鸢尾素通过调控AKT/mTOR/SREBP-1信号通路 改善非酒精性脂肪性肝病脂质代谢  

Irisin improves lipid metabolism in nonalcoholic fatty liver by regulating the AKT/mTOR/SREBP-1 signa-ling pathway

作　　者：卢超 柯雅妮 胡洁[1] LU Chao;KE Yani;HU Jie(Department of Infectious Diseases,The First Affiliated Hospital of Zhejiang Chinese Medical University(Zhejiang Provincial Hospital of Chinese Medicine),Hangzhou,Zhejiang 310006,China;Zhejiang Chinese Medical University School of Basic Medical Sciences,Hangzhou,Zhejiang 310053,China)

机构地区：[1]浙江中医院大学附属第一医院(浙江省中医院)感染科,杭州310018 [2]浙江中医药大学基础医学院,杭州310053

出　　处：《浙江中西医结合杂志》2024年第12期1086-1092,共7页Zhejiang Journal of Integrated Traditional Chinese and Western Medicine

基　　金：浙江省医药卫生科技计划项目(No.2023KY140)。

摘　　要：目的探讨鸢尾素调节AKT/mTOR/SREBP-1信号通路对小鼠非酒精性脂肪性肝病(NAFLD)细胞脂质代谢的影响。方法采用棕榈酸/油酸诱导小鼠肝细胞AML12构建NAFLD模型,以200 ng/mL的鸢尾素干预,CCK8法检测肝细胞增殖活性,油红O染色观察脂肪沉积情况,蛋白免疫印迹检测磷酸化的蛋白激酶B(p-AKT)、哺乳动物雷帕霉素靶蛋白(p-mTOR)、核糖体蛋白S6激酶(p-P70S6K)及甾醇调节元件结合蛋白1(SREBP-1)的表达,并分别使用AKT抑制剂和激动剂、mTOR抑制剂和激动剂干预,明确鸢尾素的作用位点。同时测定过氧化物酶体增殖物激活受体(PPAR)及肝型脂肪酸结合蛋白(L-FABP)、下游靶基因脂肪酸合成酶(FAS)、乙酰辅酶A羧化酶1(ACC1)和硬脂酰辅酶A去饱和酶1(SCD1)的表达情况。结果与对照组比较,棕榈酸/油酸诱导后,模型组肝细胞内脂滴[(15.39±1.85)%比(1.98±0.67)%,P<0.01]含量明显增高;与模型组比较,200 ng/mL鸢尾素组细胞内脂滴[(6.93±0.70)%比(15.39±1.85)%,P<0.01]显著降低。与对照组比较,模型组p-AKT[(9.03±0.14)比(2.96±0.12),P<0.01]、p-mTOR[(7.18±0.04)比(1.66±0.18),P<0.01]、p-P70S6K[(4.21±0.00)比(1.26±0.06),P<0.01]、SREBP-1[(3.14±0.06)比(1.02±0.10),P<0.01]蛋白表达显著升高;与模型组比较,鸢尾素组p-AKT[(5.37±0.16)比(9.03±0.14),P<0.01]、p-mTOR[(3.36±0.15)比(7.18±0.04),P<0.01]、p-P70S6K[(2.59±0.05)比(4.21±0.00),P<0.01]、SREBP-1[(2.15±0.05)比(3.14±0.06),P<0.01]蛋白表达明显下降,鸢尾素与AKT抑制剂及mTOR抑制剂有协同作用,并能被AKT激动剂及mTOR激动剂逆转(P<0.05或P<0.01)。此外,与模型组比较,鸢尾素能抑制FAS[(1.83±0.20)比(3.32±0.39),P<0.01]、ACC1[(1.93±0.30)比(3.25±0.43),P<0.01]及SCD1[(2.16±0.27)比(3.65±0.42),P<0.01]mRNA的表达,上调PPARα[(1.40±0.06)比(0.71±0.10),P<0.01],下调PPARγ[(2.42±0.02)比(3.39±0.11),P<0.01]和L-FABP[(0.96±0.05)比(1.87±0.12),P<0.01]蛋白的表达。结论鸢尾素通过抑制AKT/mTOR/SREBP-1信号�Objective To investigate the effect of irisin on lipid metabolism in cells from a mouse model of non-alcoholic fatty liver disease(NAFLD)by regulating AKT/mTOR/SREBP-1 signaling pathway.Methods An NAFLD model was developed in mouse hepatocytes AML12 using palmitic acid/oleic acid induction,followed by treatment with 200 ng/mL irisin.The proliferative activity of hepatocytes was measured using CCK8,and fat accumulation was observed through oil-red O staining.The expression levels of phosphorylated protein kinase B(pAKT), mammalian target of rapamycin(p-mTOR), ribosomal protein S6 kinase(p-P70S6K), and sterol regulatory element binding protein 1(SREBP-1) were measured by western blotting. AKT inhibitors and agonists, as well as mTOR inhibitors and agonists, were used to clarify the action site of irisin. Additionally, the expression levels of peroxisome proliferator-activated receptor (PPAR), liver-type fatty acid binding protein (L-FABP), downstream target genes fatty acid synthase (FAS), acetyl-CoA carboxylase 1 (ACC1) and stearoyl-CoA desaturase 1 (SCD1) were determined. Results Compared with the control group, the model group had a significantly higher content of intracellular lipid droplets [(15.39±1.85)% vs. (1.98±0.67)%, P<0.01] following palmitic acid/oleic acid induction. Compared with the model group, the 200 ng/mL irisin group had a significantly lower content of intracellular lipid droplets [(6.93±0.70)% vs. (15.39±1.85)%, P<0.01]. Compared with the control group, the model group showed significantly increased expression levels of p-AKT [(9.03±0.14) vs. (2.96±0.12), P<0.01], p-mTOR [(7.18±0.04) vs. (1.66±0.18), P<0.01], p-P70S6K [(4.21±0.00) vs. (1.26±0.06), P<0.01] and SREBP-1 [(3.14±0.06) vs. (1.02±0.10), P<0.01]. Compared with the model group, the irisin group showed significantly decreased expression levels of p-AKT [(5.37±0.16) vs. (9.03±0.14), P<0.01], p-mTOR [(3.36±0.15) vs. (7.18±0.04), P<0.01], p-P70S6K [(2.59±0.05) vs. (4.21±0.00), P<0.01], and SREBP-1 [(2.15±0.05) vs. (3.14

关 键 词：鸢尾素 非酒精性脂肪性肝病 AKT MTOR SREBP-1 PPAR 

分 类 号：R285.5[医药卫生—中药学]