三叉神经节卫星胶质细胞P2X7受体在偏头痛大鼠模型中作用的研究
发布时间：2018-11-16 16:53
【摘要】：目的通过大鼠腹腔注射硝酸甘油(GTN)建立急性偏头痛模型。造模成功后使用Western blot技术检测大鼠三叉神经节上P2X7受体和c-fos的表达变化;使用免疫荧光检测卫星胶质细胞上P2X7受体和胶质纤维酸性蛋白(GFAP)的表达情况。最后通过腹腔注射P2X7受体抑制剂亮蓝(BBG)观察其对三叉神经脊束核c-fos和降钙素基因相关肽(CGRP)表达的影响,来探究P2X7受体是否参与偏头痛的发病及其可能的机制。材料和方法1.Sprague-Dawley大鼠10只,随机分为2组:腹腔注射生理盐水组(n=5,2ml/Kg)和腹腔注射GTN组(n=5,10mg/Kg)。给药前使用Von Frey测痛仪测量各组大鼠后爪上的基础痛觉阈值,之后分别在给药第1、2和4小时时再次测量大鼠后爪上的痛觉阈值。使用免疫荧光观察大鼠三叉神经节P2X7受体和GFAP的表达。2.Sprague-Dawley大鼠20只,随机分为4组:正常组(n=5)和腹腔注射GTN 1小时组(n=5,10mg/Kg)、腹腔注射GTN 2小时组(n=5,10mg/Kg)和腹腔注射GTN 4小时组(n=5,10mg/Kg)。Western blot技术检测大鼠三叉神经节上P2X7受体和c-fos的表达变化。3.Sprague-Dawley大鼠20只,随机分为4组:腹腔注射生理盐水2小时组(n=5,2ml/Kg)、腹腔注射GTN 2小时组(n=5,10mg/Kg)、腹腔注射GTN+生理盐水2小时组(n=5,10mg/Kg,2ml/Kg)和腹腔注射GTN+BBG 2小时组(n=5,10mg/Kg)。使用Western blot技术检测大鼠三叉神经脊束核c-fos和CGRP的表达。结果1.痛觉阈值:腹腔注射GTN后,大鼠后爪上的痛觉阈值开始下降,在第2小时下降最为明显,明显低于生理盐水组(p0.05)。2.免疫荧光:三叉神经节上的P2X7受体和GFAP共表达。3.Western blot:腹腔注射GTN后,大鼠卫星胶质细胞P2X7受体表达开始下降,在第2小时下降最为明显,明显低于正常组(p0.05)。而三叉神经节和三叉神经脊束核c-fos和CGRP的表达明显高于正常组(p0.05)。在使用P2X7受体抑制剂BBG后c-fos和CGRP增加的更加明显(p0.05)。结论1.P2X7受体特异性表达在大鼠三叉神经节卫星胶质细胞上。2.急性偏头痛大鼠模型三叉神经节P2X7受体表达降低,提示P2X7受体在偏头痛中可能有一定作用。3.急性偏头痛大鼠腹腔注射BBG后,三叉神经脊束核c-fos和CGRP的表达进一步增高,表明卫星胶质细胞P2X7受体在偏头痛中可能具有保护作用。
[Abstract]:Objective to establish acute migraine model by intraperitoneal injection of nitroglycerin (GTN) in rats. Western blot technique was used to detect the expression of P2X7 receptor and c-fos in rat trigeminal ganglion and the expression of P2X7 receptor and glial fibrillary acidic protein (GFAP) in satellite glial cells was detected by immunofluorescence. Finally, the effects of P2X7 receptor inhibitor, brilliant blue (BBG), on the expression of c-fos and calcitonin gene-related peptide (CGRP) in trigeminal spinal tract nucleus were observed in order to explore whether P2X7 receptor is involved in the pathogenesis of migraine and its possible mechanism. Materials and methods Ten 1.Sprague-Dawley rats were randomly divided into two groups: saline group (n = 5) and GTN group (n = 10 mg / kg). The basic pain threshold on the posterior paw of rats in each group was measured by Von Frey before administration, and the pain threshold on the posterior paw of rats was measured again at the 2nd and 4th hour after administration of the drug. The expression of P2X7 receptor and GFAP in trigeminal ganglion was observed by immunofluorescence. Twenty 2.Sprague-Dawley rats were randomly divided into 4 groups: normal group (nong5) and intraperitoneal injection of GTN (10 mg / kg). The expression of P2X7 receptor and c-fos in the trigeminal ganglion was detected by intraperitoneal injection of GTN (10 mg / kg) and intraperitoneal injection of GTN (10 mg / kg). Western blot) in 20 3.Sprague-Dawley rats. They were randomly divided into 4 groups: intraperitoneal injection of normal saline for 2 hours group (n = 5), intraperitoneal injection of GTN for 2 hours group (n = 5), intraperitoneal injection of GTN saline group for 2 hours (n = 10 mg / kg), intraperitoneal injection of GTN group for 2 hours (n = 5). 2ml/Kg and intraperitoneal injection of GTN BBG for 2 hours (n = 5 10 mg / kg). Western blot technique was used to detect the expression of c-fos and CGRP in the spinal trigeminal nucleus of rats. Result 1. Pain threshold: after intraperitoneal injection of GTN, the pain threshold on the posterior claw of rats began to decrease, which was the most obvious at the 2nd hour, which was significantly lower than that in the saline group (p0.05). Immunofluorescence: P2X7 receptor and GFAP co-expressed in trigeminal ganglion. After 3.Western blot: was injected intraperitoneally with GTN, the expression of P2X7 receptor in rat satellite glial cells began to decrease, and at the 2nd hour, the expression of P2X7 receptor was significantly lower than that in normal group (p0.05). The expression of c-fos and CGRP in trigeminal ganglion and spinal trigeminal nucleus was significantly higher than that in normal group (p0.05). C-fos and CGRP increased more significantly after the use of P2X7 receptor inhibitor BBG (p0.05). Conclusion 1.P2X7 receptor is specifically expressed in rat trigeminal ganglion satellite glial cells. 2. 2. The expression of P2X7 receptor in trigeminal ganglion was decreased in acute migraine rats, suggesting that P2X7 receptor may play a role in migraine. The expression of c-fos and CGRP in spinal trigeminal nucleus of migraine was further increased after intraperitoneal injection of BBG in acute migraine rats, suggesting that the P2X7 receptor of satellite glia might play a protective role in migraine.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R747.2;R-332
[Abstract]:Objective to establish acute migraine model by intraperitoneal injection of nitroglycerin (GTN) in rats. Western blot technique was used to detect the expression of P2X7 receptor and c-fos in rat trigeminal ganglion and the expression of P2X7 receptor and glial fibrillary acidic protein (GFAP) in satellite glial cells was detected by immunofluorescence. Finally, the effects of P2X7 receptor inhibitor, brilliant blue (BBG), on the expression of c-fos and calcitonin gene-related peptide (CGRP) in trigeminal spinal tract nucleus were observed in order to explore whether P2X7 receptor is involved in the pathogenesis of migraine and its possible mechanism. Materials and methods Ten 1.Sprague-Dawley rats were randomly divided into two groups: saline group (n = 5) and GTN group (n = 10 mg / kg). The basic pain threshold on the posterior paw of rats in each group was measured by Von Frey before administration, and the pain threshold on the posterior paw of rats was measured again at the 2nd and 4th hour after administration of the drug. The expression of P2X7 receptor and GFAP in trigeminal ganglion was observed by immunofluorescence. Twenty 2.Sprague-Dawley rats were randomly divided into 4 groups: normal group (nong5) and intraperitoneal injection of GTN (10 mg / kg). The expression of P2X7 receptor and c-fos in the trigeminal ganglion was detected by intraperitoneal injection of GTN (10 mg / kg) and intraperitoneal injection of GTN (10 mg / kg). Western blot) in 20 3.Sprague-Dawley rats. They were randomly divided into 4 groups: intraperitoneal injection of normal saline for 2 hours group (n = 5), intraperitoneal injection of GTN for 2 hours group (n = 5), intraperitoneal injection of GTN saline group for 2 hours (n = 10 mg / kg), intraperitoneal injection of GTN group for 2 hours (n = 5). 2ml/Kg and intraperitoneal injection of GTN BBG for 2 hours (n = 5 10 mg / kg). Western blot technique was used to detect the expression of c-fos and CGRP in the spinal trigeminal nucleus of rats. Result 1. Pain threshold: after intraperitoneal injection of GTN, the pain threshold on the posterior claw of rats began to decrease, which was the most obvious at the 2nd hour, which was significantly lower than that in the saline group (p0.05). Immunofluorescence: P2X7 receptor and GFAP co-expressed in trigeminal ganglion. After 3.Western blot: was injected intraperitoneally with GTN, the expression of P2X7 receptor in rat satellite glial cells began to decrease, and at the 2nd hour, the expression of P2X7 receptor was significantly lower than that in normal group (p0.05). The expression of c-fos and CGRP in trigeminal ganglion and spinal trigeminal nucleus was significantly higher than that in normal group (p0.05). C-fos and CGRP increased more significantly after the use of P2X7 receptor inhibitor BBG (p0.05). Conclusion 1.P2X7 receptor is specifically expressed in rat trigeminal ganglion satellite glial cells. 2. 2. The expression of P2X7 receptor in trigeminal ganglion was decreased in acute migraine rats, suggesting that P2X7 receptor may play a role in migraine. The expression of c-fos and CGRP in spinal trigeminal nucleus of migraine was further increased after intraperitoneal injection of BBG in acute migraine rats, suggesting that the P2X7 receptor of satellite glia might play a protective role in migraine.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R747.2;R-332
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