超声造影剂对血管平滑肌细胞增殖、迁移和凋亡的影响

Effect of echo-contrast agent on the proliferation, migration and apoptosis of vascular smooth muscle cells

目的探讨超声波联合超声造影剂对平滑肌细胞增殖、迁移和凋亡的影响。方法体外培养大鼠胸主动脉血管平滑肌细胞(VSMC)。采用MTT法、Millicell小室、AnnexinV-FITC/PI双标染色和流式细胞仪,检测VSMC的增殖、迁移能力和凋亡,观察血小板衍生生长因子———BB(PDGF-BB)对VSMC增殖、迁移和凋亡的影响,频率1MHz、声强0·3W/cm2的连续波超声联合声学造影剂辐照VSMC后上述指标的变化。结果PDGF-BB对VSMC的凋亡无影响,但可促进VSMC增殖和迁移。超声联合造影剂辐照VSMC30s即可显著抑制PDGF-BB所致的VSMC增殖(P<0·05),同时细胞凋亡比例显著增高(P<0·01),辐照60s可显著抑制PDGF-BB所致的VSMC迁移(P<0·05),随着辐照时间延长,以上作用增强,辐照60s时对VSMC增殖的抑制程度最强。结论低强度超声波辐照联合超声造影剂可抑制VSMC增殖、迁移,并促进细胞凋亡。

Objective To investigate the effect of ultrasound irradiation combinded with echo - contrast agent on proliferation, migration and apoptosis of vascular smooth muscle eells（VSMC）. Methods Rat thoracic aortic VSMC were cultured in vitro. The VSMC were exposed to 1 MHz continuous waves ultrasound for 10 s- 120 s at intensity of 0.3 w/cm^2 with or without lipusome mierobubbles（ 1μl/ml）. VSMC proliferation, migration and apoptosis were observed by MTT assay, millieell chamber apparatus, Annexin V- FITC/PI double labeled staining and flow eytometrie assay respectively. Results Platelet derived growth factor BB（ PDGF- BB） did not stimulate the apoptosis of VSMC, but stimulate VSMC proliferation and migration. Ultrasound sonieation for 30 s and 60 s with microbubbles inhibited VSMC proliferation and migration induced by PDGF - BB （ P 〈 0.05） while the percentage of VSMC apoptosis increased markedly（ P 〈 0.01 ）. As the time prolonged, the extent of these effects increased too, the greatest effect of inhibiting proliferation was attained at 60 s （ P 〈 0.01 ）. Conclusion Ultrasound irradiation eombinded with echo-contrast agent can not only inhibit VSMC proliferation and migration, but also induce cell apoptosis.