中华口腔医学杂志
中華口腔醫學雜誌
중화구강의학잡지
Chinese Journal of Stomatology
2014年
2期
95-100
,共6页
许乐%祝心威%陈青峰%胡媛平%朱凌%蒋勇
許樂%祝心威%陳青峰%鬍媛平%硃凌%蔣勇
허악%축심위%진청봉%호원평%주릉%장용
牙髓炎%动物,实验%牙痛%钠通道,电压门控
牙髓炎%動物,實驗%牙痛%鈉通道,電壓門控
아수염%동물,실험%아통%납통도,전압문공
Pulpitis%Animals,laboratory%Toothache%Sodium channels,voltage-gated
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.
目的 通過檢測電壓門控鈉離子通道Nav1.9在牙髓炎動物模型牙髓中的錶達,探討Nav1.9與炎性疼痛的關繫.方法 將建立牙髓炎模型的36隻大鼠分為3箇實驗組:造模後1、3、5d組,每組12隻;以12隻健康大鼠作為正常對照組.反轉錄PCR(reverse transcription PCR,RT-PCR)法檢測各組牙髓中腫瘤壞死因子α(tumor necrosis factor-α,TNF-α)和Nav1.9 mRNA的錶達,酶聯免疫吸附測定(enzyme linked immunosorbent assay,ELISA)法和蛋白質印跡法檢測各組Navl.9蛋白的錶達,併對各組間的差異進行單因素方差分析.結果 RT-PCR結果顯示,3箇實驗組TNF-α的錶達(ld組:0.514±0.098,3d組:0.739 ±0.104,5 d組:1.238 ±0.082)均顯著高于正常對照組(0.147±0.016),各組間差異均有統計學意義(P＜0.01);3箇實驗組牙髓中Nav1.9 mRNA相對錶達量(1d組:0.296 ±0.038,3 d組:0.409±0.013,5d組:0.487 ±0.028)與正常對照組(0.223±0.020)相比均呈顯著上升趨勢,各組間差異均有統計學意義(p＜0.05).ELISA結果顯示,正常對照組、1d、3d及5d組牙髓中Nav1.9的錶達量分彆為(4.013±0.292)、(5.143±0.101)、(5.835±0.088)及(6.307 ±0.137) μg/L,各組間差異均有統計學意義(P＜0.05).蛋白質印跡法結果顯示,3箇實驗組Nav1.9蛋白相對錶達量(1d組:0.106±0.007,3d組:0.170 ±0.013,5d組:0.238±0.004)均顯著高于正常對照組(0.073±0.004)(P ＜0.05).結論 Nav1.9在疼痛牙髓中錶達增彊併隨疼痛加重而升高,提示Nav1.9與炎性牙痛感覺過程的髮生相關.
목적 통과검측전압문공납리자통도Nav1.9재아수염동물모형아수중적표체,탐토Nav1.9여염성동통적관계.방법 장건립아수염모형적36지대서분위3개실험조:조모후1、3、5d조,매조12지;이12지건강대서작위정상대조조.반전록PCR(reverse transcription PCR,RT-PCR)법검측각조아수중종류배사인자α(tumor necrosis factor-α,TNF-α)화Nav1.9 mRNA적표체,매련면역흡부측정(enzyme linked immunosorbent assay,ELISA)법화단백질인적법검측각조Navl.9단백적표체,병대각조간적차이진행단인소방차분석.결과 RT-PCR결과현시,3개실험조TNF-α적표체(ld조:0.514±0.098,3d조:0.739 ±0.104,5 d조:1.238 ±0.082)균현저고우정상대조조(0.147±0.016),각조간차이균유통계학의의(P＜0.01);3개실험조아수중Nav1.9 mRNA상대표체량(1d조:0.296 ±0.038,3 d조:0.409±0.013,5d조:0.487 ±0.028)여정상대조조(0.223±0.020)상비균정현저상승추세,각조간차이균유통계학의의(p＜0.05).ELISA결과현시,정상대조조、1d、3d급5d조아수중Nav1.9적표체량분별위(4.013±0.292)、(5.143±0.101)、(5.835±0.088)급(6.307 ±0.137) μg/L,각조간차이균유통계학의의(P＜0.05).단백질인적법결과현시,3개실험조Nav1.9단백상대표체량(1d조:0.106±0.007,3d조:0.170 ±0.013,5d조:0.238±0.004)균현저고우정상대조조(0.073±0.004)(P ＜0.05).결론 Nav1.9재동통아수중표체증강병수동통가중이승고,제시Nav1.9여염성아통감각과정적발생상관.
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.
