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目的:研究补肾益气方对皮质酮致损伤大鼠胸腺细胞周期和p16、CDK4、IL-2Rα表达的影响。方法:1×10-5M皮质酮处理新生乳鼠胸腺细胞。PI染色结合流式细胞术分析细胞周期分布;Realtime PCR和Western blotting法检测p16、CDK4、IL-2Rα的表达变化。结果:胸腺细胞经皮质酮处理后,显著阻滞于G1期;p16表达显著升高,CDK4、IL-2Rα表达降低(P<0.05)。与皮质酮组比较,补肾益气组G1期细胞数量减少,S期细胞数量升高(P<0.01);p16基因表达降低,IL-2Rα表达升高;CDK4表达无明显变化。结论:补肾益气方能抑制p16的表达,缓解皮质酮对胸腺细胞周期的抑制作用,提高胸腺依赖性免疫功能。
Objective: To study the effect of Bushengyiqi Recipe on the thymocyte cycle and the expression of p16, CDK4 and IL-2Rα in corticosterone-induced injury rats. Method: 1 × 10-5M corticosterone was used to treat neonatal thymocytes. PI staining and flow cytometry analysis of cell cycle distribution; Realtime PCR and Western blotting assay p16, CDK4, IL-2Rα expression changes. Results: Thymocytes were significantly arrested in G1 phase after corticosterone treatment. The expression of p16 was significantly increased and the expressions of CDK4 and IL-2Rα were decreased (P <0.05). Compared with corticosterone group, the number of cells in G1 phase and the number of S phase cells in Bushen Yiqi group were significantly decreased (P <0.01); the expression of p16 gene was decreased and the expression of IL-2Rα was increased; but CDK4 expression did not change significantly. Conclusion: Bushenyiqi can inhibit the expression of p16, relieve the inhibitory effect of corticosterone on thymus cell cycle, and enhance thymus-dependent immune function.