Sun Yao, Zhang Gen-Bao, Li Shu, Liu Xiao-Yu, Chen Lei, Bao Peng-Ju
Department of Pathophysiology, Institute of Snake Venom, Wannan Medical College, Wuhu, China.
School of Anesthesiology, Wannan Medical College, Wuhu, China.
J Venom Anim Toxins Incl Trop Dis. 2024 Sep 2;30:e20230099. doi: 10.1590/1678-9199-JVATITD-2023-0099. eCollection 2024.
Acid-sensing ion channel 1a (ASIC1a) plays a critical role in physiological and pathological processes. To further elucidate the biological functions of ASICs and their relationships with disease occurrence and development, it is advantageous to investigate and develop additional regulatory factors for ASICs.
In this study, cation exchange chromatography was used to separate seven chromatographic components from venom. Capillary electrophoresis was employed to detect that Ⅶ peak component containing a main protein Ⅶ-2, which could bind to ASIC1a. The analgesic effects of Ⅶ-2 protein were determined using hot plate methods, and ASIC1a expression in spinal cord tissue from rats with inflammatory pain was detected using western blot.
The purified Ⅶ-2 protein named venom-Ⅶ-2 (NNAV-Ⅶ-2) was obtained by Sephadex G-50 gel filtration, which exhibited a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis with a molecular weight of 6.7 kD. Remarkably, the NNAV-Ⅶ-2 protein demonstrated a significant analgesic effect and downregulated ASIC1a expression in the spinal cord tissue of rats with inflammatory pain.
The analgesic mechanism of the NNAV-Ⅶ-2 protein may be associated with its binding to ASIC1a, consequently downregulating ASIC1a expression in neural tissues.
酸敏感离子通道1a（ASIC1a）在生理和病理过程中发挥着关键作用。为了进一步阐明ASICs的生物学功能及其与疾病发生发展的关系，研究和开发更多ASICs的调节因子具有重要意义。
在本研究中，采用阳离子交换色谱法从毒液中分离出7个色谱组分。采用毛细管电泳检测到Ⅶ峰组分含有一种主要蛋白质Ⅶ-2，其可与ASIC1a结合。采用热板法测定Ⅶ-2蛋白的镇痛作用，采用蛋白质免疫印迹法检测炎性疼痛大鼠脊髓组织中ASIC1a的表达。
通过Sephadex G-50凝胶过滤获得纯化的Ⅶ-2蛋白，命名为毒液-Ⅶ-2（NNAV-Ⅶ-2），其在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳上呈现单一条带，分子量为6.7 kD。值得注意的是，NNAV-Ⅶ-2蛋白在炎性疼痛大鼠脊髓组织中表现出显著的镇痛作用，并下调了ASIC1a的表达。
NNAV-Ⅶ-2蛋白的镇痛机制可能与其与ASIC1a结合有关，从而下调神经组织中ASIC1a的表达。
