藿香正气滴丸干预大鼠湿困脾胃证的转录组学分析

Effect of Huoxiang Zhengqi Dripping Pills on dampness-obstructing spleen-stomach syndrome in rats based on transcriptomic analysis

目的基于转录组学技术分析藿香正气滴丸干预湿困脾胃证的潜在作用机制。方法将SD大鼠随机分为3组:正常组、模型组、藿香正气滴丸组(1 g/kg)。采用综合物理造模方法建立湿困脾胃大鼠模型,共造模20 d,造模第11天灌胃给药。测量大鼠体质量变化、观察大鼠脏器系数、结肠组织病理变化并用ELISA法检测大鼠血清免疫球蛋白G(IgG)水平。运用转录组学技术检测结肠组织中差异基因表达情况。利用RT-qPCR验证藿香正气滴丸对关键差异基因mRNA表达水平的影响。结果与正常组比较,模型组大鼠脾脏、胸腺指数及IgG水平显著降低,结肠组织变化明显。与模型组比较,藿香正气滴丸能显著回调大鼠脾脏、胸腺指数及IgG水平,能改善结肠病理变化。转录组学分析结果显示,与正常组比较,湿困脾胃证模型组差异表达基因共525个,其中有192个基因显著上调,333个基因显著下调。藿香正气滴丸共回调了113个差异表达基因。GO功能富集结果表明差异表达基因主要涉及ATP的合成、调节干扰素系统等;KEGG分析结果表明差异表达基因主要参与氧化磷酸化代谢通路、生热作用、RIG-I样受体信号通路、NOD样受体信号通路等相关信号通路;RT-qPCR验证显示,藿香正气滴丸能明显回调差异表达基因。结论藿香正气滴丸干预湿困脾胃证的作用机制可能与其调节机体能量代谢、炎症反应和改善机体免疫系统有关。

Objective To analyze the potential mechanism of Huoxiang Zhengqi Dripping Pills in intervening in dampness-obstructing spleen-stomach syndrome based on transcriptomics technology.Methods Sprague Dawley(SD)rats were randomly allocated into 3 groups,namely the control group,the model group,Huoxiang Zhengqi Dropping Pills group(1 g/kg).The rat model was established using a comprehensive physical modeling approach over a 20-day period,and the drugs were given by intragastric administration on the 11th day.The changes of body weight,organ index and pathological changes of colon tissue were measured.The level of serum immunoglobulin G(IgG)was detected by ELISA.Transcriptomics was employed to detect differentially expressed genes in colon tissues.Real-time quantitative PCR(Rt-qPCR)was performed to validate the impact of Huoxiang Zhengqi Dripping Pills on the mRNA expression level of differentially expressed genes.Results Compared with the control group,the spleen,thymus index as well as IgG levels of the model group rats were significantly reduced,and there were significant changes in the colon tissue.Compared with the model group,Huoxiang Zhengqi Dripping Pills significantly restored spleen and thymus indices,improved IgG levels,and ameliorated colon pathological changes.Transcriptomic sequencing showed that 525 differentially expressed genes were found in the model group compared with the control group,of which 192 genes were significantly up-regulated and 333 genes were significantly down-regulated.Huoxiang Zhengqi Dripping Pills collectively modulated 113 differentially expressed genes.The GO functional enrichment results indicated that differentially expressed genes mainly involve ATP synthesis and regulation of the interferon system.The KEGG analysis results indicated that differentially expressed genes are mainly involved in oxidative phosphorylation metabolic pathways,thermogenesis,RIG-I like receptor signaling pathways,NOD like receptor signaling pathways,and other related signaling pathways.RT�qPCR showed that Huoxiang Zhengqi Dripping Pills significantly reversed differentially expressed genes.Conclusion The mechanism of intervention of Huoxiang Zhengqi Dripping Pills in dampness-obstructing spleen�stomach syndrome may be related to its regulation of energy metabolism,inflammatory response,and improvement of the immune system in the body.