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; 刘志文; 杨彬; 周鑫; 谢志艳; 曾佳玉; 赵可心; 曹文宇; 钟小林

doi:10.16098/j.issn.0529-1356.2022.04.002

PDF(4008 KB)

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

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*

作者信息 +

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-yu^3* ZHONG Xiao-lin^1*

Author information +

文章历史 +

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。

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.

导出引用

陈玲刘志文杨彬周鑫谢志艳曾佳玉赵可心曹文宇钟小林. 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[J]. 解剖学报. 2022, 53(4): 412-417 https://doi.org/10.16098/j.issn.0529-1356.2022.04.002

CHEN Ling LIU Zhi-wen YANG Bin ZHOU Xin XIE Zhi-yan ZENG Jia-yu ZHAO Ke-xin CAO Wen-yu ZHONG Xiao-lin. Effect and mechanism of bromodomain-containing protein 4 inhibitor JQ1 on formaldehyde solution induced inflammatory pain in mice[J]. Acta Anatomica Sinica. 2022, 53(4): 412-417 https://doi.org/10.16098/j.issn.0529-1356.2022.04.002

中图分类号： R322.81

参考文献

［1］Wu T, Pinto HB, Kamikawa YF, et al. The BET family member BRD4 interacts with OCT4 and regulates pluripotency gene expression ［J］. Stem Cell Rep, 2015, 4(3): 390-403.

［2］Schnepp RW, Maris JM. Targeting MYCN: a good BET for improving neuroblastoma therapy ［J］. Cancer Discov, 2013, 3(3): 255-257.

［3］Tian B, Hosoki K, Liu ZQ, et al. Mucosal bromodomain-containing protein 4 mediates aeroallergen-induced inflammation and remodeling ［J］. J Allergy Clin Immunol, 2019, 143(4):1380-1394.

［4］Andrieu GP, Shafran JS, Deeney JT, et al. BET proteins in abnormal metabolism, inflammation, and the breast cancer microenvironment ［J］. J Leukoc Biol, 2018, 104(2): 265-274.

［5］White ME, Fenger JM, Carson WE. Emerging roles of and therapeutic strategies targeting BRD4 in cancer ［J］. Cell Immunol, 2019, 337: 48-53.

［6］Filippakopoulos P, Qi J, Picaud S, et al. Selective inhibition of BET bromodomains ［J］. Nature, 2010, 468(7327): 1067-1073.

［7］Leal AS, Williams CR, Royce DB, et al. Bromodomain inhibitors, JQ1 and I-BET 762, as potential therapies for pancreatic cancer ［J］. Cancer Lett, 2017, 394: 76-87.

［8］Park S, Willingham MC, Qi J, et al. Metformin and JQ1 synergistically inhibit obesity-activated thyroid cancer ［J］. Endocr Relat Cancer, 2018, 25(10): 865-877.

［9］Judith SV, Jesús AA, Xavier N, et al. BET protein inhibition regulates cytokine production and promotes neuroprotection after spinal cord injury ［J］. J Neuroinflamm ation, 2019, 16(1): 124.

［10］ Machado NJ, Sim?es AP, Silva HB, et al. Caffeine reverts memory but not mood impairment in a depression-prone mouse strain with up-regulated adenosine A 2A receptor in hippocampal glutamate synapses ［J］. Mol Neurobiol, 2017, 54(2): 1552-1563.

［11］ Hu XD, Ballo L, Pietila L, et al. BDNF-induced increase of PSD-95 in dendritic spines requires dynamic microtubule invasions ［J］. J Neurosci, 2011, 31(43): 15597-15603.

［12］ Erica K, Herre M, Ilana ZS, et al. BET protein Brd4 activates transcription in neurons and BET inhibitor Jq1 blocks memory in mice ［J］. Nat Neurosci, 2015, 18(10): 1464-1473.

［13］ Yaster M, Guan X, Petralia RS, et al. Effect of inhibition of spinal cord glutamate transporters on inflammatory pain induced by Formaldehyde solution and complete Freund’s adjuvant ［J］. Anesthesiology, 2011, 114(2): 412-423.

［14］ Hough LB, Rice FL. H3 receptors and pain modulation: peripheral, spinal, and brain interactions ［J］. J Pharmacol Exp Ther, 2011, 336(1): 30-37.

［15］ Bandopadhayay P, Bergthold G, Nguyen B, et al. BET bromodomain inhibition of MYC-amplified medulloblastoma ［J］. Clin Cancer Res, 2014, 20(4): 912-925.

［16］ Korb E, Herre M, Zucker SI, et al. BET protein Brd4 activates transcription in neurons and BET inhibitor Jq1 blocks memory in mice ［J］. Nat Neurosci, 2015, 18(10): 1464-1473.

［17］ Mele DA, Salmeron A, Ghosh S, et al. BET bromodomain inhibition suppresses TH17-mediated pathology ［J］. J Exp Med, 2013, 210(11): 2181-2190.

［18］ Jung KH, Das A, Chai JC, et al. RNA sequencing reveals distinct mechanisms underlying BET inhibitor JQ1-mediated modulation of the LPS-induced activation of BV-2 microglial cells ［J］. J Neuroinflammation, 2015, 12: 36.

［19］ Sartor GC, Powell SK, Brothers SP, et al. Epigenetic readers of lysine acetylation regulate cocaine-induced plasticity ［J］. J Neurosci, 2015, 35(45): 15062-15072.

［20］ Hsieh MC, Ho YC, Lai CY, et al. Bromodomain-containing protein 4 activates voltage-gated sodium channel 1.7 transcription in dorsal root ganglia neurons to mediate thermal hyperalgesia in rats ［J］. Anesthesiology, 2017, 127(5): 862-877.

［21］ Takahashi K, Yi H, Liu CH, et al. Spinal bromodomain-containing protein 4 contributes to neuropathic pain induced by HIV glycoprotein 120 with morphine in rats ［J］. Neuroreport, 2018, 29(6): 441-446.

［22］ Guerra B, Issinger OG. Protein kinase CK2 in human diseases ［J］. Curr Med Chem, 2008, 15(19): 1870-1886.