ABSTRACT

Background Ketamine-related cystitis (KC) has been researched in many clinical studies, but its exact mechanism is ambiguous and needs further research. Methods We established a KC rat model and analyzed physiological, biochemical, and urodynamic parameters of ketamine (KET)-related bladder injury. Bladder histologic feature, reactive oxygen species (ROS), autophagy-, apoptosis-, and endoplasmic reticulum stress (ERS)-related markers were examined by hematoxylin and eosin staining, Masson staining, ROS kit, quantitative real-time polymerase chain reaction, and western blot. In vitro, effects of 0.01, 0.1, and 1 mM KET on cell vitality, apoptosis, ROS level, autophagy-, apoptosis-, and ERS-related markers were examined again. Effects of KET-1 and salubrinal on complex formation, autophagy-, apoptosis-, and ERS-related markers were examined by Co-Immunoprecipitation and western blot. After transfection with shIRE1, complex formation, cell biological behaviors, ROS level, autophagy-, apoptosis-, and ERS-related markers were examined again. Results KET induced bladder hyperactivity and injury. KET facilitated urinary frequency, ROS production, and induced bladder histologic injury by activating autophagy-, apoptosis-, and ERS-related markers in rats. In vitro, KET (0.01, 0.1, and 1 mM) restrained cell vitality and elevated apoptosis and ROS level via activating autophagy-, apoptosis-, and ERS-related markers. Moreover, salubrinal reversed the promotion of KET-1 on complex formation, autophagy-, apoptosis-, and ERS-related marker expressions. After transfection with shIRE1, shIRE1 weakened complex formation induced by KET-1, and the effects of KET-1 on cells were offset by shIRE1. Conclusion KET enhanced autophagy and ERS in vivo and in vitro via restraining IRE1-TRAF2-ASK1-JNK pathway.

摘要

背景氯胺酮相关性膀胱炎（KC）已在许多临床研究中进行了研究，但其确切机制尚不明确，需要进一步研究。方法建立KC大鼠模型，分析氯胺酮（KET）相关膀胱损伤的生理、生化和尿动力学参数。通过苏木精和伊红染色、Masson 染色、ROS 试剂盒、定量实时聚合酶链反应和蛋白质印迹。体外，再次检查了 0.01、0.1 和 1 mM KET 对细胞活力、细胞凋亡、ROS 水平、自噬、细胞凋亡和 ERS ​​相关标志物的影响。通过免疫共沉淀和蛋白质印迹检查 KET-1 和 salubrinal 对复合物形成、自噬、凋亡和 ERS ​​相关标志物的影响。转染 shIRE1 后，再次检查复合物形成、细胞生物学行为、ROS 水平、自噬、凋亡和 ERS ​​相关标志物。结果 KET 诱发膀胱过度活动和损伤。KET 通过激活大鼠的自噬、凋亡和 ERS ​​相关标志物促进尿频、ROS 产生和诱导膀胱组织学损伤。体外、KET（0.01、0.1 和 1 mM）通过激活自噬、细胞凋亡和 ERS ​​相关标志物抑制细胞活力并提高细胞凋亡和 ROS 水平。此外，salubrinal 逆转了 KET-1 对复合物形成、自噬、凋亡和 ERS ​​相关标志物表达的促进作用。转染 shIRE1 后，shIRE1 减弱了 KET-1 诱导的复合物形成，并且 KET-1 对细胞的影响被 shIRE1 抵消。结论 KET通过抑制IRE1-TRAF2-ASK1-JNK通路在体内外增强自噬和ERS。