益气养阴活血中药对糖尿病肾病大鼠Notch/Hes1通路与血管内皮CD34、CD144的影响
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摘要
目的观察益气养阴活血中药对糖尿病肾病大鼠Notch/Hes1通路与血管内皮CD34、CD144作用机制。方法采取左肾切除术+高脂高糖饲料喂养联合腹腔注射STZ复制早期糖尿病肾病大鼠模型,将建模成功的大鼠随机分为模型组、西药贝拉普利组、中药高剂量组、中药中剂量组、中药低剂量组,设正常对照组,药物干预8周。观察24 h尿白蛋白、HE染色肾组织病理、免疫组化肾组织Hes1、Western blot法检测CD34、CD144的表达。结果与正常组比较,模型组24 h尿白蛋白、Hes1、CD34、CD144的含量比较均有明显差异(P<0.05);与模型组比较,中药高剂量组、中剂量组能降低24 h尿蛋白(P<0.05),中药各剂量组均能明显降低肾组织Hes1、CD34含量(P<0.05),中药高剂量组均能明显降低肾组织CD144含量(P<0.05);与西药组比较,中药各剂量组均能降低CD34(P<0.05),中药低剂量组降低CD144(P<0.05)。病理观察模型组大鼠出现肾小球硬化、萎缩,系膜基质增多、系膜出现增厚增宽并可见结节样增生等改变;中药高、中剂量组对上述病理改变有积极改善。结论益气养阴活血中药可以降低模型大鼠肾组织Hes1和CD34、CD144,发挥对肾功能的保护作用,延缓DN的发展。
Objective To observe the effects of a traditional Chinese medicine(TCM) capsule for replenishing qi, nourishing yin and activating blood on Notch/Hes1 signaling pathway in the renal tissue and vascular endothelial CD34 and CD144 expressions in a rat model of diabetic nephropathy. Methods Rat models of early-stage diabetic nephropathy were established by left nephrectomy and high-fat and high-sugar feeding combined with intraperitoneal injection of STZ. The rats were randomized into model group, benazepril group, and high-, moderate-, and low-dose TCM capsule groups for corresponding treatments, with 6 normal rats as the control group. After 8 weeks of drug treatment, blood glucose and 24-h urinary albumin of the rats were measured, and the renal histopathology was observed with HE staining; Hes1 expression in the renal tissue was detected with immunohistochemical staining, and the renal expressions of CD34 and CD144 were detected using Western blotting. Results Compared with the normal control group, the rat models of diabetic nephropathy showed obvious abnormalities in 24-h urinary albumin and expressions of Hes1, CD34 and CD144 d. The TCM capsule at both the high and moderate doses significantly reduced 24-h urinary albumin in the rats; the renal expressions of Hes1 and CD34 was significantly reduced in all the dose groups, and the expression of CD144 was significantly reduced in the high-dose group.Compared with benazepril group, the TCM capsule obviously reduced CD34 expression at all the 3 doses and lowered CD144 expression at the low dose. Histopathologically, the rats in the model group showed glomerular hypertrophy, increased mesenteric matrix, thickening and widening of the mesenteric membrane, and nodular hyperplasia. These pathologies were obviously alleviated by treatment with the TCM capsule at the high and moderate doses. Conclusion The Traditional Chinese medicine(TCM) capsule for replenishing qi, nourishing yin and activating blood can reduce Hes1, CD34 and CD144 in kidney tissue of model rats, play a protective role on kidney function and delay the development of DN.
Objective To observe the effects of a traditional Chinese medicine(TCM) capsule for replenishing qi, nourishing yin and activating blood on Notch/Hes1 signaling pathway in the renal tissue and vascular endothelial CD34 and CD144 expressions in a rat model of diabetic nephropathy. Methods Rat models of early-stage diabetic nephropathy were established by left nephrectomy and high-fat and high-sugar feeding combined with intraperitoneal injection of STZ. The rats were randomized into model group, benazepril group, and high-, moderate-, and low-dose TCM capsule groups for corresponding treatments, with 6 normal rats as the control group. After 8 weeks of drug treatment, blood glucose and 24-h urinary albumin of the rats were measured, and the renal histopathology was observed with HE staining; Hes1 expression in the renal tissue was detected with immunohistochemical staining, and the renal expressions of CD34 and CD144 were detected using Western blotting. Results Compared with the normal control group, the rat models of diabetic nephropathy showed obvious abnormalities in 24-h urinary albumin and expressions of Hes1, CD34 and CD144 d. The TCM capsule at both the high and moderate doses significantly reduced 24-h urinary albumin in the rats; the renal expressions of Hes1 and CD34 was significantly reduced in all the dose groups, and the expression of CD144 was significantly reduced in the high-dose group.Compared with benazepril group, the TCM capsule obviously reduced CD34 expression at all the 3 doses and lowered CD144 expression at the low dose. Histopathologically, the rats in the model group showed glomerular hypertrophy, increased mesenteric matrix, thickening and widening of the mesenteric membrane, and nodular hyperplasia. These pathologies were obviously alleviated by treatment with the TCM capsule at the high and moderate doses. Conclusion The Traditional Chinese medicine(TCM) capsule for replenishing qi, nourishing yin and activating blood can reduce Hes1, CD34 and CD144 in kidney tissue of model rats, play a protective role on kidney function and delay the development of DN.
引文
[1]Wu SL,Li XY,Zhang HM.Effects of metformin on endothelial function in type 2 diabetes[J].Exp TherMed,2014,7(5):1349-53.
[2]Krasnicki P,Dmuchowska DA,Proniewska-Skretek E,et al.Ocular haemodynamics in patients with type 2 diabetes and coronary artery disease[J].Brit J Ophthalmol,2014,98(5):675-8.
[3]Tasyurek HM,Altunbas HA,Balci MK.Incretins:their physiology and application in the treatment of diabetes mellitus[J].Diabetes Metab Res Rev,2014,30(5):354-71.
[4]Asahara T,Murohara T,Sullivan A,et al.Isolation of putative progenitor endothelial cells for angiogenesis[J].Science,1997,275(532):964-7.
[5]刘竹影,陈颖,刘倩,等.血管内皮祖细胞改善骨质疏松大鼠骨髓间充质干细胞的增殖及凋亡[J].中国组织工程研究,2016,20(14):1999-2006.
[6]Bazzoni G,Dejana E.Endothelial cell-to-cell junctions:molecular organization and role in vascular homeostasis[J].Physiol Rev,2004,84(3):869-901.
[7]Mehta D,Malik AB.Signaling mechanisms regulating endothelial permeability[J].Physiol Rev,2006,86(1):279-367.
[8]Fish JE,Wythe JD.The molecular regulation of arteriovenous specification and maintenance[J].Dev Dynam,2015,244(3,SI):391-409.
[9]Doan PL,Russell JL,Himburg HA,et al.Tie2(+)bone marrow endothelial cells regulate hematopoietic stem cell regeneration following radiation injury[J].Stem Cells,2013,31(2):327-37.
[10]鲍陶陶,杨晓春,储全根.丹蛭降糖胶囊对糖尿病大鼠主动脉超微结构,MCP-1,抵抗素mRNA表达的影响[J].中成药,2015,37(9):1888-92.
[11]郑书国,陶善珺,赵梦秋,等.丹蛭降糖胶囊对糖尿病血糖波动模型大鼠心肌纤维化的影响[J].中药材,2015,38(10):2120-4.
[12]Lu Y,Chen Y,Li R,et al.Protective effects of Danzhi jiangtang capsule on vascular endothelial damages induced by high-fat diet and palmitic acid[J].Biomed Pharmacot,2018,107(11):1631-40.
[13]贾会玉,那莎,李莉,等.丹蛭降糖胶囊对大鼠糖尿病肾病防治作用的研究[J].中华中医药杂志,2016,31(12):5244-7.
[14]高雪,安至超,何其英,等.高脂饲料喂养时间对2型糖尿病肾病大鼠模型的影响[J].中国实验动物学报,2017,26(1):1-9.
[15]徐叔云.药理实验方法学[M].北京:人民卫生出版社,1982:203.
[16]Abdul MM,Osamu I.Elewa yaser hosny Ali;nakamura teppei;Kon yasuhiro;local CD34-positive capillaries decrease in mouse models of kidney disease associating with the severity of glomerular and tubulointerstitial lesions[J].BMC Nephrol,2017,18(9):280-92.
[17]周丽娜,王玉新,方林,等.内皮祖细胞修复缺血再灌注大鼠肾损伤[J].中国组织工程研究,2014,18(32):5146-51.
[18]毕凌云,杨达胜,赵德安,等.动员自身骨髓干细胞与缺血再灌注肾损伤细胞的凋亡与增殖[J].中国组织工程研究,2013,17(49):8488-97.
[19]刘建辉,孙志平,王琴,等.糖尿病肾病患者血浆EMP与AECA的诊断价值[J].热带医学杂志,2015,15(11):1478-80.
[20]玛依努·玉苏甫.赵红丽.丹参多酚酸盐治疗早期糖尿病肾病的临床价值研究[J].医学综述,2015,21(23):4382-4.
[21]Gupta N,Wish JB.Erythropoietin mimetic peptides and erythropoietin fusion proteins for treating anemia of chronic kidney disease[J].Curr Opin Nephrol Hypertens,2018,27(5):345-50.
[22]Mansour A,Elbaky AA,Kamal S,et al.Evaluation of the procoagulant potential of endothelial microparticles CD144(VECadherin)positive in coronary syndrome patients[J].Egyp JHaematol,2014,39(3):143-8.
[23]Ji X,Wang Z,Geamanu A,et al.Inhibition of cell growth and induction of apoptosis in non-small cell lung cancer cells by deltatocotrienol is associated with notch-1 down-regulation[J].J Cell Biochem,2011,112(10):2773-83.
[24]高峰,刘淑霞,赵玉峰,等.Notch信号通路在糖尿病小鼠肾组织的表达及意义[J].中国组织化学与细胞化学杂志,2014,23(5):445-9.
[25]高家荣,庄星星,魏良兵,等.丹蛭降糖胶囊治疗2型糖尿病血管病变的作用机制研究[J].中药药理与临床,2014,30(6):143-5.
[26]罗云,方朝晖.丹蛭降糖胶囊对糖尿病动脉硬化模型大鼠胸主动脉p22phoxmRNA,p47phox mRNA表达的影响[J].新中医,2018(50):5-8.
[27]杨晓春,鲍陶陶,储全根.基于p38MAPK通路探讨丹蛭降糖胶囊对2型糖尿病模型大鼠血管病变的影响[J].中国实验方剂学杂志,2016,22(3):116-20.
[28]胡云飞,徐国兵.牡丹皮及其主要成分丹皮酚的药理作用研究进展[J].安徽医药,2014,18(4):589-92.
[29]梁裕芬.医用水蛭的生物学特性及药用价值概述[J].生物学教学,2017,42(10):2-4.
[30]Beets K,Huylebroeck D,Moya IM,et al.Robustness in angiogenesis:notch and BMP shaping waves[J].Trends Genet,2013,29(3):140-9.
[31]杜潇,张思琴,程中,等.激活Notch1信号通路对胰腺癌细胞增殖的影响[J].南方医科大学学报,2013,33(10):1494-8.
[2]Krasnicki P,Dmuchowska DA,Proniewska-Skretek E,et al.Ocular haemodynamics in patients with type 2 diabetes and coronary artery disease[J].Brit J Ophthalmol,2014,98(5):675-8.
[3]Tasyurek HM,Altunbas HA,Balci MK.Incretins:their physiology and application in the treatment of diabetes mellitus[J].Diabetes Metab Res Rev,2014,30(5):354-71.
[4]Asahara T,Murohara T,Sullivan A,et al.Isolation of putative progenitor endothelial cells for angiogenesis[J].Science,1997,275(532):964-7.
[5]刘竹影,陈颖,刘倩,等.血管内皮祖细胞改善骨质疏松大鼠骨髓间充质干细胞的增殖及凋亡[J].中国组织工程研究,2016,20(14):1999-2006.
[6]Bazzoni G,Dejana E.Endothelial cell-to-cell junctions:molecular organization and role in vascular homeostasis[J].Physiol Rev,2004,84(3):869-901.
[7]Mehta D,Malik AB.Signaling mechanisms regulating endothelial permeability[J].Physiol Rev,2006,86(1):279-367.
[8]Fish JE,Wythe JD.The molecular regulation of arteriovenous specification and maintenance[J].Dev Dynam,2015,244(3,SI):391-409.
[9]Doan PL,Russell JL,Himburg HA,et al.Tie2(+)bone marrow endothelial cells regulate hematopoietic stem cell regeneration following radiation injury[J].Stem Cells,2013,31(2):327-37.
[10]鲍陶陶,杨晓春,储全根.丹蛭降糖胶囊对糖尿病大鼠主动脉超微结构,MCP-1,抵抗素mRNA表达的影响[J].中成药,2015,37(9):1888-92.
[11]郑书国,陶善珺,赵梦秋,等.丹蛭降糖胶囊对糖尿病血糖波动模型大鼠心肌纤维化的影响[J].中药材,2015,38(10):2120-4.
[12]Lu Y,Chen Y,Li R,et al.Protective effects of Danzhi jiangtang capsule on vascular endothelial damages induced by high-fat diet and palmitic acid[J].Biomed Pharmacot,2018,107(11):1631-40.
[13]贾会玉,那莎,李莉,等.丹蛭降糖胶囊对大鼠糖尿病肾病防治作用的研究[J].中华中医药杂志,2016,31(12):5244-7.
[14]高雪,安至超,何其英,等.高脂饲料喂养时间对2型糖尿病肾病大鼠模型的影响[J].中国实验动物学报,2017,26(1):1-9.
[15]徐叔云.药理实验方法学[M].北京:人民卫生出版社,1982:203.
[16]Abdul MM,Osamu I.Elewa yaser hosny Ali;nakamura teppei;Kon yasuhiro;local CD34-positive capillaries decrease in mouse models of kidney disease associating with the severity of glomerular and tubulointerstitial lesions[J].BMC Nephrol,2017,18(9):280-92.
[17]周丽娜,王玉新,方林,等.内皮祖细胞修复缺血再灌注大鼠肾损伤[J].中国组织工程研究,2014,18(32):5146-51.
[18]毕凌云,杨达胜,赵德安,等.动员自身骨髓干细胞与缺血再灌注肾损伤细胞的凋亡与增殖[J].中国组织工程研究,2013,17(49):8488-97.
[19]刘建辉,孙志平,王琴,等.糖尿病肾病患者血浆EMP与AECA的诊断价值[J].热带医学杂志,2015,15(11):1478-80.
[20]玛依努·玉苏甫.赵红丽.丹参多酚酸盐治疗早期糖尿病肾病的临床价值研究[J].医学综述,2015,21(23):4382-4.
[21]Gupta N,Wish JB.Erythropoietin mimetic peptides and erythropoietin fusion proteins for treating anemia of chronic kidney disease[J].Curr Opin Nephrol Hypertens,2018,27(5):345-50.
[22]Mansour A,Elbaky AA,Kamal S,et al.Evaluation of the procoagulant potential of endothelial microparticles CD144(VECadherin)positive in coronary syndrome patients[J].Egyp JHaematol,2014,39(3):143-8.
[23]Ji X,Wang Z,Geamanu A,et al.Inhibition of cell growth and induction of apoptosis in non-small cell lung cancer cells by deltatocotrienol is associated with notch-1 down-regulation[J].J Cell Biochem,2011,112(10):2773-83.
[24]高峰,刘淑霞,赵玉峰,等.Notch信号通路在糖尿病小鼠肾组织的表达及意义[J].中国组织化学与细胞化学杂志,2014,23(5):445-9.
[25]高家荣,庄星星,魏良兵,等.丹蛭降糖胶囊治疗2型糖尿病血管病变的作用机制研究[J].中药药理与临床,2014,30(6):143-5.
[26]罗云,方朝晖.丹蛭降糖胶囊对糖尿病动脉硬化模型大鼠胸主动脉p22phoxmRNA,p47phox mRNA表达的影响[J].新中医,2018(50):5-8.
[27]杨晓春,鲍陶陶,储全根.基于p38MAPK通路探讨丹蛭降糖胶囊对2型糖尿病模型大鼠血管病变的影响[J].中国实验方剂学杂志,2016,22(3):116-20.
[28]胡云飞,徐国兵.牡丹皮及其主要成分丹皮酚的药理作用研究进展[J].安徽医药,2014,18(4):589-92.
[29]梁裕芬.医用水蛭的生物学特性及药用价值概述[J].生物学教学,2017,42(10):2-4.
[30]Beets K,Huylebroeck D,Moya IM,et al.Robustness in angiogenesis:notch and BMP shaping waves[J].Trends Genet,2013,29(3):140-9.
[31]杜潇,张思琴,程中,等.激活Notch1信号通路对胰腺癌细胞增殖的影响[J].南方医科大学学报,2013,33(10):1494-8.