ADAR2在神经病理性疼痛小鼠中的表达及编辑活性

Expression and editing activity of ADAR2 in mice with neuropathic pain

目的探讨RNA腺苷脱氨酶2(ADAR2)在神经病理性疼痛(NP)小鼠中的表达及编辑活性。方法40只C57BL/6J小鼠随机分为两组,每组20只。模型组小鼠坐骨神经主干均匀间隔结扎4道,构建慢性缩窄性损伤模型小鼠;假手术组仅暴露坐骨神经但不结扎。在术前及术后3、7、14、21、28 d观察小鼠行为学表现,采用逆转录PCR检测小鼠背根神经节中ADAR2 mRNA表达及编辑活性,Western blot和免疫荧光法检测ADAR2蛋白的表达。结果与假手术组比较,模型组小鼠在术后3、7、14、21、28 d机械缩足反射阈值和热缩足反射潜伏期下降(P<0.05)。术后7 d,模型组ADAR2 mRNA和蛋白表达、ADAR2选择性剪接体表达量以及选择性剪接率均高于假手术组(P<0.05)。结论ADAR2在NP小鼠中的表达及编辑活性上调,可能在NP的发生、发展中起到重要作用。

Objective To analyze the expression and editing activity of adenosine deaminase acting on RNA 2(ADAR2)in the mice with neuropathic pain(NP).Methods Forty C57 BL/6 J mice were divided into two groups with 20 mice each.The mice model with chronic constriction injury in group A was constructed by ligating the sciatic nerve trunk uniformly spaced in 4 lanes.The sciatic nerve in group C was exposed but not ligated as the control.The behavioral performance of the mice was observed before operation and on the 3^(rd),7^(th),14^(th),21^(st) and 28^(th) day after operation.The mRNA expression and editing activity of ADAR2 in the dorsal root ganglion were detected by reverse transcription PCR.The protein expression of ADAR2 was detected by Western blot and immunofluorescence.Results Compared with group C,the paw mechanical withdraw threshold and paw withdrawal latency were lower in group A on the 3^(rd),7^(th),14^(th),21^(st) and 28^(th) day after operation(P<0.05).The mRNA and protein expressions of ADAR2,ADAR2 alternative spliceosome expression and alternative splicing rate were higher in group A than those in group C on the 7^(th) day after operation(P<0.05).Conclusion The expression and editing activity of ADAR2 in NP mice are higher,which may play an important role in the occurrence and development of NP.