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目的观察肢体缺血-再灌注(LI-R)后大鼠肝脏损伤中肿瘤坏死因子α(TNF-α)、半胱天冬氨酸蛋白酶(Caspase)3表达及细胞凋亡的发生和牛磺酸预处理后对其的影响。方法Wistar大鼠建立LI-R损伤模型后,随机分为对照组、LI-R组和牛磺酸+LI-R(TR)组。测定各组动物肝组织丙二醛(MDA)、髓过氧物酶(MPO)、肝组织钙含量、TNF-α水平;免疫组化法观察Caspase-3蛋白表达;利用琼脂糖凝胶电泳观察DNA梯状条带;TUNEL法检测各组肝脏细胞的凋亡情况。结果与对照组比较,LI-R组和TR组大鼠肝脏各损伤性指标、TNF-α水平及Caspase-3蛋白表达增高,肝脏细胞DNA琼脂糖凝胶电泳可见梯状条带,TUNEL染色结果显示凋亡百分率明显升高(P<0.05)。TR组上述各项指标较LI-R组显著降低(P<0.05),肝脏细胞DNA琼脂糖凝胶电泳显示梯状条带不明显。结论细胞凋亡参与了LI-R肝损伤的发生;牛磺酸预处理可降低TNF-α、Caspase-3表达,从而减少大鼠LI-R后肝脏细胞凋亡的发生。
Objective To observe the expression of tumor necrosis factor α (TNF-α), caspase-3 and apoptosis in rat liver after limb ischemia-reperfusion (LI-R) Impact on pretreatment. Methods Wistar rats were established LI-R injury model, then randomly divided into control group, LI-R group and taurine + LI-R (TR) group. The contents of malondialdehyde (MDA), myeloperoxidase (MPO), hepatic calcium content and TNF-α in the hepatic tissue of each group were measured. The expression of Caspase-3 protein was detected by immunohistochemical method. DNA ladder; TUNEL method was used to detect the apoptosis of liver cells in each group. Results Compared with the control group, the liver injury index, the level of TNF-α and the expression of Caspase-3 in the LI-R and TR groups were increased, and the ladder cells were observed by DNA agarose gel electrophoresis. The TUNEL staining results The percentage of apoptotic cells was significantly increased (P <0.05). The above indexes in TR group were significantly lower than those in LI-R group (P <0.05). DNA agarose gel electrophoresis showed no obvious ladder-like bands in liver cells. Conclusions Apoptosis is involved in the occurrence of LI-R hepatic injury. Taurine preconditioning can reduce the expression of TNF-α and Caspase-3, thereby reducing the hepatic apoptosis after LI-R in rats.