间充质干细胞对脂多糖刺激下肺微血管内皮细胞增殖及周期的影响

Influence of MSCs on cell proliferation and cell cycle in lipopolysaccharide-stimulated pulmonary microvascular endotcells

目的探讨骨髓间充质干细胞(BM-MSC)对脂多糖(LPS)刺激下肺微血管内皮细胞(PVEC增殖及细胞周期的影响。方法实验分组:A组(正常PVEC);B组(PVEC+LPS),PVEC仅予LPS刺激;C组(PVEC/BM-MSC+LPS),将BM-MSC与PVEC共培养于Transwell体系过夜后予LPS刺激;D组(PVEC/BMMSC+PBS),同上BM-MSC与PVEC共培养后,用PBS替代LPS。用细胞计数试剂盒-8(CCK-8)法测PVEC活性、流式细胞仪测PVEC周期及qRT-PCR测细胞周期蛋白D1(CyclinD1)、细胞周期蛋白依赖激酶4(CDK4)及P27 mRNA水平。结果与A组相比,B组中PVEC增殖受抑,出现周期阻滞,且CyclinD1及CDK4受抑,P27上调(P <0.05),与BM-MSC共培养可逆转上述效应(P <0.05)。无LPS情况下,BM-MSC共培养对PVEC的增殖及细胞周期均有正性调控作用(P <0.05)。结论 BM-MSC对PVEC有保护作用,促进PVEC增殖、抵制LPS诱导的周期阻滞及调控细胞周期相关蛋白表达是可能的分子机制。

Objective To explore the influence of bone marrow derived mesenchymal stem cell(BM-MSC) on cell proliferation and cell cycle of pulmonary microvascular endothelial cells(PVECs) following lipopolysaccharide(LPS) stimulation. Methods PVECs were divided into group A(PVEC), group B(PVEC + LPS), group C( PVEC/BM-MSC + LPS) and group D( PVEC/BM-MSC + PBS). After stimulation with 0.1 mg/mL of LPS, the cell viability of PVEC was determined by cell counting kit-8(CCK-8) assay;the cell cycle of PVEC was detected by flow cytometry and the mRNA expression of CyclinD1,cyclin-dependent kinase 4(CDK4) and P27 were analyzed by qRT-PCR. Results Compared with normal PVECs,LPS stimulation caused inhibition of cell proliferation and cell cycle,suppressed the mRNA expression of CyclinD1 and CDK4,but enhanced the mRNA expression of P27 in PVECs(P < 0.05). Coculture with BM-MSCs attenuated all of these detrimental effects of LPS stimulation on PVECs(P < 0.05). Moreover,coculture with BM-MSCs had a positive effect on cell proliferation and cell cycle of PVECs without LPS stimulation(P < 0.05). Conclusion Coculture with BM-MSC has a protective effect on LPS induced injury in PVECs. The detailed mechanism might be associated with promotive proliferation, inhibition of cell cycle arrest and regulation of CyclinD1,CDK4 and P27 expression in LPS stimulated PVECs.