臭氧氧化后处理对肾脏缺血再灌注损伤的作用
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摘要
目的研究臭氧O_3氧化后处理对大鼠肾脏缺血再灌注所致肾脏慢性纤维化的作用及相关机制。方法将72例SD大鼠随机分为4组:假手术组(S组)、单纯缺血再灌注组(I/R组)、O_3氧化后处理组(I/R+O_3组)、氧气O_2后处理组(I/R+O_2组),每组4只。每组在模型复制成功后2周检测血清肌酐(Scr)和血浆尿素氮(BUN),2和10周取肾脏标本,进行苏木精-伊红染色法染色、Masson染色,以及免疫组织化学法检测转化生长因子β1(TGF-β_1)、α-平滑肌肌动蛋白(α-SMA)的表达。结果术后2周各组的血清Scr、血浆BUN基本恢复到正常范围,两组比较,差异无统计学意义(P>0.05)。术后10周Masson染色显示,肾纤维化程度,其他3组较S组重(P<0.05),I/R+O_3组较I/R组、I/R+O_2组轻(P<0.05),而I/R组与I/R+O_2组比较,差异无统计学意义(P>0.05)。与S组比较,其他3组TGF-β1、α-SMA的表达增强(P<0.05);与I/R组比较,I/R+O_3组的表达减弱(P<0.05);I/R组与I/R+O_2组比较,差异无统计学意义(P>0.05)。结论 O_3后处理可以减轻缺血再灌注损伤肾脏的慢性纤维化,其机制可能与O_3后处理抑制转化生长因子-β_1/Smad信号通路激活有关。
Objective To explore the effect of ozone oxidative post-conditioning on chronic renal fibrosis induced by ischemia reperfusion injury in rats and the mechanism. Methods Totally 72 adult male SD rats were randomly divided into four groups(18 in each): sham-operation control group(S group), ischemia reperfusion group(I/R group), ozone oxidative post-conditioning group(I/R +O_3group) and oxygen post-conditioning group(I/R+O_2group). Serum creatinine(Scr) and blood urea nitrogen(BUN) levels were evaluated2 w after successful modeling. Renal specimens were obtained in the 2th and 10 th week respectively, and the renal tissues were stained by HE and Masson staining. The protein expressions of transforming growth factor-β_1(TGF-β_1) and α-smooth muscle actin(α-SMA) were determined by immunohistochemistry analysis. Results After two weeks of treatment, Scr and BUN of each group basically returned to normal levels, and there were no significant differences among the four groups(P > 0.05). Ten weeks after operation, Masson staining revealed renal fibrosis of the I/R, I/R+O_2and I/R+O_3groups was more serious than that of the S group(P <0.05), renal fibrosis of the I/R+O_3group was milder than taht of the I/R and I/R+O_2groups(P <0.05), while there was no difference between the I/R group and the I/R+O_2group(P > 0.05). The protein expressions of TGF-β_1and α-SMA of the I/R, I/R+O_2and I/R+O_3groups were higher than those of the S group(P < 0.05);and compared with the I/R group, the expressions of TGF-β_1and α-SMA were lower in the I/R+O_3group(P <0.05). There was no difference in the expression of TGF-β_1or α-SMA between the I/R+O_2group and the I/R group(P > 0.05). Conclusions Ozone oxidative post-conditioning can relieve chronic renal fibrosis caused by ischemic reperfusion injury, and inhibition of activation of TGF-β_1/Smad signaling pathway may be one of the mechanisms.
Objective To explore the effect of ozone oxidative post-conditioning on chronic renal fibrosis induced by ischemia reperfusion injury in rats and the mechanism. Methods Totally 72 adult male SD rats were randomly divided into four groups(18 in each): sham-operation control group(S group), ischemia reperfusion group(I/R group), ozone oxidative post-conditioning group(I/R +O_3group) and oxygen post-conditioning group(I/R+O_2group). Serum creatinine(Scr) and blood urea nitrogen(BUN) levels were evaluated2 w after successful modeling. Renal specimens were obtained in the 2th and 10 th week respectively, and the renal tissues were stained by HE and Masson staining. The protein expressions of transforming growth factor-β_1(TGF-β_1) and α-smooth muscle actin(α-SMA) were determined by immunohistochemistry analysis. Results After two weeks of treatment, Scr and BUN of each group basically returned to normal levels, and there were no significant differences among the four groups(P > 0.05). Ten weeks after operation, Masson staining revealed renal fibrosis of the I/R, I/R+O_2and I/R+O_3groups was more serious than that of the S group(P <0.05), renal fibrosis of the I/R+O_3group was milder than taht of the I/R and I/R+O_2groups(P <0.05), while there was no difference between the I/R group and the I/R+O_2group(P > 0.05). The protein expressions of TGF-β_1and α-SMA of the I/R, I/R+O_2and I/R+O_3groups were higher than those of the S group(P < 0.05);and compared with the I/R group, the expressions of TGF-β_1and α-SMA were lower in the I/R+O_3group(P <0.05). There was no difference in the expression of TGF-β_1or α-SMA between the I/R+O_2group and the I/R group(P > 0.05). Conclusions Ozone oxidative post-conditioning can relieve chronic renal fibrosis caused by ischemic reperfusion injury, and inhibition of activation of TGF-β_1/Smad signaling pathway may be one of the mechanisms.
引文
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[2]邱涛,周江桥,陈晖,等.臭氧氧化预处理在肾缺血再灌注损伤中对内皮型一氧化氮合酶表达的影响[J].中华移植杂志:电子版,2013,2:83-88.
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[11]COCHRANE J,WILLIAMS B T,BANERJEE A,et al.Ischemic preconditioning at tenuates functional,metabolic,and morphologic injury from ischemic acute renal failure in the rat[J].Ren Fail,1999,21:135-145.
[12]LEE H T,EMALA C W.Protective effect s of renal ischemic preconditioning and adenosine pretreatment role of A(1)and A(3)receptors[J].AM J Physiol Renal Physiol,2000,278:380-387.
[13]HIRABARU M,MOCHIZUKIK K,TAKATSUKI M,et al.Expression of alpha smooth muscle actin in living donor liver transplant recipients[J].World J Gastroenterol,2014,20(22):7067-7074.
[14]MENG X M,NIKOLIC-PATERSON D J,LAN H Y.TGF-β:the master regulator of fibrosis[J].Nat Rev Nephrol,2016,12(6):325-338.
[15]HU N,DUAN J LI H,et al.Hydroxysafflor yellow a ameliorates renal fibrosis by suppressing TGF-β1-induced epithelialto-mesenchymal transition[J].PLo S One,2016,11(4):DOI:10.1371/journal.pone.0153409.
[16]OVERSTREET J M,SAMARAKOON R,MELDRUM K K,et al.Redox control of p53 in the transcriptional regulation of TGFbetal target genens through SMAD cooperativity[J].Cell Signal,2014,26(7):1427-1436.
[2]邱涛,周江桥,陈晖,等.臭氧氧化预处理在肾缺血再灌注损伤中对内皮型一氧化氮合酶表达的影响[J].中华移植杂志:电子版,2013,2:83-88.
[3]XING B,CHEN H,WANG L,et al.Ozone oxidative preconditioning protects the rat kidney from reperfusion injury via modulation of the TLR4-NF-κB pathway[J].Acta Cir Bras,2015,30(1):60-66.
[4]JOSE L C,YANELIS T,SILVIA M,et al.Ozone oxidative post-conditioning in acute renal failure[J].Journal of Pharmacy and Pharmacology,2009,61:221-227.
[5]AKAN M,OZBILGIN S,BOZTAS N,et al.Effect of magnesium sulfate on renal ischemia-reperfusion injury in streptozotocin-induced diabetic rats[J].Eur Rev Med Pharmacol,2016,20(8):1642-1655.
[6]ERGIN B,INCE C.Hypertonic saline improves renal oxygenation,renal function and inflammation following ischemia/reperfusion-induced acute kidney injury[J].Intensive Care Med Exp,2015,3(1):1-2.
[7]GOBE G C,JOHNSON D W.Distal tubular epithelial cells of the kidney:potential support for proximal tubular cell survival after renal injury[J].Int J Biochem Cell Biol,2007,39(9):1551-1561.
[8]MENKE J,IWATA Y,RABACAL W A,et al.CSF-1 signals directly to renal tubular epithelial cells to mediate repair in mice[J].J Clin Invest,2009,119(8):2330-2342.
[9]JIANG S,CHEN Y,ZOU J,et al.Diverse effects of ischemic pretreatments on the long-term renal damage induced by ischemia-reperfusion[J].Am J Nephrol,2009,30(5):440-449.
[10]WEI Q,XIAO X,FOGLE P,et al.Changes in metabolic profiles during acute kidney injury and recovery following ischemia/reperfusion[J].PLo S One,2014,9(9):DOI:10.1371/journal.pone.0106647.
[11]COCHRANE J,WILLIAMS B T,BANERJEE A,et al.Ischemic preconditioning at tenuates functional,metabolic,and morphologic injury from ischemic acute renal failure in the rat[J].Ren Fail,1999,21:135-145.
[12]LEE H T,EMALA C W.Protective effect s of renal ischemic preconditioning and adenosine pretreatment role of A(1)and A(3)receptors[J].AM J Physiol Renal Physiol,2000,278:380-387.
[13]HIRABARU M,MOCHIZUKIK K,TAKATSUKI M,et al.Expression of alpha smooth muscle actin in living donor liver transplant recipients[J].World J Gastroenterol,2014,20(22):7067-7074.
[14]MENG X M,NIKOLIC-PATERSON D J,LAN H Y.TGF-β:the master regulator of fibrosis[J].Nat Rev Nephrol,2016,12(6):325-338.
[15]HU N,DUAN J LI H,et al.Hydroxysafflor yellow a ameliorates renal fibrosis by suppressing TGF-β1-induced epithelialto-mesenchymal transition[J].PLo S One,2016,11(4):DOI:10.1371/journal.pone.0153409.
[16]OVERSTREET J M,SAMARAKOON R,MELDRUM K K,et al.Redox control of p53 in the transcriptional regulation of TGFbetal target genens through SMAD cooperativity[J].Cell Signal,2014,26(7):1427-1436.