Zhao Zi-gang, Niu Chun-yu, Zhang Jing, Bo Ai-hua
Department of Pathophysiology, Hebei North University, Zhangjiakou 075029, Hebei, China.
Zhongguo Wei Zhong Bing Ji Jiu Yi Xue. 2007 Dec;19(12):724-6.
To explore the effect of mesenteric lymph duct ligation in preventing apoptosis of renal tubule epithelial cells in rats by two-hits including hemorrhage and lipopolysaccharide (LPS) challenge.
Forty-five Wistar rats were randomly divided into three groups: the ligation group, the non-ligation group and sham operation group. The two-hits model was established by withdrawing the blood via the right side carotid artery of rats and intragastric administration 4 mg/kg LPS 6 hours after hemorrhage, and mesenteric lymph flow was blocked by ligating mesenteric lymph duct in ligation group. Twenty-four hours after operation, kidney was harvested for pathological examination, and the apoptosis cells rate of renal tubule epithelial cells were determined by method of terminal-deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL), the expression of apoptosis-correlating gene bcl-2 and bax protein were determined by streptavidin-biotin complex (SABC) immunohistochemical method.
After two-hits, the apoptosis rate and expression of bax protein of renal tubule epithelial cells in non-ligation group were significantly increased, and expression of bcl-2 protein was significantly lower as compared with those of sham operation group (all P<0.01). But the apoptosis rate and expression of bax protein of renal tubule epithelial cells in ligation group were significantly lower, and expression of bcl-2 protein was significantly increased as compared with non-ligation group (all P<0.01).
The results demonstrate that the ligation of mesenteric lymph duct could ameliorate the apoptosis of renal tubule epithelial cells in rats as produced by hemorrhage and LPS, and its mechanism might relate to the reduction of the down regulation of gene expression of bax protein and enhancement of the expression of bcl-2 protein after ligation of mesenteric lymph duct.
探讨肠系膜淋巴管结扎对出血合并脂多糖(LPS)二次打击致大鼠肾小管上皮细胞凋亡的防治作用。
45只Wistar大鼠随机分为结扎组、未结扎组和假手术组。采用大鼠右侧颈总动脉放血,放血6小时后经胃管给予4mg/kg LPS的方法建立二次打击模型,结扎组结扎肠系膜淋巴管以阻断肠系膜淋巴液回流。术后24小时取肾组织进行病理检查,采用脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(TUNEL)检测肾小管上皮细胞凋亡率,采用链霉亲和素-生物素复合物(SABC)免疫组化法检测凋亡相关基因bcl-2及bax蛋白表达。
二次打击后,未结扎组肾小管上皮细胞凋亡率及bax蛋白表达显著升高,bcl-2蛋白表达显著低于假手术组(均P<0.01)。与未结扎组比较,结扎组肾小管上皮细胞凋亡率及bax蛋白表达显著降低,bcl-2蛋白表达显著升高(均P<0.01)。
结果表明,结扎肠系膜淋巴管可减轻出血合并LPS所致大鼠肾小管上皮细胞凋亡,其机制可能与结扎肠系膜淋巴管后降低bax蛋白基因表达下调及增强bcl-2蛋白表达有关。