肾缺血再灌注损伤与细胞凋亡的实验研究
发布时间：2018-10-15 15:01
【摘要】：目的：探讨肾缺血再灌注损伤细胞凋亡的发生机制和吡咯烷二巯基氨甲酸(PDTC)对其保护作用。 方法：健康雄性Wistar大鼠随机分为3组：①缺血再灌注组(I／R组)：右侧肾切除加左肾动脉夹闭45分钟建立肾缺血再灌注模型；②药物干预组(PDTC组)：缺血前30分钟经大鼠尾静脉注射PDTC100mg/kg,其余步骤同I／R组；③假手术组：只麻醉开腹,钝性分离肾包膜。分别于再灌注后不同时间点(即0h、2h、8h、24h)处死取肾。检测血清中肌酐(Cr)和尿素氮(BUN)水平,HE染色观察肾脏病理学改变,免疫组化和Western blot方法检测肾组织中核因子-κB(NF-κB)、caspase-3蛋白的表达,酶联免疫吸附法(ELISA)检测肾组织匀浆中诱导型一氧化氮合酶(iNOS)活性和一氧化氮(NO)的含量,原位末端标记法(TUNEL)检测肾组织细胞凋亡的变化。 结果：I/R组血Cr和BUN水平明显高于假手术组和PDTC组(p0.05),HE染色可见肾小管上皮细胞坏死脱落,管腔扩张,肾间质炎细胞浸润,而PDTC组病理改变明显减轻。I/R组NF-κB表达于再灌注后2h明显升高且在8h达到高峰,主要表达于胞核中,再灌注后24h开始下降；iNOS、NO、caspase-3、细胞凋亡指数于再灌注后2h开始表达上调,此后一直呈上升趋势,与假手术组和PDTC组相比,差异均有统计学意义(p0.05)。PDTC组上述指标,再灌注后2h时表达上调,8h达到高峰,与假手术组比较均有显著性差异(p0.05),再灌注后24h,差异无统计学意义。 结论：肾缺血再灌注损伤可引起肾组织结构损伤和细胞凋亡,这与NF-κB引起的NO高表达有关,应用NF-κB抑制剂PDTC可发挥明显的保护作用。
[Abstract]:Aim: to investigate the mechanism of apoptosis in renal ischemia reperfusion injury and the protective effect of pyrrolidine dimercaptocarbamate (PDTC). Methods: healthy male Wistar rats were randomly divided into three groups: 1 Ischemia-reperfusion group (I / R group): right nephrectomy plus left renal artery occlusion for 45 minutes to establish renal ischemia-reperfusion model; 2Drug intervention group (PDTC group): 30 minutes before ischemia, the other steps of injection of PDTC100mg/kg, through caudal vein of rats were the same as those in group I / R, while in sham operation group, abdominal anesthesia was used only and renal capsule was obtuse. The kidneys were sacrificed at different time points after reperfusion (0 h, 2 h, 8 h and 24 h). The levels of serum creatinine (Cr) and urea nitrogen (BUN) were detected, the pathological changes of kidney were observed by HE staining, and the expression of NF- 魏 B (NF- 魏 B), caspase-3) protein in renal tissue was detected by immunohistochemistry and Western blot method. The activity of inducible nitric oxide synthase (iNOS) and the content of nitric oxide (NO) in renal homogenate were detected by enzyme-linked immunosorbent assay (ELISA), and the changes of apoptosis in renal tissue were detected by in-situ end labeling (TUNEL). Results: the levels of Cr and BUN in I / R group were significantly higher than those in sham operation group and PDTC group (p0.05), HE staining showed tubular epithelial cell necrosis and exfoliation, lumen dilatation and interstitial cell infiltration. In I / R group, the expression of NF- 魏 B increased significantly at 2 h after reperfusion and reached its peak at 8 h, mainly in the nucleus, and decreased at 24 h after reperfusion, and the apoptotic index of iNOS,NO,caspase-3, cells began to up-regulate at 2 h after reperfusion. Compared with the sham operation group and PDTC group, the difference was statistically significant (p0.05). PDTC group). The expression of P0. 05). PDTC increased at 2 h after reperfusion and reached its peak at 8 h after reperfusion. There was significant difference between sham operation group and sham operation group (p0.05), but there was no significant difference at 24 hours after reperfusion. Conclusion: renal ischemia-reperfusion injury can induce renal tissue structure damage and apoptosis, which is related to the overexpression of NO induced by NF- 魏 B. PDTC can play a protective role in renal ischemia reperfusion injury.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R363
本文编号：2272915
[Abstract]:Aim: to investigate the mechanism of apoptosis in renal ischemia reperfusion injury and the protective effect of pyrrolidine dimercaptocarbamate (PDTC). Methods: healthy male Wistar rats were randomly divided into three groups: 1 Ischemia-reperfusion group (I / R group): right nephrectomy plus left renal artery occlusion for 45 minutes to establish renal ischemia-reperfusion model; 2Drug intervention group (PDTC group): 30 minutes before ischemia, the other steps of injection of PDTC100mg/kg, through caudal vein of rats were the same as those in group I / R, while in sham operation group, abdominal anesthesia was used only and renal capsule was obtuse. The kidneys were sacrificed at different time points after reperfusion (0 h, 2 h, 8 h and 24 h). The levels of serum creatinine (Cr) and urea nitrogen (BUN) were detected, the pathological changes of kidney were observed by HE staining, and the expression of NF- 魏 B (NF- 魏 B), caspase-3) protein in renal tissue was detected by immunohistochemistry and Western blot method. The activity of inducible nitric oxide synthase (iNOS) and the content of nitric oxide (NO) in renal homogenate were detected by enzyme-linked immunosorbent assay (ELISA), and the changes of apoptosis in renal tissue were detected by in-situ end labeling (TUNEL). Results: the levels of Cr and BUN in I / R group were significantly higher than those in sham operation group and PDTC group (p0.05), HE staining showed tubular epithelial cell necrosis and exfoliation, lumen dilatation and interstitial cell infiltration. In I / R group, the expression of NF- 魏 B increased significantly at 2 h after reperfusion and reached its peak at 8 h, mainly in the nucleus, and decreased at 24 h after reperfusion, and the apoptotic index of iNOS,NO,caspase-3, cells began to up-regulate at 2 h after reperfusion. Compared with the sham operation group and PDTC group, the difference was statistically significant (p0.05). PDTC group). The expression of P0. 05). PDTC increased at 2 h after reperfusion and reached its peak at 8 h after reperfusion. There was significant difference between sham operation group and sham operation group (p0.05), but there was no significant difference at 24 hours after reperfusion. Conclusion: renal ischemia-reperfusion injury can induce renal tissue structure damage and apoptosis, which is related to the overexpression of NO induced by NF- 魏 B. PDTC can play a protective role in renal ischemia reperfusion injury.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R363
【参考文献】
相关期刊论文 前1条
1 朱同玉,欧阳嘉慧,柯嘉敏,张永康,王国民;肾脏缺血再灌注损伤后一氧化氮合酶的变化[J];复旦学报(医学版);2004年03期
，本文编号：2272915
本文链接：https://www.wllwen.com/xiyixuelunwen/2272915.html
最近更新
教材专著
