补阳还五汤抑制同型胱氨酸介导的血管平滑肌细胞增殖

Buyang Huanwu Tang Inhibits Homocysteine-mediated Smooth Muscle Cell Proliferation

目的:探讨补阳还五汤抑制同型半胱氨酸(homocysteine,Hcy)介导的血管平滑肌细胞增殖及其作用机制。方法:雄性SD大鼠补阳还五汤给药剂量为14.9 g.kg-1.d-1,给药7 d制备含药血清,取适量含药血清配制含药血清为5%,10%,20%3种培养基,取对数期细胞,按照分组将5%,10%,20%含药血清培养基预处理细胞1 h后加入Hcy 2 mmol.L-1作用24 h,流式细胞技术测定细胞周期,Western-blotting(WB)测定增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)的蛋白表达量,用荧光探针DCFH-DA试剂盒对细胞内活性氧(reactive oxygen species,ROS)的聚积量进行测定。结果:补阳还五汤能抑制血管平滑肌细胞(vascular smooth muscle cell,VSMC)细胞周期的进程,其中5%含药血清组细胞处在S+G2期为47.34%(P<0.05),10%含药血清组为38.81%(P<0.01),20%含药血清组为27.95%(P<0.01),下调细胞内PCNA的表达(P<0.01),测得补阳还五汤给药组细胞内ROS的荧光强度较Hcy组弱。结论:补阳还五汤能显著抑制Hcy介导的VSMC增殖,其作用机制可能是通过降低细胞内ROS的聚积量。

Objective： To investigate whether Buyang Huanwu decoction inhibits homocysteine （Hcy） mediated smooth muscle cell proliferation and related mechanisms. Method： Thirty male mice were treated with Buyang Buyang Huanwu decoction （14.9 g .kg-1.d-1） for 7 days to abtain Buyang Huanwu decoction contained serum, concentration of which was as 5%, 10%, 20% and added to smooth muscle ceils. After 1 hour homocysteine was added and cultured for 24 hours. Cell cycle distribution and the expression of proliferating cell nuclear antigen （PCNA） were detected by Western-blotting, DCFH-DA was used for examining the levels of reactive-oxygen species （ROS）. Result： Buyang Huanwu decoction could decrease ROS content, significantly inhibited cell cycle of VSMc, as ：20% doses make 27.95% （P 〈0.01） cells at cell cycle of S ＋ G2; and down regulated the expression of PCNA （P 〈 0.01 ） ; significantly inhibited ROS production induced by Hcy in VSMCs. Conclusion： Buyang Huanwu decoction significantly inhibit Hcy-mediated proliferation of VSMCs, and inhibition of ROS production may be related molecular mechanisms.