基于转录组测序分析壮骨止痛方治疗去势大鼠骨质疏松症的基因表达差异

Effect of Zhuanggu Zhitong Formula on Differential Genes Expression of Ovariectomized Rats with Osteoporosis Based on Transcriptome Sequencing

目的:基于转录组测序分析壮骨止痛方治疗去势大鼠骨质疏松模型的差异表达基因及相关通路。方法:将12只SD雌性大鼠随机分为实验组、模型组,每组各6只,采用国内外公认的雌性大鼠去卵巢3个月造成绝经后骨质疏松模型。造模后1周开始灌胃给药,实验组给予壮骨止痛胶囊(567 mg/kg),模型组给予同体积的蒸馏水(1 mL/150 g),每天1次,连续13周。13周后大鼠腹腔静脉取血提取外周血单个核细胞及其RNA,进行RNA-Seq测序,对表达差异的基因进行筛选及KEGG功能注释分析和GO富集分析。结果:实验共筛选出149个差异表达的基因(|log2 Fold change|≥1,q<0.05),与模型组相比,实验组上调的基因有67个,下调的基因有82个;GO富集分析显示差异表达基因主要富集在细胞部分、细胞器、细胞过程、生物调节、代谢过程、绑定分子类、催化活性等;KEGG通路富集分析显示有TNF信号通路、IL-17信号通路、沙门氏菌感染三条信号通路具有显著性富集意义。结论:发现了壮骨止痛方治疗去势大鼠骨质疏松一些关键基因和通路,如Nlrp12、Oscar、LOC100359515、Csf3r等基因,以及TNF信号通路、IL-17信号通路等通路,壮骨止痛方可能是通过这些基因和通路发挥治疗效果。

Objective:To analyze the differential genes expression and the relative signaling pathways in ovariectomized rats with osteoporosis treated by Zhuanggu Zhitong Formula based on transcriptome sequencing.Methods:12 SD rats were randomly divided into the experimental group and the model group,with 6 rats in each group.The model of menopausal osteoporosis was established by universally acknowledged ovariectomy method.One week after modelling,the experimental group was treated with Zhuanggu Zhitong Capsule(567 mg/kg),whereas the model group was given the same amount of distilled water(1 mL/150 g),once per day for 13 weeks.After 13 weeks,the peripheral blood mononuclear cells and their RNA were extracted from the abdominal vein of the rats,which then were subjected to RNA-Seq,the differentially expressed genes were screened and the GO enrichment analysis was carried out.Results:149 differentially expressed genes were identified(|log2 Fold change|≥1,q<0.05).Compared to the model group,67 up-regulated genes and 82 downregulated genes were found in the experimental group.GO enrichment analysis showed that the differentially expressed genes were mainly concentrated in cell parts,organelle,cell process,biological regulation,metabolic process,binding and catalytic activities.Enrichment analysis of KEGG pathway showed that TNF signaling pathway,IL-17 signaling pathway and Salmonella infection signaling pathway had significant enrichment.Conclusion:The current research has shown some key genes,such as Nlrp12,Oscar,LOC100359515 and Csf3r,and TNF signaling pathway and IL-17 signaling pathway,may be associated with the therapeutic effect of Zhuanggu Zhitong Formula in osteoporosis progression.