Bureau Michel, Pelletier Jérôme, Rousseau Alexandre, Bernard Geneviève, Chabaud Stéphane, Bolduc Stéphane
Centre LOEX de l'Université Laval, Génie tissulaire et régénération, Centre de recherche FRQS du CHU de Québec, Axe Médecine Régénératrice, Québec, QC.
Can Urol Assoc J. 2015 Sep-Oct;9(9-10):E613-7. doi: 10.5489/cuaj.2899. Epub 2015 Sep 9.
Ketamine is a common recreational drug. Severe lower urinary tract symptoms associated with its consumption have been reported, but little is known about the involved mechanisms. The effect of ketamine, which is excreted in urine, was evaluated by its application on an in vitro three-dimensional human tissue-engineered bladder model composed of an urothelium and a submucosa.
Human urothelial cells were cultured with medium containing various concentrations of ketamine and harvested at different times to obtain growth curves. Using this model, specific activity of caspase-3 was measured to assess the level of apoptosis induced by ketamine. Finally, a human tissue-engineered bladder model was used. Urothelial cells were plated on a stromal layer made of dermal fibroblasts and incubated at the air/liquid interface to allow their differentiation. Ketamine was then put on the mature urothelium using paper or agarose vectors for 48 hours.
The presence of ketamine increased cells' doubling times from 1.26 days for control to 1.38 days (p = 0.14) and 1.78 days (p < 0.01) for the 0.5 mM and 1.5 mM concentrations, respectively. 5 mM and 10 mM of ketamine led to decline in the major cell population. Exposure to 5 mM ketamine induced apoptosis, confirmed by a 2.5-fold increase in capase-3 specific activity from control (p = 0.03). The structure and cellular cohesion of the urothelium on the three-dimensional model, especially in the intermediate layers, were severely affected in a concentration dependant fashion with both vectors.
The presence of ketamine in the bladder directly damages the urothelium through the induction of apoptosis.
氯胺酮是一种常见的消遣性药物。已有报道称其使用会引发严重的下尿路症状，但相关机制尚不清楚。通过将氯胺酮应用于由尿路上皮和黏膜下层组成的体外三维人体组织工程膀胱模型，评估了尿液中排泄的氯胺酮的作用。
将人尿路上皮细胞与含有不同浓度氯胺酮的培养基一起培养，并在不同时间收获以获得生长曲线。使用该模型，测量半胱天冬酶 - 3的比活性以评估氯胺酮诱导的细胞凋亡水平。最后，使用人体组织工程膀胱模型。将尿路上皮细胞接种在由真皮成纤维细胞制成的基质层上，并在气液界面孵育以使其分化。然后使用纸片或琼脂糖载体将氯胺酮置于成熟的尿路上皮上48小时。
氯胺酮的存在使细胞倍增时间从对照组的1.26天分别增加到0.5 mM和1.5 mM浓度时的1.38天（p = 0.14）和1.78天（p <0.01）。5 mM和10 mM的氯胺酮导致主要细胞群体减少。暴露于5 mM氯胺酮会诱导细胞凋亡，半胱天冬酶 - 3比活性较对照组增加2.5倍，证实了这一点（p = 0.03）。两种载体均以浓度依赖性方式严重影响三维模型上尿路上皮的结构和细胞黏附，尤其是中间层。
膀胱中氯胺酮的存在通过诱导细胞凋亡直接损害尿路上皮。
