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目的研究大鼠延髓内脏带(MVZ)内星形胶质细胞与神经元对选择性性神经病理性痛的反应。方法25只成年SD大鼠,其中15只为接受左侧坐骨神经分支选择性损伤(SNI)组,5只作为假手术组,5只为对照组。在术前、术后10d、20d、30d(每时段5只)观察大鼠疼痛行为学并检测机械刺激缩足反射阈值(PWMT)的变化;应用抗Fos蛋白与抗胶质原纤维酸性蛋白(GFAP)或抗Fos蛋白与抗酪氨酸羟化酶(TH)的单一或双重免疫荧光法染色,Confocal显微镜下观察延髓MVZ内GFAP标记的星形胶质细胞的荧光强度,Fos/GFAP双标记星形胶质细胞以及Fos/TH双标记神经元的平均数。结果SNI组大鼠与对照组或假手术组相比,术后10d其痛敏增高(PWMT值降低),20d痛敏达到高峰(PWMT值最低)。免疫荧光染色显示:SNI术后MVZ内星形胶质细胞表现为激活型,GFAP免疫荧光强度明显增加;孤束核及腹外侧区的Fos/GFAP双标记星形胶质细胞及Fos/TH双标记神经元的平均值显著增高,SNI术后20d达到高峰。结论SNI术后MVZ内的星形胶质细胞和神经元均被激活。
Objective To study the response of astrocytes and neurons in medullary visceral zone (MVZ) to selective neuropathic pain in rats. Methods Twenty-five adult Sprague-Dawley rats were randomly divided into three groups: the left sciatic nerve branch selective injury (SNI) group, the five sham-operated group and the control group. The pain behavior was observed at 10 days, 20 days and 30 days after operation (5 in each time period), and the threshold of mechanical stimulus (PWMT) was measured. The anti-Fos protein and anti-glial fibrillary acidic protein GFAP) or anti-Fos protein and anti-tyrosine hydroxylase (TH) single or double immunofluorescence staining, Confocal microscopic observation of medulla oblongata MVZ GFAP-labeled astrocytes fluorescence intensity, Fos / GFAP double marker Astrocytes and the average number of Fos / TH double-labeled neurons. Results Compared with the control group or the sham operation group, the rats in the SNI group showed higher hyperalgesia (reduced PWMT value) 10 days after operation and peak pain sensitivity (lowest PWMT value) after 20 days. Immunofluorescence staining showed that astrocytes in MVZ showed activation type after SNI, and the intensity of GFAP immunofluorescence increased obviously. Fos / GFAP double labeled astrocytes and Fos / TH The average value of labeled neurons was significantly increased, peaked 20 days after SNI. Conclusion Both astrocytes and neurons in MVZ after SNI are activated.