摘要
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。
Objective To observe the effect of miR-145 on pain threshold and explore the possible underlying positive role of miR-145 in rats with diabetic neuropathic pain. Methods The total of 36 rats with diabetic neuropathic pain were randomly divided into three groups respectively with normal control group （group N） （n=12 for each group）： diabetic neuropathic pain （DNP） group （group D）, DNP-NC group （group DN） and DNP-agomiR-145 group （group agomiR-145）. The rats received agomiR-145 intrathecal injection in group agomiR-145 （10 μl, 1×106TU/ml）, or the negative control virus in group DN （10 μl, 1×106TU/ml）, or equal volume of normal saline in other two groups. Paw mechanical withdrawal threshold （MWT） and paw withdrawal latency （TWL） were measured on the day before intrathecal injection and day 1, days 3, 7 and 14 after intrathecal injection. On the days 14 after pain-related behavioral test, the RNA expression of miR-145 in the dorsal root ganglion （DRG） was detected using reverse transcription-quantitative polymerase chain reaction （RT-PCR） assay and the expression of Nav 1.8 in DRG were detected by fluorescent immunofluorescence. In addition, a dual luciferase activity assay was used to testify the target genes of miR-145. Results MWT and TWL were decreased at 1 d before intrathecal injectionin groups D, DN and agomiR-145 than that in group N （P〈0.05）. The significant increase of MWT was observed in group agomiR-145 on day 3, 7, 14 than those in group D and group DN （P〈0.05）. TWL in group agomiR-145 was increased significantly on day 7 and day 14 compared with those in groups D and DN （P〈0.05）. Compared with group N, miR-145 expression level in DRG in groups D and DN were significantly lower （P〈0.05）. In addition, the protein expression of Nav1.8 was significantly increased in group D and DN compared with that in group N （P〈0.05）. Compared with groups D and DN, miR-145 expression was increased significantly and the expression of Nav1.8
作者
颜洁
沈洁
韩翠翠
汤永峰
沈晓凤
YAN Jie, SHEN Jie, HAN Cuicui , TANG Yongfeng , SHEN Xiaofeng.(Department of Anesthesiology, Obsterics and Gynecology Hospital Affiliated to Nanjing Medical University, Nanjing 210004, Chin)
出处
《临床麻醉学杂志》
CAS
CSCD
北大核心
2018年第3期291-295,共5页
Journal of Clinical Anesthesiology
基金
国家自然科学基金青年科学基金(81400913)
