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目的探讨骨髓间质干细胞携带的血红素加氧酶-1(HO-1)基因于体外共培养体系中抑制肺动脉平滑肌细胞增殖的效果及机制。方法采用核转染技术将HO-1质粒转染至骨髓间质干细胞,共培养肺动脉平滑肌细胞和骨髓间质干细胞,评估5-羟色胺刺激后的平滑肌细胞增殖情况,检测5-羟色胺刺激后平滑肌细胞中RhoA的活性。结果 5-羟色胺可刺激肺动脉平滑肌细胞增殖,使单纯培养的平滑肌细胞总数较对照组增加2.40倍(P<0.01)。野生型骨髓间质干细胞与肺动脉平滑肌细胞共培养并不能抑制平滑肌细胞的增殖,而携带HO-1基因的干细胞则可明显抑制5-羟色胺刺激的平滑肌细胞增殖,使平滑肌细胞总数降至对照组的1.56倍(P<0.05);5-羟色胺可引起肺动脉平滑肌细胞中RhoA活性增加2.90倍,野生型干细胞与肺动脉平滑肌细胞共培养并未能改变5-羟色胺刺激的RhoA激活,但在携带HO-1基因的干细胞共培养体系中5-羟色胺诱发的RhoA活性明显降低。结论骨髓间质干细胞携带的HO-1基因可以旁分泌的形式抑制平滑肌细胞的增殖,其作用的机制可能与其产生的CO通过激活cGMP信号通路进而抑制RhoA的激活有关。HO-1可作为外源基因导入体内治疗肺动脉高压。
Objective To investigate the effect and mechanism of heme oxygenase-1 (HO-1) gene in bone marrow mesenchymal stem cells (BMSCs) carrying on proliferation inhibition of pulmonary artery smooth muscle cells in vitro co-culture system. Methods The transfected HO-1 plasmid was transfected into bone marrow mesenchymal stem cells by nuclear transfection. The pulmonary artery smooth muscle cells and bone marrow mesenchymal stem cells were co-cultured to evaluate the proliferation of smooth muscle cells stimulated by 5-HT. The effects of 5-HT on smooth muscle cells In RhoA activity. Results Serotonin stimulated the proliferation of pulmonary artery smooth muscle cells and increased the total number of cultured smooth muscle cells by 2.40-fold (P <0.01) compared with the control group. Co-culture of wild-type BMSCs with pulmonary artery smooth muscle cells could not inhibit the proliferation of smooth muscle cells, while the HO-1-carrying stem cells could significantly inhibit the proliferation of smooth muscle cells stimulated by 5-HT and the total number of smooth muscle cells decreased to the control group 1.56-fold (P <0.05). Serotonin induced a 2.90-fold increase in RhoA activity in pulmonary artery smooth muscle cells. Co-culture of wild type stem cells with pulmonary artery smooth muscle cells did not alter serotonin-stimulated RhoA activation, Serotonin-induced RhoA activity was significantly reduced in the gene-derived stem cell co-culture system. Conclusion The HO-1 gene in bone marrow mesenchymal stem cells can inhibit the proliferation of smooth muscle cells in a paracrine manner. The mechanism may be related to the CO production of BMSCs through the activation of cGMP signaling pathway and the inhibition of RhoA activation. HO-1 can be used as a foreign gene into the body to treat pulmonary hypertension.