电压门控钠离子通道Nav l.9在牙髓炎牙髓中表达的动物实验

摘要

Objective To investigate the relationship between pulpitis pain and voltage-gated sodium channel(Nav 1.9) by detecting the expression of Nav 1.9 at different time points of the rat pulpal lesion model.Methods Thirty-six SD pulpal lesions rat models were divided into three experimental groups,1 d (n =12),3 d (n =12) and 5 d group(n =12).Normal SD rats served as control(n =12).Tumor necrosis factor-α(TNF-α) and Nav 1.9 mRNA expressions were evaluated by reverse transcription PCR(RT-PCR).Nav1.9 protein expressions were analyzed by enzyme-linked immunosorbent assay (ELISA) and Western blotting.Results The expression of TNF-α in the experimental group (1 d:0.514 ± 0.098,3 d:0.739 ±0.104,5 d:1.238 ± 0.082) was higher than those in the control group (0.147 ± 0.016) (P ＜0.01).Navl.9 mRNA was up-regulated markedly in experimental groups (1 d:0.296 ± 0.038,3 d:0.409 ±0.013,5 d:0.487 ±0.028),compare with control group(0.223 ±0.020) (P ＜0.05).The ELISA results revealed that the level of Nav 1.9 in control pulp tissue was (4.013 ±0.292) μg/L,in painful pulp tissue of 1 d group was (5.143 ± 0.101) μg/L,in painful pulp tissue of 3 d group was (5.835 ±0.088) μg/L and in painful pulp tissue of 5 d group was (6.307 ± 0.137) μg/L (P ＜ 0.05).Western blotting showed the expression of Navl.9 in experimental groups(1 d:0.106 ±0.007,3 d:0.170 ±0.013,5 d:0.238 ± 0.004) was up-regulated significantly compared with control group (0.073 ± 0.004)(P ＜ 0.05).Conclusions The level of Nav 1.9 had a significant inerease in painful pulp tissue.Moreover,with the degree of pain aggravation,the expression of Nav.l.9 increased in pulp tissue.It suggests that Nav 1.9 may play an important role in the development of pulpitis pain.%目的 通过检测电压门控钠离子通道Nav1.9在牙髓炎动物模型牙髓中的表达,探讨Nav1.9与炎性疼痛的关系.方法 将建立牙髓炎模型的36只大鼠分为3个实验组:造模后1、3、5d组,每组12只;以12只健康大鼠作为正常对照组.反转录PCR(reverse transcription PCR,RT-PCR)法检测各组牙髓中肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)和Nav1.9 mRNA的表达,酶联免疫吸附测定(enzyme linked immunosorbent assay,ELISA)法和蛋白质印迹法检测各组Navl.9蛋白的表达,并对各组间的差异进行单因素方差分析.结果 RT-PCR结果显示,3个实验组TNF-α的表达(ld组:0.514±0.098,3d组:0.739 ±0.104,5 d组:1.238 ±0.082)均显著高于正常对照组(0.147±0.016),各组间差异均有统计学意义(P＜0.01);3个实验组牙髓中Nav1.9 mRNA相对表达量(1d组:0.296 ±0.038,3 d组:0.409±0.013,5d组:0.487 ±0.028)与正常对照组(0.223±0.020)相比均呈显著上升趋势,各组间差异均有统计学意义(p＜0.05).ELISA结果显示,正常对照组、1d、3d及5d组牙髓中Nav1.9的表达量分别为(4.013±0.292)、(5.143±0.101)、(5.835±0.088)及(6.307 ±0.137) μg/L,各组间差异均有统计学意义(P＜0.05).蛋白质印迹法结果显示,3个实验组Nav1.9蛋白相对表达量(1d组:0.106±0.007,3d组:0.170 ±0.013,5d组:0.238±0.004)均显著高于正常对照组(0.073±0.004)(P ＜0.05).结论 Nav1.9在疼痛牙髓中表达增强并随疼痛加重而升高,提示Nav1.9与炎性牙痛感觉过程的发生相关.