摘要
目的探讨血管内皮生长因子(VEGF)、基质金属蛋白酶9(MMP-9)及金属蛋白酶1组织抑制剂(TIMP-1)在糖尿病肾病(DN)发病中的作用以及钙离子拮抗剂硝苯地平对肾脏保护作用的机制。方法大鼠随机分为3组:健康对照(N)组、DN组和硝苯地平治疗(T)组,每组10只。采用链脲菌素诱发DN大鼠模型,治疗组以硝苯地平控释片15mg·kg^-1·d^-1干预治疗,治疗第2周和4周,检测大鼠平均动脉压(MAP)、尿蛋白量(24h)、Scr水平;用免疫组化和Western印迹方法检测大鼠肾脏VEGF、MMP-9、TIMP-1的蛋白表达水平;RT-PCR法检测VEGF、MMP-9、TIMP-1mRNA的表达。结果与N组比较,DN组大鼠尿蛋白量、Scr水平和MAP均显著上升[分别为(78.87±17.62)比(6.30±1.91)mg/24h、(203.88±18.08)比(84.98±10.09)μmol/L、(141.1±8.7)比(99.8±3.8)mmHg,P均〈0.05];肾组织VEGF、MMP-9、TIMP-1 mRNA和蛋白表达增强(P〈0.05),且MMP-9/TIMP-1比值明显下降(P〈0.05)。与DN组比较,T组尿蛋白量(24h)[(58.35±10.48)mg]、Scr水平[(165.81±15.86)μmol/L]和MAP[(102.6±4.4)mmHg]均显著下降(P均〈0.05);肾组织VEGF表达下调(P〈0.05),MMP-9、TIMP-1mRNA和其蛋白表达水平显著增高(分别P〈0.05和P〈0.01),MMP-9/TIMP-1比值显著升高(P〈0.05)。DN大鼠肾组织VEGF mRNA水平与尿蛋白量(24h)呈正相关(r=0.748,P〈0.05),与MMP-9/TIMP-1的比值呈负相关(r=-0.711,P〈0.05)。结论VEGF、MMP-9和TIMP-1可能参与了DN的发病过程,硝苯地平可能通过影响VEGF、MMP-9及TIMP-1的表达而实现对肾脏的保护作用。
Objective To investigate the role of vascular endothelial growth factor (VEGF), matrix metalloproteinase-9(MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in the progress of diabetic nephropathy(DN), and to assess the effect of nifedipine (an calcium-channel blocker) on the expression of VEGF, MMP-9 and TIMP-1, thus to elucidate the renal protection mechanism of nifedipine against DN. Methods Thirty male Wistar rats were randomly divided into three groups: normal control( N group ), diabetic nephropathy group( DN group ) and treatment group with nifedipine(T group, 15 mg·kg^-1·d^-1). At day 14 and day 28 with nifedipine, five rats of each group were sacrificed to test glomerular hypertrophy, renal function, mean artery pressure (MAP) and 24 h urinary protein excretion. The expression levels of VEGF, MMP-9 and TIMP-1 were examined by immunohistochemical technique, RT-PCR and Western blot. Results Compared with N group, 24 h urinary protein excretion, Scr and MAP increased significantly in DN group[(78.87±17.62) vs (6.30±1.91) mg/24 h,(203.88±18.08) vs (84.98±10.09) μmol/L, (141.1±8.7) vs (99.8±3.8) mm Hg, respectively, all P〈0.05], meanwhile, mRNA and protein expression levels of VEGF, MMP-9 and TIMP-1 increased significantly in DN group (P〈0.05), but the ratio of MMP-9/TIMP-1 decreased (P〈0.05 or P〈0.01). Compared with DN group, after 4 weeks with nifedipine, 24 h urinary protein excretion[( 58.35± 10.48 ) mg/24 h], Scr[ ( 165.81 ± 15.86 ) μmol/L] and MAP [(102.6 ±4.4)mm Hg] decreased significantly, VEGF expression down-regulated, the protein expression of MMP-9 and TIMP-1 up-regulated and MMP-9/TIMP-1 ratio increased (P〈 0.05 or P〈0.01) in T group. Conclusions VEGF, MMP-9 and TIMP-1 may play an important role in the progression of diabetic nephropathy. Nifedipine can suppress the expression of VEGF and increase the ratio of MMP-9/TIMP-1, thus preventing the progression of diabetic nephropathy.
出处
《中华肾脏病杂志》
CAS
CSCD
北大核心
2007年第8期534-539,共6页
Chinese Journal of Nephrology
基金
国家自然科学基金(30370663)
关键词
糖尿病肾病
血管内皮生长因子类
基质金属蛋白酶类
金属蛋白酶1组织抑制剂
钙通道阻滞药
Diabetic nephropathy
Vascular endothelial growth factors
Matrix metalloproteinases
Tissue inhibitor of metalloproteinase-1
Calcium channel blockers