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基于方剂与证候相关理论的痹证经典方剂TLR/TRAF信号通路调控的比较研究

[Comparative study of regulation of TLR/TRAF signaling pathway of classic prescriptions that deals with arthralgia syndrome based on relevent theory of prescription and syndrome].

作者信息

Xu Shijun, Li Lei, Zhang Wensheng, Wang Yongyan

机构信息

Pharmacy School of Chengdu University of Traditional Chinese Medicine, Chengdu 610075, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2010 Apr;35(8):1025-9.

Abstract

OBJECTIVE

Based on relevent theory of prescription and syndrome, to compare the gene expression differences of TLR2, TRAF6, and Faslg with adjuvant arthritis in rat spleen among Wutou decoction, Guizhi Shaoyao Zhimu decoction and Baihu Guizhi decoction.

METHOD

The experiment animal model of adjuvant arthritis in rats was established. Relative expression amount of TLR2, TRAF6, and Faslg in rats spleen was detected by SYBR Green I dye methods and implementation of fluorescence quantitative PCR technology with 18sRNA as an internal gene. 2(delta delta CT) method was used for computing and data analysis.

RESULT

TLR2, TRAF6, and Faslg gene in adjuvant arthritis rat spleen was significantly higher than those in the blank group. The various doses of Wutou decoction, Guizhi Shaoyao Zhimu decoction and Baihu Guizhi decoction can significantly inhibit or reduce the abnormally high expression of TLR2, TRAF6, and Faslg genes. The gene expression level caused by three decoctions mentioned above was followed by strong to weak as Wutou decoction Guizhi Shaoyao Zhimu decoction and Baihu Guizhi decoction with the clinical equivalent dose, but the strength of the trend to reduce the role of TRAF6 is just the opposite with the TLR2 and Faslg genes.

CONCLUSION

Wutou decoction, Guizhi Shaoyao Zhimu decoction and Baihu Guizhi decoction can reduce the abnormally high expression of TLR2, TRAF6 and Faslg in rat spleen with adjuvant arthritis, but the differences of intensity exist and remain relatively consistent with that of pharmacodynamics and regulation trends of T cell subsets. Results suggest that the suppression of TLR2/TRAF6 signal pathway and apoptosis Faslg receptor gene may be the reasons that the pharmacodynamics of three decoctions on peripheral T cell subsets in regulating intensity was different.

摘要

目的

基于方剂与证候的相关理论,比较乌头汤、桂枝芍药知母汤、白虎桂枝汤对佐剂性关节炎大鼠脾脏中TLR2、TRAF6、Faslg基因表达的差异。

方法

建立大鼠佐剂性关节炎实验动物模型。以18sRNA为内参基因,采用SYBR Green I染料法并运用荧光定量PCR技术检测大鼠脾脏中TLR2、TRAF6、Faslg的相对表达量。采用2(ΔΔCT)法进行计算和数据分析。

结果

佐剂性关节炎大鼠脾脏中TLR2、TRAF6、Faslg基因表达明显高于空白组。乌头汤、桂枝芍药知母汤、白虎桂枝汤各剂量组均能显著抑制或降低TLR2、TRAF6、Faslg基因的异常高表达。上述三方剂临床等效剂量引起的基因表达水平由强至弱依次为乌头汤、桂枝芍药知母汤、白虎桂枝汤,但降低TRAF6作用的强度趋势与TLR2、Faslg基因相反。

结论

乌头汤、桂枝芍药知母汤、白虎桂枝汤均可降低佐剂性关节炎大鼠脾脏中TLR2、TRAF6、Faslg的异常高表达,但存在强度差异,且与药效学及T细胞亚群调节趋势较为一致。结果提示,抑制TLR2/TRAF6信号通路及Faslg受体凋亡基因可能是三方剂对外周T细胞亚群调节强度药效学不同的原因。

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