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目的探讨全反式维甲酸(atRA)对糖尿病大鼠肾脏组织骨形成蛋白-7(BMP7)、转化生长因子-β1(TGFβ1)表达和细胞外基质(ECM)Ⅳ、Ⅲ型胶原代谢的影响。方法随机选择24只♂SD大鼠制作糖尿病模型,其中12只入糖尿病模型(DM)组,另12只入糖尿病模型+atRA(DM+atRA)组,此外再随机选择12只正常大鼠作为空白对照(NC)组。分别于第8、12周末各组处死6只大鼠,通过光镜、电镜观察各组大鼠肾脏病理结果;通过免疫组织化学方法观察BMP7,TGFβ1,Ⅳ、Ⅲ型胶原表达,并进行半定量分析;通过实时荧光定量PCR方法对TGFβ1mRNA、BMP7mRNA进行相对定量分析。结果 DM+atRA组大鼠肾组织BMP7表达较同时期DM组增加(P<0.01),主要表达于大鼠肾小管上皮细胞,而TGFβ1表达较同时期DM组减少(P<0.01);且DM+atRA组大鼠肾组织ECM重要成分Ⅳ、Ⅲ型胶原表达与DM组相比较显著减少(P<0.01)。同时观察到BMP7蛋白与Ⅳ型胶原蛋白表达呈负相关(r=-0.75,P<0.01),与Ⅲ型胶原蛋白表达呈负相关(r=-0.71,P<0.01),与TGFβ1蛋白表达呈负相关(r=-0.86,P<0.01)。荧光定量PCR结果表明,DM+atRA组BMP7mRNA水平升高,TGFβ1mRNA水平下降,且两者呈显著负相关(r=-0.83,P<0.01)。结论 atRA能减轻糖尿病大鼠肾脏病理损伤,延缓肾脏纤维化,这一防治作用可能与其增加BMP7表达,下调TGFβ1表达从而减少肾组织ECM沉积有关。
Objective To investigate the effects of all-trans retinoic acid (atRA) on the expression of BMP7, TGFβ1 and extracellular matrix (ECM) type Ⅳ and type Ⅲ collagen in diabetic rats Metabolic effects. Methods Twenty-four male SD rats were randomly divided into diabetic model group (DM), 12 diabetic model group (at + DM + atRA group) and 12 normal rats Control (NC) group. At the end of the 8th and 12th week, 6 rats in each group were sacrificed. The pathological results of kidney in each group were observed by light microscope and electron microscope. The expressions of BMP7, TGFβ1, Ⅳ and Ⅲ collagen were observed by immunohistochemistry. And semi-quantitative analysis; by quantitative real-time PCR method TGF 汕 1 mRNA, BMP 7mRNA relative quantitative analysis. Results The expression of BMP7 in DM + atRA group was higher than that in DM group (P <0.01), mainly in rat renal tubular epithelial cells, while the expression of TGFβ1 in DM + atRA group was lower than that in DM group ). Compared with DM group, the expressions of collagen type Ⅳ and collagen Ⅲ, an important component of ECM in the kidney of DM + atRA group were significantly decreased (P <0.01). At the same time, BMP7 protein was negatively correlated with type IV collagen expression (r = -0.75, P <0.01) and negatively correlated with type III collagen protein (r = -0.71, P <0.01) β1 protein expression was negatively correlated (r = -0.86, P <0.01). Fluorescent quantitative PCR results showed that the level of BMP7 mRNA and the level of TGFβ1 mRNA in DM + atRA group were significantly decreased (r = -0.83, P <0.01). Conclusions atRA can reduce renal pathological injury in diabetic rats and delay renal fibrosis. This preventive effect may be related to increasing BMP7 expression, decreasing TGFβ1 expression and decreasing ECM deposition in renal tissues.