缺血预适应减少大鼠肢体缺血再灌注后的肝细胞凋亡

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目的观察缺血预适应(IPC)对大鼠肢体缺血再灌注(I/R)后肝脏自由基和钙含量的改变与细胞凋亡情况以及对其变化的影响。方法实验用雄性Wistar大鼠18只,随机分为对照(Control)、缺血再灌注(IR)组和缺血预适应(IPC+IR)组,每组6只。分别测定血浆谷丙转氨酶(AST)、乳酸脱氢酶(LDH)、血浆和肝组织超氧化物歧化酶(SOD)、丙二醛(MDA)的含量,肝组织钙及线粒体钙含量;免疫组化法检测肝组织的BCL-2和BAX蛋白的表达水平;采用末端标记技术(TUNEL)检测肝细胞凋亡。结果肢体IR后血浆AST、LDH显著增多,血浆和肝SOD减少而MDA显著增加;肝组织钙及线粒体钙含量显著增多;BCL-2与BAX蛋白表达增多,而BCL-2/BAX比值显著降低;凋亡细胞显著增多。IPC显著减轻了IR后引起的MDA含量的升高,并且显著增加了SOD的含量;减轻了组织和线粒体钙超载;BCL-2的表达则明显升高而BAX表达较IR组减少,BCL-2/BAX比值升高;凋亡细胞显著减少。结论肢体IR引起肝脂质过氧化产物增多,钙超载,凋亡细胞增多。IPC可能通过减少自由基的产生及钙超载抑制细胞凋亡而减轻肢体IR后继发的肝损伤。 Objective To observe the effects of ischemic preconditioning (IPC) on the changes of free radicals and calcium contents and the apoptosis of limbs and their changes after limb ischemia / reperfusion (I / R) in rats. Methods 18 male Wistar rats were randomly divided into control group, IR group and IPC + IR group, with 6 rats in each group. The contents of AST, LDH, SOD, MDA and the content of calcium and mitochondrial calcium in liver tissue were measured respectively. The immune group The levels of BCL-2 and BAX protein in liver tissue were detected by chemiluminescence assay. Apoptosis of hepatocytes was detected by TUNEL. Results Compared with control group, the levels of AST and LDH were significantly increased in the extremity after IR and the levels of MDA and SOD in plasma and liver were significantly increased. The contents of calcium and mitochondrial calcium in liver tissue were significantly increased. The expressions of BCL-2 and BAX were increased, but the ratio of BCL-2 / BAX was significantly decreased. Apoptotic cells increased significantly. IPC significantly reduced the increase of MDA content induced by IR, and significantly increased the content of SOD; alleviated the calcium overload of tissues and mitochondria; the expression of BCL-2 was significantly increased and the expression of BAX was decreased compared with IR group; / BAX ratio increased; apoptotic cells decreased significantly. Conclusion Li limbs lead to increased hepatic lipid peroxidation products, calcium overload, apoptotic cells increased. IPC may reduce secondary liver injury after IR by reducing the production of free radicals and calcium overload and inhibiting apoptosis.
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