清脑益智法方药对烟雾病脑血管壁细胞的影响

Influence of Qingnao Yizhi Prescription (QNYZ) on cerebrovascular cells in moyamoya disease

关键词:  烟雾病  平滑肌  内皮  细胞  清脑益智方药

Key words:  moymoya disease  smooth muscle  endothelial cell  Qingnao Yizhi Prescription (QNYZ)

摘要:

[目的]观察中药清脑益智方(QNYZ)对烟雾病脑血管平滑肌细胞增殖、蛋白质合成等的影响,探讨中医药治疗烟雾病的机制。[方法]使用烟雾病患者脑血管平滑肌细胞株和人血管内皮细胞,应用细胞计数器和5-溴脱氧尿嘧啶核苷(BrdU)掺入记数观察。[结果]QNYZ在低浓度(100～200μg/mL)时仅部分抑制了血管平滑肌细胞(HCSMC)和脑血管平滑肌细胞(HMSMC)的增殖,当QNYZ的浓度增加到500μg/mL以上(500～5000μg/mL),显示出了一个完全抑制HCSMC和HMSMC增殖的作用(P<0.01);对血管壁内皮细胞在低浓度(250μg/mL)及高浓度(500、1000μg/mL)时,对其增殖均无显著影响(P>0.05)。对平滑肌细胞内DNA合成的影响上,QNYZ组的BrdU掺入率低下(P<0.01),其中在HCSMC组,不同浓度(500、1000μg/mL)的QNYZ组间BrdU掺入率无显著差异(P>0.05);而在HMSMC组,1000μg/mL浓度的QNYZ较500μg/mL的QNYZ可以显著降低BrdU掺入率,显示出一定的量效关系。[结论]QNYZ 对内皮细胞(EC)的生长无显著影响, 同时又抑制了平滑肌细胞(SMC)的增殖,起到了保护内膜完整性,维护内膜屏障的作用, 从而可能在预防烟雾病脑血管事件中发挥一定的作用。

Abstract:

[Objective] Moyamoya disease is an infrequent cerebrovascular disease with hereditary tendency and predilection in children. The influence of QNYZ on the proliferation and protein synthesis of cerebrovascular smooth muscle cell of in this disease was observed to study its therapeutic mechanism. [Methods] Human moyamoya smooth muscle cell (HMSMC) strain and human control smooth muscle cell (HCSMC) were used. The cell counter and BrdU incorporation were also chosen. [Result] QNYZ could partially inhibit the proliferation of HCSMC and HMSMC at low concentration (100～200 μg/mL) and show a complete control at high concentration (500～1 000 μg/mL) (P<0.01). It had no marked influence on the proliferation of endothelial cell at each concentration (P>0.05). For DNA synthesis in smooth muscle cell (SMC), BrdU incorporation rate was low (P<0.01) in QNYZ Group. In HCSMC Group, BrdU incorporation rate had no obvious difference between QNYZ Groups with different concentration (P>0.05). In HMSMC Group, QNYZ with the concentration of 1 000 μg/mL could more markedly decrease BrdU incorporation than that of QNYZ at 500 μg/mL with a dose-effect relationship.[Conclusion] QNYZ has no obvious influence on endothelial cell but may inhibit the proliferation of SMC.It can prevent the occurrence of cerebrovascular accident due to moyamoya disease by keeping the completion of tunica intima and maintaining the barrier effect.