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目的:探讨高糖刺激、吡格列酮(Piog)孵育对心肌成纤维细胞(CFs)胶原生成的影响及其作用机制。方法:培养1～3dSD乳鼠CFs,分为4组:Ⅰ组对照组,Ⅱ组吡格列酮(Piog-10μmol/L)干预组,Ⅲ组高糖培养组(Glu25mmol/L),Ⅳ组高糖+Piog(10μmol/L)共刺激组。分别测定各组细胞Ⅰ、Ⅲ型胶原mRNA表达;促纤维化因子TGFβ1和CTGF mRNA的表达;血管紧张素Ⅱ-1型受体(AT1-R)mRNA及蛋白的表达。结果:与Ⅰ组相比,Ⅲ组Ⅰ、Ⅲ型胶原mRNA表达明显上升;TGFβ1和CTGF mRNA的表达,AT1-R mRNA及蛋白表达显著上调(P<0.05)。Ⅳ组Piog干预后与Ⅲ组比较,Ⅰ、Ⅲ型胶原mRNA、TGFβ1 mRNA表达明显下降,AT1-R mRNA及蛋白表达显著下降(P<0.05);CTGF mRNA表达虽有下降,但差异无统计学意义。结论:高糖刺激CFs可导致Ⅰ、Ⅲ胶原合成增加,Piog干预可下调TGFβ1表达,以及AT1-R的表达、降低肾素-血管紧张素醛固酮系统活性,抑制心肌纤维化过程。
Objective: To investigate the effect of high glucose and Piog incubation on the collagen production of cardiac fibroblasts (CFs) and its mechanism. Methods: CFs of 1 ~ 3d SD neonatal rats were cultured and divided into 4 groups: control group Ⅰ, group Ⅱ Piog-10μmol / L intervention group, group Ⅲ glucose 25mmol / L, group Ⅳ high glucose + Piog (10μmol / L) costimulation group. The mRNA expression of type I and type III collagen, the expression of TGFβ1 and CTGF mRNA and the expression of AT1-R mRNA and protein were determined. Results: Compared with group Ⅰ, the mRNA expression of type Ⅰ and type Ⅲ collagen in group Ⅲ increased significantly. The expression of TGFβ1 and CTGF mRNA and the expression of AT1 - R mRNA and protein in group Ⅲ were significantly increased (P <0.05). Compared with group Ⅲ, the expressions of mRNA of type Ⅰ and type Ⅲ collagen and TGFβ1 decreased significantly (P <0.05), while the expression of AT1-R mRNA and protein decreased significantly in group Ⅳ (P <0.05), but the difference was not statistically significant significance. CONCLUSION: CFs stimulated by high glucose can lead to the increase of collagen Ⅰ and collagen Ⅲ synthesis. Piog intervention can down-regulate the expression of TGFβ1 and AT1-R, decrease the activity of renin-angiotensin aldosterone system and inhibit the process of myocardial fibrosis.