车前子水煎液对痛风性肾病大鼠的肾保护作用及机制
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摘要
目的:观察车前子水煎液对痛风性肾病大鼠的药效学指标及对肾组织半胱氨酸蛋白酶-1(cysteine-aspartic acid protease-1,Caspase-1),Nod样受体蛋白3(Nod-like receptor protein 3,NLRP3)和凋亡相关点样蛋白(fapoptosis-associated specklike protein,ASC)表达的影响,探讨车前子水煎液对痛风性肾病大鼠的肾保护作用和初步机制。方法:60只SD大鼠随机分为6组,分别为正常组,模型组,阳性药组(别嘌醇,50 mg·kg~(-1)),车前子水煎液高、中、低剂量组(1.62,0.81,0.27 g·kg~(-1)),除正常组外,其余各组采用腺嘌呤和酵母联用的方法制作痛风性肾病大鼠模型,给予车前子水煎液,观察肾脏特征;计算肾重指数;检测大鼠血清中尿酸(uric acid,UA),尿素氮(blood urea nitrogen,BUN),肌酐(creatinine,SCr),丙氨酸氨基转移酶(cereal third transaminase,ALT),天门冬氨酸氨基转移酶(aspartate transaminase,AST),碱性磷酸酶(alkalinephosphatase,ALP);采用光镜观察肾脏病理形态学变化;蛋白免疫印迹法(Western bolt)和免疫组化法(immunohistochemical methods)测定肾组织中NLRP3,ASC,Caspase-1蛋白表达量。结果:与模型组比较,别嘌醇组和车前子水煎液各剂量组的肾脏体积减小,别嘌醇组和车前子中剂量组肾重指数降低,车前子高剂量组肾重指数降低明显,别嘌醇组和车前子水煎液各剂量组血清UA,BUN,SCr,ALT,AST,ALP明显降低(P<0.05,P<0.01),肾组织病变程度均有一定减轻,其中车前子高、中剂量组最好,别嘌醇组和车前子水煎液高剂量组中NLRP3,ASC和Caspase-1蛋白的表达明显降低(P<0.05,P<0.01),车前子水煎液中剂量组中NLRP3蛋白的表达降低(P<0.05)。结论:车前子对痛风性肾病大鼠具有较好肾保护作用,可能与其下调肾组织中NLRP3,ASC和Caspase-1蛋白表达,抑制下游炎性细胞因子释放有关。
Objective: To observe pharmacodynamics index and effect of water decoction from Plantaginis Semen on Nod-like receptor protein 3( NLRP3),fapoptosis-associated speck-like protein( ASC) and cysteineaspartic acid protease-1( Caspase-1) of kidneys of rats with gouty nephropathy( GN),in order to investigate its renal protective effect and preliminary mechanism. Method: Adenine and yeast were used to make rat models of GN. The therapeutic effect and mechanism of water decoction from Plantaginis Semen on GN were studied by detecting the kidney characteristics and weight index; the levels of serum uric acid( UA),blood urea nitrogen( BUN),creatinine( SCr),cereal third transaminase( ALT),aspartate transaminase( AST),alkalinephosphatase( ALP); renal histopathological reduced under light microscope; the expressions of NLRP3,ASC,Caspase-1 were detected by Western blot and immunohistochemical methods. Result: Compared with the model group,the volumes of kidneys in allopurinol group and water decoction from Plantaginis Semen groups were smaller; the kidney weight index in allopurinol group and middle-dose water decoction from Plantaginis Semen group were lower,and the figure was significantly lower in high-dose group; the expression levels of UA,BUN,SCr,ALT,AST,ALP in serum in allopurinol group and water decoction from Plantaginis Semen groups were lower( P < 0. 05,P < 0. 01),and the pathological changes in renal tissues in GN model rats were relieved,especially in high and middle-dose water decoction from Plantaginis Semen groups. Compared with the model group,the protein expression levels of NLRP3,ASC and Caspase-1 in kidney tissues in allopurinol group and high-dose water decoction from Plantaginis Semen group were down-regulated( P < 0. 05,P < 0. 01),only the protein expression level of NLRP3 was down-regulated in middle-dose group( P < 0. 05). Conclusion: Water decoction from Plantaginis Semen may protect rats with GN by down-regulating the protein expression levels of NLRP3,ASC,Caspase-1 and inhibiting release of downstream inflammatory factors.
Objective: To observe pharmacodynamics index and effect of water decoction from Plantaginis Semen on Nod-like receptor protein 3( NLRP3),fapoptosis-associated speck-like protein( ASC) and cysteineaspartic acid protease-1( Caspase-1) of kidneys of rats with gouty nephropathy( GN),in order to investigate its renal protective effect and preliminary mechanism. Method: Adenine and yeast were used to make rat models of GN. The therapeutic effect and mechanism of water decoction from Plantaginis Semen on GN were studied by detecting the kidney characteristics and weight index; the levels of serum uric acid( UA),blood urea nitrogen( BUN),creatinine( SCr),cereal third transaminase( ALT),aspartate transaminase( AST),alkalinephosphatase( ALP); renal histopathological reduced under light microscope; the expressions of NLRP3,ASC,Caspase-1 were detected by Western blot and immunohistochemical methods. Result: Compared with the model group,the volumes of kidneys in allopurinol group and water decoction from Plantaginis Semen groups were smaller; the kidney weight index in allopurinol group and middle-dose water decoction from Plantaginis Semen group were lower,and the figure was significantly lower in high-dose group; the expression levels of UA,BUN,SCr,ALT,AST,ALP in serum in allopurinol group and water decoction from Plantaginis Semen groups were lower( P < 0. 05,P < 0. 01),and the pathological changes in renal tissues in GN model rats were relieved,especially in high and middle-dose water decoction from Plantaginis Semen groups. Compared with the model group,the protein expression levels of NLRP3,ASC and Caspase-1 in kidney tissues in allopurinol group and high-dose water decoction from Plantaginis Semen group were down-regulated( P < 0. 05,P < 0. 01),only the protein expression level of NLRP3 was down-regulated in middle-dose group( P < 0. 05). Conclusion: Water decoction from Plantaginis Semen may protect rats with GN by down-regulating the protein expression levels of NLRP3,ASC,Caspase-1 and inhibiting release of downstream inflammatory factors.
引文
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[15]金晓敏,张晓熙,郭璐,等.基于NLRP3炎性体轴土茯苓总黄酮对痛风性关节炎的作用和机制[J].中国实验方剂学杂志,2018,24(4):90-95.
[2]Roddy E,Hyon K.Epidemiology of gout[J].Rheum Dis Clin N Am,2014,40(2):155-175.
[3]鄂静,郑亚莉,曹丽,等.高尿酸血症引起慢性肾脏病的Meta分析[J].宁夏医学杂志,2015,37(3):229-231.
[4]李丹,张剑勇.痛风现代流行病学及降尿酸药物研究进展[J].风湿病与关节炎,2016,5(4):73-76.
[5]杨小梅,张小玉,杨海俊,等.舒惠荃教授治疗慢性尿酸性肾病的经验[J].实用中西医结合临床,2010,10(3):67.
[6]杨斌.清热利湿补肾法为主治疗痛风性肾病32例[J].中医临床研究,2011,3(18):84-85.
[7]张向伟,柳红芳,胡济源,等.痛风性肾病的辨机论治[J].北京中医药大学学报,2017,40(9):790-792.
[8]吴博文,田雪秋.中西医结合治疗痛风性肾病30例[J].中国医药导报,2006,3(5):76.
[9]ZHAO H,WANG Q H,SUN Y P,et al.Purification,characterization and immunomodulatory effects of Plantago depressa polysaccharides[J].Carbohydr Polym,2014,112(1):63-72.
[10]刘颖斐,王秋红,赵宏,等.车前子多糖治疗大鼠膜性肾病的实验探讨[J].中国实验方剂学杂志,2016,22(22):103-107.
[11]Tschopp J,Schroder K.NLRP3 inflammasome activation:the convergence of multiple signalling pathways on ROS production?[J].Nat Rev Immunol,2010,10(3):210-215.
[12]高双荣,马卫国,商学征,等.复方青秦对痛风性肾病大鼠肾组织病理学的影响[J].中国实验方剂学杂志,2012,18(10):183-186.
[13]WU H,ZHOU M Z,LU G,et al.Emodinol ameliorates urate nephropathy by regulating renal organic ion transporters and inhibiting immune inflammatory responses in rats[J].Biome Pharmacother,2017,96:727-735.
[14]MA C H,KANG L L,REN H M,et al.Simiao pill ameliorates renal glomerular injury via increasing Sirt1expression and suppressing NF-κB/NLRP3inflammasome activation in high fructose-fed rats[J].J Ethnopharmacol,2015,172:108-117.
[15]金晓敏,张晓熙,郭璐,等.基于NLRP3炎性体轴土茯苓总黄酮对痛风性关节炎的作用和机制[J].中国实验方剂学杂志,2018,24(4):90-95.