蠕虫来源的LNFPⅢ对周围性神经病理性疼痛的病理发生和脊髓胶质细胞活化的影响

Helminth-derived Immunomodulatory Glycan LNFP Ⅲ Impairs Pathogenesis of Peripheral Neuropathic Pain and Spinal Glial Activation

中文关键词:  周围神经 神经胶质细胞 神经病理性疼痛 LNFPⅢ

英文关键词:Peripheral nerves Neuroglial cells Neuropathic pain LNFPⅢ

中文摘要:

      目的 探讨蠕虫来源的免疫调控性糖分子乳-N-岩藻五糖Ⅲ(lacto-N-fucopentaose Ⅲ, LNFPⅢ)对成年大鼠胫神经永久性横断伤(即改良型备用性神经损伤，modified spared nerve injury，mSNI)后神经病理性疼痛的病理发生的影响，以及它对此时脊髓胶质细胞活化的影响。方法 成年雄性SD大鼠随机分为4组，即假手术组（n =6）、mSNI手术组（n =6）、mSNI手术+牛血清白蛋白（BSA）组（n =12）和mSNI手术+LNFPⅢ组（n =12）。各组动物行右侧mSNI手术或者相应的假手术，根据实验分组腹腔注射BSA或LNFPⅢ与BSA的结合物。每组取6只大鼠进行足底试验、von Frey纤维试验、针刺试验和丙酮试验，测试手术前、后手术同侧和对侧大鼠后爪足底面腓肠神经和隐神经皮肤区域特异性的疼痛反应。制备mSNI手术+BSA组和mSNI手术+LNFPⅢ组手术后7 d和14 d（各组每个时间点3只动物）的第3～第4腰椎（L3～4）脊髓节段冰冻切片，进行小胶质/巨噬细胞标志物分化抗原簇分子11b(cluster of differentiation molecule 11b, CD11b)和星形胶质细胞标志物胶质纤维酸性蛋白(glial fibrillary acidic protein, GFAP)的免疫荧光染色。结果 成年大鼠mSNI后，早期系统性给予LNFPⅢ可显著缓解手术后急性诱导期（4/5 d）和慢性转化期（7/8 d和14/15 d）中手术侧后爪足底面腓肠神经和隐神经皮肤区域特异性的机械和温度刺激诱发的病理性疼痛。同时，早期系统性给予LNFPⅢ抑制mSNI大鼠手术侧脊髓背角中术后7 d小胶质/巨噬细胞以及术后7 d和14 d星形胶质细胞的活化。 结论 早期系统性给予LNFPⅢ能抑制成年大鼠mSNI后神经病理性疼痛的病理发生（包括急性诱发和慢性转化）以及该过程中脊髓胶质细胞的活化。

英文摘要:

      Objective To investigate the effect of helminth-derived immunomodulatory glycan lacto-N-fucopentaose Ⅲ（LNFPⅢ） on the pathogenesis of neuropathic pain and spinal glial activation in the corresponding time windows after adult rat tibial nerve permanent transection (modified spared nerve injury, mSNI). Methods Ten weeks old male adult Sprague-Dawley (SD) rats weighing 250-300 g were randomly grouped into four groups: sham-operated group (n =6), mSNI group (n =6), mSNI plus bovine serum albumin (BSA) group (n =12) and mSNI plus LNFPⅢ group (n =12). Rats were subjected to surgical operation or sham operation on the right tibial nerves and were intraperitoneal injected BSA or LNEPⅢ-BSA conjugates by the group design. Animals from each group (n =6 per group) were subjected to the plantar test, von Frey hairs test, pinprick test and acetone test for critical evaluation of region-specific pain responses on the plantar sural and saphenous skin territories of ipsilateral and contralateral hindpaws after injuries. Transverse frozen sections of L3-4 spinal cords from the remaining animals of mSNI plus BSA group and mSNI plus LNFPⅢ group 7 and 14 d after injury (n =3 for each time point per group) were prepared and subjected to immunofluorescent staining of microglia/macrophage marker 〔cluster of differentiation molecule 11b (CD11b)〕 and astrocyte marker 〔glial fibrillary acidic protein (GFAP)〕, for analysis of spinal glial activation. Results After adult rat mSNI, early systematic administration of LNFPⅢ significantly but not completely attenuated region-specific pathological pain evoked by mechanical and thermal stimuli on the sural and saphenous skin territories of rat hindpaw plantar surfaces in acute (4/5 d after injuries) and subacute (7/8 d and 14/15 d after injuries) phases. Meanwhile, in the ipsilateral spinal cord dorsal horns, this early systematic treatment inhibited microglia/macrophage activation 7 d after injury and astrocyte activation 7 and 14 d after injury. Conclusion Early systematic administration of LNFPⅢ impairs the pathogenesis (acute induction and chronic transition) of neuropathic pain and spinal glial activation in the corresponding time windows after adult rat mSNI.