氯化两面针碱减轻糖尿病肾病模型大鼠的肾脏损伤
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摘要
目的观察氯化两面针碱对糖尿病肾病大鼠肾脏的保护作用,并探讨其可能作用机制。方法将大鼠随机分为对照组、糖尿病肾病模型组(DN)、氯化两面针碱(NC)20和40 mg/kg干预组。在建模成功后灌胃氯化两面针碱,1次/d,一共给药7周。试剂盒检测血清中的胰岛素与胰高血糖素;过碘酸希夫(PAS)染色法观察肾脏组织病理形态学;原位末端标记法检测肾小球细胞凋亡;免疫印迹法检测肾组织中磷酸化Smad2与Smad3表达水平、TGF-β1与Smad7蛋白表达水平。结果 DN组大鼠的胰岛素含量低于对照组(P<0.05),胰高血糖素含量高于对照组(P<0.05);给予氯化两面针碱干预后,大鼠血清中的胰岛素含量增加,胰高血糖素含量下降;低剂量组肾脏的PAS染色呈弱阳性,可见弥漫性肾小球硬化,药物高剂量组无基底膜增厚及肾小管病变;肾组织中磷酸化Smad2与Smad3表达降低(P<0.05),TGF-β1与Smad7蛋白表达降低(P<0.05)。结论氯化两面针碱对糖尿病肾病大鼠肾脏具有一定程度的保护作用,其可能是通过抑制TGF-β/Smad信号通路发挥作用。
Objective To observe the protective effects of nitidine chloride on renal injury in diabetic nephropathy rats,and to investigate the underlying mechanism.Methods All rats were divided into four groups: control group,diabetic nephropathy( DN) group,and diabetic nephropathy treated with nitidine chloride( NC)( 20 mg/kg or40 mg/kg) groups. In addition to control group,the rest of the rats were induced for diabetic nephropathy model.After the animal model was successfully established,diabetic nephropathy rats were received nitidine chloride at the dose of 20 mg/kg or 40 mg/kg by gavage once a day for 7 weeks. The level of serum insulin and glucagon was determined by commercial detection kit. The pathological morphology of kidney tissues was observed by PAS staining.Cell apoptosis in glomerular was detected by TUNEL staining. The phosphorylation of Smad2 and Smad3 and the protein expression of TGF-β1 and Smad7 were analyzed by Western blot. Results After treated with nitidine chloride,insulin level in the serum was increased,whereas the glucagon level decreased. PAS staining showed that weak positive staining and diffuse glomerular sclerosis in low dose group. Moreover,no thickening matrix structure and renal tubular lesion could be observed in high dose group. Finally,Western blot showed that the phosphoryla-tion of Smad2 and Smad3 and the protein expression of TGF-β1 and Smad7 in kidney tissues were significantly decreased after nitidine chloride administration. Conclusions Nitidine chloride prevents renal injury in diabetic nephropathy rats through inhibition of TGF-β/Smad signaling pathway.
Objective To observe the protective effects of nitidine chloride on renal injury in diabetic nephropathy rats,and to investigate the underlying mechanism.Methods All rats were divided into four groups: control group,diabetic nephropathy( DN) group,and diabetic nephropathy treated with nitidine chloride( NC)( 20 mg/kg or40 mg/kg) groups. In addition to control group,the rest of the rats were induced for diabetic nephropathy model.After the animal model was successfully established,diabetic nephropathy rats were received nitidine chloride at the dose of 20 mg/kg or 40 mg/kg by gavage once a day for 7 weeks. The level of serum insulin and glucagon was determined by commercial detection kit. The pathological morphology of kidney tissues was observed by PAS staining.Cell apoptosis in glomerular was detected by TUNEL staining. The phosphorylation of Smad2 and Smad3 and the protein expression of TGF-β1 and Smad7 were analyzed by Western blot. Results After treated with nitidine chloride,insulin level in the serum was increased,whereas the glucagon level decreased. PAS staining showed that weak positive staining and diffuse glomerular sclerosis in low dose group. Moreover,no thickening matrix structure and renal tubular lesion could be observed in high dose group. Finally,Western blot showed that the phosphoryla-tion of Smad2 and Smad3 and the protein expression of TGF-β1 and Smad7 in kidney tissues were significantly decreased after nitidine chloride administration. Conclusions Nitidine chloride prevents renal injury in diabetic nephropathy rats through inhibition of TGF-β/Smad signaling pathway.
引文
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[9] Lukas D,Yogev N,Kel JM,et al. TGF-βinhibitor Smad7 regulates dendritic cell-induced autoimmunity[J].Proc Natl Acad Sci U S A,2017,114:1480-1489.
[10] Li JH,Huang XR,Zhu HJ,et al. Advanced glycation end products activate Smad signaling via TGF-beta-dependent and independent mechanisms:implications for diabetic renal and vascular disease[J]. Faseb J,2004,18:176-178.
[2] Li JH,Zhu HJ, Huang XR, et al. Smad7 inhibits fibrotic effect of TGF-βon renal tubular epithelial cells by blocking smads activation[J]. J Am Soc Nephrol,2002,13:1464-1472.
[3] Yu SH,Dubey NK,Li WS,et al. Cordyceps militaris treatment preserves renal function in type 2 diabetic nephropathy mice[J]. PLoS One,2016,11:e0166342. doi:10.1371/journal.pone.0166342.
[4]刘华钢,李丹妮,刘丽敏.氯化两面针碱抗肿瘤作用机制研究进展[J].广西科学院学报,2009,25:187-191.
[5] Del Poeta M,Chen SF,Von Hoff D,et al. Comparison of in vitro activities of camptothecin and nitidine derivatives against fungal and cancer cells[J]. Antimicrob Agents Chemother,1999,43:2862-2868.
[6] Jie L,Pengcheng Q, Qiaoyan H, et al. Dencichine ameliorates kidney injury in induced typeⅡdiabetic nephropathy via the TGF-β/Smad signalling pathway[J].Eur J Pharmacol,2017,812:196-205.
[7] Dagher Z,Gerhardinger C,Vaz J,et al. The increased transforming growth factor-βsignaling induced by diabetes protects retinal vessels[J]. Am J Pathol,2017,187:627-638.
[8] Koga K,Yokoi H, Mori K, et al. MicroRNA-26a inhibits TGF-β-induced extracellular matrix protein expression in podocytes by targeting CTGF and is downregulated in diabetic nephropathy[J]. Diabetologia,2015,58:2169-2180.
[9] Lukas D,Yogev N,Kel JM,et al. TGF-βinhibitor Smad7 regulates dendritic cell-induced autoimmunity[J].Proc Natl Acad Sci U S A,2017,114:1480-1489.
[10] Li JH,Huang XR,Zhu HJ,et al. Advanced glycation end products activate Smad signaling via TGF-beta-dependent and independent mechanisms:implications for diabetic renal and vascular disease[J]. Faseb J,2004,18:176-178.