Ozturk Aysegul, Ozdemir Ercan, Ozkaraca Mustafa, Taskiran Ahmet Sevki, Altun Ahmet
Departments of Therapy and Rehabilitation, Vocational School of Health Services, Sivas Cumhuriyet University, Sivas, Turkey.
Departments of Physiology, Medicine Faculty, Sivas Cumhuriyet University, 58140, Sivas, Turkey.
Naunyn Schmiedebergs Arch Pharmacol. 2025 Jun 23. doi: 10.1007/s00210-025-04384-5.
Morphine is one of the drugs frequently used for severe pain in chronic diseases such as cancer, but long-term use leads to morphine tolerance. The mechanism of morphine tolerance is not yet fully understood. This study aimed to investigate the effects of the TRPV1 channel antagonist capsazepine (CPZ) on morphine tolerance and morphine-induced neurotoxicity, mitochondrial damage, and apoptosis by in vivo and in vitro methods. Thirty-six male Wistar Albino rats, aged 12-14 weeks (weight 230-250 g), were included in the study. To evaluate the effect of morphine on mitochondrial damage and apoptosis, cytochrome c, apoptosis-inducing factor (AIF), caspase-9, and caspase-3, Bax, and Bcl-2 levels were determined from tissue samples by ELISA and immunohistochemical (IHC) methods. For in vitro analysis, CPZ C6 glioma cells were treated for 1 h, and then neurotoxic morphine (4 mM) was added to the cell medium. Cell viability was measured by the XTT method. Biochemical methods and immunofluorescence staining were used to evaluate mitochondrial damage and apoptosis. The findings indicated that co-administration of CPZ with morphine significantly reduced morphine tolerance (p < 0.05). Furthermore, in vivo and in vitro tests showed that CPZ administration decreased the levels of mitochondrial markers cytochrome c and AIF and proapoptotic markers caspase-3, caspase-9, and Bax and significantly increased the expression of antiapoptotic Bcl-2 (p < 0.01). In conclusion, both in vivo and in vitro test findings demonstrated that CPZ ameliorated morphine-induced mitochondrial dysfunction and attenuated apoptosis, reducing morphine-induced toxicity and tolerance.
吗啡是常用于治疗癌症等慢性疾病严重疼痛的药物之一，但长期使用会导致吗啡耐受性。吗啡耐受的机制尚未完全明确。本研究旨在通过体内和体外方法研究瞬时受体电位香草酸亚型1（TRPV1）通道拮抗剂辣椒素（CPZ）对吗啡耐受性、吗啡诱导的神经毒性、线粒体损伤和细胞凋亡的影响。本研究纳入了36只12 - 14周龄（体重230 - 250克）的雄性Wistar白化大鼠。为评估吗啡对线粒体损伤和细胞凋亡的影响，采用酶联免疫吸附测定（ELISA）和免疫组织化学（IHC）方法从组织样本中测定细胞色素c、凋亡诱导因子（AIF）、半胱天冬酶-9、半胱天冬酶-3、促凋亡蛋白Bax和抗凋亡蛋白Bcl-2的水平。对于体外分析，CPZ处理C6胶质瘤细胞1小时，然后向细胞培养基中加入神经毒性吗啡（4 mM）。采用XTT法测定细胞活力。采用生化方法和免疫荧光染色评估线粒体损伤和细胞凋亡。结果表明，CPZ与吗啡联合使用可显著降低吗啡耐受性（p < 0.05）。此外，体内和体外试验表明，给予CPZ可降低线粒体标志物细胞色素c和AIF以及促凋亡标志物半胱天冬酶-3、半胱天冬酶-9和Bax的水平，并显著增加抗凋亡蛋白Bcl-2的表达（p < 0.01）。总之，体内和体外试验结果均表明，CPZ可改善吗啡诱导的线粒体功能障碍并减轻细胞凋亡，降低吗啡诱导的毒性和耐受性。
