丙泊酚对肢体缺血-再灌注大鼠肾损伤和肺损伤时细胞凋亡的影响

目的从细胞凋亡的角度探讨大鼠肢体缺血-再灌注后肾损伤和肺损伤的发病机制及丙泊酚对其影响的可能机制。方法复制大鼠肢体缺血-再灌注损伤(LIR)动物模型。24只SD大鼠随机分为对照组(C组)、缺血-再灌注组(IR组)、丙泊酚处理组(P组)和10%的脂肪乳对照组(T组)。采用脱氧核苷酸末端转移酶介导的DNA原位末端缺口标记技术(TUNEL)检测细胞凋亡的变化,琼脂糖凝胶电泳法进一步证实组织DNA的改变,同时用免疫组织化学的技术检测肾、肺Caspase-3、Bcl-2蛋白的表达。结果(1)C组肺组织和肾组织未见细胞凋亡现象,IR组凋亡细胞显著增多,P组凋亡细胞数较IR组和T组显著减少而多于C组(P<0.05),T组和IR组比差异无统计学意义。(2)Bcl-2表达在IR组和T组均比C组明显下降(P<0.05),P组的表达比IR组和T组增高(P<0.05)。Caspase-3在IR组和T组的表达显著高于P组和C组(P<0.05),P组和C组及IR组和T组差异有统计学意义。Caspase-3表达与凋亡细胞数呈正相关(r=0.9149,P<0.01);Caspase-3与Bcl-2的表达呈直线负相关(P<0.01)。(3)C组肾、肺组织均无凋亡梯形条带的出现,而IR组、T组有凋亡典型的梯形条带,P组梯形条带不甚清楚。结论肾、肺细胞凋亡的发生可能参与了LIR后肾、肺损伤,丙泊酚的活性成分1,6-二异丙酚通过抑制Caspase-3表达同时上调Bcl-2而减少细胞凋亡,从而减轻LIR后肾和肺的损伤。

Effects of propofol on kidney and lung apoptosis following ischemia-reperfusion of rat hind limbs

Objective To investigate the effect of propofol on apoptosis following ischemia-reperfusion(IR) of rat hind limbs. Methods Copy a rat model of limb ischemia-reperfusion. Twenty-four SD rats were randomly assigned into 4 groups with 6 rats each: in group C, the bilateral femoral artery and vein were exposed but not occluded, in group IR the bilateral femoral artery and vein were occluded for 3 h with the atraumatic micro clips and released for 4h reperfusion, the rats in group P received 10 mg/kg propofol 30 min before ischemia, and those in group T received same amount of 10% intralipid instead of propofol. Apoptosis was identified by TUNEL technique,by immunohistochemistry was detected the expression of apoptosis proteins including Caspase-3,Bcl-2. Agorose gel electrophoresis was detected to confirm the change of DNA. Results (1)There were no significant apoptotic changes in group C. IR significantly increased apoptosis index(AI) of the lung and kidney. Propofol significantly attenuated these changes induced by IR of the hind limbs in group P as compared with group IR and T. (2)Optical density of Caspase-3 protein was significantly higher in group IR and T than that in group P and C. The expression of Caspsae-3 positively correlated to AI, and negatively correlated to Bcl-2.(3)There was no DNA fragment seen in group C, which was found in group IR,T and P. Conclusion Apoptosis of the kidney and lung participate the injury induced by IR of the hind limbs. Propofol can ameliorate the development of apoptosis by inhibiting the Caspase-3 protein and up-regulating the Bcl-2 protein.