环磷酰胺诱导大鼠膀胱黏膜H2AX表达降低意义的实验研究

Experimental study on the decreasing of H2AX expression induced by cyclophosphamide in rat bladder mucosa

目的探讨H2AX在环磷酰胺诱导大鼠膀胱黏膜中表达降低的作用机制及意义。方法将12只SD大鼠随机分为对照组(n=6)和实验组(n=6),实验组大鼠给予环磷酰胺100 mg/kg腹腔注射,对照组给予相同剂量生理盐水腹腔注射。正常饲养24 h后,在无菌低温环境下剖腹取膀胱,HE染色观察膀胱形态变化,免疫组织化学方法检测H2AX蛋白的定位,蛋白印迹检测H2AX蛋白的表达量。结果 HE染色,与对照组比较,实验组大鼠膀胱黏膜肿胀,表面粗糙,呈现断续染色;免疫组化见H2AX蛋白表达在膀胱黏膜上皮细胞核中;蛋白印迹检测可见H2AX蛋白在实验组表达量显著低于对照组(P<0.05)。结论环磷酰胺可诱导大鼠膀胱黏膜H2AX蛋白表达降低,进而导致膀胱癌的发生。

Objective To explore the mechanism and significance of H2AX expression in rat bladder mucosa induced by cyclophosphamide.Methods 12 SD rats were randomly divided into control group(n=6)and treatment group(n=6),the rats in the treatment group were given the intraperitoneal injection of cyclophosphamide(100 mg/kg),the control group were given the same dose of saline.After 24 h of normal feeding,the bladders were removed by laparotomy under sterile and low temperature environment.The morphological changes of bladders were observed by HE staining,and the locations of H2AX protein were detected by immunohistochemistry,the expressions of H2AX protein were detected by Western blotting.Results Compared with the control group,the bladder mucosa in the treatment group was swollen,with rough surface and intermittent staining.Immunohistochemistry showed that H2AX protein was expressed in the nucleus of bladder epithelial cells.Western blotting showed that the expression level of H2AX protein in the treatment group was significantly lower than that in the control group(P<0.05).Conclusion Cyclophosphamide can induce the expression of H2AX protein in rat bladder mucosa.