摘要
目的探讨鞘内注射Cdk5/p35抑制剂Roscovitine对神经病理性疼痛模型大鼠痛行为及脊髓ERK1/2和PPARγ蛋白表达的影响。方法将220~250g的雄性SPF级SD大鼠52只按随机数字表法分为4组:Sham+DMSO组(二甲基亚砜,S+D组)、Sham+Roscovitine组(S+R组)、CCI+DMSO组(I+D组)、CCI+Roscovitine组(I+R组)。采用慢性坐骨神经结扎(chronic constriction injury,CCI)建立神经病理性疼痛大鼠模型。CCI术后第7~14天鞘内注射相应药物,每天1次。采用VonFrey纤维丝和全自动热辐射刺激仪测定术前1d、术后3d、7d、10d、14d时大鼠右后爪机械缩足阈值(PMWT)和热缩足潜伏期(PWTL),术后14d取大鼠脊髓腰膨大部,采用免疫印迹法测定脊髓背角Cdk5、p35、磷酸化ERK蛋白(pERK)、总ERK蛋白(ERK)、磷酸化PPARγ蛋白(pPPARγ)、总PPARγ蛋白(PPARγ)表达变化。结果鞘内注射Roscovitine7d、10d、14d后I+R组大鼠的PWMT[(4.65±0.08)g,(6.47±0.12)g,7.90±0.19)g]、PWTL[(9.22±0.33)s,(13.52±0.43)s,(12.63±0.88)s]与I+D组相比延长[PWMT:(3.71±0.06)g,(2.45±0.17)g,(2.31±0.15)g;PWTL:(8.02±0.20)s,(5.90±0.28)s,(5.40±0.38)s],均差异有统计学意义(PWMT:F分组=505.71,P<0.05,F时间×分组=15.33,P<0.05;PWTL:F分组=355,P<0.05,F时间×分组=8.589,P<0.05);免疫印迹法显示术后14d,与I+D组[(p35:(0.58±0.02);pERK:(1.12±0.13);pPPARγ:(0.77±0.16))相比,I+R组p35表达(0.46±0.04)(F=11.06,P<0.05)和pERK表达(0.79±0.11)(F=22.91,P<0.05)降低,pPPARγ表达(0.99±0.13)(F=17.62,P<0.05)升高。结论鞘内注射Roscovitin能缓解神经病理性疼痛模型大鼠的机械痛和热痛行为,其机制可能与抑制脊髓背角Cdk5/p35、ERK的活性,进而增强PPARγ的活性有关。
Objective To investigate the effects of intrathecal injection of Roscovitine on the pain behavior and the expression of pERK1/2 and pPPARγ receptor in spinal cord in rats with neuropathic pain.Methods Fifty-two male adult Sprague-Dawley rats of 220~250 g were randomly divided into 4 groups: Sham+ DMSO group(dimethyl sulfoxide,S+ D group),Sham+ Roscovitine(S+ R)group,CCI+ DMSO group(I+ D),CCI+ Roscovitine(I+ R)group.The corresponding drugs were administered by intrathecal injection from the seventh day after CCI once a day for 14 days.The right hind paw mechanical threshold value(PMWT)was measured by Von Frey filament and paw thermal withdrawal latency(PWTL)was measured by Hargreaves methods before 1 day and 3 d,7 d,10 d ,14 d after surgery.The spinal cord lumbar enlargement was taken at 14 d after surgery,and Cdk5,p35,phosphorylated ERK protein(pERK),total ERK protein(ERK),phosphorylated PPARγ protein(pPPARγ)and total PPARγ protein(PPARγ)were detecteded by Western blot.Results Roscovitine was administered by intrathecal injection alleviated mechanical allodynia and thermal hyperalgesia of CCI rats.Compared with I+ D group,PWMT in I+ R group at 7 d,10 d,14 d after administration((4.65±0.08)g,( 6.47±0.12)g,( 7.90±0.19)g,)vs((3.71±0.06)g,(2.45±0.17)g,( 2.31±0.15)g)(Fgroup=505.71,P<0.05,Ftime×group=15.33,P<0.05),PWTL((9.22±0.33)s,(13.52±0.43)s,(12.63±0.88)s)vs((8.02±0.20)s,(5.90±0.28)s,(5.40±0.38)s)(Fgroup=355,P<0.05,Ftime×group=8.589,P<0.05)were significantly increased.Compared with I+ D group(p35(0.58±0.02),pERK(1.12±0.13),pPPARγ(0.77±0.16)),p35(0.46±0.04,F=11.06,P<0.05)and pERK(0.79±0.11,F=22.91,P<0.05)in I+ R group,were significantly dereased,and the expression of pPPARγ(0.99±0.13,F=17.62,P<0.05))was significantly increased.Conclusion Intrathecal injection Roscovitine can alleviate both mechanical allodynia and thermal hyperalgesia in rats with neuropathic pain,and its mechanisms may be related to the inhibition of Cdk5/p35 and ERK activity,enhancement of PPARγ activity in the spinal dorsal horn.
作者
钟妤
陈佳林
谢玉波
Zhong Yu;Chen Jialin;Xie Yubo(Department of Anesthesiology,the First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China)
出处
《中华行为医学与脑科学杂志》
CAS
CSCD
北大核心
2018年第12期1075-1080,共6页
Chinese Journal of Behavioral Medicine and Brain Science
基金
广西自然科学基金项目(2015GXNSFAA139154,2017GXNSFBA198039).
