CAJ | 학술논문

目的:探讨脊髓星形胶质细胞TLR3在神经病理性疼痛中的作用.方法:雄性SD大鼠,体重180～250 g.实验一:72只鞘内置管大鼠随机分为:生理盐水(Ns)20 μl脊神经结扎(SNL)(A 组),错义寡聚核苷酸(MM-ODN)20μg/d+SNL(B组),反义寡聚核苷酸(AS.ODN)20μg/d+SNL (C组).实验二:72只SNL后7天大鼠随机分为:SNL+NS 20μl(D组),SNL+MM-ODN 20μg/d(E组),SNL+AS-ODN 20μg/d(F组).实验一和实验二分别于SNL前1天和后7天开始鞘注,每天一次,共7天;于术后-1、1、3、5、7、10、14天和-1、7、10、12、14天行行为学实验.实验一和实验二分别于术后7天和14天每组各随机取6只大鼠利用免疫组织化学法观察脊髓背角GFAP的表达,并于术后-1、3、7、14天和术后7、10、14天每组各随机取6只大鼠断头处死,取L4～5脊髓节段采用RT-PCR法测定TLR3和IL-6 mRNA的表达.结果:实验一:与A组比较,C组PWPT升高并持续至术后14天(P<0.05),C组各时间点PWL无明显变化(P>0.05);与A组和B组比较,C组脊髓背角GFAP免疫反应阳性产物表达相对面积分别下降46.4%(P<0.01)和45.7%(P<0.01);与A组和B组比较,C组鞘内注射AS-ODN抑制了TLR3受体和IL-6 mRNA的表达.实验二D组、E组和F组在PWPT、PWL、GFAP表达和TLR3及IL-6 mRNA表达方面的差异无统计学意义(P>0.05).结论:脊髓背角星形胶质细胞TLR3可能参与了神经病理性疼痛的发生与发展.
목적:탐토척수성형효질세포TLR3재신경병이성동통중적작용.방법:웅성SD대서,체중180～250 g.실험일:72지초내치관대서수궤분위:생리염수(Ns)20 μl척신경결찰(SNL)(A 조),착의과취핵감산(MM-ODN)20μg/d+SNL(B조),반의과취핵감산(AS.ODN)20μg/d+SNL (C조).실험이:72지SNL후7천대서수궤분위:SNL+NS 20μl(D조),SNL+MM-ODN 20μg/d(E조),SNL+AS-ODN 20μg/d(F조).실험일화실험이분별우SNL전1천화후7천개시초주,매천일차,공7천;우술후-1、1、3、5、7、10、14천화-1、7、10、12、14천행행위학실험.실험일화실험이분별우술후7천화14천매조각수궤취6지대서이용면역조직화학법관찰척수배각GFAP적표체,병우술후-1、3、7、14천화술후7、10、14천매조각수궤취6지대서단두처사,취L4～5척수절단채용RT-PCR법측정TLR3화IL-6 mRNA적표체.결과:실험일:여A조비교,C조PWPT승고병지속지술후14천(P<0.05),C조각시간점PWL무명현변화(P>0.05);여A조화B조비교,C조척수배각GFAP면역반응양성산물표체상대면적분별하강46.4%(P<0.01)화45.7%(P<0.01);여A조화B조비교,C조초내주사AS-ODN억제료TLR3수체화IL-6 mRNA적표체.실험이D조、E조화F조재PWPT、PWL、GFAP표체화TLR3급IL-6 mRNA표체방면적차이무통계학의의(P>0.05).결론:척수배각성형효질세포TLR3가능삼여료신경병이성동통적발생여발전.
Objective:To investigate the analgesic effect of TLR3 of spinal astrocytes on neuropathic pain induced by spinal nerve ligation(SNL).Methods:Male SD rats weighing 180～250 g were included in this experiment.Catheter was placed intrathecally with a tip located at lumbo-sacral segment of spinal cord for intratheeal(IT)drug administration in all animals.Neuropathic pain Was induced by ligation of right L5 spinal nerve with 4-0 silk thread.In Part I,72 animals were randomly and equally divided into 3 groups:group A,normal saline(NS)(20μl,i.t.)+SNL;group B,mismatch oligodeoxynucleotide (MM-ODN)(20 ixg/d,i.t.)+SNL;group C,antisense oligodeoxynucleotide(AS-ODN)(20μg/d,i. t.)+SNL,once a day for 7 consecutive days.In PartⅡ.on the 7th day after SNL another72 rats were randomly and equally divided into 3 groups:group D,SNL+NS(20μl,i.t.);group E,SNL+MM- ODN(20μg/d,i.t.);group F,SNL+AS-ODN(20 ixg/d,i.t.),once a day for 7 consecutive days. Pain threshold was estimated by measuring paw withdrawal pressure threshold(PWPT)and latency (PWL)with Von Frey filament stimulation at-1(1 d before operation),1,3,5,7,10,14 day in Part I and.1,7,10,12,14 day in PartⅡafter SNL,respectively.The animals were killed on the 7th(Part I)or 14th(Part Ⅱ)day after SNL and lumbar segments of spinal cord were removed and immunohisto- chemical ABC method was used to determine the expression of GFAP positive cells in the spinal dorsal horn.The rats were kiUed at-1,3,7,14 day in Part I and 7,10,14 day in PartⅡafter operation in each group and the L4～5 segment of the spinal cord was removed for determination of expression of TLR3 and IL-6 mRNA by RT-PCR.Results:Part I:In group C,PWPT Was significantly increased when com- pared with group A(P<0.05),PWL had no significant difference at different time(P>0.05).Immu- nohistochemical results indicated that the relative area of staining of GFAP immuno-reactive cells in group C decreased 46.4%and 45.7%(P<0.05)respectively,compared with that in group A and B.The expressions ofTLR3 and IL-6 mRNA were significantly inhibited in group C(P<0.05).PartⅡ:There was no significant difference in the above-mentioned five markers among group D,group E and group F (P>0.05).Conclusion:TLR3 of spinal astrocytes in the spinal dorsal horn may be involved in the de- velopment of SNL-induced neuropathic pain.