硒与苯那普利抑制单侧输尿管梗阻大鼠肾间质纤维化
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摘要
目的研究亚硒酸钠与苯那普利对单侧输尿管梗阻(UUO)大鼠肾间质纤维化(RIF)的保护作用及其机制。方法将大鼠随机分为假手术组、UUO组(结扎单侧输尿管建立UUO模型)、UUO+硒组(亚硒酸钠0.2 mg/kg·d灌胃)和UUO+苯那普利组(苯那普利10 mg/kg·d灌胃),每组18只。术后第7、14和21天,各组随机处死6只大鼠,肾组织行HE和Masson染色,观察RIF程度;免疫组化法检测结缔组织生长因子(CTGF)、转化生长因子-β1(TGF-β1)、α-平滑肌肌动蛋白(α-SMA)和Ⅲ型胶原(ColⅢ)的表达;Western blot检测CTGF和TGF-β1蛋白表达;比色法检测谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)和丙二醛(MDA)的含量。结果术后第7、14和21天,硒组和苯那普利组RIF较UUO组明显减轻(P<0.05);肾组织CTGF、TGF-β1、α-SMA、ColⅢ表达强度和MDA含量明显低于UUO组(P<0.01或P<0.05);GSH-Px和SOD含量明显高于UUO组(P<0.05)。两干预组之间各项指标无明显差异。UUO组TGF-β1、CTGF、α-SMA、ColⅢ的表达和RIF指数与MDA含量呈正相关(P<0.05),与SOD和GSH-Px含量呈负相关(P<0.05);CTGF与α-SMA、ColⅢ的表达呈正相关(P<0.05),CTGF、α-SMA和ColⅢ表达与RIF病变程度呈正相关(P<0.05)。结论亚硒酸钠与苯那普利均可减轻UUO模型大鼠RIF。
Objective To study the protective effect of the sodium selenite and benazepril on renal interstitial fibrosis( RIF) in rat model of unilateral ureteral obstruction( UUO) and its mechanism. Methods The male SD of clean grade rats were randomly divided into sham-operation group,UUO group( UUO model was established by ligating unilateral ureter),UUO + sodium selenite group group( sodium selenite 0. 2 mg/kg · d gavage),UUO +benazepril group( benazepril 10 mg/kg·d gavage),with 18 in each group.At day 7,14 and 21 after thetreatment,6 rats selected randomly from each group were killed.The extent of RIF was evaluated by HE and Masson staining of the renal tissue. The expression of connective tissue growth factor( CTGF),transforming growth factor-β1( TGF-β1),alpha smooth muscle actin( α-SMA) and Ⅲ collagen( Col Ⅲ) were detected by immunohistochemical method. Theprotein expression of CTGF and TGF-β1 were detected by Western blot. Chemical colorimetric method was used to detecte the contents of supper oxide dismutase( SOD),malondialdehyde( MDA) and glutathione peroxidase( GSH-px) in renal cortex. Results The extent of RIF and the expression of CTGF,TGF-β1,α-SMA and Col Ⅲ in renal cortex were significantly lower in sodium selenite group and benazepril group at day 7,14 and 21 after the operation compared with that in UUO group( P<0. 05 or P<0. 01). In sodium selenite group and benazepril group,the contents of SOD and GSH-px in renal cortex were higher significantly than those in UUO group at day 7,14 and 21 after the operation respectively( P<0. 05),but the MDA in renal cortex was significantly decreased( P<0. 05).There were no significant differences in the indexes between the two groups of sodium selenite and benazepril. The expression of CTGF,TGF-β1,α-SMA,Col Ⅲ and the extent of RIF were positively correlated to the level of MDA in UUO group( P<0. 05,respectively),and negatively correlated to the level of SOD and GSH-Px( P<0. 05,respectively). The expression of CTGF was positively correlated to the expression of α-SMA and Col Ⅲ in UUO group( P<0. 05).The expression of CTGF,α-SMA and ColⅢ were positively correlated to RIF in UUO group( P<0. 05). Conclusions Sodium selenite and benazepril can reduce the extent of RIF in rat model with unilateral ureteral obstruction.
Objective To study the protective effect of the sodium selenite and benazepril on renal interstitial fibrosis( RIF) in rat model of unilateral ureteral obstruction( UUO) and its mechanism. Methods The male SD of clean grade rats were randomly divided into sham-operation group,UUO group( UUO model was established by ligating unilateral ureter),UUO + sodium selenite group group( sodium selenite 0. 2 mg/kg · d gavage),UUO +benazepril group( benazepril 10 mg/kg·d gavage),with 18 in each group.At day 7,14 and 21 after thetreatment,6 rats selected randomly from each group were killed.The extent of RIF was evaluated by HE and Masson staining of the renal tissue. The expression of connective tissue growth factor( CTGF),transforming growth factor-β1( TGF-β1),alpha smooth muscle actin( α-SMA) and Ⅲ collagen( Col Ⅲ) were detected by immunohistochemical method. Theprotein expression of CTGF and TGF-β1 were detected by Western blot. Chemical colorimetric method was used to detecte the contents of supper oxide dismutase( SOD),malondialdehyde( MDA) and glutathione peroxidase( GSH-px) in renal cortex. Results The extent of RIF and the expression of CTGF,TGF-β1,α-SMA and Col Ⅲ in renal cortex were significantly lower in sodium selenite group and benazepril group at day 7,14 and 21 after the operation compared with that in UUO group( P<0. 05 or P<0. 01). In sodium selenite group and benazepril group,the contents of SOD and GSH-px in renal cortex were higher significantly than those in UUO group at day 7,14 and 21 after the operation respectively( P<0. 05),but the MDA in renal cortex was significantly decreased( P<0. 05).There were no significant differences in the indexes between the two groups of sodium selenite and benazepril. The expression of CTGF,TGF-β1,α-SMA,Col Ⅲ and the extent of RIF were positively correlated to the level of MDA in UUO group( P<0. 05,respectively),and negatively correlated to the level of SOD and GSH-Px( P<0. 05,respectively). The expression of CTGF was positively correlated to the expression of α-SMA and Col Ⅲ in UUO group( P<0. 05).The expression of CTGF,α-SMA and ColⅢ were positively correlated to RIF in UUO group( P<0. 05). Conclusions Sodium selenite and benazepril can reduce the extent of RIF in rat model with unilateral ureteral obstruction.
引文
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[10]李相友,丁国华,夏媛瑜,等.血管紧张素1-7对糖尿病大鼠肾小管间质纤维化的影响[J].中华肾脏病杂志,2012,28:798-803.
[11]Overstreet JM,Samarakoon R,Cardona-Grau D,et al.Tomor suppressor ataxia telangiectasia mutated functions downstream of TGF-β1 in orchestrating profibrotic reponses[J].FASEB J,2015,29:1258-1268.
[12]Brem AS,Gong R.Therapeutic targeting of aldosterone:a novel approach to the treatment of glomerular disease[J].Clin Sci(Lond),2015,128:527-535.
[13]Dupont JJ,Ramick MG,Farquhar WB,et al.NADPH oxidasederived reactive oxygen species contribute to impaired cutaneous microvascular function in chronic kidney disease[J].Am J Physiol Renal Physiol,2014,306:F1499-1506.
[14]Qiao X,Wang L,Wang Y,et al.Intermedin is upregulated and attenuates renal fibrosis by inhibition of oxidative stress in rats with unilateral ureteral obstruction[J].Nephrology,2015,20:820-831.
[15]Yuan XP,Liu LS,Fu Q,et al.Effects of ligustrazine on ureteral obstruction-induced renal tubulointerstitial fibrosis[J].Phytother Res,2012,26:696-703.
[2]Zhou TB,Qin YH,Lei FY,et al.Prohibitin attenuates oxidative stress and extracellular matrix accumulation in renal interstitial fibrosis disease[J].PLo S One,2013,8:e77187.doi:10.1371/journal.pone.0077187.e Collection 2013.
[3]Takeda Y,Nishikimi T,Akimoto K.Beneficial effects of a combination of Rho-kinase inhibitor and ACE inhibitor on tubulointerstitial fibrosis induced by unilateral ureteral obstruction[J].Hypertens Res,2010,33:965-973.
[4]Klahr S,Morrissey J.Obstructuve nephropathy and renal fibrosis[J].AM J Physiol Renal Physiol,2002,283:F861-875.
[5]Mezzano SA,Droguett MA,Burgos ME,et al.Overexpression of chemokines,fibrogenetic cytokines,and myofibroblasts in human membranous neph-ropathy[J].Kidney Int,2000,57:147-158.
[6]赵班,吴华,郑法雷.红细胞生成素对单侧输尿管梗阻大鼠RIF的作用及对MCP-1和TGF-β1表达的影响[J].中国血液净化杂志,2010,9:477-481.
[7]Tanino A,Okura T,Nagao T,et al.Interleukin-18 deficiency protects against renal interstitial fibrosis in aldosterone/salt-treated mice[J].Clin Sci(Lond),2016,130:1727-1739.
[8]Brown NJ.Contribution of aldosterone to cardiovascular and renal inflammation and fibrosis[J].Nat Rev Nephrol,2013,9:459-469.
[9]Liu WH,Tang NN,Zhang QD.Could mycophenolate mofetil combined with benazapril delay tubulointerstitial fibrosis in 5/6 nephrectomized rats?[J].Chin Med J(Engl),2009,122:199-204.
[10]李相友,丁国华,夏媛瑜,等.血管紧张素1-7对糖尿病大鼠肾小管间质纤维化的影响[J].中华肾脏病杂志,2012,28:798-803.
[11]Overstreet JM,Samarakoon R,Cardona-Grau D,et al.Tomor suppressor ataxia telangiectasia mutated functions downstream of TGF-β1 in orchestrating profibrotic reponses[J].FASEB J,2015,29:1258-1268.
[12]Brem AS,Gong R.Therapeutic targeting of aldosterone:a novel approach to the treatment of glomerular disease[J].Clin Sci(Lond),2015,128:527-535.
[13]Dupont JJ,Ramick MG,Farquhar WB,et al.NADPH oxidasederived reactive oxygen species contribute to impaired cutaneous microvascular function in chronic kidney disease[J].Am J Physiol Renal Physiol,2014,306:F1499-1506.
[14]Qiao X,Wang L,Wang Y,et al.Intermedin is upregulated and attenuates renal fibrosis by inhibition of oxidative stress in rats with unilateral ureteral obstruction[J].Nephrology,2015,20:820-831.
[15]Yuan XP,Liu LS,Fu Q,et al.Effects of ligustrazine on ureteral obstruction-induced renal tubulointerstitial fibrosis[J].Phytother Res,2012,26:696-703.