瘦素调节胆汁酸代谢影响胆石形成

Leptin influences cholelithiasis formation by regulating bile acid metabolism

目的探讨瘦素(leptin)和胆汁酸代谢之间的关系及其对胆石症的影响。方法收集手术治疗的89例胆囊结石患者(GS)、68例原发性肝内胆管结石患者(PIS)和72例肝脏良性疾病患者(CT)的肝脏组织及术前血清。放射免疫法测定血清瘦素水平,自动生化分析检测血清中TBA、TBIL、ALT、AST、TC、TG和γ-GGT水平。qPCR和Western blot分别检测肝脏组织长型瘦素受体(OB-Rb)、腺苷酸活化蛋白激酶-a2(AMPK-a2)、p-AMPKa2、胆盐输出泵(BSEP)的mRNA及蛋白表达量。HepG2细胞分别与瘦素和瘦素+AMPKa2抑制剂(compund C)共培养后,qPCR和Western blot分别检测细胞OB-Rb、AMPKa2、p-AMPKa2、BSEP的mRNA和蛋白表达量,自动生化分析仪检测HepG2细胞培养上清液中TBA含量。结果GS组血清瘦素水平明显高于CT和PIS组(P<0.05)。GS组和PIS组的OB-Rb、AMPKa2、p-AMPKa2和BSEP表达水平显著低于CT组(P<0.05)。瘦素刺激HepG2细胞后增加其OB-Rb、AMPKa2、p-AMPKa2和BSEP表达量。Compund C抑制瘦素对HepG2细胞中OB-Rb、AMPKa2、p-AMPKa2和BSEP的促表达作用。结论瘦素参与胆汁酸代谢的调节,其在肝脏的代谢异常可能是胆石症发生的机制之一。

Objective To further investigate the relationship between leptin and bile acid metabolism.Methods Liver tissues and preoperative serum of 89 patients with gall stone(GS),68 patients with primary intrahepatic Stone(PIS)and 72 patients with benign liver diseases(control,CT)were collected.Serum leptin levels were measured by radioimmunoassay,and the serum TBA,TBIL,ALT,AST,TC,TG and gamma-GGT levels were detected by automatic biochemical analysis.mRNA and protein expression levels of adenosine monophosphate-activated protein kinase-a2,p-AMPKa2 and bile salt export pump(BSEP)in liver samples were measured by qPCR and Western blot,respectively.After co-culture of HepG2 cells with leptin and leptin+AMPKa2 inhibitor(compund C),mRNA and protein expressions of OB-Rb,AMPKa2,p-AMPKa2 and BSEP were detected by qPCR and Western blot,respectively.TBA content in cell supernatant was detected by automatic biochemical analysis.Results Serum leptin level in GS group was significantly higher than that in CT and PIS group(P<0.05).The expression level of OB-Rb,AMPKa2,p-AMPKa2 and BSEP in GS group and PIS group were significantly lower than those in CT group(P<0.05).Leptin stimulated HepG2 cells and increased the expressions of OB-Rb,AMPKa2,p-AMPKa2 and BSEP.Compund C inhibited the effect of leptin on OB-Rb,AMPKa2,p-AMPKa2 and BSEP in HepG2 cells.Conclusions Leptin is involved in the regulation of bile acid metabolism,and its abnormal effect in liver may be one of the mechanisms of cholelithiasis.