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纸质出版:2014
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赵喆,何文娟,温惠中,阮怀珍.脊髓背角P2X7受体激活引起神经病理性疼痛的发生[J].陆军军医大学学报(原第三军医大学学报),2014,36(09):897-901.
Zhao Zhe,He Wenjuan,Wen Huizhong,Ruan Huaizhen.Activation of P2X7 receptor in spinal dorsal horn causes rat neuropathic pain[J].J Amry Med Univ (J Third Mil Med Univ),2014,36(09):897-901.
目的 观察脊髓鞘内注射P2X
7
受体激动剂[3′-O-(4-Benzoyl)benzoyl ATP,BzATP
]
及抑制剂(brilliant blue G,BBG)对大鼠痛行为及脊髓背角小胶质细胞活化的影响。 方法 大鼠脊髓蛛网膜下腔内置管,制备给药模型后大鼠分为:对照组(给予磷酸盐缓冲液PBS)、BzATP组(给予P2X
7
受体激动剂)、BBG+BzATP组(给予P2X
7
受体抑制剂+激动剂)和BBG组(给予P2X
7
受体抑制剂),分别于给药0、1、3、5、7、11、14 d通过热辐射刺激法测定大鼠热痛阈,使用von Frey纤维丝测定大鼠机械痛阈,应用免疫组织化学技术观察不同时间点脊髓腰段(L
4
~L
6
)背角OX42表达的变化,并进行分析。 结果 大鼠鞘内给予BzATP后,大鼠表现出缩足和舔足等痛觉相关行为。行为学检测结果显示,大鼠鞘内给予P2X
7
受体激动剂后,动物热痛阈和机械痛阈下降,于给药第7天达最低(
P
0.01),停药后逐渐恢复至正常水平;若先给予P2X
7
受体抑制剂后再给予激动剂,则大鼠的热痛阈和机械痛阈无明显变化。免疫荧光双标检测结果显示,脊髓背角OX42阳性小胶质细胞表达P2X
7
受体。免疫组化检测结果显示大鼠脊髓背角OX42免疫反应在给予BzATP 7 d后明显增强(
P
0.01),停止给药后恢复至正常。 结论 脊髓背角小胶质细胞P2X
7
受体激活参与了神经病理性疼痛的发生与维持。
Objective To determine the effects of intrathecal injection of P2X
7
receptor agonist
3′-O-(4-Benzoyl)benzoyl ATP (BzATP) and antagonist
Brilliant Blue G (BBG) on the activation of spinal dorsal horn microglia and pain related behaviors. Methods Totally 48 SD rats with intrathecal catheterization were
randomly divided into 4 groups
control
BzATP group
BBG+BzATP group and BBG group (n=12). The rats of above groups received an intrathecal administration of phosphorylate buffer solution (PBS)
BzATP
BBG+BzATP
and BBG
respectively since 3 d after catheterization for 7 continuous days. Pain threshold was assessed by thermal radiation and von Frey filament in 0
1
3
5
7
11
and 14 d after administration. Expression of OX42 and P2X
7
receptor in spinal dorsal horn of L
4
to L
6
was measured by immunofluoresence staining. Results After administration of BzATP
thermal hyperalgesia and mechanical allodynia appeared in the rats. The thermal withdrawl latency (TWL) and 50% paw withdrawl threshold (PWT) were decreased obviously from 1 d after injection and descended to the lowest at 7 d (
P
0.01)
then returned to the level of normal control slowly. And these results were reversed by the treatment of P2X
7
receptor antagonist
BBG. The double labeled immunofluorescence results showed that the OX42 positive microglia expressed P2X
7
receptor. Immunohistological assay showed that the reaction of OX42 was obviously increased after injection of 7 days’ treatment of BzATP (
P
0.01)
and then declined to the baseline after the end of administration. Conclusion Activation of spinal dorsal horn microglial P2X
7
receptor plays essential roles in the incidence and progress of neuropathic pain.
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