K他命及其代謝物引起膀胱上皮細胞毒性反應與作用機制之研究
柳瑞明1,2, 林宜君3, 莊恆彰1, 詹勳昊4, 許仁駿5, 林哲偉6
1衛生福利部桃園醫院 外科部 泌尿科;2國立陽明大學醫學系; 3衛生福利部桃園醫院 感染科;4佑民醫療社團法人佑民醫院 泌尿科; 5國防醫學院 生命科學研究所;6中國醫藥大學醫學系生理科
Study of ketamine- and it’s metabolite-induced toxicological effects and mechanisms in bladder epithelial cells
Jui-Ming Liu1,2, Yi-Chun Lin3 , Heng-Chang Chuang1, Hsun-Hao Chan4, Ren-Jun Hsu 5,Jhe-Wei Lin6
1Division of Urology, Department of Surgery Taoyuan General Hospital, Taoyuan, Taiwan
2Faculty of Medicine, National Yang-Ming University, Taipei, Taiwan
3Division of Infection, Department of Internal Medicine Taoyuan General Hospital, Taoyuan, Taiwan
4Division of Urology, Department of Surgery ,Yumin medical corporation Yumin hospital, Nantou, Taiwan
5 Graduate Institute of Life Sciences, National Defense Medical Center
6Physiology of section, Department of medicine, China Medical University, Taichung, Taiwan
Background
Ketamine is a common drug abuse in Juvenile delinquency. In much clinical study, it has been found that the abusers can cause some bladder-related diseases such as ulcers, congestion and disappearance of the epithelium. However, the possible mechanisms in ketamine-induced the bladder epithelial cell toxicity are still unclear. In this study, we aimed to investigate the toxicological effects in ketamine- and it’s metabolite-induced human bladder urinary epithelial cell injuries.
Methods
After human bladder urinary epithelial cells was established, cells were treated with different doses of ketamine/norketamine (the main metabolite of ketamine) for 24 h, and determined the morphological changes using microscopy and cell viability using MTT assay to obtain the level of the lethal dose (LD)50. Then, cells were exposed to LD50 dose in different times, and using Western blotting, Flow cytometry and molecular biology technologies to analysis apoptotic- and autophagic-related signaling molecule/protein changes and the expressions of upstream-regulated pathway.
Results
In this study, the results showed that the exposure to ketamine induced cytotoxicity in bladder urinary epithelial cells, which were accompanied with the production of apoptosis and autophagy events, and the media lethal dose (LD50) was approximately 2 mM. Norketamine (the main metabolite of ketamine) also could induce cytotoxicity as well as ketamine; however, the toxic potency was more than ketamine.
Conclusion
Ketamine and norketamine-induced bladder urinary epithelial cell injuries have a high relevance with apoptosis and autophagy, and norketamine may play an important role in ketamine-caused human bladder damage.