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目的探讨大鼠缺血再灌注损伤引起急性肾损伤的细胞死亡方式。方法应用光镜和电子显微镜技术、免疫组织化学染色及免疫印迹技术对缺血60 min再灌注24 h的大鼠肾脏进行了详细观察和分析。结果光镜可见皮质和髓质外带肾小管出现了①大面积细胞坏死:表现为细胞肿胀,空泡形成,崩解脱落;②坏死性凋亡:表现为细胞肿胀,核固缩;③细胞凋亡:分布于髓质外带肾小管坏死细胞之间,表现为细胞皱缩,核固缩。电镜下坏死细胞肿胀,细胞器也肿胀,崩解消失;凋亡细胞胞质皱缩,核染色质固缩边聚;坏死性凋亡呈坏死样细胞质含有一个凋亡样细胞核。免疫组化染色结果显示:PARP-1、RIPk3和caspase-3阳性细胞出现在缺血再灌注损伤肾组织内,主要分布于肾小管。免疫印迹分析结果表明:与Sham组比较肾缺血再灌注后肾组织PARP-1、RIPk3、caspase-3和TNFRa蛋白表达增强(P<0.05)。结论大鼠肾缺血60 min再灌注24 h部分肾小管上皮细胞发生了三种方式死亡,即聚合糖性死亡、坏死性凋亡和凋亡。
Objective To investigate the cell death pattern of acute kidney injury induced by ischemia-reperfusion injury in rats. Methods The kidneys of rats were observed and analyzed by light microscopy and electron microscopy, immunohistochemical staining and Western blotting in 60 min reperfusion for 60 min. Results The light microscope showed cortical and medullary tubules appeared a large area of ?? cell necrosis: the performance of cell swelling, vacuolization, disintegration shedding; ② necrotic apoptosis: the performance of cell swelling, nuclear pyknosis; ③ cells Apoptosis: distributed in medulla oblongata tubular necrosis cells, manifested as cell shrinkage, nuclear pyknosis. Under the electron microscope, the necrotic cells swollen and the organelles also swollen and disintegrated, the cytoplasm of the apoptotic cells collapsed and the nuclear chromatin was condensed. The necrotic apoptotic necrotic cytoplasm contained a apoptotic cell nucleus. The results of immunohistochemistry showed that PARP-1, RIPk3 and caspase-3 positive cells appeared in the renal tissue of ischemia-reperfusion injury and mainly distributed in the renal tubules. The results of Western blot analysis showed that compared with Sham group, the protein expressions of PARP-1, RIPk3, caspase-3 and TNFRa increased after renal ischemia-reperfusion (P <0.05). Conclusion Some renal tubular epithelial cells die in renal ischemia 60 min after reperfusion for 24 h, and there are three kinds of death, namely, polyglycosis, necrotizing apoptosis and apoptosis.