薯蓣皂苷元对非酒精性脂肪肝病模型大鼠mTOR/FASN/HIF-1α/VEGFA表达的影响  

Effect of diosgenin on mTOR/FASN/HIF-1α/VEGFA expression in rats with non-alcoholic fatty liver disease

作　　者：印国良 梁鸿艺 梁朋朋 冯亚男 陈素雯 刘湘怡 潘文超 张风霞[2] YIN Guo-liang;LIANG Hong-yi;LIANG Peng-peng;FENG Ya-nan;CHEN Su-wen;LIU Xiang-yi;PAN Wen-chao;ZHANG Feng-xia(Shandong University of Traditional Chinese Medicine,Ji'nan 250014,China;Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Ji'nan 250014,China)

机构地区：[1]山东中医药大学,山东济南250014 [2]山东中医药大学附属医院,山东济南250014

出　　处：《中国中药杂志》2023年第7期1760-1769,共10页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81573945);山东省自然科学基金面上项目(ZR2021MH054)。

摘　　要：研究薯蓣皂苷元对非酒精性脂肪肝病(non-alcoholic fatty liver disease,NAFLD)模型大鼠肝组织中哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)、脂肪酸合成酶(fatty acid synthase,FASN)、低氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)、血管内皮生长因子A(vascular endothelial growth factor A,VEGFA)表达的影响,探讨薯蓣皂苷元对NAFLD大鼠脂质生成和炎症的作用机制。40只雄性SD大鼠,随机选取8只为正常组给予普通饲料喂养,其余32只给予高脂饲料喂养以构建NAFLD模型。造模成功后随机分为高脂饮食组、薯蓣皂苷元低剂量组(150 mg·kg-1·d-1)、薯蓣皂苷元高剂量组(300 mg·kg-1·d-1)、辛伐他汀组(4 mg·kg-1·d-1),每组8只,连续灌胃给药8周。生化法检测血清中甘油三酯(triglyceride,TG)、总胆固醇(total cholesterol,TC)、低密度脂蛋白胆固醇(low-density lipoprotein cholesterol,LDL-C)、谷丙转氨酶(alanine transaminase,ALT)及谷草转氨酶(aspartate transaminase,AST)的水平;酶法检测肝脏中TG、TC的含量;酶联免疫吸附测定(enzyme-linked immunosorbent assay,ELISA)检测血清中白细胞介素1β(interleukin 1β,IL-1β)及肿瘤坏死因子α(tumor necrosis factorα,TNF-α)的水平;油红O染色检测肝脏脂质蓄积情况;苏木素-伊红(hematoxylin-eosin,HE)染色检测肝组织病理形态变化;实时荧光定量聚合酶链反应和蛋白免疫印迹法分别检测大鼠肝组织中mTOR、FASN、HIF-1α、VEGFA mRNA和蛋白的表达水平。结果显示,与正常组比较,高脂饮食组大鼠体质量以及TG、TC、LDL-C、ALT、AST、IL-1β、TNF-α水平显著升高(P<0.01),肝脏脂质蓄积显著增多(P<0.01),肝脏出现明显的脂肪变性,肝脏mTOR、FASN、HIF-1α、VEGFA mRNA水平显著升高(P<0.01),肝脏p-mTOR、FASN、HIF-1α、VEGFA蛋白水平显著升高(P<0.01)。与高脂饮食组比较,各治疗组大鼠体质量以及TG、TC、LDL-C、ALT、AST、IL-1β、TNF-α水平显著降低(P<0.05,P<0.01),�The present study aimed to investigate the effect of diosgenin on mammalian target of rapamycin(mTOR),fatty acid synthase(FASN),hypoxia inducible factor-1α(HIF-1α),and vascular endothelial growth factor A(VEGFA)expression in liver tissues of rats with non-alcoholic fatty liver disease(NAFLD)and explore the mechanism of diosgenin on lipogenesis and inflammation in NAFLD.Forty male SD rats were divided into a normal group(n=8)fed on the normal diet and an experimental group(n=32)fed on the high-fat diet(HFD)for the induction of the NAFLD model.After modeling,the rats in the experimental group were randomly divided into an HFD group,a low-dose diosgenin group(150 mg·kg-1·d-1),a high-dose diosgenin group(300 mg·kg-1·d-1),and a simvastatin group(4 mg·kg-1·d-1),with eight rats in each group.The drugs were continuously given by gavage for eight weeks.The levels of triglyceride(TG),total cholesterol(TC),low-density lipoprotein cholesterol(LDL-C),alanine transaminase(ALT),and aspartate transaminase(AST)in the serum were detected by the biochemical method.The content of TG and TC in the liver was detected by the enzyme method.Enzyme-linked immunosorbent assay(ELISA)was used to measure interleukin 1β(IL-1β)and tumor necrosis factorα(TNF-α)in the serum.Lipid accumulation in the liver was detected by oil red O staining.Pathological changes of liver tissues were detected by hematoxylin-eosin(HE)staining.The mRNA and protein expression levels of mTOR,FASN,HIF-1α,and VEGFA in the liver of rats were detected by real-time fluorescence-based quantitative polymerase chain reaction(PCR)and Western blot,respectively.Compared with the normal group,the HFD group showed elevated body weight and levels of TG,TC,LDLC,ALT,AST,IL-1β,and TNF-α(P<0.01),increased lipid accumulation in the liver(P<0.01),obvious liver steatosis,upregulated mRNA expression levels of mTOR,FASN,HIF-1α,and VEGFA(P<0.01),and increased protein expression levels of p-mTOR,FASN,HIF-1α,and VEGFA(P<0.01).Compared with the HFD group,the groups with drug tre

关 键 词：薯蓣皂苷元 非酒精性脂肪肝 脂肪生成 炎症 

分 类 号：R285.5[医药卫生—中药学]