目的 观察大鼠肢体缺血/再灌注损伤（limb ischemia/reperfusion injury， LI/RI）对小肠组织自噬的影响。 方法 采用大鼠LI/RI模型，将雄性SD大鼠48只按随机数字表法随机分为两组（每组24只）：假手术组（S组）和缺血/再灌注（ischemia/reperfusion, I/R）组，两组分别再分为再灌注0 （T0）、2（T2）、4（T4）、8 h（T8）4个亚组，每亚组6只。I/R组阻断血流3 h后再灌注，S组不阻断血流，分别于上述4个时点测定各组血清中乳酸脱氢酶（lactate dehydrogenase， LDH）活性，荧光显微镜下观察小肠细胞自噬小体，聚合酶链式反应（polymerase chain reaction， PCR）检测自噬相关基因Beclin1、Atg5的mRNA表达，蛋白免疫印迹法（Western blot）检测自噬标志蛋白LC3蛋白的表达水平。 结果 I/R组T2、T4、T8时点血清LDH分别为［（4 623±397）、（5 046±413）、（4 762±431） U/L］（P＜0.05），与S组相同时点比较，分别升高约78%、87%、81%；I/R组荧光显微镜下单位面积自噬小体与S组比较明显增加；I/R组T2、T4、T8时点Beclin1表达增加，与S组比较，分别增加约42%、69%、33%（P＜0.05）；I/R组T2、T4、T8时点Atg5表达增加，与S组比较，分别增加约80%、121%、73%（P＜0.01）；I/R组T2、T4、T8时点LC3表达增加，与S组比较，LC3Ⅱ与 LC3I的比值分别增加约169%、186%、176%（P＜0.01）。 结论 LI/RI可致大鼠小肠组织自噬水平增加。
Ｏｂｊｅｃｔｉｖｅ To investigate the effect of autophagy in small intestine indused by low limb ischemia/reperfusion injury（LI/RI） in rats. Methods The model of LI/RI was performed. Forty eight male SD rats were randomly divided into two groups： the sham-operation group（group S） and limb ischemia/reperfusion（I/R） group （group I/R）， then two groups were separated into four subgroups according to reperfusion time point 0 （T0）， 2（T2）， 4（T4）， 8 h（T8） and each subgroup contained six rats， respectively. In group I/R， bilateral limb ischemia was induced by rubber band application for 3 h. In group S， bloodstream was not blocked. Lactate dehydrogenase（LDH） activity in serum were sampled from the inferior vena and experimental animals were killed with the small intestine removed for determination of beclin1， Atg5， LC3 level at each time point. Results Compared with group S at the same time point， LDH activity in serum increased significantly at T2 ［from （2 597±493） U/L to （4 623±397） U/L （P＜0.05）］， at T4 ［from （2 679±504） U/L to（5 046±413） U/L （P＜0.05）］， at T8 ［from （2 635±489） U/L to （4 762±431） U/L （P＜0.05）］. Compared with group S， the autophagosome in small intestine increased significantly in unit area under fluorescent microscope. Compared with group S， the level of Beclin1 increased about 42%， 69% and 33% respectively from T2 to T8（P＜0.05）. Compared with group S， the level of Atg5 increased about 80%， 121% and 73% respectively from T2 to T8（P＜0.01）. Compared with group S， LC3Ⅱ/LC3I increased about 169%， 186% and 176% respectively from T2 to T8（P＜0.01）. Conclusions LI/RI may lead to increased level of autophagy in small intestine.
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