基于mTORC1/USP20/HMGCR通路探讨消木丹颗粒治疗非酒精性脂肪性肝病作用机制  

Exploration on the Mechanism of Xiaomudan Granules in the Treatment of Non-alcoholic Fatty Liver Disease Based on mTORC1/USP20/HMGCR Pathway

作　　者：黄玉 闫瑞娟 焦俊喆 闫曙光[1] 魏海梁 常占杰 李京涛 边倩 HUANG Yu;YAN Ruijuan;JIAO Junzhe;YAN Shuguang;WEI Hailiang;CHANG Zhanjie;LI Jingtao;BIAN Qian(The First Clinical Medical School of Shaanxi University of Chinese Medicine,Xianyang 712000,China;The Affiliated Hospital of Shaanxi University of Chinese Medicine,Xianyang 712000,China)

机构地区：[1]陕西中医药大学第一临床医学院,陕西咸阳712000 [2]陕西中医药大学附属医院,陕西咸阳712000

出　　处：《中国中医药信息杂志》2024年第8期110-116,共7页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：陕西省教育厅青年创新团队建设科研计划(21JP032);陕西省重点研发计划(2020ZDLSF05-15);陕西省自然科学基础研究计划(2022JQ-965);陕西省创新能力支撑计划(2022TD-55);陕西省中医药管理局委托项目(SZY-KJCYC-2023-049、SZY-KJCYC-2023-087);陕西省中医药管理局“双链融合”中青年科研创新团队(2022-SLRH-LJ-002);咸阳市重点研发计划(L2022ZDYFSF007)。

摘　　要：目的观察消木丹颗粒对非酒精性脂肪性肝病(NAFLD)大鼠胆固醇合成的影响,基于mTORC1/USP20/HMGCR通路探讨其治疗NAFLD作用机制。方法60只SD大鼠随机分为空白组、模型组、西药组(多烯磷脂酰胆碱)和中药低、中、高剂量组(消木丹颗粒)。空白组予普通饲料喂养,其余组均予高脂饲料喂养12周建立NAFLD大鼠模型。造模成功后,各给药组予相应药物灌胃,空白组和模型组予无菌蒸馏水灌胃,连续4周。记录大鼠体质量、肝质量,计算肝指数,全自动生化分析仪检测血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转氨酶(AST)、总胆固醇(TC)、三酰甘油(TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)含量,HE染色、油红O染色观察肝组织形态,实时荧光定量PCR和Western blot检测大鼠肝组织核糖体S6激酶(S6K)、泛素特异性蛋白酶20(USP20)、3-羟基-3-甲基戊二酰辅酶A还原酶(HMGCR)mRNA及p-S6K、S6K、USP20、HMGCR蛋白表达。结果与空白组比较,模型组大鼠体质量、肝质量、肝指数显著升高(P<0.01,P<0.05),肝叶体积增大,边缘变钝;血清ALT、AST、TC、TG、LDL-C含量显著升高,HDL-C含量显著降低(P<0.01);大部分肝细胞脂肪变性、有明显空泡和炎性浸润,脂滴增多,肝组织USP20、HMGCR mRNA表达显著升高(P<0.01),p-S6K、USP20、HMGCR蛋白表达显著升高(P<0.01)。与模型组比较,中药高剂量组和西药组大鼠体质量、肝质量、肝指数显著降低(P<0.01,P<0.05),肝脏外观改善;血清ALT、AST、TC、LDL-C含量降低,HDL-C含量升高(P<0.01,P<0.05);肝细胞脂肪变性和气球样变减轻,脂滴沉积减少,肝组织USP20、HMGCRmRNA及p-S6K、USP20、HMGCR蛋白表达降低(P<0.01,P<0.05)。结论消木丹颗粒可能通过mTORC1/USP20/HMGCR通路调节胆固醇合成,进而发挥治疗NAFLD作用。Objective To observe the effects of Xiaomudan Granules on cholesterol synthesis in rats with non-alcoholic fatty liver disease(NAFLD);To explore its mechanism on the treatment of NAFLD based on mTORC1/USP20/HMGCR pathway.Methods Totally 60 SD rats were randomly divided into blank group,model group,Western medicine group(polyene phosphatidylcholine)and TCM low-,medium-and high-dosage groups(Xiaomudan Granules).The blank group was fed with ordinary diet,and the other groups were fed with high-fat diet for 12 weeks to establish NAFLD rat model.After successful modeling,each administration group was given the corresponding drug intragastric administration,and the blank group and model group were given aseptic distilled water intragastric administration for 4 weeks.Body mass and liver mass of rats were recorded,liver index was calculated,and serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),total cholesterol(TC),triglyceride(TG),high density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterol(LDL-C)contents were detected by automatic biochemical analyzer,the morphological changes of liver tissue were observed by HE staining and oil red O staining,real-time fluorescence quantitative PCR and Western blot were used to detect ribosome S6 kinase(S6K),ubiquitin specific protease 20(USP20),3-hydroxy-3-methylglutaryl CoA reduction enzyme(HMGCR)mRNA and p-S6K,S6K,USP20,HMGCR protein expression in liver tissue.Results Compared with the blank group,the body mass,liver mass and liver index of rats in model group significantly increased(P<0.01,P<0.05),the volume of liver lobe increased,the edge was blunted;the contents of serum ALT,AST,TC,TG and LDL-C significantly increased,while HDL-C content significantly decreased(P<0.01);most hepatocytes showed steatosis,significant vacuole and inflammatory infiltration,increased lipid droplets,and significantly increased mRNA expression of USP20 and HMGCR in liver tissue(P<0.01)and protein expressions of p-S6K,USP20 and HMGCR(P<0.01).Compared with the model g

关 键 词：非酒精性脂肪性肝病 消木丹颗粒 mTORC1/USP20/HMGCR通路 胆固醇合成 大鼠 

分 类 号：R285.5[医药卫生—中药学]