雷洛昔芬对人主动脉瓣间质细胞增殖和凋亡的影响

Effect of raloxifene on proliferation and apoptosis of human aortic valve interstitial cells

目的探讨不同浓度的雷洛昔芬(raloxifene,RAL)对体外培养的人主动脉瓣间质细胞(AVICs)增殖和凋亡的影响。方法用Ⅱ型胶原酶裂解主动脉瓣膜,获得AVICs并进行传代培养,使用不同浓度(0 nmol/L、0.1 nmol/L、1 nmol/L、10 nmol/L、100 nmol/L、1 000 nmol/L)的RAL作用于AVICs细胞,其中0 nmol/L RAL组作为对照组,其余为实验组,培养0 d、3 d、5 d、7 d、9 d后,采用细胞增殖与毒性检测试剂盒(MTS)检测不同浓度的RAL对AVICs增殖的影响,收集第7 d的细胞用流式细胞仪检测细胞的周期和凋亡。结果与对照组相比,MTS检测结果表明10 nmol/L RAL和100 nmol/L RAL浓度组的细胞于第5、7、9 d在490 nm波长处的吸光度值(OD值)明显降低(P<0.05),收集第7 d的细胞,流式细胞检测仪检测结果显示,10 nmol/L和100 nmol/L浓度组RAL能明显降低AVICs细胞周期中S期比例(P<0.05)和细胞凋亡率(P<0.05)。结论合适浓度的RAL具有抑制AVICs增殖和凋亡的作用,为进一步研究RAL对主动脉瓣病变的作用奠定了基础。

Objective To investigate the effect of different concentrations ofraloxifene （RAL） on the proliferation and apoptosis of human aortic valve interstitial cells （AVICs） in vitro. Methods AVICs were isolated from human aortic valve by collagenase type II, and cultured in different concentrations （0 nmol/L, 0.1 nmol/L, 1 nmol/L, 10 nmol/L, 100 nmol/L and 1 000 nmol/L） of RAL. AVICs cultured in 0 nmol/L RAL were treated as the control group and those in other concentrations of RAL as the experiment groups. The proliferation and apoptosis of AVICs were evaluated by Cell Proliferation Assay （MTS assay） on day 0, 3, 5, 7 and 9. Flow cytometry was used to detect the cell cycle and apoptosis of AVICs on day 7. Results MTS results showed that the optical density value at 490 nm was much less in 10 nmol/L RAL and 100 nmol/L RAL groups （P 〈 0.05） on day 5, 7 and 9 than that in the control group. Flow cytometry results demonstrated that S-phase rate （P 〈 0.05） and cell apoptosis rate （P 〈 0.05） on day 7 were lower in the 10 nmol/L and 100 nmol/L RAL groups compared with the control group. Conclusion RAL with suitable concentration can inhibit proliferation and apoptosis of AVICs, which will lay an important foundation for further research of the role of RAL on heart valve diseases.