CAJ | 학술논문

目的动态观察细胞周期素依赖性激酶5(Cdk5)及调节亚基p35、骨架蛋白在戊四氮致痫大鼠海马各区的表达变化和苔藓纤维出芽(MFS)的情况,以探讨cdk5/p35对MFS的作用及其机制.方法 SD雄性成年大鼠240只,随机分为PTZ组和对照组;PTZ组分为PTZ第一次注射后3 d、1周、2周、4周、6周共5个亚组,每亚组24只,对照组随机分为5个亚组,与PTZ组各时间点对应.以上各亚组再分4个小组,每小组6只大鼠,分别进行(1)Timm染色并评分;(2)Cdk5/p35的免疫组化和原位杂交;(3)Cdk5活性测定;(4)tau蛋白表达及磷酸化水平(免疫组化和免疫印迹).结果 PTZ组CA3区苔藓纤维出芽最为明显,而非内分子层,苔藓纤维出芽的程度与点燃大鼠痫性发作的行为学表现一致;PTZ组Cdk5/p35 mRNA和蛋白、总tau蛋白和P-ptau(ser202)蛋白在3 d时表达增多,4周达高峰,6周时接近正常水平,与CA3区苔藓纤维出芽的过程一致.对照组无动态变化.结论 Cdk5/p35及其底物tau蛋白可能参与r苔藓纤维出芽,了解苔藓纤维出芽的机制,有助于抑制癫痫的发生.
목적 동태관찰세포주기소의뢰성격매5(Cdk5)급조절아기p35、골가단백재무사담치간대서해마각구적표체변화화태선섬유출아(MFS)적정황,이탐토cdk5/p35대MFS적작용급기궤제.방법 SD웅성성년대서240지,수궤분위PTZ조화대조조;PTZ조분위PTZ제일차주사후3 d、1주、2주、4주、6주공5개아조,매아조24지,대조조수궤분위5개아조,여PTZ조각시간점대응.이상각아조재분4개소조,매소조6지대서,분별진행(1)Timm염색병평분;(2)Cdk5/p35적면역조화화원위잡교;(3)Cdk5활성측정;(4)tau단백표체급린산화수평(면역조화화면역인적).결과 PTZ조CA3구태선섬유출아최위명현,이비내분자층,태선섬유출아적정도여점연대서간성발작적행위학표현일치;PTZ조Cdk5/p35 mRNA화단백、총tau단백화P-ptau(ser202)단백재3 d시표체증다,4주체고봉,6주시접근정상수평,여CA3구태선섬유출아적과정일치.대조조무동태변화.결론 Cdk5/p35급기저물tau단백가능삼여r태선섬유출아,료해태선섬유출아적궤제,유조우억제전간적발생.
Objective To observe the expression of eyclin-dependent kinase 5(Cdk5),p35,tau protein and the activity of Cdk5 in rat hippoeampus during pentylenetetrazole(PTZ)kindling process and their correlation with mossy fiber sprouting(MFS)so as to investigate the role of Cdk5/p35 in epileptogenesis.Methods A total of 240 healthy male SD rats were divided randomly into normal controls and pentylenetetrazole(PTZ)treatment groups.The epileptic models were established by injection of PTZ intraperitoneally.At Day 3,Weeks 1,2,4 & 6 after a daily injection of PTZ,Timm staining was scored in the CA3 region and dentate gyrus.At the same time,the mRNA and protein of Cdk5 and p35,total tau protein and its phosphorylation at set202 and Cdk5 activity were analyzed in the hilus and stratum granulosum of dentate gyms and the CA 1,CA3 regions of hippocampus.The methods of in situ hybridization,immunohistochemistry,Western blot and immuno-precipitation and liquid scintillation counter were employed respectively.Results Prominent MFS was observed in area CA3 rather than the inner molecular layer in PTZ-treated rats.And the degree of MFS progressed with the development of behavioral kindled seizures.The expressions of Cdk5/p35 mRNA and protein,tau protein and its phosphorylation at Set202 significantly increased from Day 3 to Week 4 in the PTZ treatment group.It was in accordance with the progression of MFS in area CA3.Conclusion CdkS/p35 and its substrate tau protein may be involed in M FS.Undeerstanding the molecular mechanisms of MFS may lead to therapeutic interventions for limiting epileptogenesis.