Chongqing University, College of Electrical Engineering, Chongqing 400030, China; Department of Mechanical Engineering, Department of Bioengineering, University of California Berkeley, CA 94720, USA.
Department of Mechanical Engineering, Department of Bioengineering, University of California Berkeley, CA 94720, USA; Department of Endocrinology, Luoyang Central Hospital Affiliated to Zhengzhou University, Luoyang 471009, China.
Bioelectrochemistry. 2019 Feb;125:79-89. doi: 10.1016/j.bioelechem.2018.09.007. Epub 2018 Sep 30.
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.
本研究探讨了一种联合电穿孔与电解（E2）的组织消融模式治疗肝脏后，肝脏的时间生理和分子事件。采用 E2 波形处理大鼠肝脏，分别在 1 h、3 h、6 h 和 24 h 后用 H&E、Masson 三色、TUNEL 染色和 Western blot 进行组织检查。H&E 和 TUNEL 染色显示细胞死亡始于治疗后 3 h，肝细胞再生见于治疗后 24 h。H&E 和 Masson 三色染色显示 E2 后细胞外基质和大腔隙保持完整。Western blot 显示 E2 后的以下分子事件：cleaved caspase 3 在 1 h 时下调，在 24 h 时上调（凋亡）；cleaved Caspase 1 和 cleaved GSDMD 在 6 h 时上调（细胞焦亡），RIP3 在 1 h 时上调，MLKL 在 3 h 时上调（坏死性凋亡）。细胞死亡的机制可能是由坏死性凋亡途径引发的。细胞焦亡途径也被激活。E2 诱导的细胞死亡是程序性坏死，以及关于时间分子途径的详细信息，可能对最近尝试将电穿孔介导的消融与免疫治疗相结合具有价值，因为它表明 E2 诱导的细胞死亡涉及炎症反应，并提供了可用于设计免疫佐剂注射最佳时间的相关数据。
