刺梨黄酮对诱导大鼠肾纤维化TGF-β1/Smads信号转导干预作用及机制分析
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摘要
目的:探讨观察刺梨黄酮对UUO模型大鼠肾纤维化TGF-β1/Smads信号转导干预作用,并分析其机制。方法:将75只造模成功的肾纤维化大鼠随机分为5组,即假手术组、模型对照组、氯沙坦钾组、刺梨黄酮中剂量及大剂量组,每组各15只。氯沙坦钾组给予氯沙坦钾灌胃;刺梨黄酮中剂量及大剂量组分别给予刺梨黄酮灌胃。模型组和假手术组均同时给予同等容积的生理盐水。干预14 d后,观察24 h Upro、Scr、BUN,对比各组大鼠肾脏组织TGF-β1、Smad2/3、Smad7蛋白表达水平,分析刺梨黄酮对肾纤维化大鼠作用机制。结果:干预过程中,模型组和氯沙坦钾组各有1只大鼠死亡。模型组、氯沙坦钾组、刺梨黄酮中剂量和大剂量组24 h Upro、Scr和BUN水平较假手术组显著升高(P<0.01);治疗后氯沙坦钾组、刺梨黄酮中剂量和大剂量组较模型组显著降低;与氯沙坦钾组相比,刺梨黄酮中剂量组24 h Upro、Scr和BUN表达水平无明显差异(P>0.05)。免疫组化显示模型组、氯沙坦钾组和刺梨黄酮不同剂量组中,各组TGF-β1、Smad2/3表达均明显高于假手术组(P<0.01),各治疗组的表达水平明显低于模型组(P<0.05)。各组Smad7的表达均明显低于假手术组(P<0.01),各治疗组的表达水平明显高于模型组(P<0.05)。与氯沙坦钾组比较,刺梨黄酮中剂量组TGF-β1、Smad2/3、Smad7表达水平无明显差异。结论:刺梨黄酮中剂量组可显著改善肾纤维化大鼠肾功能指标,推测和抑制TGF-β1、Smad2/3蛋白表达、激活Smad7蛋白表达有关。
Objective: To investigate the flavonoids in Rosa roxburghii Tratt on UUO rats renal fibrosis TGF beta/Smads signaling intervention effect and analyze its mechanism. Methods: A toal of 75 successfully modeling renal fibrosis rats were randomly divided into 5 groups,namely sham operation group,model control group,losartan treatment group and flavonoids in Rosa roxburghii Tratt middle dose group and high dose group,with 15 rats in each. Losartan group was given losartan orally,a dose of10 mg/( kg·d),the concentration of 20%. Rosa roxburghii Tratt middle dose and high dose groups were given flavonoids in Rosa roxburghii Tratt by intragastric administration,dose of 3G/( kg·d) and 6G/( kg·d),respectively. The concentration was 3%and 6%. Both model group and sham operation group were given equal volume of normal saline. After 14 days of intervention,observe 24 h U,SCR,BUN,conpare and analyze each group's rats' kidney tissue TGF beta 1,Smad2/3 and Smad7 protein expression levels of flavonoids from Rosa roxburghii Tratt on action mechanism of renal fibrosis in rats. Results: During the intervention,one rat died in model group and losartan potassium. Compared with sham group,model group,losartan potassium group,flavonoids dose and high dose groups' 24 h UPRO,Scr and BUN levels were significantly increased( P < 0. 01). After treatment,compared with model group,losartan potassium group,flavonoids dose and high dose groups' were significantly reduced. Compared with losartan potassium group,no significant difference in 24 h UPRO,Scr and BUN levels of expression. Immunohistochemistry showed that model group,losartan group and flavonoids different dose groups' TGF- β1 and Smad2/3 expressions were significantly higher than those of the sham group( P < 0. 01). The expression level of each treatment group significantly was lower than that of the model group( P < 0. 05). Smad7 expressions in each group were significantly lower than that of the sham group( P < 0. 01). The expression level of each treatment group was significantly higher than that of the model group( P < 0. 05). Compared with losartan potassium group,middle dose group's flavonoids TGF- β1,Smad2/3 and Smad7 expression levels had no significant difference.Conclusion: By the result of flavonoid from Rosa roxburghii Tratt middle dose group,flavonoid Rosa roxburghii Tratt can remarkably improve functional index of renal fibrosis rats. We can infer that this is sbuject to the inhibition of TGF- β,Smad2/3 protein expression and activation of Smad7 protein expression.
Objective: To investigate the flavonoids in Rosa roxburghii Tratt on UUO rats renal fibrosis TGF beta/Smads signaling intervention effect and analyze its mechanism. Methods: A toal of 75 successfully modeling renal fibrosis rats were randomly divided into 5 groups,namely sham operation group,model control group,losartan treatment group and flavonoids in Rosa roxburghii Tratt middle dose group and high dose group,with 15 rats in each. Losartan group was given losartan orally,a dose of10 mg/( kg·d),the concentration of 20%. Rosa roxburghii Tratt middle dose and high dose groups were given flavonoids in Rosa roxburghii Tratt by intragastric administration,dose of 3G/( kg·d) and 6G/( kg·d),respectively. The concentration was 3%and 6%. Both model group and sham operation group were given equal volume of normal saline. After 14 days of intervention,observe 24 h U,SCR,BUN,conpare and analyze each group's rats' kidney tissue TGF beta 1,Smad2/3 and Smad7 protein expression levels of flavonoids from Rosa roxburghii Tratt on action mechanism of renal fibrosis in rats. Results: During the intervention,one rat died in model group and losartan potassium. Compared with sham group,model group,losartan potassium group,flavonoids dose and high dose groups' 24 h UPRO,Scr and BUN levels were significantly increased( P < 0. 01). After treatment,compared with model group,losartan potassium group,flavonoids dose and high dose groups' were significantly reduced. Compared with losartan potassium group,no significant difference in 24 h UPRO,Scr and BUN levels of expression. Immunohistochemistry showed that model group,losartan group and flavonoids different dose groups' TGF- β1 and Smad2/3 expressions were significantly higher than those of the sham group( P < 0. 01). The expression level of each treatment group significantly was lower than that of the model group( P < 0. 05). Smad7 expressions in each group were significantly lower than that of the sham group( P < 0. 01). The expression level of each treatment group was significantly higher than that of the model group( P < 0. 05). Compared with losartan potassium group,middle dose group's flavonoids TGF- β1,Smad2/3 and Smad7 expression levels had no significant difference.Conclusion: By the result of flavonoid from Rosa roxburghii Tratt middle dose group,flavonoid Rosa roxburghii Tratt can remarkably improve functional index of renal fibrosis rats. We can infer that this is sbuject to the inhibition of TGF- β,Smad2/3 protein expression and activation of Smad7 protein expression.
引文
[1]杨沿浪.芜湖市健康体检人群慢性肾脏病流行病学调查及其相关危险因素分析[D].广州:南方医科大学,2014.
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[10]Glassock RJ.Urinary Chemoattractant Protein 1:A New Biomarker of Renal Fibrosis[J].Am J Nephrol,2016,43(6):451-453.
[11]刘咏梅,刘瑞华,刘文军,等.益气活血方对肾间质纤维化大鼠肾脏TGF-β/smad信号通路及结缔组织生长因子的影响[J].中西医结合学报,2010,8(12):1165-1173.
[12]Hohl M,Linz D,Fries P,et al.Modulation of the sympathetic nervou system by renal denervation prevents reduction of aortic distensibilit in atherosclerosis prone ApoE-deficient rats[J].J Transl Med,2016,14(1):167.
[13]郭碧林,朱春玲.Egr-1蛋白、MMP-2、TGF-β1在肾小管间质纤维化大鼠中的表达及意义[J].中国现代医学杂志,2013,23(7):27-33.
[14]赵瑞宝.慢性肾小球肾炎中医证型与肾间质纤维化相关指标的关系[J].中医杂志,2011,52(7):582-584,607.
[15]张汇慧,吴彩娥,范龚健,等.刺梨黄酮的精制及其抗氧化活性比较[J].南京林业大学学报(自然科学版),2015,39(3):101-105.
[16]陈丛瑾,王琪,李欣.黄酮类化合物抗氧化和抑菌生物活性研究进展[J].中国药房,2011,22(35):3346-3348.
[2]车丽双,黄荣桂.TGF-β1与CTGF在肾间质纤维化中的作用[J].医学综述,2013,19(4):624-626.
[3]詹继红,郭银雪.刺梨干粉对CKD3-4期脾肾气虚夹湿型患者氧化应激相关指标的影响[J].中国实验方剂学杂志,2014,20(23):224-226.
[4]李克剑,张悦,李靖,等.单侧输尿管梗阻法制作大鼠肾间质纤维化模型的改进[J].中国比实验动物学报,2007,15(6):410-412.
[5]蔡长景,刘持,秦晓群.巨噬细胞参与肾纤维化的分子机制及治疗研究进展[J].广东医学,2014,35(5):770-774.
[6]张新志,黄迪,吴锋,等.TGF-β1/p38MAPK通路对肾间质纤维化影响及抗纤灵冲剂干预机制的实验研究[J].中华中医药杂志,2011,26(2):245-248.
[7]Kim JH,Jeong JH,Park SH,et al.Recurrent renal cell carcinoma manifesting as a large intrathoracic fibrotic mass:A case report[J].Oncol Lett,2016,11(6):3835-3838.
[8]宋小乐.TGF-β_1/Smad信号转导通路对肾间质纤维化的影响[J].南昌大学学报(医学版),2011,51(6):92-95,98.
[9]王水华,陈帮明,刘永芳,等.排毒保肾丸对肾纤维化大鼠肾组织TGF-β1/Smad信号通路的影响[J].中医杂志,2015,56(9):792-795.
[10]Glassock RJ.Urinary Chemoattractant Protein 1:A New Biomarker of Renal Fibrosis[J].Am J Nephrol,2016,43(6):451-453.
[11]刘咏梅,刘瑞华,刘文军,等.益气活血方对肾间质纤维化大鼠肾脏TGF-β/smad信号通路及结缔组织生长因子的影响[J].中西医结合学报,2010,8(12):1165-1173.
[12]Hohl M,Linz D,Fries P,et al.Modulation of the sympathetic nervou system by renal denervation prevents reduction of aortic distensibilit in atherosclerosis prone ApoE-deficient rats[J].J Transl Med,2016,14(1):167.
[13]郭碧林,朱春玲.Egr-1蛋白、MMP-2、TGF-β1在肾小管间质纤维化大鼠中的表达及意义[J].中国现代医学杂志,2013,23(7):27-33.
[14]赵瑞宝.慢性肾小球肾炎中医证型与肾间质纤维化相关指标的关系[J].中医杂志,2011,52(7):582-584,607.
[15]张汇慧,吴彩娥,范龚健,等.刺梨黄酮的精制及其抗氧化活性比较[J].南京林业大学学报(自然科学版),2015,39(3):101-105.
[16]陈丛瑾,王琪,李欣.黄酮类化合物抗氧化和抑菌生物活性研究进展[J].中国药房,2011,22(35):3346-3348.