Mst1减轻小鼠非酒精性脂肪性肝炎的机制研究
Mechanism study of Mst1 on alleviating nonalcoholic steatohepatitis in mice
- 上海中医药大学学报 2021年35卷第3期 页码：39-44
- 作者机构：
1.上海中医药大学附属第七人民医院胃肠疾病诊疗部（上海 200137）
- 作者简介：
常玲，女，硕士，主治医师，主要从事消化系统疾病的基础与临床研究
陈国雁，副主任医师，硕士生导师；E-mail：Ben13501908035@163.com
- 基金信息：上海市青年科技英才扬帆计划项目(21YF1434700);上海市浦东新区卫生系统学科带头人培养计划资助项目(PWRd2016-12);上海中医药大学附属第七人民医院人才培养计划项目(XX2020-19)
DOI：10.16306/j.1008-861x.2021.03.008
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常玲, 王智博, 徐晖, 等. Mst1减轻小鼠非酒精性脂肪性肝炎的机制研究[J]. 上海中医药大学学报, 2021,35(3):39-44.
CHANG Ling, WANG Zhibo, XU Hui, et al. Mechanism study of Mst1 on alleviating nonalcoholic steatohepatitis in mice[J]. Academic Journal of Shanghai University of Traditional Chinese Medicine, 2021,35(3):39-44.
常玲, 王智博, 徐晖, 等. Mst1减轻小鼠非酒精性脂肪性肝炎的机制研究[J]. 上海中医药大学学报, 2021,35(3):39-44. DOI： 10.16306/j.1008-861x.2021.03.008.
CHANG Ling, WANG Zhibo, XU Hui, et al. Mechanism study of Mst1 on alleviating nonalcoholic steatohepatitis in mice[J]. Academic Journal of Shanghai University of Traditional Chinese Medicine, 2021,35(3):39-44. DOI： 10.16306/j.1008-861x.2021.03.008.
摘要
目的：,2,研究哺乳动物不育系20样激酶1（Mst1）减轻小鼠非酒精性脂肪性肝炎（NASH）的作用及机制。,方法：,2,以野生型C57BL/6小鼠（WT）和全身性Mst1敲除小鼠（Mst1,-/-,）为实验对象，构建蛋氨酸－胆碱缺乏（MCD）饮食诱导的小鼠NASH模型，并以蛋氨酸－胆碱充足（MCS）饲料为对照。饲料干预4周后，取血，收集肝组织。全自动生化分析仪检测血清丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）水平；试剂盒检测肝组织总胆固醇（TC）、三酰甘油（TG）、丙二醛（MDA）水平及超氧化物歧化酶（SOD）活性；行特殊染色后光镜下观察肝组织形态学变化；免疫组化检测肝组织α-平滑肌肌动蛋白（α-SMA）表达；PCR检测肝组织炎症因子白介素1β（,IL-1β,）、肿瘤坏死因子α（,TNF-α,）、白介素6（,IL-6,）、趋化因子,CCL-2,的mRNA表达水平；Western blot检测Nod样受体蛋白3（NLRP3）、凋亡相关斑点样蛋白（ASC）、剪切型天冬氨酸特异性半胱氨酸蛋白酶1（cleaved-Caspase-1）的蛋白表达。,结果：,2,MCD饮食诱导4周后，与野生型小鼠（WT+MCD）相比，Mst1敲除小鼠（Mst1,-/-,+MCD）血清ALT、AST水平及肝组织TC、TG含量明显降低（,P,<,0.01）；病理染色观察显示，Mst1敲除小鼠肝细胞脂肪空泡减少，炎症细胞浸润及肝脏纤维化明显减轻；进一步检测发现，Mst1敲除小鼠肝组织SOD活性明显升高（,P,<,0.01），MDA含量明显下降（,P,<,0.01），同时肝组织炎症因子,IL-1β,、,TNF-α,、,IL-6,和,CCL-2,的mRNA表达水平明显降低（,P,<,0.05，,P,<,0.01），NLRP3、ASC和cleaved-Caspase-1的蛋白表达水平显著下调（,P,<,0.05，,P,<,0.01）。,结论：,2,Mst1缺乏能够减轻MCD饮食诱导的NASH小鼠肝损伤，其作用机制可能与增强肝细胞抗氧化能力、抑制肝脏NLRP3炎症小体活化有关。
Abstract
Objective:,2,To study the effects and mechanisms of mammalian sterile 20-like kinase 1(Mst1) on nonalcoholic steatohepatitis(NASH) in mice.,Methods:,2,Wild type C57BL/6 mice(WT) and systemic Mst1 knockout mice(Mst1,-/-,) were used as experimental objects, the methionine-choline deficiency(MCD) diet induced NASH model was established in mice, and methionine-choline sufficiency(MCS) diet was taken as the control.After feed intervention for 4 weeks, the blood was taken and the liver tissue was collected.The serum levels of alanine aminotransferase(ALT) and aspartate aminotransferase(AST) were detected by automatic biochemical analyzer, and the contents of total cholesterol(TC), triglyceride(TG), malondialdehyde(MDA) and the activity of superoxide dismutase(SOD) in liver tissue were detected by kits.After staining, the morphological changes of liver tissue were observed under light microscope.The expression of α-smooth muscle actin(α-SMA) was detected by immunohistochemistry.The mRNA expressions of inflammatory cytokines interleukin-1β(,IL-1β,), tumor necrosis factor-α(,TNF-α,), interleukin-6(,IL-,6) and ,CCL-2, were detected by PCR.The protein expressions of Nod-like receptor protein 3(NLRP3), apoptosis-associated speck-like protein containing a CARD(ASC) and cleaved cysteinyl aspartate specific proteinase-1(cleaved-Caspase-1) were detected by Western blot.,Results:,2,After MCD diet induction for 4 weeks, compared with the wild-type mice(WT+MCD), the serum levels of ALT and AST and the contents of TC and TG in liver tissue were significantly decreased in Mst1 knockout mice(Mst1,-/-,+MCD) (,P,<,0.01) .Pathological staining showed that the fat vacuoles in hepatocytes were reduced and the inflammatory cell infiltration and liver fibrosis were obviously alleviated in Mst1 knockout mice.Further, the activity of SOD in liver tissue of Mst1 knockout mice was increased significantly(,P,<,0.01), and the content of MDA was decreased significantly(,P,<,0.01) .Moreover, the mRNA expression levels of ,IL-1β,TNF-α,IL-6, and ,CCL-2, in liver tissue of Mst1 knockout mice were significantly decreased(,P,<,0.05,P,<,0.01), and the protein expression levels of NLRP3, ASC and cleaved-Caspase-1 were significantly decreased(,P,<,0.05,P,<,0.01) .,Conclusion:,2,Mst1 deficiency can alleviate the liver injury in MCD diet induced NASH mice, and its mechanism may be related to enhancing the antioxidant capacity of hepatocytes and inhibiting the activation of NLRP3 inflammasome.
关键词
哺乳动物不育系20样激酶1（Mst1）非酒精性脂肪性肝炎氧化应激炎症小体小鼠
Keywords
mammalian sterile 20-like kinase 1 (Mst1)nonalcoholic steatohepatitisoxidative stressinflammasomemouse
references
RINELLA M E. Nonalcoholic fatty liver disease：a systematic review[J]. JAMA，2015，313(22)：2263-2273.
ZHOU J，ZHOU F，WANG W，et al. Epidemiological Features of NAFLD From 1999 to 2018 in China[J]. Hepatology，2020，71(5)：1851-1864.
WANG F S，FAN J G，ZHANG Z，et al. The global burden of liver disease：the major impact of China[J]. Hepatology，2014，60(6)：2099-2108.
YOUNOSSI Z，ANSTEE Q M，MARIETTI M，et al. Global burden of NAFLD and NASH：trends，predictions，risk factors and prevention[J]. Nat Rev Gastroenterol Hepatol，2018，15(1)：11-20.
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，2020，77(8)：1806-1821.
MAI W，XU Y，XU J，et al. Berberine Inhibits Nod-Like Receptor Family Pyrin Domain Containing 3 Inflammasome Activation and Pyroptosis in Nonalcoholic Steatohepatitis via the ROS/TXNIP Axis[J]. Front Pharmacol，2020，11：185.
ANSTEE Q M，GOLDIN R D. Mouse models in non-alcoholic fatty liver disease and steatohepatitis research[J]. Int J Exp Pathol，2006，87(1)：1-16.
XU Y，GUO W，ZHANG C，et al. Herbal Medicine in the Treatment of Non-Alcoholic Fatty Liver Diseases-Efficacy，Action Mechanism，and Clinical Application[J]. Front Pharmacol，2020，11：601.
LIANG Z，CHEN X，SHI J，et al. Efficacy and safety of traditional Chinese medicines for non-alcoholic fatty liver disease：a systematic literature review of randomized controlled trials[J]. Chin Med，2021，16(1)：9.
LING P，LU T J，YUAN C J，et al. Biosignaling of mammalian Ste20-related kinases[J]. Cell Signal，2008，20(7)：1237-1247.
ZHOU D，CONRAD C，XIA F，et al. Mst1 and Mst2 maintain hepatocyte quiescence and suppress hepatocellular carcinoma development through inactivation of the Yap1 oncogene[J]. Cancer Cell，2009，16(5)：425-438.
QIAO Y，LI T，ZHENG S，et al. The Hippo pathway as a drug target in gastric cancer[J]. Cancer Lett，2018，420：14-25.
ZHOU D，ZHANG Y，WU H，et al. Mst1 and Mst2 protein kinases restrain intestinal stem cell proliferation and colonic tumorigenesis by inhibition of Yes-associated protein（Yap）overabundance[J]. Proc Natl Acad Sci U S A，2011，108(49)：E1312-E1320.
MRIDHA A R，WREE A，ROBERTSON A，et al. NLRP3 inflammasome blockade reduces liver inflammation and fibrosis in experimental NASH in mice[J]. J Hepatol，2017，66(5)：1037-1046.
WREE A，EGUCHI A，MCGEOUGH M D，et al. NLRP3 inflammasome activation results in hepatocyte pyroptosis，liver inflammation，and fibrosis in mice[J]. Hepatology，2014，59(3)：898-910.
YUE S，ZHU J，ZHANG M，et al. The myeloid heat shock transcription factor 1/β-catenin axis regulates NLR family，pyrin domain-containing 3 inflammasome activation in mouse liver ischemia/reperfusion injury[J]. Hepatology，2016，64(5)：1683-1698.
SCHREUDER T C，VERWER B J，VAN NIEUWKERK C M，et al. Nonalcoholic fatty liver disease：an overview of current insights in pathogenesis，diagnosis and treatment[J]. World J Gastroenterol，2008，14(16)：2474-2486.
LI X，SHI Z，ZHU Y，et al. Cyanidin-3-O-glucoside improves non-alcoholic fatty liver disease by promoting PINK1-mediated mitophagy in mice[J]. Br J Pharmacol，2020，177(15)：3591-3607.
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