P2X3受体在神经病理性疼痛中上调对JNK和P38活化的影响

发布时间：2018-04-26 17:03

本文选题：C-Jun + N　；参考：《第二军医大学》2015年硕士论文

【摘要】：目的：通过慢性背根神经节压迫(CCD)的大鼠神经病理性疼痛(NP)模型,观察P2X3受体激活后能否引起JNK和P38在DRG上表达和活化的改变,进一步探讨P2X3受体在NP中的作用机制。方法：1.用12只Sprague-Dawley (SD)雄性大鼠随机构建NP模型(CCD)组(n=6)及其假手术(SHAM)组(n=6)：在手术前1d,术后第1、4、7、10、14、21、28和35d测量双侧后足的50%机械缩腿痛阈阈值(50%MPWT)。2.检测CCD大鼠术后28d内的P2X3受体在DRG上的表达趋势：用15只SD雄性大鼠随机分成未处理大鼠(NAIVE)组(n=3)和构建CCD模型(n=12)；CCD大鼠随机在术后第7d (POD07) (n=3)、术后第14d (POD14) (n=3)、术后第21d(POD21) (n=3)或术后第28d (POD28) (n=3)快速取材L4、L5 DRG,应用蛋白定量检测P2X3受体水平。3.检测CCD大鼠和SHAM大鼠术后给予P2X3受体激动剂后引发的痛行为之一抬腿的持续时间变化：将36只SD雄性大鼠随机构建CCD组(18只)和SHAM组(18只),18只大鼠分别又随机在术后第14d给予足底注射:H2O(n=6),p,y-meATP 100nmol (n=6)和β,γ-meATP 500nmol (n=6),体积均为501μl；检测各组给药后每分钟抬腿时间(PLTPM)。4.检测CCD大鼠和SHAM大鼠术后JNK、pJNK (JNK磷酸化活化型)、P38和pP38(P38磷酸化活化型)在DRG上的表达变化：将9只SD雄性大鼠随机分成NAIVE组(n=3)、SHAM组(n=3)和CCD组(n=3)；快速取材L4和L5 DRG,应用蛋白定量检测JNK、pJNK、P38和pP38。5.检测CCD大鼠和SHAM大鼠术后给予P2X3受体激动剂后P2X3、JNK、 pJNK、P38和pP38在DRG上的表达变化：将18只SD雄性大鼠随机构建CCD组(9只)和SHAM组(9只),9只大鼠分别又随机在术后第14d给予足底注射：H2O(n=3), β,γ-meATP 100nmol (n=3)和β,γ-meATP 500nmol (n=3),体积均为50μl；快速取材L4和L5 DRG,应用蛋白定量检测P2X3、JNK、pJNK、P38和pP38。结果：1.CCD模型大鼠术后第1、4、7、10、14、21、28、35d CCD组的手术侧50%MPWT比较自身手术对侧或SHAM组的手术侧后足均显著下降(P0.001)；2.CCD模型大鼠DRG上P2X3在POD14组与NAIVE组比较表达增强(P0.05)；3.在18只SHAM大鼠,给予β,γ-meATP 100nmol后第1min与给予H20比较PLTPM明显增加(P0.01),在给予β,γ-meATP 500nmol后第1min与给予H2O比较PLTPM显著增加(P0.001)；在18只CCD大鼠,给予β,γ-meATP 100nmol后第1min与给予H2O比较PLTPM显著增加(P0.001)；给予β,γ-meATP 500nmol后第1、2、3、5min与给予H20比较PLTPM显著增加(P0.001),第6min PLTPM增加(P0.05)；此外,给予β,γ-meATP 500nmol的CCD大鼠和给予β,γ-meATP 500nmol的SHAM大鼠相比在给药后第2min PLTPM显著增加(P0.001)。4. NAIVE组、SHAM组和CCD组组间比较DRG上JNK、pJNK、P38和pP38都没有统计学意义的变化。5.检测CCD大鼠和SHAM大鼠术后给予P2X3受体激动剂后P2X3、JNK、 pJNK、P38和pP38在DRG上的表达变化：(1)P2X3：给予H2O的CCD大鼠和给予H20的SHAM大鼠相比表达增加(P0.05),给予β,γ-meATP 100nmol的CCD组和给予β,γ-meATP 100nmol的SHAM组比较明显增加(P0.01)；此外,给予β,γ-meATP 100nmol的CCD组与给予H2O的CCD组比较增加(P0.05)。(2)JNK:给予β,γ-meATP 100nmol的CCD组和给予β,γ-meATP 100nmol的SHAM组比较明显增加(P0.01),给予β,γ-meATP 500nmol的CCD组和给予β,γ-meATP 500nmol的SHAM组比较增加(P0.05);此外在CCD大鼠,给予β,γ-meATP 100nmol和给予β,γ-meATP 500nmol均比较给予H2O表达增加(P0.05)。(3)pJNK:给予β,γ-meATP 500nmol的CCD组和给予β,γ-meATP 500nmol的SHAM组比较减少(P0.05)。(4)P38:多组间比较均无统计学意义。(5)pP38:给予β,γ-meATP 100nmol的CCD组和给予β,γ-me ATP 100nmol的SHAM组比较减少(P0.05)。结论：在CCD模型大鼠中P2X3受体激活后发生了JNK在DRG上表达总量的增加,而磷酸化活化量的减少；P38在DRG上表达无变化,而磷酸化活化量也减少；这提示NP中P2X3受体在DRG上激活可能会引起JNK和P38通路下调。
[Abstract]:Objective: To observe the changes in the expression and activation of JNK and P38 on DRG after P2X3 receptor activation in rats with chronic dorsal root ganglion compression (CCD), and to further explore the mechanism of the action of P2X3 receptor in NP. Method: 1. the NP model (CCD) was randomly constructed in 12 male rats with Sprague-Dawley (SD). ) and the sham operation (SHAM) group (n=6): before operation 1D, 1,4,7,10,14,21,28 and 35d after operation, the 50% mechanical contraction threshold (50%MPWT).2. of bilateral hind legs (50%MPWT).2. was used to detect the expression trend of P2X3 receptor within 28d in CCD rats: 15 male rats were randomly divided into untreated rats and constructed models. CD rats were randomly assigned to 7d (POD07) (n=3) after operation, 14d (POD14) (n=3) after operation, 21d (POD21) (n=3) or 28d (POD28) after operation. 36 SD male rats were randomly constructed in group CCD (18) and group SHAM (18 rats), and 18 rats were randomly injected at the foot of 14d after operation: H2O (n=6), P, y-meATP 100nmol (n=6), beta, and gamma -meATP 500nmol (501 mu). JNK, pJNK (JNK phosphorylation activation), P38 and pP38 (P38 phosphorylation activation type) expression on DRG: 9 male SD rats were randomly divided into NAIVE group (n=3), SHAM group (n=3) and groups. The expression changes of post P2X3, JNK, pJNK, P38 and pP38 on DRG: 18 male SD rats were randomly constructed in group CCD (9) and SHAM group (9), and 9 rats were randomly injected into the foot to be injected into the foot. The results of quantitative detection of P2X3, JNK, pJNK, P38, and pP38.: 1.CCD model rats were compared to the operative side of the 1,4,7,10,14,21,28,35d CCD group after operation, and the operation side of the side or SHAM group was significantly decreased (P0.001). The 2.CCD model rats were compared with those in the group, and 3. in 18 rats, After beta, 1min and H20 were significantly increased after gamma -meATP 100nmol (P0.01), and the comparison between 1min and H2O was significantly increased after giving beta and -meATP 500nmol (P0.001). Compared with H20, PLTPM was significantly increased (P0.001) and 6min PLTPM increased (P0.05). Moreover, the CCD rats given beta, gamma -meATP 500nmol and beta, gamma -meATP 500nmol SHAM rats were significantly increased. The changes of expression of P2X3, JNK, pJNK, P38 and pP38 on DRG after the operation of P2X3 receptor agonists were detected by.5. in CCD rats and SHAM rats after operation: (1) P2X3. P0.01); in addition, the CCD group that gave beta, gamma -meATP 100nmol increased (P0.05) compared with the CCD group giving H2O. (2) JNK: gave beta, the CCD group of gamma -meATP 100nmol and beta, gamma -meATP groups were significantly increased. D rats were given beta, gamma -meATP 100nmol and beta, and gamma -meATP 500nmol increased H2O expression (P0.05). (3) pJNK: gave beta, CCD group of gamma -meATP 500nmol and beta, gamma -meATP groups decreased. (4) there was no statistical significance between groups. (5) The SHAM group of -me ATP 100nmol decreased (P0.05). Conclusion: after the activation of P2X3 receptor in the CCD model rats, the total amount of JNK in DRG increased, and the activation amount of phosphorylation was reduced; P38 in DRG was not changed, and the activation amount of phosphorylation was also reduced. The road is down.

【学位授予单位】：第二军医大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R614

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