基于转录组测序技术研究大建中汤治疗大鼠脾阳虚证的作用机制

Mechanism ofDa-Jian-Zhong decoction for treatment of spleen-Yang deficiency syndrome based on RNA-sequenced technique

目的:研究大建中汤对大鼠脾阳虚证的治疗作用和潜在作用机制。方法:大鼠随机分成正常组,脾阳虚证模型组,附子理中丸阳性组和大建中汤低、中、高剂量组。除正常组外,其余各组采用“饮食失节+劳倦+苦寒泻下”法复制脾阳虚证大鼠模型,并给予相应药物灌胃2周。检测各组大鼠体重、肛温、运动能力等指标,并对大鼠回肠和结肠组织病理变化进行检测。进一步提取回肠总RNA进行转录组测序,从而对差异基因进行GO和KEGG富集分析。结果:与模型组相比,大建中汤给药组大鼠体重、进食量、肛温以及脏器指数均明显升高( P <0.001),且运动能力接近正常组。大鼠回肠和结肠组织病理变化(以炎性浸润为主)得到改善,以低、中剂量效果较好。转录组测序结果显示,大鼠回肠组织的B细胞受体信号通路、NF-κB信号通路等免疫系统相关的信号通路明显上调。结论:大建中汤能明显改善脾阳虚证大鼠症状,其作用机制可能与提高机体免疫功能有关。

Objective: To investigate the effect and its potential mechanism of Da-Jian-Zhong decoction(DJZD) on rats with Spleen-Yang deficiency(SYD) syndrome. Methods: The SD rats were randomly divided into normal group, SYD model group, Fuzi lizhong pill positive group and three doses of DJZD treatment(low, middle and high). Except for the normal group, the other rats were given with “indiscipline in diet + excessive fatigue + intragastric administration of senna water extracts” for establishing SYD syndrome. After treatment for 2weeks, the macroscopic characterization indexes including rats body weight, rectal temperature, exercise ability, and the histopathological changes of colon and ileum were measured. Additionally, the transcriptome analysis for rats ileum tissue was performed using RNA-sequencing(RNA-seq) technology and the differentially expressed genes(DEGs) were further analyzed by using KEGG and GO enrichment. Results: Compared with the model group, the rats body weight, rectal temperature, and organ index in the DJZD treatment group were significantly increased( P <0.001), and the exercise capacity was similar to the normal group. In addition, the pathological changes of rats ileum and colon tissues were improved, mainly focused on the inflammatroy infiltration, with medium and low doses as the better effect. RNA-seq results indicated that the signaling pathways related with immune system, such as B cell receptor signaling pathway and NF-κB signaling pathway were markedly up-regulated after DJZD treatment. Conclusion: DJZD possess an obviously improving effects for the rats with spleen-Yang deficiency syndrome, and its mechanism may be related with the increased immune system function.