解剖学报

解剖学报 ›› 2022, Vol. 53 ›› Issue (4): 412-417.doi: 10.16098/j.issn.0529-1356.2022.04.002

• 神经生物学 • 上一篇    下一篇

溴结构域蛋白4抑制剂JQ1在甲醛诱导小鼠炎性痛中的作用及机制

陈玲1 刘志文2 杨彬2 周鑫3 谢志艳3 曾佳玉3 赵可心3 曹文宇3* 钟小林1*   

  1. 1. 南华大学附属第一医院内分泌与代谢疾病研究所，湖南 衡阳 421001; 2. 南华大学附属第二医院麻醉科，湖南 衡阳 421001; 3. 南华大学衡阳医学院应用解剖学与生殖医学研究所，湖南 衡阳 421001
  • 收稿日期:2020-10-22 修回日期:2021-01-15 出版日期:2022-08-06 发布日期:2022-09-11
  • 通讯作者: 曹文宇;钟小林 E-mail:zhxl20130623@163.com
  • 基金资助:
    湖南省自然科学青年基金

Effect and mechanism of bromodomain-containing protein 4 inhibitor JQ1 on formaldehyde solution induced inflammatory pain in mice

CHEN  Ling1 LIU  Zhi-wen2  YANG  Bin2  ZHOU  Xin3  XIE  Zhi-yan3  ZENG  Jia-yu3  ZHAO  Ke-xin3  CAO  Wen-yu3* ZHONG  Xiao-lin1*   

  1. 1.Department of Endocrinology and Metabolism, the First Affiliated Hospital of University of South China, Hu’nan Hengyang 421001, China; 2.Institute of Clinical Anesthesiology, the Second Affiliated Hospital of University of South China, Hu’nan Hengyang 421001, China;  3.Clinical Anatomy and Reproductive Medicine Application Institute, Hengyang Medical School, University of South China, Hu’nan Hengyang 421001, China
  • Received:2020-10-22 Revised:2021-01-15 Online:2022-08-06 Published:2022-09-11
  • Contact: CAO Wen-yu;ZHONG Xiao-lin E-mail:zhxl20130623@163.com

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摘要：

目的  探讨溴结构域蛋白4（BRD4）抑制剂JQ1在甲醛诱导炎性痛模型小鼠脊髓中的表达变化及作用。   方法  32只ICR小鼠随机分为生理盐水组、甲醛注射5 min、30 min和60 min组，每组8只，Western blotting（n=4/组）和Real-time PCR（n=4/组）检测各组小鼠脊髓BRD4的蛋白和mRNA的表达水平；66只小鼠随机分为甲醛注射组，溶媒（DMSO）加甲醛注射组，以及6.25、12.5、25 和50 mg/kg JQ1注射加甲醛溶液组，每组11只，观察BRD4抑制剂JQ1对每组小鼠自发性疼痛的影响（n=11/组）；免疫组织化学（n=3/组）、Real-time PCR（n=4/组）或Western blotting（n=4/组）的方法检测25 mg/kg JQ1对模型小鼠脊髓即早基因c-fos及谷氨酸受体2(GluR2)表达的影响。   结果  甲醛注射后30 min和60 min，小鼠脊髓BRD4的蛋白（P<0.01）和mRNA水平明显升高（P<0.05）；行为学结果显示，25 mg/kg和50 mg/kg JQ1处理组小鼠第Ⅱ相痛反应持续时间与溶媒（DMSO）组相比均明显缩短（P<0.05）； 免疫组织化学及Real-time PCR结果显示，25 mg/kg JQ1干预明显减少小鼠脊髓c-fos的阳性细胞数（P<0.05) 和mRNA水平（P<0.05），Western blotting结果显示，25 mg/kg JQ1干预明显降低甲醛诱导的小鼠脊髓GluR2（P<0.001）的蛋白表达。   结论  BRD4在炎性痛中枢敏化中起重要作用，JQ1可能是通过抑制疼痛中枢敏化的形成从而缓解炎性疼痛。

关键词: 溴结构域蛋白4, 炎性痛, 中枢敏化, 脊髓, 免疫印迹法, 小鼠

Abstract:

Objective  To explore the expression and role of bromodomain-containing protein 4(BRD4) in spinal cord of mice which suffered inflammatory pain induced by formaldehyde solution.    Methods  Thirty-two ICR mice were randomly divided into normal saline group and formaldehyde injection 5 minutes, 30 minutes and 60 minutes groups, with 8 mice in each group. The expression of BRD4 protein and mRNA in spinal cord of mice in each group were detected by Western blotting (n=4/group) and Real-time PCR (n=4/group); 66 mice were randomly divided into formaldehyde injection group, vehicle (DMSO) plus formaldehyde injection group and 6.25, 12.5, 25 and 50 mg/kg JQ1 injection plus formaldehyde solution group, with 11 mice in each group. The effect of BRD4 inhibitor JQ1 on spontaneous pain in each group was observed (n=11/group); Immunohistochemistry (n=3/group), Real-time PCR (n=4/group) or Western blotting (n= 4/group) were used to detect the effects of 25 mg/kg JQ1 on the expression of c-fos and glutamate receptor 2(GluR2) in the spinal cord of model mice.    Results  The result  showed that levels of BRD4 protein (P<0.01) and mRNA in spinal cord increased significantly 30 min and 60 min after formaldehyde solution injection (P<0.05). The behavioral test showed that both 25 mg/kg and 50 mg/kg JQ1 administration could reduce the second phase spontaneous pain compared with the solvent (DMSO) group (P<0.05). Furthermore, immunohistochemistry and Real-time PCR result  showed that 25 mg/kg JQ1 injection could significantly reduce positive numbers (P<0.01) and high mRNA expression of c-fos in mouse spinal cord induced by formaldehyde solution (P<0.05), and the Western blotting result  showed that 25 mg/kg JQ1 administration could significantly reduce the expression of glutamate receptor GluR2 (P<0.001).     Conclusion  BRD4 may play an important role in the induction of central sensitization of inflammatory pain, and JQ1 may alleviate inflammatory pain behavior by inhibiting the formation of central sensitization of pain.

Key words: Bromodomain-containing protein 4, Inflammatory pain, Central sensitization, Spinal cord,  , Western blotting,  , Mouse

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