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Modulatory effect of curcumin on ketamine-induced toxicity in rat thymocytes: Involvement of reactive oxygen species (ROS) and the phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) pathway.
Biomolecules and Biomedicine ( IF 3.4 ) Pub Date : 2018-11-07 , DOI: 10.17305/bjbms.2018.2607
Svetlana Pavlovic 1 , Zorica Jovic , Radmila Karan , Dane Krtinic , Gorana Rankovic , Mladjan Golubovic , Jelena Lilic , Voja Pavlovic
Affiliation  

Ketamine is a widely used anesthetic in pediatric clinical practice. Previous studies have demonstrated that ketamine induces neurotoxicity and has a modulatory effect on the cells of the immune system. Here, we evaluated the potential protective effect and underlying mechanisms of natural phenolic compound curcumin against ketamine-induced toxicity in rat thymocytes. Rat thymocytes were exposed to 100 µM ketamine alone or combined with increasing concentrations of curcumin (0.3, 1, and 3 μM) for 24 hours. Cell viability was analyzed with CCK-8 assay kit. Apoptosis was analyzed using flow cytometry and propidium iodide as well as Z-VAD-FMK and Z-LEHD-FMK inhibitors. Reactive oxygen species (ROS) production and mitochondrial membrane potential [MMP] were measured by flow cytometry. Colorimetric assay with DEVD-pNA substrate was used for assessing caspase-3 activity. Involvement of phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway was tested with Wortmannin inhibitor. Ketamine induced toxicity in cells, increased the number of hypodiploid cells, caspase-3 activity and ROS production, and inhibited the MMP. Co-incubation of higher concentrations of curcumin (1 and 3 μM) with ketamine markedly decreased cytotoxicity, apoptosis rate, caspase-3 activity, and ROS production in rat thymocytes, and increased the MMP. Application of Z-VAD-FMK (a pan caspase inhibitor) or Z-LEHD-FMK (caspase-9 inhibitor) with ketamine effectively attenuated the ketamine-induced apoptosis in rat thymocytes. Administration of Wortmannin (a PI3K inhibitor) with curcumin and ketamine significantly decreased the protective effect of curcumin on rat thymocytes. Our results indicate that ketamine-induced toxicity in rat thymocytes mainly occurs through the mitochondria-mediated apoptotic pathway and that the PI3K/Akt signaling pathway is involved in the anti-apoptotic effect of curcumin.

中文翻译:

姜黄素对氯胺酮诱导的大鼠胸腺细胞毒性的调节作用:涉及活性氧(ROS)和磷酸肌醇3激酶(PI3K)/蛋白激酶B(Akt)途径。

氯胺酮是儿科临床实践中广泛使用的麻醉剂。先前的研究表明,氯胺酮可诱发神经毒性,并对免疫系统的细胞具有调节作用。在这里,我们评估了天然酚类化合物姜黄素对氯胺酮诱导的大鼠胸腺细胞毒性的潜在保护作用及其潜在机制。将大鼠胸腺细胞单独暴露于100 µM氯胺酮中或与浓度不断增加的姜黄素(0.3、1和3μM)混合暴露24小时。用CCK-8测定试剂盒分析细胞活力。使用流式细胞仪和碘化丙啶以及Z-VAD-FMK和Z-LEHD-FMK抑制剂分析细胞凋亡。通过流式细胞仪测量活性氧(ROS)的产生和线粒体膜电位[MMP]。用DEVD-pNA底物进行比色测定可评估caspase-3活性。用Wortmannin抑制剂测试了磷酸肌醇3激酶(PI3K)/蛋白激酶B(Akt)信号通路的参与。氯胺酮诱导细胞毒性,增加二倍体细胞数量,caspase-3活性和ROS产生,并抑制MMP。将高浓度姜黄素(1和3μM)与氯胺酮共同孵育可显着降低大鼠胸腺细胞的细胞毒性,凋亡率,caspase-3活性和ROS产生,并增加MMP。Z-VAD-FMK(泛半胱天冬酶抑制剂)或Z-LEHD-FMK(caspase-9抑制剂)与氯胺酮一起使用可有效减弱氯胺酮诱导的大鼠胸腺细胞凋亡。将Wortmannin(一种PI3K抑制剂)与姜黄素和氯胺酮一起给药可显着降低姜黄素对大鼠胸腺细胞的保护作用。我们的结果表明,氯胺酮诱导的大鼠胸腺细胞毒性主要通过线粒体介导的凋亡途径发生,而PI3K / Akt信号传导途径参与姜黄素的抗凋亡作用。
更新日期:2020-08-21
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