香烟尘粒对血管平滑肌细胞增殖的作用

Effects of Dimethylsulfoxide (DMSO)-Soluble Particles from Cigarette Smoke on the Proliferation of Vascular Smooth Muscle Cell

为探讨香烟尘粒对血管平滑肌细胞是否有促增殖作用及其作用机制 ,以兔主动脉平滑肌细胞为实验对象 ,选用细胞存活率 >90 %的小剂量 (1∶10 0 0、2∶10 0 0和 3∶10 0 0 )香烟尘粒 ,分别加入培养基中 ,采用四甲基偶氮唑盐微量酶反应比色法、流式细胞仪分析、细胞蛋白测定和放射免疫法观察香烟尘粒对血管平滑肌细胞增殖及内皮素 1分泌的影响。结果发现 ,与对照组 (不加香烟尘粒组 )相比 ,1∶10 0 0、2∶10 0 0和 3∶10 0 0香烟尘粒组细胞蛋白吸光度值无明显差异 (P >0 .0 5 ) ;1∶10 0 0和 2∶10 0 0香烟尘粒组血管平滑肌细胞增殖明显 (P <0 .0 1) ;流式细胞仪分析结果显示 ,2∶10 0 0香烟尘粒组G1 期细胞数明显减少 (P <0 .0 5 ) ,S期细胞数显著增多 (P <0 .0 1)。 1∶10 0 0、2∶10 0 0和 3∶10 0 0香烟尘粒组培养基中内皮素 1含量均显著增高 ,尤以 2∶10 0 0组增高最明显 (P <0 .0 1)。结果表明 ,1∶10 0 0和 2∶10 0 0香烟尘粒有促进血管平滑肌细胞增殖的作用 ,其促增殖作用可能与内皮素

Aim To ascertain whether Dimethylsulfoxide (DMSO)-soluble particles from cigarette smoke (DSP) stimulate the proliferation of vascular smooth muscle cells (VSMC). Methods Rabbit aortic vascular smooth muscle cells were used as target cell. The proliferation of VSMC was assayed by the measurement of MTT and flow cytometry. Cellular protein assay (ELISA) was used to observe the change of protein content. The amount of endothelin-1 (ET-1) secreted by VSMC was measured by radioimmunoassay (RIA). Results In contrast to the control group (without DSP), protein absorbance of VAMC added 1∶1000, 2∶1000 and 3∶1000 DSP did not make significant difference (P>0.05). VSMC added 1∶1000 and 2∶1000 DSP showed proliferation significantly (P<0.01), and yet 3∶1000 DSP group did not. The amount of ET-1 in the medium of DSP groups were significantly increased, especially in 2∶1000 DSP group (P<0.01) and the result of flow cytometry showed that in 2∶1000 DSP group, the cell numbers of G 1 phase were decreased and cell numbers of S phase were increased markedly (P<0.01). Conclusion 1∶1000 and 2∶1000 DSP can stimulate significantly the proliferation of VSMC, and ET-1 produced by VSMC may play an important role in the proliferation of VSMC.