丹苘软胶囊干预大鼠非酒精性脂肪肝细胞模型基因芯片的组学分析

Gene Chipset Analysis in Rats with Non-alcoholic Fatty Liver Disease Intervened by Danqing Soft Capsule

目的:通过筛选mRNA(Clariom^(TM) S Assay)芯片及miRNA芯片的显著差异性基因,预测microRNA相关靶基因,探究丹苘软胶囊改善肝细胞脂肪代谢的调控机制。方法:提取丹苘软胶囊灌胃SD大鼠的含药血清,并对BRL大鼠肝细胞取对数生长期细胞进行实验,以阴性对照血清做对照,提取细胞总RNAs,采用Clariom S表达谱芯片及microRNA芯片检测细胞mRNA及miRNA的表达水平。运用生物信息学方法预测miRNA的靶基因,对脂肪代谢相关基因进行富集分析和通路分析。结果:经筛选和预测得到5629个预测靶基因参与20种生物学过程,7种细胞成分,有20种分子作用,神经营养素信号通路、丙型肝炎、PI3K-Akt信号通路被富集;rno-miR-26b-5p、rno-miR-129b-5p、rno-miR-21b-5p为调控网络中心;IPA数据库共检出31个与实验有关的脂肪代谢基因。结论:丹苘软胶囊在治疗非酒精性脂肪肝(nonalcoholic fatty liver disease,NAFLD)的机制可能与rno-miR-26b-5p、rno-miR-129b-5p、rno-miR-21b-5p,神经营养素信号通路、丙型肝炎、PI3K-Akt信号通路相关性较高。

Objective: To predict and investigate the regulatory mechanism of Danqing Soft Capsule improving fat metabolism of hepatocyte based on screening the difference between mRNA(Clariom^TM S Assay)and miRNA expression. Methods:We extracted blood-serum of the SD rats which received intragastric administration of Danqing Soft Capsule. BRL rat hepatocytes extracted logarithmic growth phase cells to perform experiments. Then Danqing Soft Capsule serum was used as experimental group and negative control serum was used as control group, respectively.Total RNAs were extracted and differently expressed miRNAs were detected by Clariom S expression profile chip for expressions level of mRNA and miRNA.Bioinformatics method was used to predict target genes of differential miRNAs regulation. Enrichment of functions and signaling pathways of the target genes was conducted by fat metabolism related genes. Result: In screening and predicting,5629 target genes were involved in 19 kinds of biological processes,11 kinds of cellular components,and 10 kinds of molecular functions.Neurotrophin signaling pathway, Hepatitis C and PI3 K-Akt signaling pathway were enriched.The rno-miR-26 b-5 p, rno-miR-129 b-5 p, rno-miR-21 b-5 p were the center of the molecular network. According to IPA database, 31 genes with fat metabolism effective interaction were screened out. Conclusion: The results shows rno-miR-26 b-5 p, rno-miR-129 b-5 p, rno-miR-21 b-5 p and neurotrophin signaling pathway. Hepatitis C and PI3 K-Akt signaling pathway may play a role in the differentiation process of non-alcoholic fatty liver induced by Danqing Soft Capsule.