發(fā)布時(shí)間：2018-08-22 08:55

【摘要】：目的:觀察缺血后處理對(duì)大鼠肢體缺血再灌注(LIR)后腎組織細(xì)胞凋亡的影響,探討其可能機(jī)制。方法:30只SD大鼠隨機(jī)分為對(duì)照組、缺血再灌注組(IR組)和缺血后處理加再灌注組(I-postC組),每組10只。建立大鼠肢體缺血再灌注(LIR)模型,即橡皮帶環(huán)繞大鼠雙后肢根部阻斷血流4h再恢復(fù)血流灌注4h。對(duì)照組大鼠僅松弛環(huán)繞橡皮帶不阻斷血流,I-postC組大鼠則在再灌注前附加反復(fù)3次缺血5min-再灌注5min操作,即缺血后處理。全自動(dòng)生化分析儀測(cè)各組大鼠血漿肌酐(Cr)、尿素氮(BUN)和C反應(yīng)蛋白(CRP)水平;免疫組織化學(xué)法檢測(cè)大鼠腎組織凋亡相關(guān)蛋白Bcl-2和Bax的表達(dá),采用自動(dòng)圖像分析系統(tǒng)統(tǒng)計(jì)其定量結(jié)果并計(jì)算Bcl-2/Bax比值;TUNEL染色后在激光共聚焦顯微鏡下觀察腎組織細(xì)胞凋亡情況,電鏡下觀察腎組織超微結(jié)構(gòu)。結(jié)果:與對(duì)照組比較,IR組和I-postC組大鼠血漿Cr、BUN和CRP水平均明顯增高(P0.01);與IR組比較,I-postC組大鼠血漿Cr、BUN和CRP水平均明顯降低(P0.01)。與對(duì)照組比較,IR組和I-postC組大鼠腎組織中Bax和Bcl-2表達(dá)水平明顯升高(P0.05或P0.01),Bcl-2/Bax比值降低(P0.05);與IR組比較,I-postC組大鼠腎組織中Bax表達(dá)水平降低(P0.05),Bcl-2表達(dá)水平升高(P0.01),Bcl-2/Bax比值升高(P0.05)。激光共聚焦顯微鏡下觀察,與對(duì)照組比較,IR組大鼠腎組織中凋亡細(xì)胞明顯增多;與IR組比較,I-postC組大鼠腎組織中凋亡細(xì)胞明顯減少。透射電鏡下觀察,對(duì)照組大鼠腎組織細(xì)胞結(jié)構(gòu)清晰完整;IR組大鼠腎組織中腎近曲小管上皮細(xì)胞核固縮,溶酶體和致密顆粒沉積增多,線粒體數(shù)目減少,部分線粒體嵴斷裂或模糊,腎小球足突細(xì)胞突起不規(guī)則、融合,有空泡現(xiàn)象,粗面內(nèi)質(zhì)網(wǎng)擴(kuò)張;與IR組比較,I-postC組大鼠腎組織中腎小管上皮細(xì)胞及腎小球損傷有一定程度改善。結(jié)論:LIR可誘發(fā)大鼠腎組織細(xì)胞凋亡,缺血后處理可抑制肢體缺血再灌注后的腎細(xì)胞凋亡,對(duì)改善大鼠腎功能有一定作用。
[Abstract]:Aim: to observe the effect of post-ischemic treatment on apoptosis of renal tissue after limb ischemia reperfusion (LIR) in rats and to explore its possible mechanism. Methods Thirty Sprague-Dawley rats were randomly divided into control group, ischemia reperfusion group (IR group) and post-ischemic plus reperfusion group (I-postC group) with 10 rats in each group. The (LIR) model of limb ischemia-reperfusion was established in rats, that is, the rubber band around the roots of the hind limbs blocked the blood flow for 4 h and then recovered the blood flow for 4 h. In the control group, the rats in the I-postC group were treated with repeated 5 min-reperfusion 5min for 3 times before reperfusion, that is, after ischemia. The levels of plasma creatinine (Cr), urea nitrogen (BUN) and C-reactive protein (CRP) were measured by automatic biochemical analyzer and the expression of apoptosis-related proteins Bcl-2 and Bax in renal tissue of rats were detected by immunohistochemical method. The quantitative results were analyzed by automatic image analysis system and the Bcl-2/Bax ratio was calculated by Tunel staining. The apoptosis of renal tissue was observed under laser confocal microscope and the ultrastructure of renal tissue was observed under electron microscope. Results: compared with the control group, the plasma levels of Cr bun and CRP in IR group and I-postC group were significantly increased (P0.01), and those in I-postC group were significantly lower than those in IR group (P0.01). Compared with the control group, the expression of Bax and Bcl-2 in renal tissue of IR group and I-postC group were significantly increased (P0.05 or P0.01) and the ratio of Bcl-2 / Bax was decreased (P0.05), and the expression of Bax in renal tissue of I-postC group was lower than that of IR group (P0.05). The expression level of Bcl-2 was increased (P0.01) and the ratio of Bcl-2 / Bax was increased (P0.05). Compared with the control group, the apoptotic cells in renal tissue of IR group were significantly increased, and those in I-postC group were significantly decreased compared with IR group under confocal laser microscope. Under transmission electron microscope, the renal cell structure of the control group was clear and intact. In IR group, the renal proximal convoluted tubule epithelium was pyknosis, lysosome and dense granules were increased, and the number of mitochondria was decreased. Some mitochondria cristae were broken or blurred, glomerular podocytes were irregular, fused, empty vesicles, rough endoplasmic reticulum dilated, and renal tubular epithelial cells and glomerular damage in I-postC group were improved to some extent compared with IR group. Conclusion: WLIR can induce renal cell apoptosis in rats, and post ischemic treatment can inhibit the apoptosis of renal cells after limb ischemia reperfusion, which can improve the renal function of rats to some extent.
【作者單位】： 華北理工大學(xué)基礎(chǔ)醫(yī)學(xué)院病理生理學(xué)系;
【基金】：河北省衛(wèi)計(jì)委醫(yī)學(xué)研究重點(diǎn)項(xiàng)目資助課題(20130060)
【分類號(hào)】：R692

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