重组人脂联素基因对非酒精性脂肪性肝病大鼠模型作用的探讨

目的研究重组脂联素(adiponectin,AdipoQ)基因在大鼠体内的表达情况,探讨其对非酒精性脂肪性肝病(Nonalcoholic fatty liver disease,NAFLD)模型大鼠肝细胞脂肪变性的治疗作用,为人AdipoQ在NAFLD的基因治疗方面的应用奠定基础。方法建立大鼠NAFLD实验模型。将携带重组质粒pEGFP-N1-AdipoQ的减毒沙门氏菌通过口服饲喂途径,注入NAFLD大鼠体内,采用荧光显微镜检测肝组织内重组AdipoQ基因表达情况;同时动态测定大鼠血浆中游离脂肪酸(free fatty acid,FFA),甘油三酯(triglycercide,TG)含量,并采用肝组织切片HE及苏丹Ⅲ染色研究其对肝细胞脂肪变性的抑制情况。结果以携带重组质粒pEGFP-N1-Adi-poQ的减毒沙门氏菌灌胃NAFLD大鼠后,荧光显微镜下可见肝组织内EGFP-AdipoQ融合蛋白的表达;与对照组相比,其血浆中FFA,TG含量降低(p0.05),肝细胞脂肪变性程度明显减轻。结论减毒沙门氏菌能够将重组AdipoQ基因导入大鼠体内并表达;重组脂联素能够使肝细胞脂肪变性程度减轻,对NAFLD有治疗作用。

Study On Human Adiponectin Gene In Nonalcoholic Fatty Liver Disease

【Objective】 To research the expression of recombinant AdipoQ in NAFLD rats,and to investigate the fatty degeneration inhibitory effect of AdipoQ to NAFLD rats.To lay the foundation for the NAFLD gene therapy with AdipoQ.【Methods】（1） NAFLD rat model was induced by feeding with high-fat diet.（2）Attenuated Salmonella containing pEGFP-N1-AdipoQ was introduced into NAFLD rats by gavage,the expression of EGFP-AdipoQ was observed with fluorescence microscope;the levels of FFA and TG in serum were measured,and the fatty degeneration of hepatocytes were dected in pathology.【Results】（1） The NAFLD rat model was constructed successfully,compared with normal group,the levels of FFA and TG were higher,the hepatocytes presented fatty degeneration.（2）The expression of fusion protion of AdipoQ could be seen through fluorescence microscope in hepatic tissue after Attenuated Salmonella containing pEGFP-N1-AdipoQ was introduced into NAFLD rats;compared with control groups,the levels of FFA and TG in serum were lower,the fatty degeneration of hepatocytes decreased significantly.【Conclusion】 The NAFLD rat model could be establisded by feeding with high-fat diet.The gene of AdipoQ carried by Attenuated Salmonella could be expressed in NAFLD rats,the recombinant AdipoQ could inhibite the fatty degeneration of hepatocytes signficantly.