藏药四味姜黄方对STZ诱导糖尿病肾病大鼠模型的剂量配比关系的初步研究
|
摘要
目的开展藏药四味姜黄方的剂量配比关系研究,探索其处方剂量配比关系对糖尿病肾病(DN)大鼠的保护作用及其机制。方法除正常组外,其余大鼠均使用60 mg/kg体质量的链脲佐菌素(STZ)一次性腹腔内注射法复制糖尿病模型,待DN模型成功后将其分为模型组、二甲双胍阳性组、四味姜黄方8个不同剂量配比组(采用均匀设计法),按给药剂量表每天灌胃给予相应药物1次,连续给药4周。使用血糖仪测空腹血糖(FBG),化学法检测血清尿素氮(BUN)、尿酸(UA)、肌酐(Scr)和总蛋白(TP)浓度,ELISA试剂盒检测转化生长因子-β1(TGF-β1)和血管内皮生长因子(VEGF)水平,电镜下观察肾脏病理形态学变化及测定肾小球基底膜厚度。采用主成分分析(PCA)、多元逐步回归分析(MSRA)等多元分析方法,对配比剂量-药效学结果进行剂量配比关系分析。结果各给药组的各项药效学指标均有不同程度的改善。MSRA结果表明BUN、肾指数、FBG、肾小球基底膜厚度、VEGF、Scr和UA 7项指标水平与剂量配比存在多元一次线性关系或多元二次非线性关系,其回归方程有显著性意义(P<0.05);TGF-β1和TP水平与剂量配比间无明显线性或非线性关系,其回归方程无显著性意义(P>0.05);在均匀设计取值范围内的全局寻优比较,大鼠模型的优化配比剂量为姜黄∶小檗皮∶余甘子∶蒺藜=1∶2∶1∶2。结论四味姜黄方各剂量配比对DN的各项指标均有不同程度的改善,这可能与其降低BUN、肾指数、FBG、肾小球基底膜厚度、VEGF、Scr和UA等指标的水平有关。
AIM To investigate the dose-proportion relation of Tibetan medicine Siwei Jianghuang Prescription( SJP) for protective effects on diabetic nephropathy( DN),and the underlying mechanism. METHODS Diabetes mellitus rats induced by intraperitoneal injection of streptozotocin( STZ)( 60 mg/kg) were randomly divided into model group,metformin support group,and eight SJP groups with dose-proportion variation( with reference to the uniform design method) for corresponding drug administration once a day,for four weeks. Measurement of fasting blood glucose( FBG) by a blood glucose meter,the concentrations of blood urea nitrogen( BUN),uric acid( UA),serum creatinine( SCr) and total protein( TP) by chemical methods,serum transforming growth factor-β1( TGF-β1) and vascular endothelial growth factor( VEGF) by ELISA kits were conducted,the pathological morphology observation and glomerular basement membrane thickness detection by electron microscope were accomplished as well. Principal components analysis( PCA) and multivariate progressive regression analysis( MSRA)were employed to analyze the relationship between the dose-proportion to pharmacodynamics. RESULTS The resultant indexes revealed variant pharmacological improvement in each treatment group. MSRA results showed that the levels of BUN,renal index,FBG,glomerular basement membrane thickness,VEGF,Scr, and UA had correlative relations with a multiple linear or a multiple non-linear in all groups,which regression equation had a statistical significance( P < 0. 05); TGF-β1 level and total protein index with the dose-proportion had no linear or non-linear relation,which the regression equation statistical showed non-significance( P > 0. 05). In the global optimization comparison around the range of uniform design,the optimal dosage of the rats model was Curcumae Longae Rhizoma ∶ Berberidis dictyophyllae Cortex ∶ Phyllanthi Fructus ∶ Tribuli Fructus = 1 ∶ 2 ∶ 1 ∶ 2.CONCLUSION Siwei Jianghuang Prescription shows better therapeutic effects on DN,which may be related to reducing the levels of BUN,renal index,FBG,glomerular basement membrane thickness,VEGF,Scr and UA.
AIM To investigate the dose-proportion relation of Tibetan medicine Siwei Jianghuang Prescription( SJP) for protective effects on diabetic nephropathy( DN),and the underlying mechanism. METHODS Diabetes mellitus rats induced by intraperitoneal injection of streptozotocin( STZ)( 60 mg/kg) were randomly divided into model group,metformin support group,and eight SJP groups with dose-proportion variation( with reference to the uniform design method) for corresponding drug administration once a day,for four weeks. Measurement of fasting blood glucose( FBG) by a blood glucose meter,the concentrations of blood urea nitrogen( BUN),uric acid( UA),serum creatinine( SCr) and total protein( TP) by chemical methods,serum transforming growth factor-β1( TGF-β1) and vascular endothelial growth factor( VEGF) by ELISA kits were conducted,the pathological morphology observation and glomerular basement membrane thickness detection by electron microscope were accomplished as well. Principal components analysis( PCA) and multivariate progressive regression analysis( MSRA)were employed to analyze the relationship between the dose-proportion to pharmacodynamics. RESULTS The resultant indexes revealed variant pharmacological improvement in each treatment group. MSRA results showed that the levels of BUN,renal index,FBG,glomerular basement membrane thickness,VEGF,Scr, and UA had correlative relations with a multiple linear or a multiple non-linear in all groups,which regression equation had a statistical significance( P < 0. 05); TGF-β1 level and total protein index with the dose-proportion had no linear or non-linear relation,which the regression equation statistical showed non-significance( P > 0. 05). In the global optimization comparison around the range of uniform design,the optimal dosage of the rats model was Curcumae Longae Rhizoma ∶ Berberidis dictyophyllae Cortex ∶ Phyllanthi Fructus ∶ Tribuli Fructus = 1 ∶ 2 ∶ 1 ∶ 2.CONCLUSION Siwei Jianghuang Prescription shows better therapeutic effects on DN,which may be related to reducing the levels of BUN,renal index,FBG,glomerular basement membrane thickness,VEGF,Scr and UA.
引文
[1]Forbes J M,Cooper M E.Mechanisms of diabetic complications[J].Physiol Rev,2013,93(1):137-188.
[2]陈娟,张明华,章丽,等.牡丹皮苷/酚组分对糖尿病肾病大鼠肾损伤的保护作用及其机制研究[J].中国中药杂志,2016,41(11):1990-1998.
[3]Li J J,Kwak S J,Jung D S,et al.Podcyte biology in diabetic nephropathy[J].Kidney Int Suppl,2007(106):S36-S42.
[4]王智森,陆付耳.中藏医对糖尿病病因病机的认识[J].中西医结合研究,2011,3(6):313-316.
[5]田华,母传贤,尤丽菊,等.姜黄素对糖尿病肾病大鼠肾脏的保护作用及其机制[J].吉林大学学报:医学版,2013,39(4):747-750,866.
[6]张燕,孟宪丽,岳丽珺,等.藏药小檗皮对糖尿病模型小鼠血糖水平影响的初步研究[J].现代生物医学进展,2013,13(19):3619-3622,3607.
[7]胡炜.余甘子提取物对糖尿病大鼠肌肉和脂肪组织胰岛素信号通路的影响[J].中国组织工程研究,2012,16(11):2007-2010.
[8]张素军,冯尚彩.蒺藜皂苷对正常和2型糖尿病大鼠餐后血糖水平的影响[J].实用药物与临床,2012,15(1):1-3.
[9]赖先荣,吴秦西,向永臣,等.均匀设计优选藏药如达胃宁的组方[J].中国药房,2010,21(19):1734-1736.
[10]李晓萍李进,黄学宽.楂芪降糖汤对糖尿病肾病大鼠肾脏TGF-β1及VEGF表达的影响[J].中成药,2013,35(3):450-455.
[11]芝敏,胡兰英,徐佩茹.2型糖尿病肾病大鼠模型的建立及其肾脏病理特点[J].新疆医学,2012,42(12):13-17.
[12]赖先荣,周邦华,杜明胜,等.6种黄连饮片中6种生物碱的RP-HPLC含量测定及与“治消渴”药效学的谱-效关系分析[J].中国中药杂志,2016,41(24):4579-4586.
[13]赵磊,李继海,朱大洲,等.5种鹿茸营养成分的主成分分析[J].光谱学与光谱分析,2010,30(9):2571-2575.
[14]万星火,檀亦丽.主成分分析原始数据的预处理问题[J].中国卫生统计,2005,22(5):327-329.
[15]代晓琳,马学强,王甜甜.改进的主成分分析法对拟南芥基因的分析[J].微型机与应用,2015,34(14):51-53,57.
[16]Lan T,Liu W,Xie X,et al.Berberine suppresses high glucose-induced TGF-β1 and fibronectin synthesis in mesangial cells through inhibition of sphingosine kinase 1/AP-1 pathway[J].Eur J Pharmacol,2012,697(1-3):165-172.
[17]Tufro A,Veron D.VEGF and Podocytes in diabetic nephropathy[J].Semin Nephrol,2012,32(4):385-393.
[18]李晓萍,李进,黄学宽.楂芪降糖汤对糖尿病肾病大鼠肾脏TGF-β1及VEGF表达的影响[J].中成药,2013,35(3):450-455.
[19]刘圣,余娜,张小力,等.小檗碱对早期糖尿病肾病大鼠肾组织TGF-β1/Sno N表达失衡及其Smad信号通路的调控作用[J].中国中药杂志,2012,37(23):3604-3610.
[2]陈娟,张明华,章丽,等.牡丹皮苷/酚组分对糖尿病肾病大鼠肾损伤的保护作用及其机制研究[J].中国中药杂志,2016,41(11):1990-1998.
[3]Li J J,Kwak S J,Jung D S,et al.Podcyte biology in diabetic nephropathy[J].Kidney Int Suppl,2007(106):S36-S42.
[4]王智森,陆付耳.中藏医对糖尿病病因病机的认识[J].中西医结合研究,2011,3(6):313-316.
[5]田华,母传贤,尤丽菊,等.姜黄素对糖尿病肾病大鼠肾脏的保护作用及其机制[J].吉林大学学报:医学版,2013,39(4):747-750,866.
[6]张燕,孟宪丽,岳丽珺,等.藏药小檗皮对糖尿病模型小鼠血糖水平影响的初步研究[J].现代生物医学进展,2013,13(19):3619-3622,3607.
[7]胡炜.余甘子提取物对糖尿病大鼠肌肉和脂肪组织胰岛素信号通路的影响[J].中国组织工程研究,2012,16(11):2007-2010.
[8]张素军,冯尚彩.蒺藜皂苷对正常和2型糖尿病大鼠餐后血糖水平的影响[J].实用药物与临床,2012,15(1):1-3.
[9]赖先荣,吴秦西,向永臣,等.均匀设计优选藏药如达胃宁的组方[J].中国药房,2010,21(19):1734-1736.
[10]李晓萍李进,黄学宽.楂芪降糖汤对糖尿病肾病大鼠肾脏TGF-β1及VEGF表达的影响[J].中成药,2013,35(3):450-455.
[11]芝敏,胡兰英,徐佩茹.2型糖尿病肾病大鼠模型的建立及其肾脏病理特点[J].新疆医学,2012,42(12):13-17.
[12]赖先荣,周邦华,杜明胜,等.6种黄连饮片中6种生物碱的RP-HPLC含量测定及与“治消渴”药效学的谱-效关系分析[J].中国中药杂志,2016,41(24):4579-4586.
[13]赵磊,李继海,朱大洲,等.5种鹿茸营养成分的主成分分析[J].光谱学与光谱分析,2010,30(9):2571-2575.
[14]万星火,檀亦丽.主成分分析原始数据的预处理问题[J].中国卫生统计,2005,22(5):327-329.
[15]代晓琳,马学强,王甜甜.改进的主成分分析法对拟南芥基因的分析[J].微型机与应用,2015,34(14):51-53,57.
[16]Lan T,Liu W,Xie X,et al.Berberine suppresses high glucose-induced TGF-β1 and fibronectin synthesis in mesangial cells through inhibition of sphingosine kinase 1/AP-1 pathway[J].Eur J Pharmacol,2012,697(1-3):165-172.
[17]Tufro A,Veron D.VEGF and Podocytes in diabetic nephropathy[J].Semin Nephrol,2012,32(4):385-393.
[18]李晓萍,李进,黄学宽.楂芪降糖汤对糖尿病肾病大鼠肾脏TGF-β1及VEGF表达的影响[J].中成药,2013,35(3):450-455.
[19]刘圣,余娜,张小力,等.小檗碱对早期糖尿病肾病大鼠肾组织TGF-β1/Sno N表达失衡及其Smad信号通路的调控作用[J].中国中药杂志,2012,37(23):3604-3610.