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目的探讨尾加压素Ⅱ(urotensinⅡ,UⅡ)对乳鼠心肌成纤维细胞(cardiac myofibroblasts,CFs)增殖及胶原合成的影响。方法体外培养CFs作为实验模型,不同浓度UⅡ处理细胞后,通过ELISA法检测各组培养细胞上清中TGF-β1的含量变化,分别利用CKK-8细胞增殖法及Western blot分析UⅡ受体拮抗剂(SB-611812)及PKA特异性阻断剂(KT5720)对UⅡ诱导的CFs增殖及胶原蛋白col-Ⅰ、col-Ⅲ表达的影响。结果 UⅡ10~(-10),10~(-9),10~(-8) mol·L~(-1)处理细胞后,CFs培养上清中TGF-β1的含量及各组CFs的吸光度值与对照组相比明显增加(P<0.05),而在10-7 mol·L~(-1) UⅡ处理细胞后,上述参数与对照组比较无显著差异。1 mol·L~(-1) SB-611812+10-8 mol·L~(-1) UⅡ组和1 mol·L~(-1) KT5720+10~(-8) mol·L~(-1) UⅡ组的TGF-β1的含量及CFs的吸光度值均显著高于对照组(P<0.05),但显著低于10-8 mol·L~(-1) UⅡ组(P<0.05),且两组col-Ⅰ、col-Ⅲ型胶原蛋白表达均低于10~(-8) mol·L~(-1) UⅡ组。结论 UⅡ上调TGF-β1水平促进了CFs的增殖,诱导细胞表达胶原蛋白col-Ⅰ、col-Ⅲ,这一过程可能涉及cAMP-PKA信号转导通路。
Objective To investigate the effects of urotensin Ⅱ (UⅡ) on the proliferation and collagen synthesis of neonatal rat cardiac myofibroblasts (CFs). Methods CFs were cultured in vitro as an experimental model. After the cells were treated with different concentrations of UⅡ, the content of TGF-β1 in the culture supernatant of each group was detected by ELISA. The effects of UⅡ receptor antagonist (SB-611812) and PKA-specific inhibitor (KT5720) on the proliferation of CFs induced by UⅡ and the expression of col-Ⅰ and col-Ⅲ. Results After treatment with UⅡ10 -10, -10 -9 and 10 -8 mol·L -1, the content of TGF-β1 in the supernatant of CFs and the absorbance of CFs (P <0.05). Compared with the control group, the above parameters were not significantly different after 10-7 mol·L -1 UⅡ treatment. 1 mol·L -1 SB-611812 + 10-8 mol·L -1 UⅡgroup and 1 mol·L -1 KT5720 + 10 -8 mol·L -1 U - 1) The content of TGF-β1 and the absorbance of CFs in UⅡ group were significantly higher than those in control group (P <0.05), but significantly lower than those in UMI group (10-8 mol·L -1 UⅡgroup, P <0.05) The expression of col-Ⅰ and col-Ⅲ collagen in the two groups were lower than those in UⅡ group of 10 -8 mol·L -1. Conclusion Upregulation of TGF-β1 by UⅡ can promote the proliferation of CFs and induce the expression of col-Ⅰ and col-Ⅲ in cells. This process may involve the cAMP-PKA signal transduction pathway.