【摘要】 目的 觀察百草枯中毒大鼠腎組織中血紅素氧合酶1(HO1)的表達(dá),探討其病理生理機(jī)制。 方法 SD大鼠126只隨機(jī)分為空白對(duì)照組、中毒組和褪黑素組,各42只。中毒組、褪黑素組予以百草枯(25 mg/kg)腹腔注射染毒,對(duì)照組予以等量生理鹽水腹腔注射,15 min后褪黑素組予以褪黑素(10 mg/kg)腹腔注射,對(duì)照組、中毒組予以等量生理鹽水腹腔注射。于1、3、6、12 h,1、2、3、5 d時(shí)各組隨機(jī)取6只處死。蘇木精〖CD3/5〗伊紅（HE）染色觀察各組腎組織病理學(xué)變化,采用免疫組識(shí)化學(xué)和RTPCR觀察腎組織HO1蛋白和mRNA表達(dá)。 結(jié)果 ①與對(duì)照組相比,中毒組染毒后3 h即可見(jiàn)充血、水腫及空泡變性等病理變化,1 d達(dá)頂峰,病理?yè)p傷評(píng)分3.30±0.31(Plt;0.05),其后緩解趨勢(shì)不明顯；而褪黑素組病理變化明顯減輕且緩解趨勢(shì)明顯,1 d時(shí)病理?yè)p傷評(píng)分2.70±0.26,與中毒組相比差異有統(tǒng)計(jì)學(xué)意義(Plt;0.05)。②與對(duì)照組相比,中毒組染毒3 h在皮質(zhì)部腎小管上皮細(xì)胞的細(xì)胞膜及細(xì)胞漿HO1呈陽(yáng)性表達(dá),免疫組識(shí)化學(xué)評(píng)分(IHS)3.33±1.75,HO1 mRNA表達(dá)增強(qiáng),與對(duì)照組相比差異有統(tǒng)計(jì)學(xué)意義(Plt;0.05),1 d達(dá)頂峰,HIS為7.00±2.00,之后減弱,5 d仍有陽(yáng)性表達(dá),但與對(duì)照組相比差異無(wú)統(tǒng)計(jì)學(xué)意義(Pgt;0.05)；褪黑素組HO1表達(dá)較中毒組明顯增強(qiáng),IHS評(píng)分6 h~3 d差異具有統(tǒng)計(jì)學(xué)意義(Plt;0.05),5 d不再有統(tǒng)計(jì)學(xué)意義(Pgt;005)。 結(jié)論 HO1在百草枯中毒大鼠腎組織中呈高表達(dá),褪黑素能明顯改善百草枯中毒腎臟病理?yè)p傷,增強(qiáng)HO1表達(dá)可能是其作用途徑之一,而氧化損傷可能是百草枯中毒腎損傷病理生理機(jī)制之一。【Abstract】 Objective To investigate the expression of heme oxygenase1 (HO1) in paraquartinduced renal injury in rats and the protective effects of melatonin, and explore possible mechanism of paraquartinduced renal injury. Methods One hundred and twentysix adult healthy SpragneDawley rats were randomly divided into three groups and 42 in each group: control group (A), paraquart group (B), and melatonin group (C). The rats in group B and group C were treated with paraquart (25 mg/kg) intraperitoneally, the rats in group A were treated with the same dose of normal saline. In 15 minutes, the rats in group C were given melatonin intraperitoneally at a dose of 10 mg/(kg·d) and the rats in group A and B were treated intraperitoneally with the same dose of normal saline. Six rats in each group were randomly sacrificed at one, three, 12 hours and one, tou, three, five days respectively. Renal histopathological changes were observed under light microscope by HE staining. The protein and mRNA expressions of HO1 were evaluated by immunohistochemical staining and RTPCR respectively. Results ①I(mǎi)n group B, there were obvious lesions in the renal tubule of cortical part, including edema, congestion and vacuolar degeneration. These pathologic changes gradually reached the peak on the first day and did not relieved till the end of this study, the pathologic injury score was 3.30±0.31, and there was a statistical significance between group B and group A (Plt;0.05). The aforementioned pathological lesion was more palliative in group C, the pathologic injury score was 2.70±0.26 at the first day; Compared with group B, there was a statistical significance. ②In group A, there was no or weak expression of HO1 and HO1 mRNA. At the third hour, the expression of HO1 in group B was observed in the membrane and cytoplasm of renal tubular epithelial cell of cortical part. Immunohistochemistry score (IHS) was 3.33±1.75 and the expression of HO1 mRNA increased, there was a statistical significance between group B and group A (Plt;0.05). It reached the peak on the first day, IHS was 7.00±2.00, but there was no statistical difference between group B and group A on the fifth day (Pgt;0.05); Compared with group B, the expression in group C was enhanced obviously, IHS were higher obviously on the six hour till to the third day (Plt;0.05), but there were no statistical differences on the fifth day (Pgt;0.05). Conclusion The expression of HO1 in the paraquartdamaged kidney increases and melatonin surely has an protective effect by increasing the expression of HO1, which suggests that oxidative injury might be the main mechanism of paraquartinduced renal injury.
目的 以基因工程干細(xì)胞治療的方式來(lái)改善缺血-再灌注狀況，使移植肝臟更好地耐受缺血-再灌注，以減少移植術(shù)后的并發(fā)癥，延長(zhǎng)移植器官存活期。方法 利用已建立的Wistar大鼠冷缺血原位肝移植(OLT)模型，術(shù)中經(jīng)受體大鼠門(mén)靜脈輸入IL-13基因修飾的或未修飾的肝卵圓細(xì)胞（HOC）即轉(zhuǎn)染組和未轉(zhuǎn)染組，分別于術(shù)后1、3、7及14 d檢測(cè)受體大鼠肝臟功能變化、組織學(xué)變化、膽管上皮細(xì)胞(BEC)的增殖情況、α-平滑肌肌動(dòng)蛋白(α-SMA)的表達(dá)、肝組織中HO-1 mRNA的表達(dá)，并觀察移植大鼠術(shù)后的存活情況。結(jié)果 轉(zhuǎn)入IL-13基因的HOC對(duì)冷保存損傷的肝臟有一定的保護(hù)作用； 術(shù)后7 d，肝移植組和未轉(zhuǎn)染組的α-SMA表達(dá)較轉(zhuǎn)染組明顯（Plt;0.05）； 術(shù)后3 d，未轉(zhuǎn)染組和轉(zhuǎn)染組的BEC增殖指數(shù)明顯低于肝移植組（Plt;0.05）。轉(zhuǎn)染組術(shù)后各時(shí)相點(diǎn)的HO-1 mRNA表達(dá)水平均明顯高于相應(yīng)時(shí)相點(diǎn)的其他各組的水平(Plt;0.05)； 移植術(shù)中經(jīng)門(mén)靜脈輸入HOC對(duì)缺血性膽管損傷所誘導(dǎo)的BEC增殖有一定的抑制作用，對(duì)BEC具有一定的保護(hù)作用； 肝移植組生存率明顯低于假手術(shù)組和轉(zhuǎn)染組(Plt;0.05)。結(jié)論 IL-13的持續(xù)表達(dá)能促進(jìn)術(shù)后移植肝內(nèi)HO-1 mRNA的表達(dá)，這有利于對(duì)供肝的保護(hù)，有利于受體大鼠術(shù)后肝功能的恢復(fù)。促進(jìn)HO-1 mRNA的表達(dá)是IL-13對(duì)BEC的具有保護(hù)作用的機(jī)理之一。