Klotho蛋白对缺血-再灌注肾损伤大鼠氧化应激的影响
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摘要
目的:探讨Klotho蛋白是否通过调节缺血-再灌注肾损伤大鼠模型的一氧化氮(NO)合成从而影响其氧化应激反应。方法:120只Wistar雄性大鼠随机分为正常组+生理盐水(NOR+NS)组、假手术组+生理盐水(Sham+NS)组、假手术组+Klotho(Sham+Klotho)组、缺血再灌注手术+生理盐水(I/R+NS)组、手术组+Klotho(IR+Klotho)组,建立缺血-再灌注急性肾损伤模型,分别于造模成功后1h、5h、12h、24h各组处死大鼠6只,行HE染色观察肾脏形态学改变,比色法测定血浆NO,肾组织过氧化物酶(MPO)和超氧化物歧化酶(SOD)水平,ELISE法检测血浆Klotho蛋白。结果:缺血再灌注24h后,I/R+NS组肾脏较多肾小管上皮空泡变性,而I/R+Klotho组空泡变性明显减少; I/R+NS组Klotho蛋白的表达明显低于其他各组(P<0.01),NO及SOD活力低于其他组(P<0.05),MPO、血清肌酐(SCr)和血尿素氮(BUN)明显高于其他组(P<0.05);I/R+Klotho组NO、MPO、SOD、SCr及BUN较I/R+NS组均有所改善(P<0.05)。结论:Klotho蛋白可能通过调节NO合成抵抗氧化应激,减轻肾缺血-再灌注损伤。
Objective:To research the effect of Klotho protein on oxidative stress in ischemia-reperfusion renal injury rats,and regulating of nitric oxide(NO) synthesis. Methodology:One hundred twenty male Wistar rats were randomly divided into NOR+NS group,Sham+NS group,Sham+Klotho group,I/R+NS group and I/R+Klotho group. After making models of acute ischemia-reperfusion kidney injury,six rats were put to death separately at 1 hour,5 hours,12 hours,and 24 hours after operation per group. Morphological changes in the kidney were observed by HE staining,plasma levels of NO,Tissue levels of peroxidase(MPO) and superoxide dismutase(SOD) in kidney were detected by colorimetric method,Klotho protein were detected by the ELISE method. Results:24 hours after ischemia-reperfusion injury,I/R+NS group rats had more vacuolar degeneration of renal tubular epithelial cells,however I/R+Klotho group rats had little tubular epithelial changes. In I/R+NS group,expression of Klotho protein, NO and SOD activities was significantly lower than other groups, also MPO,Scr and BUN were significantly higher than other groups. In I/R+Klotho group, the levels of NO,MPO,SOD,Scr and BUN were improved compared with I/R+NS group(P<0.05). Conclusion:Klotho protein may alleviate renal ischemia-reperfusion injury by regulating NO synthesis against oxidative stress.
Objective:To research the effect of Klotho protein on oxidative stress in ischemia-reperfusion renal injury rats,and regulating of nitric oxide(NO) synthesis. Methodology:One hundred twenty male Wistar rats were randomly divided into NOR+NS group,Sham+NS group,Sham+Klotho group,I/R+NS group and I/R+Klotho group. After making models of acute ischemia-reperfusion kidney injury,six rats were put to death separately at 1 hour,5 hours,12 hours,and 24 hours after operation per group. Morphological changes in the kidney were observed by HE staining,plasma levels of NO,Tissue levels of peroxidase(MPO) and superoxide dismutase(SOD) in kidney were detected by colorimetric method,Klotho protein were detected by the ELISE method. Results:24 hours after ischemia-reperfusion injury,I/R+NS group rats had more vacuolar degeneration of renal tubular epithelial cells,however I/R+Klotho group rats had little tubular epithelial changes. In I/R+NS group,expression of Klotho protein, NO and SOD activities was significantly lower than other groups, also MPO,Scr and BUN were significantly higher than other groups. In I/R+Klotho group, the levels of NO,MPO,SOD,Scr and BUN were improved compared with I/R+NS group(P<0.05). Conclusion:Klotho protein may alleviate renal ischemia-reperfusion injury by regulating NO synthesis against oxidative stress.
引文
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4 Chen YJ,Gong CL,Tan F,et al.Pretreatment with dexmedetomidine ameliorates renal inflammation and oxidative stress in rats with lipopolysaccharide-induced sepsis and acute kidney injury.Nan Fang Yi Ke Da Xue Xue Bao,2015,35(10):1472-1475.
5 Ogawa T,Nussler AK,Tuzuner E,et al.Contribution of nitric oxide to the protective effects of ischemic preconditioning in ischemia-reperfused rat kidneys.J Lab Clin Med,2001,138(1):50-58.
6 Goligorsky MS,Brodsky SV,Noiri E.Nitric oxide in acute renal failure:NOS versus NOS.Kidney Int,2002,61(3):855-861.
7 陈军,摇焦占,李广平,等.MPO、NO在造影剂肾病中的表达及其与氧化应激的关系.中外医学研究杂志,2012,10(6):1-3.
8 王羿,柏帅,徐旖旎,等.高交感活性诱导的大鼠心肌损伤模型中氧化应激的受体调控机制.中国病理生理杂志,2014,30(6):1029-1033.
9 Kuro-o M,Matsumura Y,Aizawa H,et al.Mutation of the mouse klotho gene leads to a syndrome resembling ageing.Nature,1997,390(6655):45-51.
10 Mitobe M,Yoshida T,Sugiura H,et al.Oxidative stress decreases klotho expression in a mouse kidney cell line.Nephron Exp Nephrol,2005,101(2):e67-74.
11 Ikushima M,Rakugi H,Ishikawa K,et al.Anti-apoptotic and anti-senescence effects of Klotho on vascular endothelial cells.Biochem Biophys Res Commun,2006,339(3):827-832.
12 Hu MC,Shi M,Zhang J,et al.Klotho deficiency is an early biomarker of renal ischemia-reperfusion injury and its replacement is protective.Kidney Int,2010,78(12):1240-1251.
2 Koyama D,Sato Y,Aizawa M,et al.soluble a Klotho as a candidates for the biomarker of aging.Biochem biophys Res Common,2015,267(4):1019-1025.
3 Chertow GM,Burdick E,Honour M,et al.Acute kidney injury,mortality,length of stay,and costs in hospitalized patients.J Am Soc Nephrol,2005,16(11):3365-3370.
4 Chen YJ,Gong CL,Tan F,et al.Pretreatment with dexmedetomidine ameliorates renal inflammation and oxidative stress in rats with lipopolysaccharide-induced sepsis and acute kidney injury.Nan Fang Yi Ke Da Xue Xue Bao,2015,35(10):1472-1475.
5 Ogawa T,Nussler AK,Tuzuner E,et al.Contribution of nitric oxide to the protective effects of ischemic preconditioning in ischemia-reperfused rat kidneys.J Lab Clin Med,2001,138(1):50-58.
6 Goligorsky MS,Brodsky SV,Noiri E.Nitric oxide in acute renal failure:NOS versus NOS.Kidney Int,2002,61(3):855-861.
7 陈军,摇焦占,李广平,等.MPO、NO在造影剂肾病中的表达及其与氧化应激的关系.中外医学研究杂志,2012,10(6):1-3.
8 王羿,柏帅,徐旖旎,等.高交感活性诱导的大鼠心肌损伤模型中氧化应激的受体调控机制.中国病理生理杂志,2014,30(6):1029-1033.
9 Kuro-o M,Matsumura Y,Aizawa H,et al.Mutation of the mouse klotho gene leads to a syndrome resembling ageing.Nature,1997,390(6655):45-51.
10 Mitobe M,Yoshida T,Sugiura H,et al.Oxidative stress decreases klotho expression in a mouse kidney cell line.Nephron Exp Nephrol,2005,101(2):e67-74.
11 Ikushima M,Rakugi H,Ishikawa K,et al.Anti-apoptotic and anti-senescence effects of Klotho on vascular endothelial cells.Biochem Biophys Res Commun,2006,339(3):827-832.
12 Hu MC,Shi M,Zhang J,et al.Klotho deficiency is an early biomarker of renal ischemia-reperfusion injury and its replacement is protective.Kidney Int,2010,78(12):1240-1251.