Toll样受体信号转导通路在肾阳虚证中的改变及右归丸干预的影响

Effects of Yougui Pills on Toll-like receptor signaling pathway in kidney yang deficiency syndrome patients

目的:从Toll样受体信号转导通路的角度探讨肾阳虚证患者的免疫功能失调机制及右归丸的干预作用。方法:分别采集15例肾阳虚证患者右归丸干预前后的外周血,另以10例健康体检者为健康对照组,提取外周血白细胞,抽提mRNA,逆转录后以TLR信号通路实时定量PCR芯片进行杂交,采用ΔCt方法计算RNA的表达量,筛选肾阳虚证及右归丸干预作用相关的TLR信号通路差异表达基因。结果:筛选出与肾阳虚证相关差异表达基因28个,与右归丸干预作用相关差异表达基因38个(P<0.05)。二者中包含相同差异基因10个,其中8个在肾阳虚证中表达下调的基因经右归丸治疗后显著上调(P<0.05),2个在肾阳虚证中表达上调的基因经右归丸治疗后显著下调(P<0.05),而且TLR1、TLR8、TOLLIP、MYD88、CLEC4E、HRAS、LY86和LY96的表达量经右归丸干预后向健康对照组趋近。结论:TLR家族基因介导的信号转导异常可能是肾阳虚证免疫功能失调发生发展的信号转导机制之一。右归丸可借助免疫信号转导途径调控肾阳虚证患者的免疫功能,其机制可能主要通过上调TLR1和TLR8介导的MYD88依赖信号途径的表达,进而引起下游通路信号分子传递而实现。

Objective:To explore the effect of Yougui Pills on Toll-like receptor signaling pathway in kidney yang deficiency syndrome patients.Methods:Peripheral blood samples were collected from 15 patients with kidney-yang deficiency before and after the intervention of Yougui Pills.Another 10 healthy people were taken as control group.Leukocyte was extracted from peripheral blood and RNA was extracted.After retroviral transcription,TLR signal pathway was hybridized by real-time quantitative PCR chip.The expression of RNA was calculated byΔCt method.Differentially expressed genes of TLR signaling pathway related to kidney-yang deficiency syndrome and Yougui Pills intervention were screened.Results:Twenty-eight differentially expressed genes related to kidney-yang deficiency syndrome and 38 differentially expressed genes related to Yougui Pills intervention were screened out(P<0.05).There were 10 differentially expressed genes in both groups.Eight of them were significantly up-regulated after treatment with Yougui Pills(P<0.05).Two genes up-regulated after treatment with Yougui Pills were significantly down-regulated(P<0.05).The expression of TLR1,TLR8,TOLLIP,MYD88,CLEC4 E,HRAS,LY86 and LY96 tended to approach the healthy control group after intervention with Yougui Pills.Conclusion:Abnormal signal transduction mediated by TLR family genes may be one of the signal transduction mechanisms for the occurrence and development of immune dysfunction in kidney yang deficiency syndrome.Yougui Pills can regulate the immune function of patients with kidney yang deficiency by means of immune signal transduction pathway.Its mechanism may be realized by up-regulating the expression of MYD88 dependent signal pathway mediated by TLR1 and TLR8,and then inducing signal molecule transduction in downstream pathway.