新型PDE4抑制剂Roflupram改善阿尔茨海默病大鼠的认知障碍及神经炎症  

The new PDE4 inhibitor Roflupram ameliorated cognitive deficits and neuroinflammation in a rat model of Alzheimer's disease

作　　者：王灿茂 程玉芳[1] 吴金刚[1] 甘丹娜 蒋毅萍[1] 徐江平[1] 

机构地区：[1]南方医科大学药学院药理学系,广州广东510515

出　　处：《中国药理学通报》2015年第3期327-333,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金委员会-广东省人民政府联合基金资助项目(No U1032006);国家自然科学基金资助项目(No81373384)

摘　　要：目的验证新型PDE4抑制剂罗氟普兰(roflupram)能否改善Aβ25-35诱导的阿尔茨海默病大鼠的学习记忆障碍,及可能的机制是否与缓解小胶质细胞激活引起的炎症相关。方法 SD大鼠双侧海马CA1区微量注射Aβ25-35(10μg)造模。动物分组包括:假手术对照组、Aβ25-35注射组、Aβ25-35注射+盐酸多奈哌齐组、Aβ25-35注射+Rolipram组、Aβ25-35注射+Roflupram低、中、高剂量。连续灌胃给药14 d后,进行Morris水迷宫实验,20 d后进行避暗实验。采用Western blot法检测海马c AMP下游信号分子(p-PKA、p-CREB),前致炎因子(i NOS、COX-2、TNF-α、IL-1β),胶质细胞激活标记物(GFAP、Iba-1),炎症相关蛋白(p-p38、核内NF-κB p65)的蛋白水平变化,并用PCR的方法分析海马内i NOS、COX-2、TNF-α和IL-1βmRNA水平的变化。结果行为学结果显示,Roflupram能明显改善由Aβ25-35造成大鼠的空间学习记忆能力和被动回避学习记忆能力障碍。分子生物学分析的结果如下:与Aβ25-35注射组比较,各药物处理后均可以使海马内p-CREB、p-PKA蛋白表达水平不同程度的升高,使NF-κB p65(核内)、p-p38、i NOS、COX-2、TNF-α、IL-1β、GFAP和Iba-1的蛋白水平不同程度的降低,i NOS、COX-2、TNF-α和IL-1βmRNA水平下降。结论 Roflupram能够改善Aβ25-35诱导痴呆大鼠的学习记忆功能障碍,该效应与抑制小胶质细胞的活化、减少炎性因子的表达,从而缓解神经炎症有关。Aim To investigate whether the new PDE4 inhibitor Roflupram can ameliorate Aβ25-35 induced cognitive deficits through the anti-neuroinflammation effects. Methods The Alzheimer ’ s disease model was produced by microinjection of amyloid-β（ Aβ） 25-35 fibrils into the bilateral hippocampal CA1 area of male rats. Then the rats were randomly divided into sham-operated control group, Aβ25-35 microinjected group, Aβ25-35 microinjected followed by donepezil hydrochlo-ride or Rolipram treated group, Aβ25-35 microinjected combined Roflupram treated group with different doses. Treatment was given （ i. g. ） 24 h after microinjection surgery operation for consecutive 23 days. Morris water maze and passive avoidance test were used to test the behavior performance after drug treatments of 14 and 20 days. The protein level of cAMP downstream signa-ling molecules （ p-PKA, P-CREB ） , proinflammatory cytokines （ iNOS, COX-2, TNF-α, IL-1β）, microglia and astrocyte activation marker（ Iba-1,GFAP） and in-flammation related protein （ nuclear NF-κB p65 , p-p38） in hippocampus were detected by Western blot. The mRNA level of proinflammatory cytokines was measured by semi-quantitative PCR. Results The re- suhs of behavioral manifestations indicated that Roflu- pram could significantly ameliorate the spatial learning and memory ability and the passive avoidance learning and memory ability impairment induced by microinjec- tion of AI325_35. Biochemical analysis revealed that the hippocampus protein level of p-CREB and p-PKA of rats was significantly decreased induced by microinjec- tion of Aβ25 -35, while the level of NF-KBp65 （ Nucle- us）, p-p38, iNOS, COX-2, TNF-a, IL-1β, GFAP and Iba-1 was increased, and these effects were re- versed by each kind of drug treatment. Semi-quantita- tive PCR test showed that each treatment could reduce the mRNA level of proinflammatory cytokines compared to the Aβ25_35 microinjection animal. Conclusion The new PDE4 inhibitor Roflupram can ameliorate cog- nitive

关 键 词：Roflupram 阿尔茨海默病 PDE4抑制剂 神经炎症 Β-淀粉样蛋白 认知 小胶质细胞 

分 类 号：R-332[医药卫生] R322.81