酮康唑抑制氧化亚油酸代谢物介导的外周炎性疼痛机制

Mechanism of Ketoconazole Inhibits Oxidized Linoleic Acid Metabolite-mediated Peripheral Inflammatory Pain

目的探讨细胞色素P450(CYP)抑制剂酮康唑抑制氧化亚油酸代谢物(OLAM)介导的外周炎性疼痛的机制。方法通过弗罗因德佐剂(CFA)模型研究OLAM在炎症过程中作用,CYP酶在OLAM对热痛觉过敏过程中的影响进行研究。结果培养的三叉神经节(TG)神经元在使用递质进行预处理后,随后进行亚油酸(LA)刺激,[Ca2+]i呈现3倍的增长,至(0.156±0.01)。酮康唑的预处理消除LA对[Ca2+]i增强的影响(P<0.05)。再悬浮的提取物引起降钙素与基因相关肽(CGRP)释放增至(980.39±21.04)%,与瞬时受体电位香草酸亚型-1(TRPV1)拮抗剂I-RTX共同处理提取物的CGRP释放减弱至(200.31±10.01)%,CFA/酮康唑+递质处理提取物的CGRP释放减弱至(498.03±13.02)%。在对照组(未发炎)后足中注射LA不显著延长自发性伤害性反应行为的时间。在CFA炎症建立后24 h,足底注射LA产生剂量依赖的伤害性镇痛效应,自发性伤害性反应行为时间上调至(148.92±13.42)s。使用阻断剂的预处理可减少被唤起的非综合行为。与野生型小鼠相比,在基因敲除TRPV1的小鼠中,LA-激发的疼痛行为显著减少,非综合行为的时间为(5.31±0.41)s。与接受对照抗山羊IgG抗体(HBSS)组比较,足底注射抗9和抗13-HODE抗体对LA诱导的伤害性行为减少约72%。注射LA前30 min用酮康唑预处理大鼠发炎后足,酮康唑对LA的抑制作用达到70%,伤害性行为时间下调至(11.28±4.10)s。结论OLAM对外周产生炎症性痛觉,而CYP酶在调节OLAM对炎症性热痛觉过敏的作用方面起着至关重要的作用。

Objective To explore the mechanism of cytochrome P450 inhibitor ketoconazole inhibiting peripheral inflammatory pain mediated by oxidized linoleic acid metabolites(OLAM).Methods The role of OLAM in inflammatory process and the effect of cytochrome P450 enzyme on hyperalgesia were studied by using Freund's adjuvant model.Results The cultured TG neurons were pretreated with a transmitter,followed by linoleic acid(LA)stimulation,and[Ca2+]i showed a 3-fold increase to(0.156±0.01).Ketoconazole pretreatment eliminated the effect of LA on[Ca2+]i enhancement(P<0.05).The application of the resuspended extract on cultured TG neurons resulted in an increase in calcitonin gene related peptide(CGRP)release to(980.39±21.04)%,a significant decrease in CGRP release to(200.31±10.01)%in the extract treated with TRPV1 antagonist I-RTX,and(498.03±13.02)%in the CFA/ketoconazole+transmitter treated extract.In the control group(uninflamed),injection of LA did not significantly increase the duration of spontaneous injurious reactivity.24 h after the establishment of CFA inflammation,the plantar injection of LA produced a dose-dependent analgesic effect,and the time of spontaneous injurious response was up-regulated to(148.92±13.42)s.Pretreatment with blockers can reduce the nonsynthetic behavior evoked.Compared with wild-type mice,la-induced pain was significantly reduced in the mice with the gene deletion TRPV1(5.31±0.41)s for the noncompre-hensive behavior).Plantar injection of anti-9 and anti-13-hode antibodies reduced LA induced injurious behavior by about 72%,compared to the control group receiving anti-goat IgG or vector(HBSS).Ketoconazole was used to pretreat inflamed rat hindpaw 30 min before injection of LA,and the inhibitory effect of ketoconazole on LA was observed to reach 70%,and the time of harmful behavior was reduced to(11.28±4.10)s.Conclusion OLAM can induce peripheral inflammatory hyperalgesia,and cytochrome P450 plays an important role in regulating the role of OLAM in inflammatory hyperalgesia.