K他命膀胱炎自嗜作用經由粒線體以及內質網的分子機轉
阮雍順1,2,3,4、 李懿倫4,5、吳文正1,2,3、 張美玉3、 莊淑棉6
高雄醫學大學 泌尿科1，醫學研究所2，轉譯醫學中心6高雄醫學大學; 高雄市立小港醫院 泌尿科3；高雄醫學大學附設中和紀念醫院 泌尿科4；； 衛生署立新營醫院 外科部 泌尿科5；
The molecular mechanism of autophagy via mitochondrial and endoplasmic reticulum pathway in ketamine-induced ulcerative cystitis in animal model
Yung-Shun Juan, Yi-Lun Lee, Wen-Jeng Wu, Mei-Yu Jang, Shu-Mien Chuang4*,
1Department of Urology, 4Graduate Institute of Medical Science, 6Translational Research Center, Cancer Center, Kaohsiung Medical University, Kaohsiung, Taiwan; 2Department of Urology, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan;3Department of Urology, Kaohsiung Municipal Hsiao-Kang Hospital, Kaohsiung, Taiwan;5Department of Urology, SinYing Hospital, Department of Health, Executive Yuan, Kaohsiung, Taiwan;
Abstract
Introduction and Objective: To study the mechanism of bladder overactivity in ketamine-induced cystitis (KIC), we will elucidate whether ketamine-induced autophagy are mediated via the mitochondrial and endoplasmic reticulum (ER) pathway.
Methods: A ketamine-associated ulcerative cystitis model by intraperitoneally injecting ketamine (30 mg/kg) in rat was developed. Cystometry (CMG) and micturition frequency/volume studies were recorded for bladder function. Transmission electron microscopy (TEM) study was performed to evaluate the bladder morphology and evaluated the distribution of autophagosome and autolysosome. Quantitative real-time polymerase chain reaction analysis and western blot analyses were carried out to examine the expressions of autophagy-associated protein﹝mTOR (the mammalian target of rapamycin), Atg 12 (autophagy related gene 12), Atg 7, LC3 (Atg 8; microtubule-associated protein 1 light chain 3), Beclin 1 (Atg 6) , VPS 34 (vacuolar protein protein sorting 34) in bladder.
Results: Ketamine-treated rats showed bladder hyperactivity, however, ketamine with rapamycin treatment improved bladder function compared with ketamine. The bladder ultrastructure showed intact mitochondria and no autophagosome-like structure in the control group. However, in the ketamine group, some swelling and degraded mitochondria engulfed by autolysosome as well as apoptotic nuclear condensation and shrinkage. Whereas in ketamine with rapamycin group, some swelling and degraded mitochondria engulfed by double-membrane autophagosome. The mRNA and protein levels of autophagy related proteins (mTOR, Atg 12, Atg 7, Beclin1, LC3, and VPS 34) were elevated significantly after treatment. Additionally, the expression of the anti-apoptotic protein BCL-2 and BCL-XL in association with mitochondria in bladder tissues were significantly decreased in the ketamine and rapamycin treated groups. In contrast, the expressions of the pro-apoptotic proteins BAD, tBID, and BAX in association with mitochondria were significantly increased.
Conclusions: Autophagy and mitoptosis has important impacts on ketamine-induced bladder dysfunction and was correlated with mitochondria- and ER-dependent pathways. Therefore, this ketamine-induced ulcerative cystitis rat model could be useful for further investigating the associated mechanisms and for developing or refining therapeutic approaches especially for ketamine abusers.