三叉神经脊束核小胶质细胞在大鼠口面部癌性疼痛调节过程中的作用评价

doi:10.19438/j.cjoms.2021.05.004

摘要/Abstract

摘要： 目的探讨三叉神经脊束核尾侧亚核（Vc）小胶质细胞在口腔癌性疼痛调控中的作用。方法健康成年雄性Wistar大鼠（280~300 g）,采用舌黏膜下注射Walker 256细胞悬液,制备舌肿瘤疼痛模型。实验一,将大鼠随机分为2组（n=10）,即对照组（Sham组）和肿瘤组（Tumor组）。观察肿瘤生长,通过测定大鼠机械缩头反应阈值（HWMT）观察口面部机械痛行为,通过免疫荧光技术检测Vc小胶质细胞的增殖、活化情况。实验二,将大鼠随机分为4组（n=10）,即对照+空白组（Sham+Veh组）、对照+抑制剂组（Sham+Mino组）、肿瘤+空白组（Tumor+Veh组）、肿瘤+抑制剂组（Tumor+Mino组）。检测Vc小胶质细胞的增殖、活化情况,利用qRT-PCR检测Vc中Iba-1、IL-6、IL-1β及TNF-α的mRNA表达水平。测定HWMT,观察大鼠口面部机械痛行为变化。采用SPSS 19.0 软件包对数据进行统计学处理。结果与Sham组相比,随着肿瘤不断增长,Tumor组于第5天时开始出现口面部机械痛（P<0.05）,Vc小胶质细胞显著增殖、活化,直至第10天仍处于口面部机械痛敏和小胶质细胞活化状态。给予米诺环素后,与Tumor+Veh组相比,Tumor+Mino组大鼠的Vc小胶质细胞增殖活化显著受到抑制,Iba-1、IL-6、IL-1β及TNF-α mRNA水平显著降低（P<0.01）,HWMT显著升高（P<0.05）,机械痛敏显著减轻。结论小胶质细胞增殖活化促进炎症因子释放,参与口面部癌性疼痛的发生、发展过程。抑制小胶质细胞活化,可显著减轻口面部癌性疼痛。

关键词: 癌性疼痛, 三叉神经脊束核尾侧亚核, 小胶质细胞, 米诺环素

Abstract: PURPOSE: To explore the role of microglia activation in the trigeminal spinal subnucleus caudalis (Vc) in the development of orofacial cancer-induced pain. METHODS: Adult male Wistar rats(280-300 g) were injected with Walker 256 cell suspension under the tongue mucosa to establish tongue tumor pain model. In experiment 1, the rats were randomly divided into control group (Sham) and tumor group (Tumor) (n=10). Head withdrawal mechanical threshold (HWMT) was measured with Von Frey Hair, and immunofluorescence technology was used to detect the proliferation and activation of microglia in the Vc. In experiment 2, the rats were randomly divided into four groups(n=10), i.e., Sham + Veh (intraperitoneal injection with PBS in group Sham), Sham + Mino (intraperitoneal injection with minocycline in group Sham), Tumor + Veh (intraperitoneal injection with PBS in group Tumor), Tumor + Mino (intraperitoneal injection with minocycline in group Tumor). The proliferation and activation of microglia in the Vc were detected, the mRNA expression level of Iba-1, IL-6, IL-1β and TNF-α in Vc were detected by qRT-PCR, and the orofacial mechanical pain of rats was measured by HWMT. SPSS 19.0 software package was used for data analysis. RESULTS: As the tumors continued to grow, HWMT in the tumor-bearing rats were decreased initially on day 5(P<0.05) versus the sham group and continued to decline until day 10. Marked proliferated and activated microglia were observed on day 5 and day 10 compared with the sparsely ramified microglia in the sham group. Treatment with minocycline (Tumor + Mino) inhibited the proliferation and activation of microglia and the mRNA expression level of Iba-1, IL-6, IL-1β and TNF-α in the Vc. HWMT was significantly higher in tumor-bearing rats receiving minocycline (Tumor + Mino) than in tumor-bearing rats receiving vehicle (Tumor + Veh). CONCLUSIONS: Activation of microglia promoted the release of inflammatory factors, which played a role in the development of orofacial cancer pain. Inhibition of microglia activation could significantly alleviate orofacial cancer-induced pain.

Key words: Cancer-induced pain, Trigeminal spinal subnucleus caudalis, Microglia, Minocycline

中图分类号:

R739.8

王慧, 孙晋虎. 三叉神经脊束核小胶质细胞在大鼠口面部癌性疼痛调节过程中的作用评价[J]. 中国口腔颌面外科杂志, 2021, 19(5): 405-410.

WANG Hui, SUN Jin-hu. The role of microglia in the trigeminal spinal subnucleus caudalis in regulation of orofacial cancer-induced pain in rats[J]. China J Oral Maxillofac Surg, 2021, 19(5): 405-410.

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