健肝消脂方对非酒精性脂肪性肝病胰岛素抵抗、脂质沉积的影响及相关mRNA差异表达初步分析

Preliminary Analysis of the Effect of Jiangan Xiaozhi Recipe on Insulin Resistance and Lipid Deposition and Different Expressions of Related mRNA in Non-alcoholic Fatty Liver Disease

目的通过观察动物模型空腹胰岛素、空腹血糖、肝脏脂质沉积情况,计算胰岛素抵抗(IR)相关指标,并使用基因芯片技术观察相关mRNA表达谱差异,以深入阐释健肝消脂方治疗非酒精性脂肪性肝病(NAFLD)的分子机制。方法使用高糖高脂饲料复制NAFLD大鼠模型,造模成功后使用随机数字表法分为治疗组(Z组)及模型组(M组)。Z组根据10.86g/kg的生药量予健肝消脂方灌胃给药,M组给予等体积的蒸馏水灌胃,均每日给药1次,连续60天。给药结束后检测两组空腹胰岛素水平、空腹血糖水平,计算胰岛素敏感性及胰岛素抵抗指数(HOMA-IR),肝脏病理切片染色后观察肝脏脂质沉积情况,采用基因芯片技术分析实验动物肝脏相关mRNA表达谱差异。结果与M组比较,Z组大鼠空腹胰岛素水平明显下降(P<0.05),胰岛素敏感性明显升高(P<0.05),HOMA-IR明显降低(P<0.05),肝脏脂质沉积明显改善;两组肝脏mRNA表达存在明显差异(P<0.05),其中上调的基因有52个,下调的基因有13个;对基因功能进行分析可知,生物途径(pathway)富集可得出排序前十的pathway富集结果,GO富集可得到8个GO分子功能(molecular function)富集结果,排序前十的GO生物过程(biological process)富集结果及2个GO细胞成分(cellular component)富集结果。结论健肝消脂方可以改善IR及肝脏脂质沉积,其作用机制可能与该方影响肝脏基因表达有关。其分子机制可能是该方能够调节干扰素调节因子(IRF)7、GO分子功能GO:0016597、GO生物过程GO:0006952,并且影响了血管内皮生长因子(VEGF)信号通路、丝裂原活化蛋白激酶(MAPK)信号通路、Toll样体信号通路。同时健肝消脂方可能在分子层面上对肝纤维化、丙型肝炎病毒在肝脏内的复制等产生一定影响。

Objective To further explain the molecular mechanism of Jiangan Xiaozhi Recipe in the treatment of non-alcoholic fatty liver disease(NAFLD),by investigating the fasting insulin,fasting blood glucose and liver lipid deposition in animal models and calculating insulin resistance(IR)related indicators,and observing the differences of related mRNA expression profiles by using gene chip technology.Methods NAFLD rat models were reproduced with high-sugar and high-fat diet.After successful establishment,the models were divided into treatment group(Z group)and model group(M group),according to random number table.Group Z was given Jiangan Xiaozhi Recipe orally once a day for 60 consecutive days according to the dosage of 10.86 g/kg.Group M was given distilled water of equal volume once a day for 60 consecutive days.Fasting insulin and fasting blood glucose levels were measured after intervention.Insulin sensitivity and insulin resistance index(HOMA-IR)were calculated.Liver pathological sections were stained to observe liver lipid deposition.Gene chip technology was used to analyze the different expressions of related mRNA in experimental animals'livers.Results Compared with group M,fasting insulin level of rats in group Z decreased significantly(P<0.05),insulin sensitivity increased significantly(P<0.05),HOMA-IR decreased significantly(P<0.05),and liver lipid deposition improved significantly(P<0.05).There were significant differences in liver gene expression between the two groups(P<0.05),including 52 up-regulated genes and 13 down-regulated genes.The results of pathway enrichment in the first ten rankings can be obtained.The results of molecular function enrichment can be obtained by GO enrichment.The biological process enrichment of GO in the first ten rankings and cell component enrichment of two GO cells can be obtained by GO enrichment.Conclusion Its molecular mechanism may be that Jiangan Xiaozhi Recipe can effect vascular endothelial growth factor(VEGF)signaling pathway,mitogen-activated protein kinase(MAPK)signaling pathway,and Toll-like receptor signaling pathway,meanwhile adjusted interferon regulatory factor 7(IRF7),GO:0016597 of GO molecular function and GO:0006952 of GO biological process.At the same time,Jiangan Xiaozhi Recipe may have an effect on liver fibrosis and hepatitis C virus at the molecular level.