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为支架磁感应热疗预防和治疗PTCA术后血管再狭窄提供基础研究数据,对临床常用的316L型不锈钢冠脉支架进行磁感应诱导升温,探讨加热对兔血管平滑肌细胞(VSMCs)的增殖、迁移、凋亡及周期的影响。将平滑肌细胞置于恒温水浴槽中,待其达到预定温度(43、47℃)后持续10min,观察细胞形态变化,采用MTT法检测加热对血管平滑肌细胞增殖活性的影响,流式细胞术检测VSMC细胞周期和细胞凋亡,划痕实验检测加热对细胞迁移能力的影响,免疫细胞化学法检测不同温度作用后血管平滑肌细胞中增殖细胞核抗原(PCNA)的表达。结果发现,支架在交变磁场下可以升温至热疗所需温度,加热后细胞增殖受到了显著抑制,43℃组细胞存活率为(83.23±2.87)%,47℃组细胞存活率仅为(37.58±0.78)%。细胞的凋亡率显著增加,47℃组细胞凋亡率为(87.37±2.95)%,与对照组相比具有极显著差异,而43℃组的凋亡率为(6.00±0.26)%,与对照组相比无统计学差异。细胞周期也受到抑制,被阻滞在S期。加热后随着时间的推移,47℃组细胞的迁移能力受到显著抑制,而加热对43℃组细胞的迁移能力无显著影响。免疫细胞化学检测显示,随着温度的升高,PCNA的表达受到明显抑制。研究表明,临床常用冠脉支架在交变磁场下升温可行。加热显著抑制了细胞的增殖并促进了细胞凋亡,使细胞周期阻滞在S期,且抑制了细胞的迁移。加热能显著抑制细胞PCNA的表达,这些可能是加热抑制细胞增殖和迁移、促进凋亡的机制之一。
To provide basic data for the prevention and treatment of vascular restenosis after PTCA with magnetic induction hyperthermia, and to induce induction of heating on the commonly used 316L stainless steel coronary stent. To investigate the effect of heating on the proliferation and migration of rabbit vascular smooth muscle cells (VSMCs) Death and the impact of the cycle. The smooth muscle cells were placed in a thermostatic water bath until the desired temperature (43, 47 ℃) for 10min to observe the morphological changes of cells. The effects of heat on the proliferation of vascular smooth muscle cells were detected by MTT assay. Flow cytometry Cell cycle and apoptosis. The scratch test was used to detect the effect of heating on cell migration. The expression of proliferating cell nuclear antigen (PCNA) in vascular smooth muscle cells was detected by immunocytochemistry. The results showed that the stent could be heated to the required temperature for hyperthermia under alternating magnetic field and the cell proliferation was significantly inhibited after heating. The survival rate of the cells at 43 ℃ was (83.23 ± 2.87)%, the survival rate at 47 ℃ was only 37.58 ± 0.78)%. (87.37 ± 2.95)% at 47 ℃, which was significantly different from that of the control group (6.00 ± 0.26)%, while the apoptosis rate at 43 ℃ was (6.00 ± 0.26)% and No significant difference between the control group. Cell cycle is also inhibited, was blocked in the S phase. With the passage of time, the migration ability of cells at 47 ℃ was significantly inhibited, while heating had no significant effect on the migration ability of cells at 43 ℃. Immunocytochemistry showed that with the increase of temperature, the expression of PCNA was significantly inhibited. Studies have shown that clinically common coronary stents in the alternating magnetic field heating feasible. Heating significantly inhibited cell proliferation and promoted cell apoptosis, arresting cell cycle in S phase and inhibiting cell migration. Heating can significantly inhibit the expression of PCNA, which may be one of the mechanisms of heating to inhibit cell proliferation and migration and promote apoptosis.