摘要
目的:通过观察吗啡耐受对大鼠痛行为的影响和脊髓后角胶质细胞纤维酸性蛋白阳性产物表达的变化,探讨吗啡耐受对大鼠脊髓后角星形胶质细胞的影响。方法:实验于2003-11/2004-09在同济医院麻醉研究室完成。①造模:SD大鼠18只,随机分为对照组和吗啡耐受组,每组9只。在大鼠L3～5背部纵形切口,将细聚乙烯导管向头端插入蛛网膜下腔2cm,置管后第4天,对照组和吗啡耐受组分别经导管注射生理盐水10μL和吗啡10μL(20μg),1次/d,连续5d。吗啡耐受组注射吗啡后第6天,鞘内注射吗啡10μL(5μg)进行吗啡激发实验。②观察星形胶质细胞激活状态:应用免疫组织化学方法观察星形胶质细胞特异性标志物胶质纤维酸性蛋白染色情况,同时计算平均吸光度值和积分吸光度值,表示星形胶质细胞内胶质纤维酸性蛋白表达的强度(A值越大表示胶质纤维酸性蛋白表达越强)。③测定热伤害性缩腿反应潜伏期及非伤害性机械刺激的敏感程度:置管后第3天分别测定缩腿反应潜伏期和机械性触痛痛阈作为基础值。给药第6天吗啡激发试验前测定机械性触痛痛阈,吗啡激发实验后测定缩腿反应潜伏期,并计算最大镇痛效率。结果:18只大鼠均进入结果分析。①置管第3天及激发实验前缩足总数的变化:置管第3天的两组大鼠缩足阳性总次数基本相近犤(0.11±0.33)次犦,第6天吗啡激发试验前,对照组的缩足总数变化轻微,而吗啡耐受组缩足阳性总次数增多犤(10.66±1.41)次,(P<0.01)犦。②吗啡激发后的最大镇痛效率:对照组显著高于耐受组犤(59.20±4.00)%,(8.86±1.42)%,(P<0.01)犦。③脊髓胶质细胞胶质纤维酸性蛋白阳性表达产物相对面积、光密度和积分光密度,对照组分别少于吗啡耐受组犤3.417±0.268比6.530±0.423(P<0.01),0.357±0.024比0.520±0.025(P<0.01),1.689±0.303比2.310±0.314(P<0.01)犦。结论:脊髓吗啡耐受导致触诱发痛,激活脊髓后角星形胶质细胞,脊髓星形胶质细胞可能在吗啡耐受的形成中发挥重要作用。
AIM： Through the observation of the effects of morphine tolerance on the ache behavior of rats and the changes of expression of die positive product of fiber acidity protein of glial cells on the spinal cord posterior horn, to explore the effects of morphine tolerance on astrocytes of spinal cord posterior horn in rats. METHODS： The experiment was done in the Research Room of Anesthesiology, Tongji Hospital between Novemher 2003 antt September 2004. ①Build models： The 18 SD rats were assigned randomly into control group and morphine tolerance group with 9 rats in each group. The longitudinal incision was cut on the L3-5 hack of rats. Catheterization of micropolyetylen was achieved on 2 cm subarachnoid space. At the 4^th day of putting the tube, the rats in the control group antt morphine tolerance group were treated with 10μL saline and 10μL （20μg）morphine through tube injection, once a day for 5 days. At tbe 6^th day after injecting the morphine, the rats in the morphine toleranee group were treated with 10μL （5μg）morphine by the intrathecal injection to perform morphine excitation experiment. ②To observe the activating status of astroeyte： The specific sign, glial fibriliary acidic protein staining of astrocyte was observed by the immunohistochemical method. Meanwhile, the average absorbance （A） and integra A was calculated to show the expressive intensity of the glial fibriliary acidic protein in the astrocyte （the bigger A value showed that the expression of glial fibriliary acidic protein was more strong）. ③ To detect the response latency of hot injured drawing the legs and sensitive degree of non-injured mechanical irritation： At the 3^ed day of putting tube, the response latency of drawing the legs and mechanical pain threshold were tested respectively as basic value. At the 6^th day of administration, the mechanical pain threshold was detected before morphine excitation trial, and the response latency of drawing the legs was tested after the morphine excitation trial, and the maximum analgesia efficiency was calculated. RESULTS： Totally 18 rats were involved in the result analysis.① The changes of the drawing leg number at the 3^rd day of putting tube and before excitation trial： The total positive number of times of drawing leg at the 3^rd day of putting tube in the two groups was nearly the same [（0.11±0.33） times]. The total number of times of drawing leg had slightly changes in the control group at the 6^th day before the morphine excitation trial, while the total positive number of times of drawing leg in the morphine tolerance group increased [（ 10.66±1.41 ）times, （P 〈 0.01 ）].②The maximum analgesia efficiency after the morphine excitation： It was significantly higher in the control group than that in the tolerance group [（59.20±4.00）%, （8.86±1.42）%, （ P 〈 0.01 ）].③ The relative area, oculus dehter and integra oculus dehter of the positively expressive product of glial fibriliary acidic protein of glial cells on spinal cord were less in the control group than that in the mor phine tolerance group, respectively [3.417±0.268 vs 6.530±0.423 （P 〈 0.01）, 0.357±0.024 vs 0.520±0.025 （P 〈 0.01）, 1.689±0.303 vs 2.310±0.314 （P 〈 0.01）. CONCLUSION： The morphine tolerance in spinal cord induces the ache, activates the astroeytes in spinal dorsal horn. The astrocytes in the spinal cord maybe play an important role in the fomation of morphine tolerance.
出处
《中国临床康复》
CAS
CSCD
北大核心
2005年第32期135-137,F0003,共4页
Chinese Journal of Clinical Rehabilitation
作者简介
简道林，男，1958年生，湖北省宜昌市人，汉族，华中科技大学同济医学院在读博士，主任医师，主要从事麻醉和疼痛治疗研究．
