沙苑子苷A对脂肪变性L02细胞HNF4α调控的TG转运通路的影响  

Effect of Complanatoside A on TG transport pathway regulated by HNF4αin steatotic L02 cells

作　　者：刘双巧 李丽 邵紫萱 高晶 贾岚 刘佳琪 扈觐玺 王景霞 LIU Shuangqiao;LI Li;SHAO Zixuan;GAO Jing;JIA Lan;LIU Jiaqi;HU Jinxi;WANG Jingxia(School of Chinese Medicine,Beijing University of Chinese Medicine,Beijing 102488,China;Beijing University of Chinese Medicine Third Affiliated Hospital,Beijing 100029,China)

机构地区：[1]北京中医药大学中医学院,北京102488 [2]北京中医药大学附属第三医院

出　　处：《北京中医药大学学报》2022年第10期1044-1051,共8页Journal of Beijing University of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(No.81673617)。

摘　　要：目的探讨脂质代谢过程中沙苑子苷A对脂肪变性L02细胞肝细胞核因子4α(HNF4α)调控甘油三酯(TG)转运通路的影响。方法体外培养L02细胞,以油酸钠∶棕榈酸钠=2∶1的比例配制1 mmol/L的游离脂肪酸(FFA),干预L02细胞24 h后建立细胞脂肪变性模型。将L02细胞分为空白组、模型组、沙苑子苷A低浓度组(25μmol/L)、沙苑子苷A中浓度组(50μmol/L)和沙苑子苷A高浓度组(100μmol/L)。细胞贴壁培养24 h后,空白组给予含有10%胎牛血清(FBS)的高糖DMEM培养基(即完全培养基)培养48 h;模型组给予完全培养基培养24 h后,再使用含有1 mmol/L FFA的完全培养基培养24 h;沙苑子苷A各浓度组给予相应浓度的沙苑子苷A预处理24 h后,再使用含有1 mmol/L FFA和不同浓度沙苑子苷A的完全培养基共同培养L02细胞24 h。采用微板法检测各组TG、总胆固醇(TC)、丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)含量,酶联免疫吸附(ELISA)法检测各组瘦素(LEP)、脂联素(ADPN)、抵抗素(Retn)、白细胞介素-2(IL-2)、白细胞介素-10(IL-10)、载脂蛋白B100(ApoB100)含量,Western blotting法检测各组HNF4α、核因子κB(NF-κB)/p65蛋白的表达。结果与模型组相比,沙苑子苷A低浓度组TC、Retn含量降低(P<0.05),IL-10含量升高(P<0.05);沙苑子苷A中浓度组TC、ALT、Retn含量降低(P<0.05),ADPN、IL-10含量升高(P<0.05);沙苑子苷A高浓度组TG、TC、ALT、AST、Retn、ApoB100含量降低(P<0.05),ADPN、IL-10含量升高(P<0.05);沙苑子苷A低、中、高浓度组LEP、IL-2含量降低,但差异无统计学意义(P>0.05)。与模型组相比,沙苑子苷A高浓度组HNF4α蛋白表达上调(P<0.05),NF-κB/p65蛋白表达下调(P<0.05)。结论沙苑子苷A对TG转运通路的影响可能是通过刺激HNF4α转录调控ApoB100的装配和分泌,激活脂肪因子促进脂肪酸氧化,以此加速脂质转运和抑制脂质生成,从而发挥沙苑子维持脂质代谢稳态和减轻肝细胞炎症损伤Objective To investigate the effect of Complanatoside A on triglyceride(TG)transport pathway regulated by hepatocyte nuclear factor 4α(HNF4α)in steatotic L02 cells during lipid metabolism.Methods L02 cells were cultured in vitro,and 1 mmol/L free fatty acid(FFA)was prepared with a ratio of sodium oleate:sodium palmitate=2∶1,and a cellular steatosis model was established after 24 h of intervention in L02 cells.The experiment included the blank group,model group(0μmol/L),low mass concentration group of Complanatoside A(25μmol/L),middle mass concentration group of Complanatoside A(50μmol/L)and high mass concentration group of Complanatoside A(100μmol/L).After the cells were cultured for 24 h,the blank group was cultured for 48 h in high glucose DMEM medium(complete medium)containing 10%fetal bovine serum;the model group was cultured in complete medium for 24 h,and then cultured for 24 h in complete medium containing 1 mmol/L FFA;each mass concentration group of Complanatoside A was pretreated with the corresponding concentration of Complanatoside A for 24 h,and then L02 cells were cultured for 24 h in complete medium containing 1 mmol/L FFA and different concentrations of Complanatoside A.TG,total cholesterol(TC),glutamic-pyruvic transaminase(ALT)and glutamic oxaloacetic transaminase(AST)in each group were measured with the microplate method.Leptin(LEP),adiponectin(ADPN),resistin(Retn),interleukin-2(IL-2),interleukin-10(IL-10)and apolipoprotein B100(ApoB100)in each group were measured with the ELISA method.The expression of HNF4αand nuclear factor kappa-B(NF-κB)/p65 proteins in each group were measured with the Western blotting method.Results Compared with the model group,TC and Retn in the low mass concentration group of Complanatoside A decreased(P<0.05),and IL-10 increased(P<0.05);TC,ALT,and Retn in the middle mass concentration group of Complanatoside A decreased(P<0.05),and ADPN and IL-10 increased(P<0.05);TG,TC,ALT,AST,Retn and ApoB100 in the high mass concentration group of Complanatoside A decrease

关 键 词：沙苑子 沙苑子苷A L02细胞 脂肪变性 脂质代谢 甘油三酯转运 肝细胞核因子4Α 

分 类 号：R285.5[医药卫生—中药学]