六味地黄丸对糖尿病肾病大鼠肾脏组织RhoA/ROCK1表达的影响
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摘要
目的:观察六味地黄丸对糖尿病肾病(diabetic kidney disease,DKD)大鼠肾脏组织RhoA/ROCK1表达的影响。方法:将36只SD雄性大鼠按照随机数字表法分为空白组、模型组、六味地黄丸组(六味组)各12只,采用一次性腹腔注射链脲佐菌素(45 mg·kg~(-1))的方法建立DKD大鼠模型。造模成功后,空白组、模型组灌服生理盐水,六味组给予六味地黄丸。8周后检测24小时尿蛋白定量、血清白蛋白(serum albumin,ALB)、天门冬氨酸氨基转移酶(aspartate aminotransferase,ALT)、血肌酐(serum creatinine,Scr)等水平;测量大鼠体质量、肾脏质量,计算肥大指数;苏木精-伊红染色、过碘酸-雪夫氏染色观察肾脏模型组织;酶联免疫吸附法测定检测肾脏组织RhoA、ROCK1的水平。结果:与空白组比较,模型组24小时尿蛋白定量升高,ALB下降,体质量下降,肥大指数升高,差异均有统计学意义(P<0.05);与模型组比较,六味组24小时尿蛋白定量显著下降,ALB显著升高,体质量升高,肥大指数降低,且RhoA、ROCK1的表达水平显著下降,差异均有统计学意义(P<0.05)。空白组肾小球、肾小管、肾间质结构大小等均正常完整;六味组与模型组均有不同程度的肾脏病理改变,其中六味组较模型组病理改变减轻。结论:六味地黄丸可能通过影响DKD大鼠肾脏组织RhoA、ROCK1的表达,降低蛋白尿,减轻肾脏病理损害,且对肝肾功能无明显影响。
Objective:To observe effect of liuwei dihuang pill on expression of RhoA/ROCK1 in renal tissues of diabetic kidney disease( DKD) rats.Methods:36 SD male rats were randomly divided into blank group,model group and liuwei dihuang pill group(liuwei group),12 in each group.The DKD rat model was produced by a single intraperitoneal injection of streptozotocin(45 mg·kg-1).After the successful modeling,the blank group and the model group were given normal saline,and the liuwei group was given liuwei dihuang pill. After 8 weeks,24-hour urine protein quantitation,serum albumin( ALB),aspartate aminotransferase(ALT),serum creatinine(Scr) were measured.The body and kidney weight of the rats were measured and the hypertrophy index was calculated.Kidney model tissue was observed through Hematoxylin-Eosin and Periodic Acid-Schiff dyeing.Renal tissue RhoA,ROCK1 level were measured through enzyme linked immunosorbent assay.Results:Compared with the blank group,the 24-hour urine protein level increased and ALB decreased in the model group. The body weight decreased and the hypertrophy index increased.The differences were statistically significant( P < 0. 05). Compared with the model group,the 24-hour urine protein decreased significantly in the liuwei group.The ALB and the body weight increased significantly.The hypertrophy index decreased.The expression level of RhoA and ROCK1 were significantly decreased.The differences were statistically significant( P< 0.05).The glomeruli,renal tubules,and renal interstitial structures in the blank group were normal and intact.There were different degrees of renal pathological changes in the liuwei group and the model group,and compared with the model group,the pathological changes in the liuwei group were alleviated.Conclusion:liuwei dihuang pill may reduce proteinuria and renal pathological damage by affecting the expression of RhoA and ROCK1 in renal tissue of of DKD rats,and has no significant effect on liver and kidney function.
Objective:To observe effect of liuwei dihuang pill on expression of RhoA/ROCK1 in renal tissues of diabetic kidney disease( DKD) rats.Methods:36 SD male rats were randomly divided into blank group,model group and liuwei dihuang pill group(liuwei group),12 in each group.The DKD rat model was produced by a single intraperitoneal injection of streptozotocin(45 mg·kg-1).After the successful modeling,the blank group and the model group were given normal saline,and the liuwei group was given liuwei dihuang pill. After 8 weeks,24-hour urine protein quantitation,serum albumin( ALB),aspartate aminotransferase(ALT),serum creatinine(Scr) were measured.The body and kidney weight of the rats were measured and the hypertrophy index was calculated.Kidney model tissue was observed through Hematoxylin-Eosin and Periodic Acid-Schiff dyeing.Renal tissue RhoA,ROCK1 level were measured through enzyme linked immunosorbent assay.Results:Compared with the blank group,the 24-hour urine protein level increased and ALB decreased in the model group. The body weight decreased and the hypertrophy index increased.The differences were statistically significant( P < 0. 05). Compared with the model group,the 24-hour urine protein decreased significantly in the liuwei group.The ALB and the body weight increased significantly.The hypertrophy index decreased.The expression level of RhoA and ROCK1 were significantly decreased.The differences were statistically significant( P< 0.05).The glomeruli,renal tubules,and renal interstitial structures in the blank group were normal and intact.There were different degrees of renal pathological changes in the liuwei group and the model group,and compared with the model group,the pathological changes in the liuwei group were alleviated.Conclusion:liuwei dihuang pill may reduce proteinuria and renal pathological damage by affecting the expression of RhoA and ROCK1 in renal tissue of of DKD rats,and has no significant effect on liver and kidney function.
引文
[1]PERCO P,MAYER G.Molecular,histological,and clinical phenotyping of diabetic nephropathy:valuable complementary information[J].Kidney Int,2018,93(2):308-310.
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[3]李志杰,张悦,陆海英,等.六味地黄丸对糖尿病肾病大鼠肾脏组织TGF-β1-smad通路的影响[J].时珍国医国药,2017,28(8):1811-1814.
[4]杨浠.六味地黄丸治疗糖尿病肾病的机理研究进展[J].中西医结合心血管病电子杂志,2018,6(4):22.
[5]刘兴国,杜冬琛,付明洁.六味地黄丸合生脉汤加减辅助治疗早中期糖尿病肾病效果及血清Cys-C水平变化[J].山东医药,2018,58(2):73-75.
[6]张丽芬,吕仁和,黄文政.链脲佐菌素糖尿病肾病大鼠模型的建立及稳定性评价[J].中国比较医学杂志,2014,24(4):8-12,18.
[7]徐叔云,卞如濂,陈修.药理实验方法学[M].4版.北京:人民卫生出版社,2010:206-207.
[8]MOON J,LEE C J,LEE S H,et al.The Impact of Diabetes Mellitus on Vascular Biomarkers in Patients with End-Stage Renal Disease[J].Yonsei Med J,2017,58(1):75-81.
[9]TAN H,ZHONG Y,SHEN X,et al.Erythropoietin promotes axonal regeneration after optic nerve crush in vivo by inhibition of Rho A/ROCK signaling pathway[J].Neuropharmacology,2012,63(6):1182-1190.
[10]ZHANG L,JI T,WANG Q,et al.Calcium-Sensing Receptor Stimulation in Cultured Glomerular Podocytes Induces TRPC6-Dependent Calcium Entry and Rho A Activation[J].Cell Physiol Biochem,2017,43(5):1777-1789.
[11]陆世龙,王龙龙,黄国东.中医药治疗早期糖尿病肾病的研究进展[J].辽宁中医杂志,2016,43(5):1101-1103.
[12]柳红芳,张向伟,张先慧.糖尿病肾病的审因论治[J].中医杂志,2016,57(19):1646-1648.
[13]张向伟,柳红芳,张先慧.糖尿病肾病病机层次分析与辨治[J].中医杂志,2017,58(5):390-393.
[14]李志杰,张悦.六味地黄丸(汤)治疗肾脏病的研究进展[J].中成药,2017,39(5):1024-1028.
[15]冯蕾,李兴波.六味地黄丸联合二甲双胍治疗2型糖尿病肾阴亏虚的疗效研究[J].药物评价研究,2017,40(8):1130-1133.
[16]谭颖颖,屈直,张琪.六味地黄丸含药血清减低高糖诱导的肾小管上皮细胞的氧化损伤和凋亡的研究[J].时珍国医国药,2015,26(7):1566-1569.
[17]张家丽,徐钊熠,马秀宁.六味地黄汤加减治疗慢性肾炎110例临床疗效分析[J].中国医药指南,2016,14(35):195-196.
[18]王久香.六味地黄丸联合氯沙坦钾治疗早期糖尿病肾病的临床研究[J].现代药物与临床,2015,30(6):674-677.
[19]张亚男.六味地黄丸对狼疮肾炎患者糖基化终末产物受体、单核细胞趋化蛋白1及趋化因子Fractalkine的影响[J].临床肾脏病杂志,2017,17(8):499-502.
[20]陈丽,周忠志,何泽云,等.六味地黄丸对慢性肾脏病研究的新进展[J].湖南中医药大学学报,2013,33(8):104-107.
[21]司美君,叶增纯,赵文波,等.2型糖尿病合并慢性肾脏病患者的肾脏病理与临床表现特点[J].中华肾脏病杂志,2016,32(6):401-405.
[2]洪金妮,黎巍威,王学美.糖尿病肾病中足细胞损伤的信号通路及中药的干预研究进展[J].中药药理与临床,2017,33(3):215-219.
[3]李志杰,张悦,陆海英,等.六味地黄丸对糖尿病肾病大鼠肾脏组织TGF-β1-smad通路的影响[J].时珍国医国药,2017,28(8):1811-1814.
[4]杨浠.六味地黄丸治疗糖尿病肾病的机理研究进展[J].中西医结合心血管病电子杂志,2018,6(4):22.
[5]刘兴国,杜冬琛,付明洁.六味地黄丸合生脉汤加减辅助治疗早中期糖尿病肾病效果及血清Cys-C水平变化[J].山东医药,2018,58(2):73-75.
[6]张丽芬,吕仁和,黄文政.链脲佐菌素糖尿病肾病大鼠模型的建立及稳定性评价[J].中国比较医学杂志,2014,24(4):8-12,18.
[7]徐叔云,卞如濂,陈修.药理实验方法学[M].4版.北京:人民卫生出版社,2010:206-207.
[8]MOON J,LEE C J,LEE S H,et al.The Impact of Diabetes Mellitus on Vascular Biomarkers in Patients with End-Stage Renal Disease[J].Yonsei Med J,2017,58(1):75-81.
[9]TAN H,ZHONG Y,SHEN X,et al.Erythropoietin promotes axonal regeneration after optic nerve crush in vivo by inhibition of Rho A/ROCK signaling pathway[J].Neuropharmacology,2012,63(6):1182-1190.
[10]ZHANG L,JI T,WANG Q,et al.Calcium-Sensing Receptor Stimulation in Cultured Glomerular Podocytes Induces TRPC6-Dependent Calcium Entry and Rho A Activation[J].Cell Physiol Biochem,2017,43(5):1777-1789.
[11]陆世龙,王龙龙,黄国东.中医药治疗早期糖尿病肾病的研究进展[J].辽宁中医杂志,2016,43(5):1101-1103.
[12]柳红芳,张向伟,张先慧.糖尿病肾病的审因论治[J].中医杂志,2016,57(19):1646-1648.
[13]张向伟,柳红芳,张先慧.糖尿病肾病病机层次分析与辨治[J].中医杂志,2017,58(5):390-393.
[14]李志杰,张悦.六味地黄丸(汤)治疗肾脏病的研究进展[J].中成药,2017,39(5):1024-1028.
[15]冯蕾,李兴波.六味地黄丸联合二甲双胍治疗2型糖尿病肾阴亏虚的疗效研究[J].药物评价研究,2017,40(8):1130-1133.
[16]谭颖颖,屈直,张琪.六味地黄丸含药血清减低高糖诱导的肾小管上皮细胞的氧化损伤和凋亡的研究[J].时珍国医国药,2015,26(7):1566-1569.
[17]张家丽,徐钊熠,马秀宁.六味地黄汤加减治疗慢性肾炎110例临床疗效分析[J].中国医药指南,2016,14(35):195-196.
[18]王久香.六味地黄丸联合氯沙坦钾治疗早期糖尿病肾病的临床研究[J].现代药物与临床,2015,30(6):674-677.
[19]张亚男.六味地黄丸对狼疮肾炎患者糖基化终末产物受体、单核细胞趋化蛋白1及趋化因子Fractalkine的影响[J].临床肾脏病杂志,2017,17(8):499-502.
[20]陈丽,周忠志,何泽云,等.六味地黄丸对慢性肾脏病研究的新进展[J].湖南中医药大学学报,2013,33(8):104-107.
[21]司美君,叶增纯,赵文波,等.2型糖尿病合并慢性肾脏病患者的肾脏病理与临床表现特点[J].中华肾脏病杂志,2016,32(6):401-405.