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探讨Rho/ROCK通路在5-羟色胺(5-HT)诱导的大鼠肺动脉平滑肌细胞(PASMCs)增殖中的作用以及间尼索地平(m-Nis)对此的影响。组织贴块法原代培养大鼠PASMCs;噻唑蓝(MTT)比色法观察1μmol·L-15-HT作用不同时间对大鼠PASMCs增殖的影响以及m-Nis的干预作用;Western blot及RT-PCR等方法检测5-HT作用下大鼠PASMCs增殖中Rho A、ROCK1 m RNA和p-MYPT1蛋白表达的变化以及m-Nis对此的影响,深入探讨其作用机制。结果显示:1μmol·L-1 5-HT作用12~72 h可明显诱导大鼠PASMCs增殖(P<0.05或P<0.01);m-Nis(1×10-5、1×10-6、1×10-7和1×10-8 mol·L-1)均可明显抑制5-HT诱导的大鼠PASMCs增殖,并呈现一定的浓度依赖性(P<0.05或P<0.01);另外,不同浓度m-Nis还明显抑制了5-HT诱导的Rho A和ROCK1 m RNA的表达以及ROCK的活化(P<0.05或P<0.01)。结果表明Rho/ROCK通路在5-HT诱导的大鼠PASMCs增殖中起重要作用,m-Nis明显抑制了5-HT诱导的大鼠PASMCs增殖,该作用可能与其抑制Rho/ROCK通路有关。
To investigate the role of Rho / ROCK pathway in 5-HT (5-HT) -induced proliferation of rat pulmonary artery smooth muscle cells (PASMCs) and the effect of m-Nis on it. PASMCs were primarily cultured by tissue patch method. The effects of 1μmol·L-15-HT on the proliferation of PASMCs and the intervention of m-Nis were observed by MTT colorimetric assay. Western blot and RT- PCR and other methods to detect 5-HT rat PASMCs proliferation Rho A, ROCK1 m RNA and p-MYPT1 protein expression changes and m-Nis on this effect in-depth discussion of its mechanism. The results showed that: 1μmol·L-1 5-HT could induce the proliferation of PASMCs significantly (P <0.05 or P <0.01) in 12-72 h; m-Nis (1 × 10-5, 1 × 10-6, × 10-7 and 1 × 10-8 mol·L-1) could significantly inhibit the proliferation of PASMCs induced by 5-HT in a concentration-dependent manner (P <0.05 or P <0.01). In addition, Concentration m-Nis also significantly inhibited the expression of Rho A and ROCK1 m RNA induced by 5-HT and the activation of ROCK (P <0.05 or P <0.01). The results showed that Rho / ROCK pathway plays an important role in 5-HT-induced proliferation of rat PASMCs. M-Nis significantly inhibits 5-HT-induced proliferation of PASMCs in rats, which may be related to the inhibition of Rho / ROCK pathway.