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目的 围生期肾小管上皮细胞 (RTEs)急性损伤是否与窒息所致肾组织早期低氧环境直接相关 ,始终是国内外学术界争论的焦点。该实验以处于细胞增殖周期关键性调控点G1/S界面的RTEs作为体外低氧损伤的靶细胞 ,进行低氧对G1/S细胞增殖周期影响的研究 ,拟探讨低氧与RTEs急性损伤的关系。方法 以新生猪G1/S期RTEs作为研究对象 ,采用氰化钠体外低氧损伤制成细胞缺氧模型 (缺氧组 ) ,同时设干预组 (斑螯素 +氰化钠 )和未处理对照组。应用流式细胞术DNA检测法 ,比较缺氧组、干预组和对照组细胞在恢复 0 ,6 0 ,12 0 ,180minG1、S期细胞分布率以及凋亡细胞比率 ,同时应用免疫细胞化学比较三组细胞 p2 1的表达。 结果 恢复 0 ,6 0min ,缺氧组S期细胞分布率明显低于干预组 (1.4 %± 2 .5 %vs 98.3%± 1.6 % ;0 .5 %± 0 .9%vs 99.0 %± 1.0 % ;P <0 .0 1) ;在恢复 12 0、180min缺氧组凋亡细胞的比率 (33.6 %± 0 .8% ,37.5 %± 1.2 % )明显高于干预组 (2 0 .9%±1.7% ,2 2 .5 %± 1.1% )和对照组 (2 5 .6 %± 1.1,2 3.6 %± 1.4 % ) (均P <0 .0 1) ;G1期细胞分布率与相应细胞内p2 1的表达呈正线性关系 (r =0 .6 4 ,P <0 .0 1) ,而与凋亡细胞比率则无相关关系 (r =0 .342 ,P >0 .0 5 )。结论围生期RTEs急性损伤与低?
Aims Whether the acute injury of peritoneal tubular epithelial cells (RTEs) is directly related to asphyxia-induced early hypoxia in renal tissues has always been the focus of academic debate both at home and abroad. In this study, RTEs, which is the key regulatory point G1 / S of cell cycle, were used as target cells for hypoxia in vitro to study the effect of hypoxia on the proliferation cycle of G1 / S cells and to explore the relationship between hypoxia and acute injury of RTEs . Methods The neonatal pigs with G1 / S phase RTEs were selected as research objects. The hypoxia injury was induced by sodium hypochlorite in vitro (hypoxia group), and the intervention group (cantharidin + sodium cyanide) and untreated control group group. Flow cytometry DNA assay was used to compare the percentage of cells in G1, S phase and the percentage of apoptotic cells recovered in 0, 60, 120, 180 minutes in hypoxia, intervention and control groups. Immunocytochemistry was used to compare three Group of cells p21 expression. The results showed that at 0, 60 min, the distribution of cells in S phase in hypoxia group was significantly lower than that in intervention group (1.4% ± 2.5% vs 98.3% ± 1.6%; 0.5% ± 0.9% vs 99.0% ± 1.0% ; P <0.01). The percentage of apoptotic cells in the hypoxia group after resuscitation at 120,180 minutes (33.6% ± 0.8%, 37.5% ± 1.2%) was significantly higher than that in the intervention group (20.9% ± 1.7%, 22.5% ± 1.1%) and control group (25.6 ± 1.1, 3.6% ± 1.4%) (all P <0.01) The expression of p21 was positively correlated (r = 0.64, P <0.01), but not to the percentage of apoptotic cells (r = 0.342, P> 0.05). Conclusions Perinatal RTEs acute injury and low?