This study examined the temporal physiological and molecular events following the treatment of the liver with a tissue ablation modality that combined electroporation with electrolysis (E2). Rat liver was treated with an E2 waveform and the tissue examined, 1 h, 3 h, 6 h and 24 h with: H&E, Masson Trichrome, TUNEL stains and Western blot. H&E and TUNEL stains have shown that cell death began to be evident 3 h and hepatocyte regeneration was seen 24 h after treatment. H&E and Masson trichrome have shown that the extracellular matrix and the large lumens, appeared intact after E2. Western blot has shown the following molecular events after E2: cleaved caspase 3–downgraded at 1 h, upgraded at 24 h (apoptosis); cleaved Caspase 1 and cleaved GSDMD–upgraded at 6 h (pyroptosis), RIP3–upgraded at 1 h, MLKL–upgraded at 3 h (necroptosis). The mechanism of cell death was possible initiated by necroptosis pathway. Pyroptosis pathway was also activated. The observation that cell death from E2 was by programed necrosis and the details on the temporal molecular pathways, may have value for the recent attempt to combine electroporation mediated ablation with immunological treatment, by demonstrating that the cell death from E2 involves an inflammatory response and by providing data that could be used to design the optimal timing for the injection of immunological adjuvants.

这项研究检查了组织消融方式结合电穿孔与电解（E2）对肝脏进行治疗后的时间生理和分子事件。用E2波形处理大鼠肝脏，并在1小时，3小时，6小时和24小时使用H＆E，Masson Trichrome，TUNEL染色和Western blot检测组织。H＆E和TUNEL染色表明，治疗后3小时开始出现细胞死亡，并且在24小时后观察到肝细胞再生。H＆E和Masson trichrome已表明，E2后，细胞外基质和大管腔完整无缺。Western印迹显示E2发生以下分子事件：裂解的半胱天冬酶3在1 h降级，在24 h升高（凋亡）。切割Caspase 1并切割GSDMD-在6 h升高（焦化），RIP3-在1 h升高，MLKL-在3 h升高（坏死）。细胞死亡的机制可能是由坏死病途径引发的。细胞凋亡途径也被激活。观察到E2引起的细胞死亡是由程序性坏死引起的，以及有关时间分子途径的细节，对于证明电穿孔介导的消融与免疫治疗相结合的最新尝试可能具有价值，这表明E2引起的细胞死亡涉及炎性反应，并且通过提供可用于设计注射免疫佐剂的最佳时机的数据。