脊髓A1型星形胶质细胞在外周炎性痛中的动态变化  

Changes in spinal A1 astrocyte polarization during peripheral inflammatory pain

作　　者：李倩[1] 李玲玲[1] 李爽 黄楚天 周君梅[1] LI Qian;LI Lingling;LI Shuang;HUANG Chutian;ZHOU Junmei(Department of Central Laboratory,Shanghai Children’s Hospital,Shanghai Jiaotong University,Shanghai 200040,China)

机构地区：[1]上海市儿童医院,上海交通大学附属儿童医院中心实验室,上海200040

出　　处：《中国实验动物学报》2021年第5期578-584,共7页Acta Laboratorium Animalis Scientia Sinica

基　　金：国家自然科学基金(81500946)。

摘　　要：目的研究外周炎性痛小鼠痛行为以及脊髓A1型星形胶质细胞的动态变化,为阐明炎性痛的病理机制提供实验基础及理论依据。方法雄性C57BL/6小鼠16只,分为对照组8只和炎性痛组8只。小鼠右侧足底注射完全弗氏佐剂(CFA)建立外周炎性痛模型,对照组右侧足底注射相同体积生理盐水。在造模前以及造模后第1、3、5、7天检测小鼠机械刺激缩足阈值(MWT)和辐射热刺激缩爪潜伏期(PWL)的变化。以逆转录聚合酶链反应检测A1、A2型星形胶质细胞标志物在炎性痛不同时程脊髓内的动态表达变化。用免疫荧光法检测小鼠脊髓背角GFAP形态以明确星形胶质细胞激活,同时统计脊髓背角A1型星形胶质细胞标志物C3与GFAP共定位水平。结果炎性痛小鼠造模后患侧MWT、PWL均明显下降,小鼠出现机械性痛觉超敏与热痛觉过敏;且在造模后第3天,脊髓背角GFAP表达开始升高,表达量为1.84±0.10 vs 1.08±0.22(P<0.05);炎性痛第7天,A1型星形胶质细胞标志物Serping1、H2-T23的mRNA水平均显著升高(P<0.05),A2型星形胶质细胞标志物S100a10、Ptx3的mRNA水平降低(P<0.05);炎性痛组小鼠脊髓背角星形胶质细胞内C3的表达增加(P<0.05)。结论炎性痛小鼠脊髓反应性星形胶质细胞向A1型极化增加,提示A1型星形胶质细胞可能参与了炎性痛的发生发展。Objective Chronic inflammatory pain is a common disease that severely disrupts the quality of life of patients.The latest research shows that reactive astrocytes can be polarized to two different phenotypes,A1 and A2 astrocytes.Type A1 astrocytes can secrete pro-inflammatory cytokines and promote neuroinflammation,and type A2 astrocytes can secrete neurotrophic factors and promote tissue repair.This paper aimed to study pain behavior and changes to A1 astrocytes in the spinal cord of mice with peripheral inflammatory pain,to elucidate the pathological mechanisms of peripheral inflammatory pain.Methods A total of 16 male C57 BL/6 mice were divided into the control group and inflammatory pain group(eight mice per group).The inflammatory pain model was established by injection of complete Freund’s adjuvant(CFA)into the plantar surface of the right hind paw.The control group were injected with an equivalent volume of saline.The mechanical withdrawal threshold(MWT)and the radiant heat stimulating paw withdrawal latency(PWL)of mice were measured before and 1,3,5 and 7 days after the CFA injection.Reverse transcription polymerase chain reaction was used to detect the expression of A1 and A2 astrocyte markers.Immunohistochemistry was used to detect the expression of glial fibrillary acidic protein(GFAP)in the spinal dorsal horn to identify the process of astrocyte activation.Finally,the co-localization of A1 astrocyte marker C3 and GFAP in the spinal dorsal horn was detected.Results The ipsilateral MWT and PWL had both significantly decreased after CFA injection,indicating that the mice developed mechanical hyperalgesia and thermal hyperalgesia.At the third day after CFA injection,the expression levels of spinal GFAP were significantly increased compared to the day 0(D0)group(1.84±0.10 vs.1.08±0.22,respectively,P<0.05).On the 7 th day after CFA injection,the mRNA levels of A1 astrocyte markers Serping1 and Lcn2 were increased vs.the D0 group(P<0.05),and mRNA levels of A2 astrocyte markers S100 a10 and Ptx3 were decre

关 键 词：炎性痛 A1型星形胶质细胞 反应性星形胶质细胞 脊髓 小鼠 

分 类 号：Q95-33[生物学—动物学]