补肾及健脾复方对皮质酮大鼠T细胞凋亡信号相关基因群调控模式的对比研究

目的:探讨补肾及健脾复方对皮质酮大鼠T细胞凋亡信号相关基因表达的调控模式.方法:采用TUNEL标记的流式细胞技术,对皮质酮大鼠模型及各治疗组激活诱导的T细胞凋亡进行定量分析;采用荧光实时定量PCR技术对各组大鼠T细胞凋亡信号相关基因群mRNA表达水平进行定量与比较;采用分光光度法检测caspase-8、caspase-3的活性.结果:(1)与正常组相比,模型组T细胞凋亡百分率明显增高;右归饮组及补肾益寿胶囊组T细胞凋亡百分率与皮质酮模型组相比明显降低;四君子汤与模型组相比T细胞凋亡百分率无显著性差异.(2)模型组促凋亡的Fas、FasL、TNR1、Bax基因表达上调,抗凋亡的TNFR2、Bcl-2、cIAP1、cIAP2基因表达下调;两个补肾复方均可下调FasL、TNFR1 mRNA表达水平,并可上调Bcl-2、cIAPl、cIAP2 mRNA表达水平;四君子汤仅对Bcl-2,cIAP2 mRNA表达水平有显著上调作用.(3)模型组caspase-8、caspase-3活性显著增高,两个补肾复方均可降低caspase-8、caspase-3的活性,四君子汤对caspase-8、caspase-3的活性无显著下调作用.结论:两个补肾复方下调促凋亡基因FasL、TNFR1的转录,上调抗凋亡基因Bcl-2、cIAPl、cIAP2的转录,并且降低caspase-8、caspase-3的活性,从而降低皮质酮大鼠过度的T细胞凋亡,是补肾法特有的对皮质酮大鼠T细胞凋亡相关基因群的调控模式.

Comparative study of kidney-tonifying recipes and spleen-invigorating recipe on regulating pattern of the expression of T lymphocyte apoptosis signaling-related genes in corticosterone-treated rats

Objective:To investigate the regulating pattern of kidney-tonifying recipes and spleen-invigorating recipe on the expression of T lymphocyte apoptosis signaling-related genes in corticosterone-treated rats. Methods: We analyzed the activation-induced T lymphocyte apoptpsis rate through TUNEL-labelled flow-cytometry assay, quantified the mRNA expression level of T cell apoptosis-regulating genes by using the fluoresecence real-time PCR, analyzed the activity of caspase-8 and caspase-3 by colorimetric assay. Results:(1) T lymphocyte apoptosis rate of corticosterone-treated rats was higher than that of the normal control rats; T lymphocyte apoptosis rates of Youguiyin treated rats and Bushen Yishou Capsule treated rats were lower than that of the corticosterone-treated group; T lymphocyte apoptosis rate of Sijunzi Decoction treated rats displayed no significance compared with that of the corticosterone-treated group. (2) Pro-apoptotic genes such as Fas, FasL, TNFR1, and Bax displayed increased expression level in corticosterone-treated group compared with normal control group. Two kidney-tonifying recipes down-regruated the expression level of FasL and TNFR1, at the same time, both recipes up-regulated the expression level of Bcl-2. The spleen-invigorating recipe had up-regulating effect on the expression level of Bcl-2. And all these results had statistical significance. (3) The activities of caspase-8 and caspase-3 were increased in corticosterone-treated rats compared with normal control; both kidney-tonifying recipes had significant effect on decreasing the activities of these two apoptosis executioners. Conclusion: The excessive apoptosis of T cell in corticosterone-treated rats could be effectively modulated with two kidney-tonifying recipes. Down-regulating the expression of FasL and TNFR1 and up-regulating the expression of Bcl-2 as well as decreasing the activities of caspase-8 and caspase-3 may be the unique regulating pattern of tonify-ing-kidney method with respect to T cell apoptosis related genes in corticosterone-treated rats.