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目的:观察缺血预适应(IPC)对大鼠肢体缺血再灌注(LIR)后肝脏损伤和细胞凋亡的影响,以进一步探讨IPC对肢体缺血再灌注后肝脏功能的保护作用。方法:将实验用雄性Wistar大鼠18只,随机分为对照组、缺血再灌注(IR)组和缺血预适应加再灌注(IPC+IR)组,每组6只。测定各组动物血浆中丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、透明质酸(HA)和乳酸脱氢酶(LDH);检测肝组织细胞的DNA双链百分率;利用原位脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(TUNEL法)检测各组肝细胞凋亡情况;采用免疫组织化学方法检测各组动物肝组织Bcl-2和Bax比值及Caspase-3。结果:大鼠LIR后血浆中ALT、AST、HA、LDH含量均明显增加;LIR后肝组织细胞DNA双链百分率降低,凋亡细胞百分率增高。Bcl-2/Bax蛋白比值减小,Caspase-3蛋白表达明显增强。IPC+IR组上述各指标较IR组改善,但与对照组比较差异仍有统计学意义(P<0.05或P<0.01)。结论:肢体缺血再灌注可以导致肝脏的损伤,缺血预适应可以减轻损伤的发生,其保护效应可能与其抑制肝细胞凋亡有关。
OBJECTIVE: To observe the effect of ischemic preconditioning (IPC) on liver injury and apoptosis after limb ischemia-reperfusion (LIR) in rats to further explore the protective effect of IPC on liver function after limb ischemia-reperfusion. Methods: Eighteen male Wistar rats were randomly divided into control group, ischemia reperfusion group and IPC + IR group, with 6 rats in each group. The levels of ALT, AST, HA and LDH in the plasma of each group were determined. The DNA double-stranded Percentage of hepatocytes were detected by in situ deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) assay. Immunohistochemistry was used to detect the ratio of Bcl-2 to Bax And Caspase-3. Results: After LIR, plasma ALT, AST, HA, LDH levels were significantly increased; DNA double-stranded percentage decreased and the percentage of apoptotic cells increased after LIR. Bcl-2 / Bax protein ratio decreased, Caspase-3 protein expression was significantly enhanced. The above indexes in IPC + IR group were better than those in IR group, but there was still significant difference compared with control group (P <0.05 or P <0.01). Conclusion: Limb ischemia-reperfusion can lead to liver injury. Preconditioning with ischemia can reduce the occurrence of injury and its protective effect may be related to the inhibition of hepatocyte apoptosis.