白藜芦醇对肾纤维化大鼠肾组织ORP150、GRP78、GRP94蛋白表达的影响
|
摘要
目的探讨白藜芦醇对单侧输尿管梗阻(UUO)大鼠肾组织内质网应激分子伴侣氧调节蛋白150(ORP150)和葡萄糖调节蛋白78(GRP78)、GRP94表达的影响。方法采用UUO大鼠模型,利用随机数字表将大鼠分为假手术组、模型组、依那普利组和白藜芦醇高、中、低剂量组,每组各10只;手术后2周将大鼠处死并收集血清和肾组织,检测肌酐(Scr)、血尿素氮(BUN)水平;采用HE染色观察大鼠肾组织的损害程度并计算损伤参数;采用Masson染色检测大鼠肾间质胶原沉积面积值;采用TUNEL法检测大鼠肾组织中原位细胞凋亡情况;Western blotting法测定大鼠肾组织ORP150、GRP78、GRP94蛋白表达水平。结果与模型组比较,各治疗组大鼠血清中Scr、BUN均显著降低,肾小管损伤程度明显减轻,肾间质胶原相对面积降低,肾组织中ORP150蛋白表达水平显著升高(P<0.05、0.01),原位细胞凋亡指数和GRP78、GRP94蛋白表达水平显著降低(P<0.05、0.01)。白藜芦醇高剂量组和依那普利组比较各检测指标均差异显著(P<0.05)。结论白藜芦醇对UUO大鼠肾功能具有保护作用,能够减轻肾脏肾盂积水,降低肾间质胶原沉积面积,诱导内质网应激过程中的关键信号分子ORP150的表达,下调分子伴侣GRP78、GRP94的表达,减缓凋亡的发展,抑制因细胞凋亡沉积而带来的肾间质纤维化进程。
Objective To assess the effect of resveratrol on the expression of endoplasmic reticulum stress molecular partners150-KD oxygen-regulated protein(ORP150) in renal tissues of rats with unilateral ureteral obstruction(UUO). Methods The UUO rat model of renal interstitial fibrosis was established. The rats were randomly divided into sham operation group, model group, enalapril group, the high-dose, medium-dose and low-dose groups of resveratrol, each group of 10. After 14 d of surgery, rats were sacrificed to collect serum and kidney tissue to detect the level of serum Scratinine(Scr) and blood urea nitrogen(BUN); Pathological changes were stained by HE to evaluate of renal tubular damage index, renal interstitial collagen deposition area was detected by Masson staining, apoptosis of in situ cell in renal tissue was determined by TUNEL assay, and using the western blot for the detection of protein expression of ORP150, GRP78, and GRP94 in renal tissue. Results After comparison of the treatment groups with model group, BUN and Scr levels in serum were significantly reduced in high-dose resveratrol group, the damage degree of renal tubules was significantly reduced, the relative area of the renal interstitial collagen decreased, In addition, the expression of ORP150 was increased(P < 0.05, 0.01), GRP78 and GRP94 proteins expression were significantly reduced,(P < 0.05, 0.01). Moreover, the resveratrol high-dose group seems more effective than enalapril groups in reversing the phenotype(P < 0.05). Conclusion Resveratrol was able to protect renal function, alleviate hydronephrosis, reduce interstitial injury of renal tubular, induce the expression of the key endoplasmic reticulum stress signal molecule ORP150 and the down-regulated molecular partner GRP78 and GRP94, delay the apoptosis of renal tubular epithelial cell of rats after UUO and inhibit renal interstitial fibrosis caused by apoptosis deposition.
Objective To assess the effect of resveratrol on the expression of endoplasmic reticulum stress molecular partners150-KD oxygen-regulated protein(ORP150) in renal tissues of rats with unilateral ureteral obstruction(UUO). Methods The UUO rat model of renal interstitial fibrosis was established. The rats were randomly divided into sham operation group, model group, enalapril group, the high-dose, medium-dose and low-dose groups of resveratrol, each group of 10. After 14 d of surgery, rats were sacrificed to collect serum and kidney tissue to detect the level of serum Scratinine(Scr) and blood urea nitrogen(BUN); Pathological changes were stained by HE to evaluate of renal tubular damage index, renal interstitial collagen deposition area was detected by Masson staining, apoptosis of in situ cell in renal tissue was determined by TUNEL assay, and using the western blot for the detection of protein expression of ORP150, GRP78, and GRP94 in renal tissue. Results After comparison of the treatment groups with model group, BUN and Scr levels in serum were significantly reduced in high-dose resveratrol group, the damage degree of renal tubules was significantly reduced, the relative area of the renal interstitial collagen decreased, In addition, the expression of ORP150 was increased(P < 0.05, 0.01), GRP78 and GRP94 proteins expression were significantly reduced,(P < 0.05, 0.01). Moreover, the resveratrol high-dose group seems more effective than enalapril groups in reversing the phenotype(P < 0.05). Conclusion Resveratrol was able to protect renal function, alleviate hydronephrosis, reduce interstitial injury of renal tubular, induce the expression of the key endoplasmic reticulum stress signal molecule ORP150 and the down-regulated molecular partner GRP78 and GRP94, delay the apoptosis of renal tubular epithelial cell of rats after UUO and inhibit renal interstitial fibrosis caused by apoptosis deposition.
引文
[1]Lin B,Shao L,Luo O,et al.Prevalence of chronic kidney disease and its association with metabolic disease:A cross-sectional survey in Zhejiang province,Eastern China[J].BMC Nephrol,2015,doi:10.1186/1471-2369-15-36.
[2]朱可可,杨波.肾间质纤维化发病机制研究进展[J].广东医学,2015,36(2):324-326.
[3]顾铜,李均.肾小管上皮细胞凋亡与肾纤维化及中药组分的干预进展[J].中国老年学杂志,2014,34(17):5011-5014.
[4]邹朝霞,李均.内质网应激与肾脏纤维化[J].海南医学,2017,28(4):615-617.
[5]Li J H,Qu X L,Ricardo S D,et al.Resveratrol inhibits renal fibrosis in the obstructed kidney[J].Am J Pathol,2010,177(3):1065-1071.
[6]李先宽,李赫宇,李帅,等.白藜芦醇研究进展[J].中草药,2016,47(14):2568-2578.
[7]Polycarpou E,Meira L B,Carrington S,et al.Resveratrol3-O-D-glucuronide and resveratrol 4′-O-D-glucuronide inhibitcolon cancer cell growth:evidence for a role of A3adenosine receptors,cyclin D1 depletion,and G1 cell cycle arrest[J].Mol Nutr Food Res,2013,57(10):1708-1717.
[8]刘顺,李赫宇,赵玲.白藜芦醇降血尿酸、抗炎作用研究进展[J].药物评价研究,2016,39(2):304-307.
[9]陈薪薪,仝欢,陈宇,等.白藜芦醇对小鼠纤维化TGF-β1/Smads信号转导通路的干预研究[J].中华中医药学刊,2016,34(5):1224-1227.
[10]Liang J,Tian S F,Han J X,et al.Resveratrol as a therapeutic agent for renal fibrosis induced by unilateral ureteral obstruction[J].Ren Fail,2014,36(2):285-291.
[11]Radford G,Donadio J.Predicting renal outcome in Ig A nephropathy[J].J Am Soc Nephrol,1997,8(2):199-207.
[12]Zhang L,Wang F,Wang L,et al.Prevalence of chronic kidney disease in China:A cross-sectional survey[J].Lancet,2012,379(9818):815-822.
[13]Bando Y.The functional role of stress proteins in ER stress mediated cell death[J].Anat Sci Int,2012,87(1):14-23.
[14]Wang Y,Wu Z,Li D,et al.Involvement of oxygen-regulated protein 150 in AMP-activated protein kinase-mediated alleviation of lipid-induced endoplasmic reticulum stress[J].J Biol Chem,2011,286(13):11119-11131.
[15]Kusaczuk M,Cechowska-Pasko M.Molecular chaperone ORP150 in ER stress-related diseases[J].Curr Pharm Design,2013,19(15):2807-2818.
[16]于杜娟.NSCLC中GRP78/94及肺癌相关因子蛋白表达及临床意义[D].长春:吉林大学,2013.
[2]朱可可,杨波.肾间质纤维化发病机制研究进展[J].广东医学,2015,36(2):324-326.
[3]顾铜,李均.肾小管上皮细胞凋亡与肾纤维化及中药组分的干预进展[J].中国老年学杂志,2014,34(17):5011-5014.
[4]邹朝霞,李均.内质网应激与肾脏纤维化[J].海南医学,2017,28(4):615-617.
[5]Li J H,Qu X L,Ricardo S D,et al.Resveratrol inhibits renal fibrosis in the obstructed kidney[J].Am J Pathol,2010,177(3):1065-1071.
[6]李先宽,李赫宇,李帅,等.白藜芦醇研究进展[J].中草药,2016,47(14):2568-2578.
[7]Polycarpou E,Meira L B,Carrington S,et al.Resveratrol3-O-D-glucuronide and resveratrol 4′-O-D-glucuronide inhibitcolon cancer cell growth:evidence for a role of A3adenosine receptors,cyclin D1 depletion,and G1 cell cycle arrest[J].Mol Nutr Food Res,2013,57(10):1708-1717.
[8]刘顺,李赫宇,赵玲.白藜芦醇降血尿酸、抗炎作用研究进展[J].药物评价研究,2016,39(2):304-307.
[9]陈薪薪,仝欢,陈宇,等.白藜芦醇对小鼠纤维化TGF-β1/Smads信号转导通路的干预研究[J].中华中医药学刊,2016,34(5):1224-1227.
[10]Liang J,Tian S F,Han J X,et al.Resveratrol as a therapeutic agent for renal fibrosis induced by unilateral ureteral obstruction[J].Ren Fail,2014,36(2):285-291.
[11]Radford G,Donadio J.Predicting renal outcome in Ig A nephropathy[J].J Am Soc Nephrol,1997,8(2):199-207.
[12]Zhang L,Wang F,Wang L,et al.Prevalence of chronic kidney disease in China:A cross-sectional survey[J].Lancet,2012,379(9818):815-822.
[13]Bando Y.The functional role of stress proteins in ER stress mediated cell death[J].Anat Sci Int,2012,87(1):14-23.
[14]Wang Y,Wu Z,Li D,et al.Involvement of oxygen-regulated protein 150 in AMP-activated protein kinase-mediated alleviation of lipid-induced endoplasmic reticulum stress[J].J Biol Chem,2011,286(13):11119-11131.
[15]Kusaczuk M,Cechowska-Pasko M.Molecular chaperone ORP150 in ER stress-related diseases[J].Curr Pharm Design,2013,19(15):2807-2818.
[16]于杜娟.NSCLC中GRP78/94及肺癌相关因子蛋白表达及临床意义[D].长春:吉林大学,2013.