鞘内注射JWH-015抑制炎性介质介导的NR2B活化对神经病理性疼痛的影响

发布时间：2018-04-07 17:18

本文选题：CB2　切入点：JWH-015　出处：《青岛大学》2017年硕士论文

【摘要】：目的:我们将研究CB2受体激动剂JWH-015能否抑制CCI引起的神经病理疼痛。同时,我们观察CB2受体和NR2B在DRG,脊髓背角中激活的时间过程,CB2受体激动剂JWH-015和抑制剂AM630对大鼠热痛阈的影响及对DRG和脊髓NR2B的变化。方法:将大鼠造出坐骨神经结扎(CCI)模型,然后在将大鼠分别鞘内置管成功后,将48只成熟雄性SD大鼠随机分为4组,假手术组(sham组),仅暴露坐骨神经;CCI组,将大鼠坐骨神经结扎,仅注射DMSO液体,10μg/天,持续7天;F组,CCI后鞘内注射CB2激动剂JWH-015,10μg/天,持续7天;P组,CCI后在注射JWH-015前半小时注射CB2受体抑制剂AM630。在术后3天、7天、14天分别测定大鼠的热痛阈,并用免疫组化和western blot测定NB2B及炎性因子的表达情况。结果:1:与假手术组相比较,CCI组术后各时间点热痛阈显著下降(p0.05),F组与CCI组相比,CCI组术后各时间点热痛阈热痛阈显著增高(p0.05)。然而在P组中,预先注射CB2受体拮抗剂AM630后给予CB2受体激动剂后,各组大鼠搁时间点的热痛阈值较假手术组无明显变化,这表明预先鞘内注射AM-630后,CB2受体激动剂JWH-015的效应失效。2:CCI对大鼠脊髓背角中TNF-α和IL-1β的影响:坐骨神经结扎后脊髓背角TNF-α和IL-1β的表达明显增多,两种实验方法都说明了这一现象,免疫组化检测CCI后脊髓背角中TNF-α和IL-1β的表达与假手术组相比,CCI后各组大鼠TNF-α和IL-1β的表达明显增加(P0.05),Western blot结果同样显示CCI后脊髓TNF-α和IL-1β表达明显增多。然而在CCI模型后鞘内注射JWH-015,能够抑制CCI模型后TNF-α和IL-1β的表达,且这种效应在预注射AM630后失效。3:CCI对大鼠脊髓背角中NR2B的影响:坐骨神经结扎后脊髓背角NR2B的表达明显增多,Western blot检测CCI后脊髓背角中NR2B的表达与假手术组相比,CCI后各组大鼠NR2B的表达明显增加(P0.05),然而在CCI模型后鞘内注射JWH-015,能够抑制CCI模型后NR2B的表达,且这种效应在预注射AM630后失效。结论:1.CCI后大鼠脊髓TNF-α、IL-1β和NR2B较假手术组明显升高,炎性反应明显增强,TNF-α、IL-1β和NR2B都参与了神经病理性疼痛。2.鞘内注射JWH-015能有效减轻CCI模型后所引起的神经病理性疼痛,鞘内注射JWH-015的大鼠DRG和脊髓TNF-α、IL-1β和NR2B明显下降。3.预先鞘内注射AM-630后,JWH-015的效应失效。4.JWH-015是通过抑制炎性介质介导的NR2B活化从而进一步抑制CCI模型后所导致的神经病理性疼痛。
[Abstract]:Aim: to investigate whether CB2 receptor agonist JWH-015 can inhibit neuropathic pain induced by CCI.At the same time, we observed the effects of CB2 receptor and NR2B on the thermal pain threshold and the changes of DRG and NR2B in the spinal cord of rats during the time course of activation of JWH-015 and AM630, the agonist of CB2 receptor in the dorsal horn of spinal cord.Methods: the rat model of rat sciatic nerve ligation (CCI) was established, and then 48 adult male SD rats were randomly divided into 4 groups, sham group (sham group) and CCI group exposed only to sciatic nerve (CCI).The sciatic nerve of rats was ligated, only 10 渭 g / day of DMSO fluid was injected, 10 渭 g / day of CB2 agonist JWH-015C was injected intrathecally in group F for 7 days, and AM630 was injected into CB2 receptor inhibitor AM630 half an hour before JWH-015 injection in group P after 7 days of JWH-015 injection.The thermal pain threshold was measured on the 3rd day, 7th day and 14th day after operation, and the expression of NB2B and inflammatory factors were detected by immunohistochemistry and western blot.Results: compared with the sham-operated group, the thermal pain threshold of the CCI group was significantly lower than that of the CCI group at different time points. The thermal pain threshold of the CCI group was significantly higher than that of the CCI group.However, in group P, the thermal pain threshold of rats in each group was not significantly changed compared with that in sham operation group after CB2 receptor agonist was given to AM630, a CB2 receptor antagonist, before and after injection of CB2 receptor agonist.This indicated that the effect of JWH-015, the agonist of CB2 receptor, failed after intrathecal injection of AM-630. 2: CCI effect on TNF- 伪 and IL-1 尾 in rat spinal dorsal horn: the expression of TNF- 伪 and IL-1 尾 in spinal dorsal horn increased significantly after ligation of sciatic nerve, which was demonstrated by both experimental methods.However, intrathecal injection of JWH-015 at the back of CCI model inhibited the expression of TNF- 伪 and IL-1 尾 in CCI model.The effect of AM630 on the expression of NR2B in the spinal dorsal horn of rats after ligation of sciatic nerve was significantly increased. The expression of NR2B in the dorsal horn of spinal cord after CCI was detected by Western blot compared with that in the sham operation group.The expression of NR2B increased significantly in each group. However, intrathecal injection of JWH-015 at the back of CCI model could inhibit the expression of NR2B after CCI model.And this effect failed after AM630 was preinjected.Conclusion 1. The levels of TNF- 伪 IL-1 尾 and NR2B in the spinal cord of rats after CCI were significantly higher than those in the sham operation group, and the inflammatory response was significantly enhanced. Both TNF- 伪 and IL-1 尾 and NR2B were involved in neuropathic pain.Intrathecal injection of JWH-015 could effectively alleviate the neuropathic pain induced by CCI model, and the DRG and TNF- 伪 IL-1 尾 and NR2B in spinal cord of rats with intrathecal injection of JWH-015 decreased significantly.Effect failure of JWH-015 after intrathecal injection of AM-630. 4. JWH-015 further inhibits neuropathic pain after CCI model by inhibiting NR2B activation mediated by inflammatory mediators.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R614

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