摘要
目的研究10-11易位甲基胞嘧啶双加氧酶1(TET1)去甲基化调控μ阿片受体对神经病理性疼痛(neuropathic pain,NPP)吗啡耐受的影响及机制。方法采用坐骨神经慢性压迫性损伤模型,背根神经节(dorsal root ganglia,DRG)显微注射表达全长TET1 mRNA的单纯疱疹病毒,行为学检测TET1过表达对痛觉过敏及吗啡耐受的影响,Western blot检测DRG内μ阿片受体(μ-opioid receptor,MOR)的表达,DNA blot方法检测DRG全基因5-甲基胞嘧啶(5mC)和5-羟甲基胞嘧啶(5hmC)水平,亚硫酸氢盐测序和Tab-seq技术检测MOR启动子区域5mC和5hmC水平。结果与对照组相比,TET1过表达明显缓解神经损伤导致的痛觉过敏和吗啡耐受,增强吗啡的镇痛效果。TET1过表达对DRG整体的甲基化水平没有影响,但可以调节DRG内MOR启动子区域的甲基化水平,增加MOR的表达。结论TET1过表达通过调节MOR启动子区域甲基化水平,增加MOR的表达,从而改善神经损伤导致的痛觉过敏和吗啡耐受。
Aim To identify the effect of ten-eleven translocation methylcytosine dioxygenase 1(TET1)on hyperalgesia and morphine tolerance in neuropathic pain(NPP)and the underlying mechanism.Methods The chronic constriction injury(CCI)model was used.Herpes simplex virus expressing full-length TET1 mRNA(HSV-TET1)was microinjected into DRG to overexpress TET1.The effects of TET1 overexpression on the pain sensitivity and morphine tolerance in CCI rats were examined by behavioral testing.The expression ofμ-opioid receptor(MOR)in DRG was detected by Western blot.The total level of 5-methylcytosine(5mC)and 5-hydroxymethylcytosine(5hmC)in DRG gene was measured by DNA blot.The levels of 5mC and 5hmC in MOR promoter region were detected by bisulfite sequencing and Tab-seq technique.Results Compared with control group(sham group),TET1 overexpression could significantly alleviate hyperalgesia and improve morphine tolerance induced by nerve injury,and enhance the analgesic effect of morphine on CCI rats.Overexpression of TET1 did not affect the overall methylation level of DRG,but it could regulate the methylation level of MOR promoter region in DRG and increase the expression of MOR.Conclusions Overexpression of TET1 can improve pain allergy and morphine tolerance induced by nerve injury by regulating the methylation level of MOR promoter region and rescuing the expression level of MOR.
作者
吴强
林静
刘民强
刘丽
何仁亮
WU Qiang;LIN Jing;LIU Min-qiang;LIU Li;HE Ren-liang(Dept of Anesthesiology,the Third People’s Hospital of Shenzhen,Shenzhen Guangdong 518000,China;Supply Center,the Third People’s Hospital of Shenzhen,Shenzhen Guangdong 518000,China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2020年第6期803-808,共6页
Chinese Pharmacological Bulletin
基金
广东省自然科学基金资助项目(No 2018A0303130341)。
