【摘要】：目的：探討腎缺血再灌注損傷細(xì)胞凋亡的發(fā)生機(jī)制和吡咯烷二巰基氨甲酸(PDTC)對其保護(hù)作用。 方法：健康雄性Wistar大鼠隨機(jī)分為3組：①缺血再灌注組(I／R組)：右側(cè)腎切除加左腎動脈夾閉45分鐘建立腎缺血再灌注模型；②藥物干預(yù)組(PDTC組)：缺血前30分鐘經(jīng)大鼠尾靜脈注射PDTC100mg/kg,其余步驟同I／R組；③假手術(shù)組：只麻醉開腹,鈍性分離腎包膜。分別于再灌注后不同時間點(diǎn)(即0h、2h、8h、24h)處死取腎。檢測血清中肌酐(Cr)和尿素氮(BUN)水平,HE染色觀察腎臟病理學(xué)改變,免疫組化和Western blot方法檢測腎組織中核因子-κB(NF-κB)、caspase-3蛋白的表達(dá),酶聯(lián)免疫吸附法(ELISA)檢測腎組織勻漿中誘導(dǎo)型一氧化氮合酶(iNOS)活性和一氧化氮(NO)的含量,原位末端標(biāo)記法(TUNEL)檢測腎組織細(xì)胞凋亡的變化。 結(jié)果：I/R組血Cr和BUN水平明顯高于假手術(shù)組和PDTC組(p0.05),HE染色可見腎小管上皮細(xì)胞壞死脫落,管腔擴(kuò)張,腎間質(zhì)炎細(xì)胞浸潤,而PDTC組病理改變明顯減輕。I/R組NF-κB表達(dá)于再灌注后2h明顯升高且在8h達(dá)到高峰,主要表達(dá)于胞核中,再灌注后24h開始下降；iNOS、NO、caspase-3、細(xì)胞凋亡指數(shù)于再灌注后2h開始表達(dá)上調(diào),此后一直呈上升趨勢,與假手術(shù)組和PDTC組相比,差異均有統(tǒng)計學(xué)意義(p0.05)。PDTC組上述指標(biāo),再灌注后2h時表達(dá)上調(diào),8h達(dá)到高峰,與假手術(shù)組比較均有顯著性差異(p0.05),再灌注后24h,差異無統(tǒng)計學(xué)意義。 結(jié)論：腎缺血再灌注損傷可引起腎組織結(jié)構(gòu)損傷和細(xì)胞凋亡,這與NF-κB引起的NO高表達(dá)有關(guān),應(yīng)用NF-κB抑制劑PDTC可發(fā)揮明顯的保護(hù)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.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R363

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 朱同玉,歐陽嘉慧,柯嘉敏,張永康,王國民;腎臟缺血再灌注損傷后一氧化氮合酶的變化[J];復(fù)旦學(xué)報(醫(yī)學(xué)版);2004年03期

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