摘要
目的:观察超声波辐照对培养的血管平滑肌细胞(VSMCs)增殖和凋亡的影响,筛选防止介入治疗后血管再狭窄的辐照参数,并探讨其机理。方法:单声源压电陶瓷超声辐照系统声强连续可调,台盼蓝排斥实验分析辐照后即刻细胞存活率;MTT比色法描述辐照后5天VSMCs的生长动力学变化;BrdU掺入法检测辐照对VSMCs S期DNA复制的影响;ELISA法检测辐照后VSMCs凋亡情况。结果:1.超声辐照对培养的VSMCs有直接杀伤作用,该作用随辐照强度和辐照时间的增加而增加,相同辐照强度和时间下,低频超声该作用更显著。2.超声辐照组较对照组VSMCs生长曲线平缓,0.705MHz组以0.5W/cm2,5min辐照最为明显。3.BrdU掺入法测定S期细胞增殖发现,0.705MHz,0.5W/cm2,5min辐照组BrdU%低于对照组(40.5%±6.8%vs55.8%±3.3%,P<0.05)。4.0.705MHz,0.5W/cm2,5min辐照的凋亡富积因子与阳性对照相比为1.851±0.087vs1.924±0.074,P=0.836。结论:0.705MHz,0.5W/cm2,5min超声辐照在无明显杀伤细胞的情况下诱导培养的VSMCs凋亡,阻止VSMCs S期DNA复制,提示USE是一种潜在的治疗介入治疗后血管再狭窄的手段。
Objective ：To observe the effects of ultrasound exposure （USE） on proliferation and apoto,sis of vuscalur smooth muscle cells （VSMCs） and to screen out the optimmum parameters of USE and （to）discuss its mechanism. Methods：Ultrasound device was made of mono -source of piezoelectric ceramics. Guided by the result of Trypan Blue cell count, the optimum exposure dosage was determined by MTT cell growth curve in 5 days. BrdU incorporation was used to show changes of S phase DNA synthesis after USE. Apoptcsis was detected using ELISA 4 hours later after USE. Results ： USE had direct cytotoxic effect on cultured VSMCs. The effect was intensified as the duration or intensity increased. At the same duration and intensity level, the effect was stronger in lower frequency group. Growth curves in 5 days were flattened in USE groups especially in 0. 705MHz,0.5w/cm^2, 5min exposure. BrdU% resuits indicated that with 0. 705MHz, 0.5W/cm^2, 5min USE, BrdU% against control was 40.5 ＋- 6.8 % vs 55.8 ＋. 3.3 %, P 〈 0.05. Cell ~ath Detection ELISA assay showed apoptosis enrichment factor in 0. 705MHz, 0.5W/cm^2, 5min group comparing with positive control was 1. 851 ± 0. 087 vs 1. 924 ± 0. 074, P =0. 836. Conclusion ： After 0. 705MHz, 0.5W/cm^2, 5min USE, VSMCs apoptosis was induced and S phase I）NA synthesis decreased without obvious immediate cell death, which suggests USE had the potential as an anti - restenosis therapy.
出处
《重庆医科大学学报》
CAS
CSCD
2005年第5期636-639,共4页
Journal of Chongqing Medical University
基金
国家自然科学基金(30040030)
关键词
血管平滑肌细胞
超声
细胞增殖
细胞凋亡
Vascular smooth muscle cells
Ultrasound
Proliferation
Apoptosis
