目的观察微小RNA-145(miR-145)对糖尿病神经病理性痛大鼠痛阈的影响,并探讨其可能机制。方法取正常大鼠12只作为对照组(N组),糖尿病神经痛模型制备成功的SD大鼠36只,随机分为三组:糖尿病神经痛组(D组)、糖尿病神经痛-空载组(DN组)和糖...目的观察微小RNA-145(miR-145)对糖尿病神经病理性痛大鼠痛阈的影响,并探讨其可能机制。方法取正常大鼠12只作为对照组(N组),糖尿病神经痛模型制备成功的SD大鼠36只,随机分为三组:糖尿病神经痛组(D组)、糖尿病神经痛-空载组(DN组)和糖尿病神经痛-miR-145过表达组(agomiR-145组),每组12只。N组和D组大鼠鞘内分别注射等剂量生理盐水10μl,DN组大鼠鞘内注射阴性对照病毒(10μl,1×106TU/ml),agomiR-145组大鼠鞘内注射agomiR-145(10μl,1×106TU/ml)。于鞘内注药前1 d和注药后1、3、7、14 d测定四组大鼠机械缩足痛阈(MWT)和热缩足反射潜伏期(TWL)。然后采用RT-PCR法检测大鼠背根神经节(DRG)中miR-145mRNA相对表达量,免疫荧光检测大鼠背根神经节Nav1.8阳性细胞荧光相对强度。荧光素酶报告实验验证miR-145的靶基因。结果注药前1 d D组、DN组和agomiR-145组MWT明显低于、TWL明显短于N组(P<0.05);注药后3、7、14 d agomiR-145组MWT明显高于D组和DN组(P<0.05);注药后7、14 d agomiR-145组大鼠TWL明显长于D组和DN组(P<0.05)。D组和DN组DRG中miR-145 mRNA相对表达量明显低于N组和agomiR-145组,电压门控钠离子通道1.8(Nav1.8)阳性细胞明显多于N组和agomiR-145组(P<0.05)。荧光素酶报告实验验证miR-145作用于Nav 1.8 3'-非翻译区并调控Nav1.8的表达。结论 miR-145通过下调背根神经节Nav 1.8表达缓解糖尿病大鼠机械痛敏和热痛敏。展开更多
Neuropathic pain was produced by chronic constriction injury of the sciatic nerve in rats. Behaviora tests showed that the thresholds for thermal and mechanical hyperalgesia were significantly reduced in neuropathic p...Neuropathic pain was produced by chronic constriction injury of the sciatic nerve in rats. Behaviora tests showed that the thresholds for thermal and mechanical hyperalgesia were significantly reduced in neuropathic pain rats 3 28 days following model induction. The results of immunohistochemistry, western blot assays and reverse transcription-PCR showed that Nay1.7 protein and mRNA expression was significantly increased in the injured dorsal root ganglia. These findings indicated that Nay1.7 might play an important role in the model of chronic neuropathic pain展开更多
文摘目的观察微小RNA-145(miR-145)对糖尿病神经病理性痛大鼠痛阈的影响,并探讨其可能机制。方法取正常大鼠12只作为对照组(N组),糖尿病神经痛模型制备成功的SD大鼠36只,随机分为三组:糖尿病神经痛组(D组)、糖尿病神经痛-空载组(DN组)和糖尿病神经痛-miR-145过表达组(agomiR-145组),每组12只。N组和D组大鼠鞘内分别注射等剂量生理盐水10μl,DN组大鼠鞘内注射阴性对照病毒(10μl,1×106TU/ml),agomiR-145组大鼠鞘内注射agomiR-145(10μl,1×106TU/ml)。于鞘内注药前1 d和注药后1、3、7、14 d测定四组大鼠机械缩足痛阈(MWT)和热缩足反射潜伏期(TWL)。然后采用RT-PCR法检测大鼠背根神经节(DRG)中miR-145mRNA相对表达量,免疫荧光检测大鼠背根神经节Nav1.8阳性细胞荧光相对强度。荧光素酶报告实验验证miR-145的靶基因。结果注药前1 d D组、DN组和agomiR-145组MWT明显低于、TWL明显短于N组(P<0.05);注药后3、7、14 d agomiR-145组MWT明显高于D组和DN组(P<0.05);注药后7、14 d agomiR-145组大鼠TWL明显长于D组和DN组(P<0.05)。D组和DN组DRG中miR-145 mRNA相对表达量明显低于N组和agomiR-145组,电压门控钠离子通道1.8(Nav1.8)阳性细胞明显多于N组和agomiR-145组(P<0.05)。荧光素酶报告实验验证miR-145作用于Nav 1.8 3'-非翻译区并调控Nav1.8的表达。结论 miR-145通过下调背根神经节Nav 1.8表达缓解糖尿病大鼠机械痛敏和热痛敏。
基金This study was supported by the National Natural Science Foundation of China(The mechanism of Na_v 1.7 (encoded by SCN9A gene) activated by chronic neuropathic pain and the experimental study of analgesia by SCN9A RNAi), No.81171059
文摘Neuropathic pain was produced by chronic constriction injury of the sciatic nerve in rats. Behaviora tests showed that the thresholds for thermal and mechanical hyperalgesia were significantly reduced in neuropathic pain rats 3 28 days following model induction. The results of immunohistochemistry, western blot assays and reverse transcription-PCR showed that Nay1.7 protein and mRNA expression was significantly increased in the injured dorsal root ganglia. These findings indicated that Nay1.7 might play an important role in the model of chronic neuropathic pain
