鞘内移植人前脑啡肽原基因修饰的永生化大鼠星形胶质细胞株对大鼠慢性神经痛的镇痛作用  

Analgesic effect of intrathecal transplant of immortalized rat astrocyte strain genetically modified by human preproenkephalin gene on rat chronic neuropathic pain

作　　者：安珂[1] 田玉科[1] 杨辉[1] 王贤裕[1] 金小高[1] 高峰[1] 徐颖[1] 田学愎[1] 王鹏[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院麻醉学教研室

出　　处：《中华医学杂志》2005年第38期2711-2714,共4页National Medical Journal of China

基　　金：国家自然科学基金资助项目(30170905)

摘　　要：目的探讨鞘内移植人前脑啡肽原基因修饰的永生化大鼠星形胶质细胞株(IAST/hPPE)对大鼠慢性神经痛的镇痛作用。方法成年雄性SD大鼠40只,随机分为四组:未处理组、SNI组、SNI+IAST组和SNI+IAST/hPPE组;除未处理组外,建立右侧下肢保留性神经损伤模型(SNI);1周后于脊髓L4-6水平鞘内移植与5′溴-2-脱氧尿苷(B rdU)共培养的永生化大鼠星形胶质细胞株(IAST)或IAST/hPPE细胞株;每周测定各组大鼠左右两侧50%缩足阈值(PWT)的差值及腹腔注射纳洛酮对其镇痛效应的影响;免疫组织化学和放射免疫法检测脊髓组织中亮氨酸脑啡肽(L-EK)的含量变化以及移植细胞B rdU的表达。结果SNI后1周大鼠右足即出现痛觉异常现象,与未处理组相比,其余三组的左右侧50%PWT的差值明显增大(均P<0.01);SNI+IAST/hPPE组痛觉异常症状明显减轻,而SNI组和SNI+IAST组未见明显变化;SNI+IAST/hPPE组左右侧50%PWT差值与SNI组和SNI+IAST组相比明显减小,P<0.01;SNI+IAST/hPPE组的镇痛效应可被纳洛酮拮抗;与未处理组相比,其余3组每毫克脊髓L-EK的含量明显升高(均P<0.01),SNI+IAST/hPPE组(108.1 pg/mg±12.5 pg/mg)明显高于SNI组和SNI+IAST组(均P<0.01),而SNI组和SNI+IAST组相比差异无统计学意义;移植6周后,脊髓背角移植细胞的B rdU免疫染色阳性,表明细胞仍然存活。结论鞘内移植IAST/hPPE细胞可以抑制慢性神经痛大鼠的痛觉异常行为,该效应与移植细胞持续分泌的脑啡肽作用于阿片受体有关。Objective To observe the analgesic effect of intrathecal transplant of immortalized rat astrocyte genetically modified by human preproenkephalin gene （IAST/hPPE）on chronic neuropathic pain. Methods 40 adult male Sprague-Dawley rats were randomly divided into four groups, 10 rats for each group. Naive group, SNI group, SNI ＋ IAST group and SNI ＋ IAST/hPPE group. The immortalized rat astrocyte （IAST） or IAST/hPPE co-incubated with bromodeoxyuridine（BrdU） in vitro were transplanted in the lumbar 4 to 6 subarachnoid space near the spinal cord 1 week after right side spared nerve injury （ SNI ）. All animals were tested for bilateral 50% hindpaw withdrawal threshold（PWT） to a graded series of Von Frey hairs stimulation once a week from one week before SNI to six weeks after transplant, the difference value for right 50% PWT minus left 50% PWT was calculated and the effect of intraperitoneal naloxone on the analgesic efficacies was also observed. The content of L-EK in the spinal cord of L4-6 and was determined using immunohistochemistry and radioimmunoassay, and the expression of BrdU in grafts was determined using immunohistochemistry. Results Allodynia-like behaviour after 1 week following SNI was observed.As compared with Naive group, the difference value for the right 50% PWT minus left 50% PWT in the other three groups was higher significantly （ P 〈 0.01 ） . The tactile allodynia induced by SNI was significantly alleviated during the 1 to 6 week period after transplantation of IAST/hPPE cells, but transplants of lAST cells had no effect on the allodynia-like behaviour. The difference value for the right 50% PWT minus left 50% PWT in SNI ＋ IAST/hPPE group was lower significantly than that in the SNI and SNI ＋ IAST group （P 〈0.01 ） , but there was no significant difference between SNI and SNI ＋ IAST group （ P 〉 0. 05 ） . The efficacies were reversed by intraperitoneal naloxone in SNI ＋ IAST/hPPE group. The content of L-EK in the lumbar spinal cord in IAST/hPPE group

关 键 词：脑啡肽类 星形细胞 移植 神经痛 多瘤病毒转化 慢性神经痛 前脑啡肽原 SD大鼠 移植细胞 星形胶质 

分 类 号：R614[医药卫生—麻醉学]