红芪多糖对非酒精性脂肪肝病细胞模型FXR-SHP-SREBP-1c信号通路的影响  

Effects of Hedysarum polysaccharide on FXR-SHP-SREBP-1c signaling pathway in non-alcoholic fatty liver disease cell model

作　　者：张磊 金智生 杨晓轶 李佳蔚 李亚玲 金彩云 陈彦旭 ZHANG Lei;JIN Zhi-sheng;YANG Xiao-yi;LI Jia-wei;LI Ya-ling;JIN Cai-yun;CHEN Yan-xu(School of Basic Medicine,ansu University of Chinese Medicine,Lanzhou 730000,Gansu Province,China;Key Laboratory of Dunhuang Medicine,Ministry of Educationcansu University of Chinese Medicine,Lanzhou 730000,Gansu Province,China;Clinical College of Traditional Chinese Medicine,Gansu University of Chinese Medicine,Lanzhou 730000,Gansu Province,China;Traditional Chinese Medicine Syndrome Research Base,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,Fujian Province,China)

机构地区：[1]甘肃中医药大学基础医学院,甘肃兰州730000 [2]甘肃中医药大学敦煌医学与转化教育部重点实验室,甘肃兰州730000 [3]甘肃中医药大学中医临床学院,甘肃兰州730000 [4]福建中医药大学中医证研究基地,福建福州350122

出　　处：《中国临床药理学杂志》2024年第2期200-204,共5页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金地区基金资助项目(81660777);甘肃省高等学校青年博士基金资助项目(2022QB-096);甘肃省科技计划(自筹经费)基金资助项目(21JR1RA274);敦煌医学与转化教育部重点实验室开放课题基金资助项目(DHYX20-04)。

摘　　要：目的研究红芪多糖(HPS)对非酒精性脂肪肝病细胞模型法尼醇X受体(FXR)-小异二聚体伴侣(SHP)-甾醇调节元件结合蛋白1c(SREBP-1c)信号通路的影响。方法用1.2 mmol·L^(-1)脂肪酸培养LO-2细胞复制非酒精性脂肪肝病细胞模型。将细胞分为正常组(细胞正常培养)、模型组(建立非酒精性脂肪肝病细胞模型)、阳性对照组(造模后用50μmol·L^(-1)硫辛酸处理),实验组(造模后用80 mg·L^(-1)HPS处理),各组细胞均培养24 h。用GPO-PAP酶法检测细胞三酰甘油(TG)、胆固醇(TC)含量和谷草转氨酶(GOT)、谷丙转氨酶(GPT)活性,用蛋白质印迹(Western blot)法和逆转录聚合酶链反应(RT-PCR)检测肝细胞FXR-SHP-SREBP-1c信号通路蛋白和mRNA的表达水平。结果正常组、模型组、对照组、实验组的肝细胞TG含量分别为(2.91±1.13)、(6.81±1.32)、(3.72±0.52)和(4.67±0.62)mmol·gprot^(-1),TC含量分别为(23.66±4.92)、(67.96±5.56)、(29.41±4.22)和(54.34±3.96)mmol·gprot^(-1),GOT活性分别为(249.10±11.59)、(322.63±28.81)、(288.89±19.14)和(266.91±8.77)U·gprot^(-1),GPT活性分别为(58.83±16.88)、(134.55±22.96)、(89.63±15.81)和(77.37±7.25)U·gprot^(-1),FXR mRNA表达水平分别为1.01±0.16、2.09±0.12、1.83±0.17和1.45±0.15,SHP mRNA表达水平分别为1.00±0.11、0.51±0.15、0.64±0.14和0.70±0.14,SREBP-1c mRNA表达水平分别为1.00±0.08、1.57±0.19、1.37±0.13和1.21±0.15,FXR蛋白表达水平分别为1.00±0.02、1.63±0.03、1.42±0.02和1.25±0.03,SHP蛋白表达水平分别为1.00±0.02、0.23±0.01、0.54±0.21和0.62±0.02,SREBP-1c蛋白表达水平分别为1.00±0.03、4.08±0.05、1.99±0.02和1.48±0.01。上述指标,模型组与正常组比较,实验组与模型组比较,在统计学上差异均有统计学意义(均P<0.05)。结论HPS可能通过调控FXR-SHP-SREBP-1c信号通路,减少肝细胞脂质合成,从而保护肝细胞。Objective To study the effect of Hedysarum polysaccharides(HPS)on the farnisol X receptor(FXR)-small heterodimer chaperone(SHP)-sterol regulatory element-binding protein 1c(SREBP-1c)signaling pathway in the non-alcoholic fatty liver disease cell model.Methods The cells were cultured with 1.2 mmol·L^(-1)fatty acids to construct the non-alcoholic fatty liver disease cell model.The cell were divided into normal group(complete medium),model group(1.2 mm ol·L^(-1)fatty acid solution),positive control group(1.2 mmol·L^(-1)fatty acid solution+50μmol·L^(-1)alpha-lipoic acid)and experimental group(1.2 mmol·L^(-1)fatty acid solution+80 mg·L^(-1)HPS),culture for 24 h.The content of triglyceride(TG)and total cholesterol(TC),the activity of glutamate transaminase(GOT)and glutamate transaminasewas(GPT)detected by GPO-PAP enzyme method;the apoptosis rate was detected by flow cytometry;the expressions of FXR,SHP,SREBP-1c protein and mRNA in hepatocytes were detected by Western blot and reverse transcription-polymerase chain reaction(RT-PCR).Results The contents of TG in hepatocytes of normal group,model group,positve control group and experimental group were(2.91±1.13),(6.81±1.32),(3.72±0.52)and(4.67±0.62)mmol·gprot^(-1);the contents of TC in these four groups were(23.66±4.92),(67.96±5.56),(29.41±4.22)and(54.34±3.96)mmol·gprot^(-1);the activity of GOT in these four groups were(249.10±11.59),(322.63±28.81),(288.89±19.14)and(266.91±8.77)U·gprot^(-1);the activity of GPT in these four groups were(58.83±16.88),(134.55±22.96),(89.63±15.81)and(77.37±7.25)U·gprot^(-1),respectively;FXR mRNA expression levels were 1.01±0.16,2.09±0.12,1.83±0.17 and 1.45±0.15,respectively;SHP mRNA expression levels were 1.00±0.11,0.51±0.15,0.64±0.14 and 0.70±0.14,respectively;SREBP-1c mRNA were 1.00±0.08,1.57±0.19,1.37±0.13 and 1.21±0.15;the expression levels of FXR protein were 1.00±0.02,1.63±0.03,1.42±0.02 and 1.25±0.03,respectively;the expression levels of SHP protein were 1.00±0.02,0.23±0.01,0.54±0.21 and 0.

关 键 词：红芪多糖 非酒精性脂肪肝病 法尼醇X受体 脂肪变性 信号通路 

分 类 号：R28[医药卫生—中药学]