摘要
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）20μl＋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＋NS20μl（D组）,SNL＋MM-ODN20μg/d（E组）,SNL＋AS-ODN20μg/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。
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 intrathecal （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 ） （ 20 μl, i. t. ） ＋ SNL; group B, mismatch oligodeoxynucleotide （ MM-ODN） （20 μg/d, i. t. ） ＋ SNL; group C, antisense oligodeoxynucleotide （AS-ODN） （20 μg/d, i. t. ） ＋ SNL, once a day for 7 consecutive days. In Part Ⅱ ,on the 7th day after SNL another 72 rats were randomly and equally divided into 3 groups： group D, SNL ＋ NS（20 μl,i. t. ） ; group E, SNL ＋ MM- ODN （20 μg/d,i, t. ） ; group F, SNL ＋ AS-ODN （20μg/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 Part Ⅱ after 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 immunohistochemical ABC method was used to determine the expression of GFAP positive cells in the spinal dorsal horn. The rats were killed at -1, 3, 7, 14 day in Part I and 7, 10, 14 day in Part Ⅱ after 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 compared with group A （ P 〈 0. 05 ）, PWL had no significant difference at different time （ P 〉 0.05 ）. Immunohistochemical 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 of TLR3 and IL-6 mRNA were significantly inhibited in group C （P 〈 0.05）. Part Ⅱ ： 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 development of SNL-induced neuropathic pain.
出处
《中国疼痛医学杂志》
CAS
CSCD
2009年第4期230-235,共6页
Chinese Journal of Pain Medicine
