二氢杨梅素对糖尿病肾病大鼠肾纤维化的影响
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摘要
目的:探讨二氢杨梅素(DMY)对糖尿病肾病(DN)大鼠的保护作用及其对肾纤维化的影响。方法:采用大剂量(55 mg·kg~(-1))链脲佐菌素(STZ)腹腔注射建立DN模型,以血糖>16.7 mmol·L~(-1)为建模成功。将大鼠随机分为正常组、模型组和低、中、高剂量DMY组(125、250、500 mg·kg~(-1))。DMY组采用灌胃给药,模型组采用生理盐水灌胃,持续12周。生化检测各组大鼠24 h尿蛋白(24 h-Pro)、血尿素氮(BUN)和血肌酐(Scr)的含量。HE染色观察各组大鼠肾脏形态学变化,PAS染色观察各组肾组织基底膜厚度的变化,Masson染色观察各组肾小球硬化和肾间质纤维化程度。Western blot检测各组大鼠肾组织TGF-β1、Smad2、Smad7表达水平。结果:与正常组比较,DN模型组24 h-Pro、BUN、Scr的含量明显升高(P<0.01)。与模型组比较,各剂量DMY组24 h-Pro、BUN、Scr的含量下降(P<0.01)。病理结果显示模型组大鼠肾组织可见肾小球体积增大,胶原纤维增生,基底膜增厚。与模型组比较,各剂量DMY组胶原纤维不同程度减少。与正常组比较,模型组大鼠TGF-β1、Smad2蛋白表达水平增加,Smad7蛋白表达水平下降(P<0.05)。DMY干预后,能明显抑制大鼠肾组织中TGF-β1、Smad2蛋白表达上调和Smad7蛋白下调(P<0.05)。结论:DMY能改善DN大鼠肾脏组织纤维化,其机制可能与抑制TGF-β1/Smads通路过度激活有关。
Objective:To investigate the protective effect of Dihydromyricetin(DMY)on diabetic nephropathy(DN)rats and its effect on renal fibrosis.Methods:DN model was established by intraperitoneal injection of streptozotocin with high dose(55 mg·kg~(-1)).The model was successfully established with blood sugar>16.7 mmol·L~(-1).Rats were randomly divided into normal group,model group,low,medium and high dose DMY group(125,250,500 mg·kg~(-1)).DMY group was given intragastric administration,while model group was given normal saline intragastricadministration for12 weeks.The levels of 24-hour urinary protein(24 h-Pro),blood urea nitrogen(BUN)and Serum creatinine(Scr)in rats of each group were detected by biochemical method.HE staining was used to observe the morphological changes of kidneys of rats in each group,PAS staining was used to observe the changes of renal basement membrane thickness in each group,Masson staining was used to observe the degree of glomerulosclerosis and renal interstitial fibrosis in each group.The expression levels of TGF-β1,Smad2 and Smad7 in kidney tissues of rats in each group were detected by Western blot.Results:Compared with the normal group,the levels of 24 h-Pro,BUN and Scr in DN model group were significantly higher(P<0.01).Compared with model group,the levels of 24 h-Pro,BUN and Scr in DMY group decreased(P<0.01).Pathological results showed that glomerular volume increased,collagen fibers proliferated and basement membrane thickened in the model group.Compared with the model group,collagen fibers in DMY group decreased in different degrees.Compared with the normal group,the expression levels of TGF-β1 and Smad2 increased and Smad7decreased in the model group(P<0.05).DMY could significantly inhibit the up-regulation of TGF-β1,Smad2 and down-regulation of Smad7 in renal tissue of rats(P<0.05).Conclusions:DMY can improve renal fibrosis in rats with DN,and its mechanism may be related to the inhibition of excessive activation of TGF-β1/Smads pathway.
Objective:To investigate the protective effect of Dihydromyricetin(DMY)on diabetic nephropathy(DN)rats and its effect on renal fibrosis.Methods:DN model was established by intraperitoneal injection of streptozotocin with high dose(55 mg·kg~(-1)).The model was successfully established with blood sugar>16.7 mmol·L~(-1).Rats were randomly divided into normal group,model group,low,medium and high dose DMY group(125,250,500 mg·kg~(-1)).DMY group was given intragastric administration,while model group was given normal saline intragastricadministration for12 weeks.The levels of 24-hour urinary protein(24 h-Pro),blood urea nitrogen(BUN)and Serum creatinine(Scr)in rats of each group were detected by biochemical method.HE staining was used to observe the morphological changes of kidneys of rats in each group,PAS staining was used to observe the changes of renal basement membrane thickness in each group,Masson staining was used to observe the degree of glomerulosclerosis and renal interstitial fibrosis in each group.The expression levels of TGF-β1,Smad2 and Smad7 in kidney tissues of rats in each group were detected by Western blot.Results:Compared with the normal group,the levels of 24 h-Pro,BUN and Scr in DN model group were significantly higher(P<0.01).Compared with model group,the levels of 24 h-Pro,BUN and Scr in DMY group decreased(P<0.01).Pathological results showed that glomerular volume increased,collagen fibers proliferated and basement membrane thickened in the model group.Compared with the model group,collagen fibers in DMY group decreased in different degrees.Compared with the normal group,the expression levels of TGF-β1 and Smad2 increased and Smad7decreased in the model group(P<0.05).DMY could significantly inhibit the up-regulation of TGF-β1,Smad2 and down-regulation of Smad7 in renal tissue of rats(P<0.05).Conclusions:DMY can improve renal fibrosis in rats with DN,and its mechanism may be related to the inhibition of excessive activation of TGF-β1/Smads pathway.
引文
[1]谢晓东.百令胶囊对糖尿病肾脏疾病氧化应激水平及肾功能的影响[J].赣南医学院学报,2018,38(11):1103-1104.
[2]Lan H Y.Transforming growth factor-beta/Smad signalling in diabetic nephropathy[J].Clin Exp Pharmacol Physiol,2012,39(8):731-738.
[3]Hills C E,Squires P E.The role of TGF-beta and epithelial-to mesenchymal transition in diabetic nephropathy[J].Cytokine Growth Factor Rev,2011,22(3):131-139.
[4]陈亚丽,尹跃霏,李赟,等.二氢杨梅素药理作用研究进展[J].中国新药杂志,2019,28(2):173-178.
[5]Liu L,Wan J,Lang H,et al.Dihydromyricetin delays the onset of hyperglycemia and ameliorates insulin resistance without excessive weight gain in Zucker diabetic fatty rats[J].Mol Cell Endocrinol,2017,439:105-115.
[6]吕慧婕,许拓,何剑琴,等.二氢杨梅素与糖尿病肾病的研究进展[J].临床与病理杂志,2018,38(10):2233-2237.
[7]周海云,王文清,施春阳,等.二氢杨梅素药理及药物相互作用研究进展[J].中草药,2018,49(14):3411-3418.
[8]甘露,邓之婧,王献哲,等.二氢杨梅素对小鼠局灶性脑缺血/再灌注损伤的抗凋亡作用及机制研究[J].中国当代医药,2018,25(14):4-8.
[9]逯凤肖,王恩花,秦湉,等.二氢杨梅素对糖尿病小鼠降糖作用研究[J].中药药理与临床,2016,32(3):45-48.
[10]王莹.二氢杨梅素保护Apo E(-/-)动脉粥样硬化小鼠的作用及其机制[J].中成药,2018,40(3):511-516.
[11]许妍妍.二氢杨梅素对四氯化碳致肝纤维化大鼠防护作用及其机制研究[J].世界临床药物,2017,38(8):522-527.
[12]李加林,郭小华,吴艳娇,等.二氢杨梅素对高糖诱导的肾小球系膜细胞增殖及纤维连接蛋白堆积的影响[J].中国药房,2017,28(34):4784-4787.
[13]陈红,公欣华,陈丽君,等.二氢杨梅素激活AMPK通路预防糖尿病肾病的作用研究[J].现代生物医学进展,2017,17(34):6613-6619+6612.
[14]Tervaert T W,Mooyaart A L,Amann K,et al.Pathologic classification of diabetic nephropathy[J].J Am Soc Nephrol,2010,21(4):556-563.
[15]吴素珍,李加林,吴冰,等.转化生长因子-β在糖尿病肾病肾小球硬化中的作用[J].赣南医学院学报,2017,37(2):312-316+337.
[16]Chang A S,Hathaway C K,Smithies O,et al.Transforming growth factor-beta1 and diabetic nephropathy[J].Am J Physiol Renal Physiol,2016,310(8):689-696.
[17]Zhao Y,Yin Z,Li H,et al.MiR-30c protects diabetic nephropathy by suppressing epithelial-to-mesenchymal transition in db/db mice[J].Aging Cell,2017,16(2):387-400.
[18]Loeffler I,Liebisch M,Allert S,et al.FSP1-specific SMAD2 knockout in renal tubular,endothelial,and interstitial cells reduces fibrosis and epithelial-to-mesenchymal transition in murine STZ-induced diabetic nephropathy[J].Cell Tissue Res,2018,372(1):115-133.
[19]Hills C E,Squires P E.TGF-beta1-induced epithelialto-mesenchymal transition and therapeutic intervention in diabetic nephropathy[J].Am J Nephrol,2010,31(1):68-74.
[20]Chen S,Iglesias-De La Cruz M C,Jim B,et al.Reversibility of established diabetic glomerulopathy by antiTGF-beta antibodies in db/db mice[J].Biochem Biophys Res Commun,2003,300(1):16-22.
[2]Lan H Y.Transforming growth factor-beta/Smad signalling in diabetic nephropathy[J].Clin Exp Pharmacol Physiol,2012,39(8):731-738.
[3]Hills C E,Squires P E.The role of TGF-beta and epithelial-to mesenchymal transition in diabetic nephropathy[J].Cytokine Growth Factor Rev,2011,22(3):131-139.
[4]陈亚丽,尹跃霏,李赟,等.二氢杨梅素药理作用研究进展[J].中国新药杂志,2019,28(2):173-178.
[5]Liu L,Wan J,Lang H,et al.Dihydromyricetin delays the onset of hyperglycemia and ameliorates insulin resistance without excessive weight gain in Zucker diabetic fatty rats[J].Mol Cell Endocrinol,2017,439:105-115.
[6]吕慧婕,许拓,何剑琴,等.二氢杨梅素与糖尿病肾病的研究进展[J].临床与病理杂志,2018,38(10):2233-2237.
[7]周海云,王文清,施春阳,等.二氢杨梅素药理及药物相互作用研究进展[J].中草药,2018,49(14):3411-3418.
[8]甘露,邓之婧,王献哲,等.二氢杨梅素对小鼠局灶性脑缺血/再灌注损伤的抗凋亡作用及机制研究[J].中国当代医药,2018,25(14):4-8.
[9]逯凤肖,王恩花,秦湉,等.二氢杨梅素对糖尿病小鼠降糖作用研究[J].中药药理与临床,2016,32(3):45-48.
[10]王莹.二氢杨梅素保护Apo E(-/-)动脉粥样硬化小鼠的作用及其机制[J].中成药,2018,40(3):511-516.
[11]许妍妍.二氢杨梅素对四氯化碳致肝纤维化大鼠防护作用及其机制研究[J].世界临床药物,2017,38(8):522-527.
[12]李加林,郭小华,吴艳娇,等.二氢杨梅素对高糖诱导的肾小球系膜细胞增殖及纤维连接蛋白堆积的影响[J].中国药房,2017,28(34):4784-4787.
[13]陈红,公欣华,陈丽君,等.二氢杨梅素激活AMPK通路预防糖尿病肾病的作用研究[J].现代生物医学进展,2017,17(34):6613-6619+6612.
[14]Tervaert T W,Mooyaart A L,Amann K,et al.Pathologic classification of diabetic nephropathy[J].J Am Soc Nephrol,2010,21(4):556-563.
[15]吴素珍,李加林,吴冰,等.转化生长因子-β在糖尿病肾病肾小球硬化中的作用[J].赣南医学院学报,2017,37(2):312-316+337.
[16]Chang A S,Hathaway C K,Smithies O,et al.Transforming growth factor-beta1 and diabetic nephropathy[J].Am J Physiol Renal Physiol,2016,310(8):689-696.
[17]Zhao Y,Yin Z,Li H,et al.MiR-30c protects diabetic nephropathy by suppressing epithelial-to-mesenchymal transition in db/db mice[J].Aging Cell,2017,16(2):387-400.
[18]Loeffler I,Liebisch M,Allert S,et al.FSP1-specific SMAD2 knockout in renal tubular,endothelial,and interstitial cells reduces fibrosis and epithelial-to-mesenchymal transition in murine STZ-induced diabetic nephropathy[J].Cell Tissue Res,2018,372(1):115-133.
[19]Hills C E,Squires P E.TGF-beta1-induced epithelialto-mesenchymal transition and therapeutic intervention in diabetic nephropathy[J].Am J Nephrol,2010,31(1):68-74.
[20]Chen S,Iglesias-De La Cruz M C,Jim B,et al.Reversibility of established diabetic glomerulopathy by antiTGF-beta antibodies in db/db mice[J].Biochem Biophys Res Commun,2003,300(1):16-22.