Abstract
Objective To evaluate the effects of hypoxia/reoxygenation (H/R) on apoptosis and gap junction in isolated renal tubular epithelial cells of rats.Methods The renal tubular epithelial cells were seeded in 6-well plates at a density of 1 × 105 cells/ml and randomly divided into 2 groups (n =18 wells each) using a random number table:control group (group C) and H/R group.In group C,the cells were cultured in a regular incubator with 21％ oxygen at 37 ℃.In group H/R,the cells were incubated in an anaerobic chamber for 24 h and then returned to a regular incubator and incubated for 4 h.Hoechst33258 staining method and flow cytometry were used to detect the cell apoptosis.Gap junction function was determined by specific parachute assay.Gap function protein connexin 43(Cx43) expression was measured by Western blot.Results Compared with group C,the apoptosis rate was significantly increased and gap junction function and Cx43 expression were decreased in group H/R (P ＜ 0.05).Conclusion H/R promotes apoptosis in isolated renal tubular epithelial cells through destroying intercellular gap junction in rats. Key words: Cell hypoxia ; Oxygen ; Kidney tubules ; Epithelial cells ; Apoptosis ; Connexin 43
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