自噬抑制剂3-甲基腺嘌呤对肝肺综合征大鼠肺动脉平滑肌细胞表型转化及增殖的影响

Effect of autophagy inhibitor 3-methyladenine on phenotype transformation and proliferation of pulmonary arterial smooth muscle cells

目的:探讨3-MA对PASMCs表型转化及增殖的影响。方法 :体外培养SD大鼠PASMCs,设立空白对照组和低(2.0 mmol/L)、中(4.0 mmol/L)及高剂量组(8.0 mmol/L)。Western blot法检测不同浓度3-MA对PASMCs中自噬水平标志蛋白LC3Ⅱ、OPN以及Vimentin表达水平的影响;MTT法检测3-MA对PASMCs增殖的影响。结果:不同剂量3-MA作用于PASMCs后,细胞自噬水平和增殖能力降低,呈剂量依赖性;LC3Ⅱ、OPN以及Vimentin相对表达量均明显低于对照组(P<0.05)。结论:3-MA可以下调大鼠PASMCs的LC3Ⅱ表达,抑制PASMCs合成表型标志蛋白OPN、Vimentin的表达,显著降低PASMCs增殖能力。

Objective To evaluate the effect of autophagy inhibitor 3-methyladenine （3-MA） on phenotype transformation and proliferation of rat pulmonary arterial smooth muscle cells （PASMCs）. Methods Cultured PASMCs were treated with different concentrations of 3-MA （low-dose group, 2.0 mmol/L; middle-dose group, 4.0 mmol/L; high-dose group, 8.0 mmol/L; control group, 0 mmol/L ）. The protein expression of LC3 I, OPN and Vimentin was detected by Western blotting. Cell proliferation was detected by MTT assay. Results The autophagy of PASMCs was decreased with the increase of the concentration of 3-MA. Compared with the control group, significantly down-regulated protein expression of LC3 Ⅱ, OPN and Vimentin was observed in 3-MA-treated cells, with significantly lower proliferation activity. Conclusion The autophagy inhibitor 3-MA significantly down-regulated the expression of LC31], OPN and Vimentin in PASMCs, with inhibiting the proliferation of PASMCs.