高强度聚焦超声经线粒体途径诱导在体子宫肌瘤细胞发生凋亡的机制研究

Research of the mechanism of high intensity focused ultrasound in inducing apoptosis of invivo uterine myoma cell through mitochondrial pathway

目的:研究高强度聚焦超声(HIFU)治疗在体子宫肌瘤后肌瘤组织的细胞形态学、肌瘤细胞凋亡率及肌瘤组织中凋亡蛋白表达,从细胞凋亡角度探讨HIFU治疗在体子宫肌瘤有效性的机制。方法:取HIFU治疗后靶区的子宫肌瘤组织、靶区周边肌瘤组织和未治疗组的肌瘤组织标本。光镜下和电镜下观察3组标本的细胞形态学,Annexin V-FITC/PI流式细胞学检测细胞凋亡率,Western blot法检测3组肌瘤组织中Bax、Bcl-2和Caspase-3蛋白表达。结果:靶区组织出现凝固性坏死,靶区周边组织见明显凋亡征象,未治疗组未出现病理改变。靶区周边组肌瘤细胞的凋亡率较靶区组和未治疗增加,差异有统计学意义(P<0.05)。靶区周边肌瘤组织中Bax和Caspase-3蛋白表达量较靶区组和未治疗组增加,差异有统计学意义(P<0.05)。未治疗组肌瘤组织中Bcl-2蛋白表达量较靶区组和靶区周边组增加,但差异无统计学意义(P>0.05)。结论:HIFU消融在体子宫肌瘤后靶区组织出现了凝固性坏死,同时诱导靶区周边肌瘤细胞的凋亡,其机制可能是上调Bax和Caspase-3蛋白高表达,通过线粒体介导的内源性信号通路启动细胞的凋亡。

Objective： To observe and detect the cell morphology changes,the alterations in apoptosis rate of myoma cell and the differences in apoptin expression of invivo uterine myoma treated by High Intensity Focused Ultrasound（ HIFU）. To explore the effective mechanism of HIFU from the angulation of apoptosis.Methods： Three specimens were taken from the ablation group,the surrounding of the ablation group and no treatment group.To observe the cell morphology changes of each group under optical microscopy and electron microscope. Annexin V-FITC/PI double staining flow cytometry was performed to detect.To compare the alterations in apoptosis rate of myoma cell among the ablation group,the surrounding of the ablation group and no treatment group.Western blot was used to detect the differences in protein expressions of Bax,Bcl-2 and Caspase-3 among the ablation group,the surrounding of the ablation group and no treatment group. Results： Coagulative necrosis were detected in ablation group,apoptosis signs were seen in the surrounding of the ablation group,no abnormal changes occurred in no treatment group.Compared with the ablation group and no treatment group,the apoptosis rate of myoma cell in the surrounding of ablation group was increased,the difference between groups was statistically significant（ P 0. 05）. Compared with the ablation group and no treatment group,the protein expressions of bax and caspase-3 in the surrounding of ablation group was increased,the difference was statistically significant（ P0.05）.The protein expression of Bcl-2 in no treatment group was increased compared to ablation group and the surrounding of ablation group,but the difference between groups was no statistically significant（ P0.05）.Conclusion：After the invivo myoma was ablated by HIFU,coagulative necrosis was caused in the ablation group and apoptosis of myoma cell was induced in the surrounding of ablation group.The possible mechanism may be high expression of Bax and caspase-3 protein was induced in the surrounding of ablation group which mediated apoptosis of myoma cell through the mitochondria pathway.