内源性大麻素2-AG通过调控神经胶质细胞上的大麻素受体参与调控福尔马林诱导的大鼠炎性持续性痛

发布时间：2018-04-17 19:04

本文选题：内源性大麻素2-AG + 大麻素受体　；参考：《兰州大学》2014年博士论文

【摘要】：目的：鞘内注射内源性大麻素2-AG和大麻素受体CB1和CB2拮抗剂后,通过行为学和形态学实验方法,观察大麻素受体和脊髓背角神经细胞在2-AG抑制福尔马林诱导的炎性持续性痛过程中扮演的角色。方法：通过大鼠足底注射福尔马林(5%,50μl),建立炎性持续性痛动物模型,形态学观察大麻素受体亚型(CB1和CB2)在脊髓背角的分布情况,进一步观察这两个受体亚型与神经细胞(星形胶质细胞、小胶质细胞和神经元)的共存情况。行为学观察鞘内注射内源性大麻素2-AG后,对福尔马林诱导的自发性伤害性行为(舔咬爪和缩腿反射)的抑制效应；鞘内注射大麻素CB1、CB2受体拮抗剂(AM281或者AM630)后,观察是否翻转了内源性大麻素2-AG的抑制效应。结果：鞘内注射内源性大麻素2-AG(1,10,100μg；注射剂量10μl)后,发现,随着2-AG药物浓度的增加,抑制效应逐渐增强,但是不成剂量依赖关系,各处理组之间存在显著性差异(比如对照组和不同剂量内源性大麻素2-AG组)(舔咬爪,F(3,287)=64.963,P0.001；缩腿反射,F(3,287)=131.255,P0.001),时间点之间也存在显著性差异(F(11,287)=14.580,P0.001；F(11,287)=11.250,,P0.001),药物和时间之间存在交互作用(F(33,287)=5.663,P0.001；F(33,287)=1.737,P=0.010)。内源性大麻素2-AG诱导的抑制效应可以被预先鞘内注射大麻素受体CB1受体拮抗剂AM281翻转,各处理组之间存在显著性差异(舔咬爪,F(3,215)=58.322,P0.001；缩腿反射,F(2,215)=139.472,P0.001),时间点之间也存在显著性差异(F(11,215)=19.150,P0.001；F(11,215)=16.816,P0.001),药物和时间之间存在交互作用(F(22,215)=7.530,P0.001；F(22,215)=2.670,P0.001)。同样,内源性大麻素2-AG诱导的抑制效应可以被预先鞘内注射大麻素受体CB2受体拮抗剂AM630翻转,各处理组之间存在显著性差异(舔咬爪,F(2,215)=57.085,P0.001；缩腿反射,F(2,215)=84.265,P0.001),时间点之间也存在显著性差异(F(11,215)=19.032,P0.001；F(11,215)=14.019,P0.001),药物和时间之间存在交互作用(F(22,215)=4.260,P0.001；F(22,215)=3.300,P0.001)。形态学的研究结果显示,在完全空白大鼠(Naive),腰段脊髓背角的CB1受体主要在神经元上表达,在小胶质细胞有少量表达,而在星形胶质细胞未见表达。在大鼠足底注射福尔马林后,腰段脊髓背角的CB1受体仍主要在神经原上表达,在小胶质细胞有少量表达,同时在星形胶质细胞也开始表达。在完全空白大鼠脊髓背角CB2受体不表达,但是在足底注射福尔马林后,CB2受体表达增多,并且主要与星形胶质细胞和小胶质细胞共表达,跟神经元有少量共表达。结论：研究结果提示,内源性大麻素2-AG主要通过直接或者间接的作用于脊髓背角神经胶质(星形胶质细胞和小胶质细胞)上的大麻素受体CB1和CB2发挥对福尔马林诱导的伤害性行为抑制效应,而CB2受体可能在此过程中起主导作用。
[Abstract]:Objective: to study the behavior and morphology of endogenous cannabinoid 2-AG and cannabinoid receptor CB1 and CB2 antagonists after intrathecal injection.To observe the role of cannabinoid receptor and spinal dorsal horn neurons in the inhibition of formalin-induced inflammatory persistent pain by 2-AG.Methods: the inflammatory persistent pain model was established by injection of Rhizoma Formalinus formalin into the plantar of rats. The distribution of cannabinoid receptor subtypes CB1 and CB2 in the dorsal horn of spinal cord was observed by morphology.The coexistence of these two receptor subtypes with neurons (astrocytes, microglia and neurons) was further observed.To observe the inhibitory effect of intrathecal injection of endogenous cannabinoid 2-AG on formalin induced spontaneous nociceptive behavior (licking claw and leg reflex), and after intrathecal injection of cannabinoid CB1 and CB2 receptor antagonist AM281 or AM630.To observe whether the inhibitory effect of endogenous cannabinoid 2-AG was reversed.Results: after intrathecal injection of endogenous cannabinoid 2-AGN (10100 渭 g, dose 10 渭 l), it was found that the inhibitory effect was increased with the increase of 2-AG concentration, but not in a dose-dependent manner.The inhibitory effect induced by endogenous cannabinoid 2-AG can be reversed by intrathecal injection of cannabinoid receptor CB1 receptor antagonist AM281.There were significant differences among the treatment groups (licking claw nip, 58.322, P 0.001; leg reflexes, 139.472, P 0.001; F11215, 19.150, P 0.001, F11215, 16.816, P0.001, P 0.001, and the interaction between the drug and the time, F2221530, P0.001, 2.670, P0.001, P 0.001, P 0.001, P 0.001, P 0.001), and the interaction between the drug and the time was also found in the treatment group, and there was also a significant difference between the two groups (P 0.001), and there was also a significant difference between the two groups (P < 0.05), and there was also a significant difference between the two groups in the treatment time, and there was also a significant difference between the two groups. There was an interaction between the two drugs.Similarly, the inhibitory effect induced by endogenous cannabinoid 2-AG can be reversed by pre-intrathecal injection of cannabinoid receptor CB2 receptor antagonist AM630.The results of morphological study showed that the CB1 receptor in the lumbar spinal dorsal horn was mainly expressed in neurons, a little in microglia, but not in astrocytes.After formalin injection into the plantar of rats, the expression of CB1 receptor in the dorsal horn of lumbar spinal cord was still mainly expressed on neurogen, a little on microglia and also on astrocytes.There was no expression of CB2 receptor in the spinal dorsal horn of rats with complete blank, but the expression of CB2 receptor was increased after formalin injection into the plantar, mainly co-expressed with astrocytes and microglial cells, and a little coexpression with neurons.Conclusion: the results of the study suggest that,Endogenous cannabinoid 2-AG inhibits formalin induced nociceptive behavior mainly by directly or indirectly acting on cannabinoid receptors (CB1 and CB2) on spinal dorsal horn glia (astrocytes and microglia).CB2 receptors may play a leading role in this process.
【学位授予单位】：兰州大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R965

【共引文献】