PCSK9 gene on methionine choline deficiency diet (MCD)-induced nonalcoholic steato-hepatitis (NASH) in mice. Methods Sixteen C57BL/6J wild-type mice were selected and randomly divided into the hPCSK9^D374Y group and the control GFP group. MCD was fed for 6 weeks, and then the serum level of hepatic triglyceride, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) was examined. Oil Red O and Sirius Red staining microscopy were used to identify hepatic lipid infiltration and fibrosis severity. F4/80-positive cell infiltration was analyzed using immunohistochemistry. Lipid synthesis and inflammatory response-related proteins were detected by Western blot and related mRNA expression was analyzed by RT-qPCR. Results Hepatic hPCSK9 protein and mRNA were significantly up-regulated, LDLR protein expression was down-regulated, and serum level of ALT and AST was significantly elevated in the hPCSK9^D374Y group of mice(P＜0.05). The degree of hepatic steatosis and fibrosis increased and F4/80-positive cells were significantly increased (P＜0.01). FASN and SCD1 proteins were significantly up-regulated and PPARα was down-regulated in the hPCSK9^D374Y group; The expression of TLR4 and p-P65 was elevated, whereas the expression of IκBα was decreased (P＜0.001). RT-qPCR results showed a significant increase of mRNA coding inflammatory factors TNF-α, IL-1β, IL-6, and MCP-1, and a significant up-regulation of fibrosis-associated mRNAs (collagen Ⅰα and collagen Ⅲα) was found (P＜0.001). Conclusions Functionally acquired mutation in the PCSK9 gene (hPCSK9^D374Y) exacerbates MCD-induced hepatic steatosis, inflammatory response and fibrosis in mice.

Key words: non-alcoholic steatohepatitis, proprotein convertase subtilisin/kexin type 9 (PCSK9), methionine choline deficiency diet, inflammatory factor

中图分类号:

R575.5

阿比旦·阿卜杜如苏力, 陈小翠, 崔元峰, 托罗娜依·米吉提, 邓李惠, 陈邦党. 人源PCSK9^D374Y加重蛋氨酸胆碱缺乏饮食诱导的小鼠非酒精性脂肪性肝炎[J]. 基础医学与临床, 2025, 45(5): 637-643.

Abidan·ABUDURUSULI, CHEN Xiaocui, CUI Yuanfeng, Tuoluonayi·MIJITI, DENG Lihui, CHEN Bangdang. Human PCSK9^D374Y exacerbates methionine choline deficiency diet-induced nonalcoholic steatohepatitis in mice[J]. Basic & Clinical Medicine, 2025, 45(5): 637-643.

参考文献 13

[1]	Younossi ZM. Non-alcoholic fatty liver disease-a global public health perspective[J]. J Hepatol, 2019, 70: 531-544.
[2]	李想,李晔.非酒精性脂肪性肝病中自噬与铁死亡相互作用的研究进展[J]. 基础医学与临床,2023, 43: 1294-1298.
[3]	Diehl AM, Day C. Cause, pathogenesis, and treatment of nonalcoholic steatohepatitis[J]. N Engl J Med, 2017, 377: 2063-2072.
[4]	Estes C, Razavi H, Loomba R, et al. Modeling the epidemic of nonalcoholic fatty liver disease demonstrates an exponential increase in burden of disease[J]. Hepatology (Baltimore, Md), 2018, 67: 123-133.
[5]	Bertot LC, Adams LA. Trends in hepatocellular carcinoma due to non-alcoholic fatty liver disease[J]. Expert Rev Gastroenterol Hepatol, 2019, 13: 179-187.
[6]	Raza S, Rajak S, Upadhyay A, et al. Current treat-ment paradigms and emerging therapies for NAFLD/NASH[J]. Front Biosci(Landmark Edition), 2021, 26: 206-237.
[7]	Wargny M, Ducluzeau PH, Petit JM, et al. Circulating PCSK9 levels are not associated with the severity of hepatic steatosis and NASH in a high-risk population[J]. Atherosclerosis, 2018, 278: 82-90.
[8]	Tsuchida T, Lee YA, Fujiwara N, et al. A simple diet- and chemical-induced murine NASH model with rapid progression of steatohepatitis, fibrosis and liver cancer[J]. J Hepatol, 2018, 69: 385-395.
[9]	Grewal T, Buechler C. Emerging insights on the diverse roles of proprotein convertase subtilisin/kexin type 9 (PCSK9) in chronic liver diseases: cholesterol metabolism and beyond[J]. Int J Mol Sci, 2022, 23:1070.doi:10.3390/ijms23031070.
[10]	秦可欣,陈小翠,崔元峰,等.PPARα基因敲除诱导小鼠肝脏脂质蓄积的作用研究[J].新疆医科大学学报, 2023,46:1267-1272.
[11]	Wang Q, Ou Y, Hu G, et al. Naringenin attenuates non-alcoholic fatty liver disease by down-regulating the NLRP3/NF-κB pathway in mice[J]. Br J Pharmacol, 2020, 177: 1806-1821.
[12]	Rex J, Lutz A, Faletti L E, et al. IL-1β and TNFα differentially influence NF-κB activity and fasl-induced apoptosis in primary murine hepatocytes during LPS-induced inflammation[J]. Front Physiol, 2019, 10: 117.doi:10.3389/fphys.2019.00117.
[13]	Cai J, Zhang XJ, Li H. Role of innate immune signaling in non-alcoholic fatty liver disease[J]. Trends Endocrinol Metab: TEM, 2018, 29: 712-722.

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人源PCSK9^D374Y加重蛋氨酸胆碱缺乏饮食诱导的小鼠非酒精性脂肪性肝炎

Human PCSK9^D374Y exacerbates methionine choline deficiency diet-induced nonalcoholic steatohepatitis in mice

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人源PCSK9D374Y加重蛋氨酸胆碱缺乏饮食诱导的小鼠非酒精性脂肪性肝炎

Human PCSK9D374Y exacerbates methionine choline deficiency diet-induced nonalcoholic steatohepatitis in mice

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人源PCSK9^D374Y加重蛋氨酸胆碱缺乏饮食诱导的小鼠非酒精性脂肪性肝炎

Human PCSK9^D374Y exacerbates methionine choline deficiency diet-induced nonalcoholic steatohepatitis in mice