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目的:探讨柔肝通络汤对大脑中动脉阻断(MACO)所致脑缺血/再灌注损伤大鼠脑组织X-连锁凋亡蛋白(X-linked inhibitor of apoptosis protein,XIAP)、Smac/DIABLO表达的影响。方法:将126只健康雄性SD大鼠分为假手术组、模型组及柔肝通络汤高、中、低剂量组。采用线栓法复制大鼠局灶性脑缺血/再灌注损伤模型,各组分别予生理盐水和不同剂量柔肝通络汤灌胃干预7 d,采用蛋白免疫组化染色(SP)法分别观察缺血再灌注后3 h、6 h、12 h、24 h、48 h各组大鼠脑组织XIAP、Smac/DIABLO的蛋白表达量及其关系曲线。结果:XIAP及Smac/DIABLO在MCAO模型中广泛表达,明显高于假手术组(P<0.05);高、中剂量柔肝通络汤组XIAP蛋白表达量显著高于模型组(P<0.05,P<0.01),Smac/DIABLO蛋白表达量明显低于模型组(P<0.05,P<0.01);两者蛋白表达动态趋势呈负相关(r=-0.999,P<0.001)。结论:XIAP及其负性调控因子Smac/DIABLO参与脑缺血/再灌注损伤后神经元凋亡的调控;柔肝通络汤对脑缺血再灌注神经元损伤具有保护作用,上调XIAP的表达、下调Smac表达是其神经保护作用的机制之一。
Objective: To investigate the effects of “Rougan Tongluo Decoction” on the expression of X-linked inhibitor of apoptosis protein (XIAP), Smac / Smad2 in cerebral ischemia-reperfusion injury induced by middle cerebral artery occlusion Impact of DIABLO expression. Methods: 126 healthy male SD rats were divided into sham operation group, model group and Rougan Tongluo Decoction high, medium and low dose groups. The model of focal cerebral ischemia / reperfusion injury was established by thread occlusion. The rats in each group were treated with normal saline and different doses of Rougan Tongluo Decoction for 7 days. The protein expression was detected by immunohistochemistry (SP) The expression of XIAP and Smac / DIABLO in brain tissue of rats in each group were observed at 3 h, 6 h, 12 h, 24 h, 48 h after ischemia-reperfusion. Results: XIAP and Smac / DIABLO were widely expressed in MCAO model and significantly higher than those in sham operation group (P <0.05). XIAP protein expression in high and medium dose Rougan Tongluo Decoction group was significantly higher than that in model group (P <0.05, P <0.01). The protein expression of Smac / DIABLO was significantly lower than that of the model group (P <0.05, P <0.01). The dynamic trend of protein expression was negatively correlated (r = -0.999, P <0.001). CONCLUSION: XIAP and its negative regulator Smac / DIABLO are involved in the regulation of neuronal apoptosis after cerebral ischemia / reperfusion injury. Rougan Tongluo decoction has a protective effect on neuronal injury induced by cerebral ischemia-reperfusion and up-regulates the expression of XIAP , Down-regulation of Smac expression is one of the mechanisms of neuroprotection.