目的：探讨Src激酶抑制剂PP2对大鼠星形胶质细胞缺氧/复氧损伤的保护作用。方法实验分为正常对照组，缺氧/复氧组（H/R；缺氧8 h/复氧24 h），PP2+缺氧/复氧组（PP2+H/R；缺氧前给予10µmol/L PP2作用24 h）。MTT法检测各组星形胶质细胞存活率，流式细胞术测定各组星形胶质细胞凋亡率，Western blotting法检测各组星形胶质细胞中Src激酶、Bax及Bcl-2的蛋白表达。结果 MTT检测结果显示H/R损伤后细胞存活率降低，而在使用PP2预处理后细胞存活率增加（P<0.01）；流式细胞术实验结果显示H/R损伤后细胞凋亡增加，而在使用PP2预处理后细胞凋亡显著减少（P<0.01）；Western blotting法检测结果显示H/R损伤后Src激酶表达增加，用PP2预处理后Src激酶表达降低。H/R损伤后凋亡相关蛋白Bax/Bcl-2的比值增加，在用PP2预处理后Bax/Bcl-2的比值显著降低（P<0.01）。结论 PP2可对星形胶质细胞H/R损伤起保护作用，其机制可能与抑制Src激酶的蛋白表达和激活抗凋亡机制有关。%Objective To investigate the effect of Src kinase inhibitor PP2 on hypoxia/reoxygenation (H/R) injury in rat astrocytes in vitro. Methods In vitro cultured rat astrocytes were exposed to hypoxia for 8 h followed by reoxygenation for 24 h with or without pretreatment with PP2 (10 µmol/L) for 24 h before H/R injury. MTT assay and flow cytometry were used to detect the viability and apoptosis of the exposed astrocytes, respectively, and the protein expressions of Src, Bax, and Bcl-2 in the cells were determined using Western blotting. Results PP2 pretreatment significantly increased the viability and decreased the apoptosis rate of rat astrocytes exposed to H/R injury (P<0.01). Western blotting showed that H/R injury caused increased expression of Src kinase, which was lowered by PP2 pretreatment. The ratio of Bax/bcl-2 in the astrocytes increased after H/R injury, and was significantly decreased by PP2 pretreatment (P<0.01). Conclusion PP2 protects rat astrocytes from H/R injury possibly by inhibiting the expression of Src kinase and activating the anti-apoptotic mechanisms in the cells.
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