刺芒柄花素对单侧输尿管梗阻大鼠ASK1、JNK蛋白表达的影响
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摘要
目的探讨刺芒柄花素对单侧输尿管梗阻(UUO)大鼠凋亡信号调节激酶1(ASK1)、c-Jun氨基末端激酶(JNK)蛋白表达的影响。方法制备单侧输尿管梗阻诱导肾间质纤维化动物模型,将大鼠随机分为假手术组、模型组、依那普利组及刺芒柄花素高、中、低剂量(100、50、25 mg/kg)组;术后14 d处死大鼠并采集血清检测血清肌酐(Scr)和血尿素氮(BUN)水平;采用HE染色观察大鼠肾脏的病理改变及评定肾小管损伤指数;Masson染色观察肾间质胶原沉积的面积;采用Western blotting免疫印迹法检测肾组织ASK1、JNK蛋白表达水平;采用TUNEL法检测肾小管上皮细胞的凋亡。结果与模型组比较,各治疗组大鼠血清Scr、BUN水平均显著降低(P<0.05、0.01);肾小管损伤指数、肾间质胶原分布相对面积均显著降低(P<0.05、0.01);肾组织ASK1、JNK蛋白表达水平显著降低(P<0.05、0.01);肾小管上皮细胞凋亡指数显著降低(P<0.05、0.01)。刺芒柄花素高剂量组各项指标优于依那普利组。结论刺芒柄花素能够抑制UUO模型大鼠ASK1、JNK蛋白表达水平,阻止细胞凋亡,从而减缓梗阻性肾病病理进程的发生和发展。
Objective To study the effect of formononetin on the expression of ASK1 and JNK on the protein level in rat after unilateral ureteral obstruction(UUO). Methods Rats were then randomly divided into control group, model group, the enalapril group, and the high, medium, and low dose groups of formononetin(100, 50, and 25 mg/kg). Renal interstitial fibrosis(RIF) rats model was established by unilateral ureteral obstruction except the control group. The rats were sacrificed 14 d after surgery, and blood samples were collected to detect serum creatinine(Scr) and blood urea nitrogen(BUN) levels. HE staining was used to observe renal pathologic change and determine renal tubular damage index. The area percentage of RIF was detected by Masson staining. Expressions of ASK1 and JNK protein in kidney were determined by Western blotting. Tubular epithelial cell apoptosis was detected by TUNEL assay. Results Serum Scr, BUN level, tubulointerstitial injury index, RIF, the expressions of ASK1 and JNK protein, and apoptotic index were significantly decreased in the treatment groups when compared with the model group(P < 0.05, 0.01). The high dose group of formononetin was more effective than enalapril group. Conclusion Formononetin inhibited the expressions of ASK1 and JNK protein in UUO rats model. Ultimately renal tubular epithelial cell apoptosis was suppressed and the progression of obstructive nephropathy pathologic process was retarded.
Objective To study the effect of formononetin on the expression of ASK1 and JNK on the protein level in rat after unilateral ureteral obstruction(UUO). Methods Rats were then randomly divided into control group, model group, the enalapril group, and the high, medium, and low dose groups of formononetin(100, 50, and 25 mg/kg). Renal interstitial fibrosis(RIF) rats model was established by unilateral ureteral obstruction except the control group. The rats were sacrificed 14 d after surgery, and blood samples were collected to detect serum creatinine(Scr) and blood urea nitrogen(BUN) levels. HE staining was used to observe renal pathologic change and determine renal tubular damage index. The area percentage of RIF was detected by Masson staining. Expressions of ASK1 and JNK protein in kidney were determined by Western blotting. Tubular epithelial cell apoptosis was detected by TUNEL assay. Results Serum Scr, BUN level, tubulointerstitial injury index, RIF, the expressions of ASK1 and JNK protein, and apoptotic index were significantly decreased in the treatment groups when compared with the model group(P < 0.05, 0.01). The high dose group of formononetin was more effective than enalapril group. Conclusion Formononetin inhibited the expressions of ASK1 and JNK protein in UUO rats model. Ultimately renal tubular epithelial cell apoptosis was suppressed and the progression of obstructive nephropathy pathologic process was retarded.
引文
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[11]de Borst M H,Prakash J,Sandovici M,et al.c-Jun NH2-terminal kinase is crucially involved in renal tubulo-interstitial inflammation[J].J Pharm Exp Ther,2009,331(3):896-905.
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[13]Kanellis J,Ma F R R,Dowling J P,et al.JNK signalling in human and experimental renal ischaemia/reperfusion injury[J].Nephrol Dial Transplant,2010,25(9):2898-2908.
[14]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.
[2]Inagi R,Ishimoto Y,Nangaku M.Proteostasis in endoplasmic reticulum-new mechanisms in kidney disease[J].Nat Rev Nephrol,2014,10(7):369-378.
[3]Tanjore H,Lawson W E,Blackwell T S.Endoplasmic reticulum stress as a pro-fibrotic stimulus[J].BBA-Mol Basis Dis,2013,1832(7):940-947.
[4]Sakauchi C,Wakatsuki H,Ichijo H,et al.Pleiotropic properties of ASK1[J].BBA-Gen Subjects,2016,1861(1):3030-3038.
[5]王健,孙瑜,陈磊等.芒柄花素的现代研究进展[J].山西中医学院学报,2017,18(5):74-77.
[6]Huang D,Wang C Y,Meng Q,et al.Protective effects of formononetin against rhabdomyolysis-induced acute kidney injury by upregulating Nrf2 in vivo and in vitro[J].Royal Soc Chem,2016,112(6):110874-110883.
[7]袁媛.大鼠单侧输尿管结扎模型和饥饿模型尿液蛋白质组研究[D].北京:北京协和医学院,2015.
[8]陈娜子,姜潮,李校堃.内质网应激与疾病[J].中国生物工程杂志,2016,36(1):76-85.
[9]Gong J,Wang X Z,Chen J J,et al.Molecular signal networks and regulating mechanisms of the unfolded protein response[J].J Zhejiang Univ-Sci B,2017,18(1):1-14.
[10]Ma F Y,Tesch G H,Nikolic-Paterson D J.ASK1/p38signaling in renal tubular epithelial cells promotes renal fibrosis in the mouse obstructed kidney[J].Am J PhysiolRenal Physiol,2014,307(11):1263-1273.
[11]de Borst M H,Prakash J,Sandovici M,et al.c-Jun NH2-terminal kinase is crucially involved in renal tubulo-interstitial inflammation[J].J Pharm Exp Ther,2009,331(3):896-905.
[12]Lan R,Geng H,Polichnowski A J,et al.PTEN loss defines a TGF-β-induced tubule phenotype of failed differentiation and JNK signaling during renal fibrosis[J].Am J Physiol Renal Physiol,2012,302(9):F1210-F1223.
[13]Kanellis J,Ma F R R,Dowling J P,et al.JNK signalling in human and experimental renal ischaemia/reperfusion injury[J].Nephrol Dial Transplant,2010,25(9):2898-2908.
[14]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.