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目的:探讨脉冲高强度聚焦超声(pulsed high intensity focused ultrasound,PHIFU)联合微泡(microbubbles,MBs)以非热效应导致兔肝VX2移植瘤细胞凋亡。方法:建立兔肝VX2移植瘤模型,将36只荷瘤兔随机分为对照组、PHIFU辐照组和PHIFU+MBs辐照组。PHIFU辐照参数为频率0.87 MHz、空间峰值声强(spatial peak intensity,ISP)5900 W/cm2、脉冲重复频率100 Hz、占空比15%、辐照时间90 s,MBs采用0.1 m L/kg Sono Vue。HIFU辐照后1 d处死动物,取部分肿瘤组织采用TUNEL法检测细胞凋亡,免疫组化染色检测PCNA抗原,剩余肿瘤组织进行2,3,5-氯化三苯基四氮唑(2,3,5-triphenyl tetrazolium chloride,TTC)染色观察。结果:对照组、PHIFU组和PHIFU+MBs组经TTC染色后,肉眼可见肿瘤组织被均匀红染,表明各组均无肉眼可见的凝固性坏死形成。TUNEL法检测发现,对照组发现少量阳性染色的肿瘤细胞,PHIFU组和PHIFU+MBs组见较多细胞核棕褐色染色的阳性细胞。PCNA检测结果表明,对照组靶区见大量着色部位在肿瘤细胞核的阳性细胞,而PHIFU组和PHIFU+UCA组阳性染色细胞少。PHIFU辐照组和PHIFU+UCA辐照组凋亡指数(apoptosis index,AI)比对照组高,增殖指数(proliferating index,PI)比对照组低(P=0.000);PHIFU+MBs辐照组AI最高,PI最低(P=0.000)。结论:PHIFU联合MBs可促进兔肝VX2移植瘤细胞凋亡。
Objective: To investigate the non-thermal effects of pulsed high intensity focused ultrasound (PHIFU) combined with microbubbles (MBs) on the apoptosis of VX2 transplanted hepatocytes in rabbits. Methods: Rabbit liver VX2 xenograft models were established. Thirty-six tumor-bearing rabbits were randomly divided into control group, PHIFU irradiation group and PHIFU + MBs irradiation group. The irradiation parameters of PHIFU were 0.87 MHz, spatial peak intensity (ISP) 5900 W / cm2, pulse repetition frequency 100 Hz, duty cycle 15%, irradiation time 90 s and MBs 0.1 m L / kg Sono Vue. Animals were sacrificed on day 1 after irradiation with HIFU. TUNEL method was used to detect the apoptosis of some tumor tissues. PCNA antigen was detected by immunohistochemical staining. The remaining tumor tissues were subjected to 2,3,5-triphenyltetrazolium chloride (2, 3,5-triphenyl tetrazolium chloride (TTC) staining. Results: After TTC staining, the control group, the PHIFU group and the PHIFU + MBs group were stained with naked eye uniformly, which showed that no coagulation necrosis was observed in all groups. TUNEL assay found that a small amount of positive staining tumor cells were found in the control group, and more positive cells were observed in PHIFU group and PHIFU + MBs group. The results of PCNA showed that a large number of positive cells in the nucleus of the tumor were seen in the target area of the control group, but fewer in the PHIFU group and the PHIFU + UCA group. The apoptotic index (AI) in PHIFU irradiation group and PHIFU + UCA irradiation group was higher than that in control group, and the proliferation index (PI) was lower than that in control group (P = 0.000) Highest, lowest PI (P = 0.000). Conclusion: PHIFU combined with MBs can promote the apoptosis of VX2 transplanted tumor in rabbit liver.