高频脉冲电场肾动脉消融可行性探究

Study on the feasibility of renal artery ablation with high-frequency pulsed electric field

目的探究高频脉冲电场消融在肾动脉去神经术中的可行性及安全性。方法使用脉冲电场消融仪在电压500 V和700 V,脉冲个数500,脉冲模式为单脉冲、长脉冲、双向脉冲5-5-5-5、双向脉冲5-3-3-5、双向脉冲7-3-3-7下消融人体神经母细胞瘤细胞SH-SY5Y和大鼠胸大动脉平滑肌细胞A7r5,MTT法检测细胞存活率。4只巴马香猪经皮股动脉穿刺植入电极并消融肾动脉交感神经系统,术后取消融组织制作切片,染色后镜下观察,验证消融效果。结果显微镜下可见A7r5细胞和SH-SY5Y细胞分别在电压500 V和700 V时接近全部凋亡。电压500 V时,SH-SY5Y细胞存活率均高于同类型脉冲下A7r5细胞(P<0.01),A7r5细胞单脉冲与长脉冲细胞存活率差异无统计学意义,但低于双向脉冲5-5-5-5(P<0.05),双向脉冲5-5-5-5、5-3-3-5、7-3-3-7细胞存活率依次升高(P<0.05);电压700 V时,A7r5细胞双向脉冲5-3-3-5消融后细胞存活率高于双向脉冲5-5-5-5(P<0.05),其余脉冲模式下细胞存活率差异无统计学意义。猪肾动脉消融中,双向脉冲5-5-5-5模式下发生轻微抽搐,双向脉冲5-3-3-5、7-3-3-7的消融过程平稳。消融后肾动脉HE染色可见动脉局灶外膜、中膜断裂,管壁纤维化,局部内皮受损,血栓形成。Masson染色可见动脉血管壁纤维化,Warthin-starry银染色可见交感神经组织呈浅褐色,神经细胞死亡。结论高频脉冲电场对平滑肌细胞A7r5和神经细胞SH-SY5Y消融效果良好,高频脉冲电场可消融猪肾动脉交感神经,双向不对称脉冲5-3-3-5可有效避免肌肉收缩,为最优参数组。

Objective To explore the feasibility and safety of high-frequency pulse electric field ablation in renal artery denervation.Methods Human neuroblastoma cell SH-SY5Y and rat thoracic artery smooth muscle cell A7r5 cells were ablated under the voltage of 500 V and 700 V,the number of pulses 500 and pulse mode:single pulse,long pulse,bidirectional pulse 5-5-5-5,bidirectional pulse 5-3-3-5 and bidirectional pulse 7-3-3-7.The cell survival rate was detected by MTT method.Electrodes were implanted by percutaneous femoral artery puncture,and renal artery sympathetic nervous system was ablated in four Bama miniature pigs.Surgical tissue melting sections were removed and observed under the microscope to verify the ablation effect.Results Under the microscope,A7r5 cells and SH-SY5Y cells were nearly completely apoptotic at 500 V and 700 V,respectively.When the voltage was 500 V,the survival rate was higher in SHSY5Y cells than that of A7r5 cells under the same pulse(P<0.01).There was no significant difference in the survival rate of A7r5 cells between single pulse and long pulse,but it was lower than that of bidirectional pulse 5-5-5-5(P<0.05).The survival rate increased gradually in bidirectional pulse 5-5-5,5-3-3-5 and 7-3-3-7(P<0.05).When the voltage was 700 V,the cell survival rate was higher in A7r5 cells after two-way pulse 5-3-3-5 ablation than that of two-way pulse 5-5-5-5(P<0.05).There was no significant difference in cell survival rate between other pulse modes.During the ablation of porcine renal artery,slight convulsions occurred under the bidirectional pulse 5-5-5-5 mode.The ablation process of bidirectional pulse 5-3-3-5 and 7-3-3-7 was stable.After ablation,the staining of renal artery showed focal rupture of adventitia and media,wall fibrosis,local endothelial damage and thrombosis.Masson staining showed arterial wall fibrosis,and silver nitrate staining showed light brown nerve tissue and nerve cell death.Conclusion High frequency pulsed electric field has good ablation effect on smooth muscle cells A7r5 and nerve cells SH-SY5Y.High frequency pulsed electric field can ablate porcine renal artery sympathetic nerve.Bidirectional asymmetric pulse 5-3-3-5 can effectively avoid muscle contraction,which is the optimal parameter array.