补气健脾益肾法对糖尿病肾病大鼠肾小管上皮细胞转分化及肾间质纤维化影响
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摘要
目的:分析补气健脾益肾法对糖尿病肾病(DN)大鼠肾小管上皮细胞的间充质转分化(EMT)及肾间质纤维化影响。方法:选取SPF级Wistar雄性大鼠45只,随机数字表法将分成三组,正常组、模型组和观察组,每组各15只,观察组和模型组制备DN模型,观察组大鼠使用补气健脾益肾法灌胃,剂量为1.4g/kg,正常组和模型组大鼠使用同剂量生理盐水灌胃,每天1次,共灌胃8周。末次给药后大鼠置入代谢笼内,采集其24小时排泄尿液,计算其24小时尿量,全自动生化分析仪检测24小时尿蛋白、血尿素氮及血肌酸酐含量,血糖仪检测其血糖含量,肾组织常规行HE染色,在光镜下查看大鼠肾脏组织的病理状况,免疫组化检测肾组织内转化生长因子-β(TGF-β)、骨成形蛋白(BMP-7)及α-平滑肌肌蛋白(α-SMA)表达状况。结果:模型组大鼠24h尿蛋白、肾质量及24h尿量比正常组上升,观察组24h尿蛋白、肾质量及24h尿量比模型组下降,差异均有统计学意义(P<0.05);模型组及观察组血糖、模型组血尿素氮、血肌酐酸含量比正常组上升,观察组血尿素氮、血肌酐酸比模型组下降,差异均有统计学意义(P<0.05);模型组大鼠BMP-7表达量比正常组下降,TGF-β、α-SMA表达量比正常组上升,观察组BMP-7表达量比模型组上升,TGF-β、α-SMA表达量比模型组下降,差异均有统计学意义(P<0.05)。结论:补气健脾益肾法可降低DN大鼠尿蛋白含量,抑制肾小球EMT和肾间质纤维化发生发展。
Objective:To analyze the effect of Buqi Jianpi Yishen Recipe on mesenchymal transdifferentiation(EMT) and renal interstitial fibrosis in renal tubular epithelial cells of diabetic nephropathy(DN) rats.Methods:A total of 45 male Wistar male rats of SPF grade were randomly divided into three groups:normal group,model group and observation group,with 15 rats in each group.The observation group and the model group were prepared with DN model.Buqi Jianpi Yishen Recipe was intragastrically administered at a dose of 1.4 g/kg.The rats in the normal group and the model group were intragastrically administered with the same dose of normal saline once a day for a total of eight weeks.After the last administration,the rats were placed in a metabolic cage,and the urine was collected for 24 hours to calculate the urine volume.The 24 h urine protein,blood urea nitrogen and blood creatinine content were detected by automatic biochemical analyzer,and the blood glucose level was detected by blood glucose meter.Renal tissue was routinely stained with HE.The pathological condition of rat kidney tissue was observed under light microscope.Immunohistochemistry was used to detect the expression status of transforming growth factor-β(TGF-β),bone forming protein(BMP-7) and α-smooth muscle myoprotein(α-SMA).Results:The 24 h urine protein,kidney mass and 24 h urine volume of the model group were higher than those of the normal group.The 24 h urine protein,kidney mass and 24 h urine volume of the observation group were significantly lower than the model group(P<0.05).The blood glucose and blood creatinine levels in the model group and the observation group were higher than those in the normal group.The blood urea nitrogen and serum creatinine ratio in the observation group decreased,and the difference was statistically significant(P<0.05).The expression of BMP-7 in rats was lower than that in the normal group.The expression of TGF-β and α-SMA was higher than that in the normal group.The expression of BMP-7 in the observation group was higher than that in the model group,and the expression of TGF-β and α-SMA was higher than that in the model group.The difference was statistically significant(P<0.05).Conclusion:Buqi Jianpi Yishen method can reduce the urine protein content of DN rats and inhibit the development of glomerular EMT and renal interstitial fibrosis.
Objective:To analyze the effect of Buqi Jianpi Yishen Recipe on mesenchymal transdifferentiation(EMT) and renal interstitial fibrosis in renal tubular epithelial cells of diabetic nephropathy(DN) rats.Methods:A total of 45 male Wistar male rats of SPF grade were randomly divided into three groups:normal group,model group and observation group,with 15 rats in each group.The observation group and the model group were prepared with DN model.Buqi Jianpi Yishen Recipe was intragastrically administered at a dose of 1.4 g/kg.The rats in the normal group and the model group were intragastrically administered with the same dose of normal saline once a day for a total of eight weeks.After the last administration,the rats were placed in a metabolic cage,and the urine was collected for 24 hours to calculate the urine volume.The 24 h urine protein,blood urea nitrogen and blood creatinine content were detected by automatic biochemical analyzer,and the blood glucose level was detected by blood glucose meter.Renal tissue was routinely stained with HE.The pathological condition of rat kidney tissue was observed under light microscope.Immunohistochemistry was used to detect the expression status of transforming growth factor-β(TGF-β),bone forming protein(BMP-7) and α-smooth muscle myoprotein(α-SMA).Results:The 24 h urine protein,kidney mass and 24 h urine volume of the model group were higher than those of the normal group.The 24 h urine protein,kidney mass and 24 h urine volume of the observation group were significantly lower than the model group(P<0.05).The blood glucose and blood creatinine levels in the model group and the observation group were higher than those in the normal group.The blood urea nitrogen and serum creatinine ratio in the observation group decreased,and the difference was statistically significant(P<0.05).The expression of BMP-7 in rats was lower than that in the normal group.The expression of TGF-β and α-SMA was higher than that in the normal group.The expression of BMP-7 in the observation group was higher than that in the model group,and the expression of TGF-β and α-SMA was higher than that in the model group.The difference was statistically significant(P<0.05).Conclusion:Buqi Jianpi Yishen method can reduce the urine protein content of DN rats and inhibit the development of glomerular EMT and renal interstitial fibrosis.
引文
[1] 毕慧欣,刘瑞洪,尹友生,等.热休克蛋白47在单侧输尿管梗阻大鼠肾小管上皮细胞转分化中的作用[J].中国中西医结合肾病杂志,2017,18(3):205～208
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[12] 成芳,涂珍珍,周利利,等.雷帕霉素对肾小管上皮细胞生物学行为的影响及其治疗糖尿病肾病分子机制的初步研究[J].安徽医科大学学报,2017,52(1):1～6
[13] 陈红淑,杨元宵,周小杰,等.肾气丸通过TGF-β1/Smads/ILK信号途径干预肾小管上皮细胞间充质转分化的分子机制[J].中华中医药杂志,2016,13(6):2102～2105
[14] Xu J,Li P X,Wu J,et al.Involvement of the Hippo pathway in regeneration and fibrogenesis after ischaemic acute kidney injury:YAP is the key effector.[J].Clinical Science,2016,130(5):349～363
[15] 陈雨思,李均.肝细胞生长因子抗肾纤维化的机制[J].海南医学,2016,27(20):3364～3366
[16] 苗永辉,赵宗江,杨冠男,等.糖肾平含药血清对高糖诱导肾小管上皮细胞增殖作用及RhoA/ROCK信号通路影响的研究[J].世界科学技术-中医药现代化,2017,9(6):1050～1056
[2] 何平红,吴欣,董蓉,等.胰岛素样生长因子1受体拮抗剂通过抑制肾小管上皮-间质细胞转分化改善糖尿病肾病小鼠肾脏纤维化[J].中华肾脏病杂志,2018,3(1):17～22
[3] Yi Y,Li S,Nie Y,et al.Effect of astragalus injection on renal tubular epithelial transdifferentiation in type 2diabetic mice[J].Bmc Complementary Alternative Medicine,2016,16(1):222～225
[4] 刘爱林,张皎月.罗格列酮对糖尿病肾病大鼠肾小管上皮整合素连接激酶和α-平滑肌肌动蛋白表达的影响[J].实用药物与临床,2016,19(9):1085～1088
[5] 雷作熹,罗仁,董晓蕾,等.STZ诱导糖尿病肾病大鼠模型的建立[J].中国实验动物学报,2005,13(3):163～165
[6] Huang H,Zheng F,Dong X,et al.Allicin inhibits tubular epithelial-myofibroblast transdifferentiation under high glucose conditions in vitro[J].Experimental Therapeutic Medicine,2017,13(1):254～262
[7] 杨冠男,赵宗江,张新雪,等.糖肾宁对糖尿病肾病KKAy小鼠的肾脏保护作用及其对Notch/Snail1信号转导通路的影响[J].世界科学技术-中医药现代化,2017,11(6):1057～1065
[8] 王文文,张赛,朱晓卉,等.桑叶总黄酮对1型糖尿病小鼠肾间质纤维化的防治作用及机制[J].中国药理学通报,2017,33(9):1278～1285
[9] 肖静.肾小管上皮细胞-间充质转分化在肾间质纤维化中的机制[J].国际儿科学杂志,2017,44(10):304～308
[10] Ibrahim Z S,Alkafafy M E,Ahmed M M,et al.Renoprotective effect of curcumin against the combined oxidative stress of diabetes and nicotine in rats[J].Molecular Medicine Reports,2016,13(4):3017～3026
[11] 靳晶,王全胜.糖尿病肾病上皮细胞向间充质细胞的转分化[J].临床肾脏病杂志,2017,17(2):68～74
[12] 成芳,涂珍珍,周利利,等.雷帕霉素对肾小管上皮细胞生物学行为的影响及其治疗糖尿病肾病分子机制的初步研究[J].安徽医科大学学报,2017,52(1):1～6
[13] 陈红淑,杨元宵,周小杰,等.肾气丸通过TGF-β1/Smads/ILK信号途径干预肾小管上皮细胞间充质转分化的分子机制[J].中华中医药杂志,2016,13(6):2102～2105
[14] Xu J,Li P X,Wu J,et al.Involvement of the Hippo pathway in regeneration and fibrogenesis after ischaemic acute kidney injury:YAP is the key effector.[J].Clinical Science,2016,130(5):349～363
[15] 陈雨思,李均.肝细胞生长因子抗肾纤维化的机制[J].海南医学,2016,27(20):3364～3366
[16] 苗永辉,赵宗江,杨冠男,等.糖肾平含药血清对高糖诱导肾小管上皮细胞增殖作用及RhoA/ROCK信号通路影响的研究[J].世界科学技术-中医药现代化,2017,9(6):1050～1056