IL-36Ra抑制炎性痛小鼠脊髓A1型星形胶质细胞极化  

IL-36Ra inhibited A1 astrocyte polarization in spinal cord of mice subjected to inflammatory pain

作　　者：李倩[1] 朱怡文[2,3] 李玲玲 LI Qian;ZHU Yi-wen;LI Ling-ling(Department of Central Laboratory,Shanghai Children′s Hospital,Shanghai Jiaotong University,Shanghai 200040,China;Shanghai Institute of Medical Genetics,Shanghai Jiaotong University,Shanghai 200040,China;NHC Key Laboratory of Medical Embryogenesis and Developmental Molecular Biology&Shanghai Key Laboratory of Embryo and Reproduction Engineering,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 [2]上海交通大学医学遗传研究所,上海200040 [3]国家卫健委医学胚胎分子生物学重点实验室,上海市胚胎与生殖工程重点实验室,上海200040

出　　处：《中国药理学通报》2022年第1期85-90,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81500946)。

摘　　要：目的评价不同剂量IL-36Ra对炎性痛小鼠痛行为以及脊髓A1型星形胶质细胞极化的影响。方法雄性C57BL/6小鼠32只,采用足底注射完全弗氏佐剂(CFA)建立炎性痛模型。随机将小鼠分为CFA+生理盐水组、CFA+IL-36Ra 50 ng组、CFA+IL-36Ra 100 ng组以及CFA+IL-36Ra 200 ng组,每组8只。各组小鼠在造模后d 1~d 7,每日1次鞘内给药。同时,分别在造模前以及造模后d 1、3、5、7检测4组小鼠机械刺激缩足阈值(mechanical withdraw threshold, 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IL-36Ra对CFA小鼠脊髓A1、A2型星形胶质细胞标志物表达水平的影响。以免疫组化法检测各组小鼠脊髓背角A1型星形胶质细胞标志物C3与GFAP共表达水平的变化。结果炎性痛小鼠造模后患侧MWT、PWL均明显下降,IL-36Ra(100、200 ng)可显著改善小鼠的机械性痛觉超敏与热痛觉过敏;且在治疗7 d后,IL-36Ra可降低CFA小鼠脊髓星形胶质细胞激活标志物GFAP、Lcn2的表达水平,抑制星形胶质细胞激活;同时,IL-36Ra(100、200 ng)可下调CFA小鼠脊髓A1型星形胶质细胞标志物Serping1、 H2-T23的mRNA水平,但IL-36Ra各个剂量对A2型星形胶质细胞标志物表达没有明显作用;此外,IL-36Ra还可抑制脊髓背角A1型星形胶质细胞标志物C3表达。结论 IL-36Ra可能通过抑制CFA小鼠脊髓A1型星形胶质细胞极化,进而改善小鼠炎性痛痛行为。Aim To evaluate the effects of different doses of IL-36 Ra on pain behavior and the polarization of spinal A1 astrocytes in mice with inflammatory pain. Methods A total of 32 male C57 BL/6 mice were divided into four groups: CFA+Saline group, CFA+IL-36 Ra 50 ng group, CFA+IL-36 Ra 100 ng group and CFA+IL-36 Ra 200 ng group by random grouping. The inflammatory pain model was established by injection of complete Freund’s adjuvant(CFA) into the plantar surface of the right hind paw of mice. The drugs were given daily from the 1 st day to the 7 th day after CFA injection in each group by intrathecal injection. The changes in the mechanical withdrawal threshold(MWT) and the radiant heat stimulating paw withdrawal latency(PWL) of the mice were detected before and 1, 3, 5 and 7 days after the CFA injection. Reverse transcription polymerase chain reaction was used to detect the expression changes of A1 and A2 astrocyte markers after IL-36 Ra treatment. Immunohistochemistry was used to test the effect of IL-36 Ra on the co-expression level of A1 astrocyte marker C3 and GFAP in the spinal dorsal horn. Results MWT and PWL of the ipsilateral paw significantly decreased after the CFA injection, and IL-36 Ra(100 ng, 200 ng) treatment could significantly improve the mechanical allodynia and thermal hyperalgesia of CFA mice. After treatment for 7 days, IL-36 Ra 200 ng successfully reversed the increase of GFAP and Lcn2 expression in the spinal cord of CFA mice, which demonstrated IL-36 Ra could inhibit the activation of astrocytes. IL-36 Ra significantly inhibited the expression of A1 astrocyte maker Serping1, H2-T23 in spinal cord but showed no effects on the expression of A2 astrocytes marker with each dose. Furthermore, IL-36 Ra inhibited the expression of C3 within the astrocytes in the spinal dorsal horn of CFA mice. Conclusion IL-36 Ra attenuates the inflammatory pain via inhibiting the polarization of A1 reactive astrocytes in the spinal cord of mice with inflammatory pain.

关 键 词：炎性痛 IL-36Ra A1型星形胶质细胞 脊髓 小鼠 A2型星形胶质细胞 

分 类 号：R-332[医药卫生] R322.81