CAJ | 학술논문

目的：探讨蛇床子素( osthole,Ost)对感染APP( amy-loid precursor protein)基因的神经元突触的保护作用及其机制。方法将培养的神经元分为对照组(感染GFP组)、模型组(感染APP组)和给药组( Ost+APP组)。通过感染含有突变位点的APP基因来模拟阿尔茨海默病, CCK-8法检测神经元的存活能力,免疫荧光化学法对突触素-1( synap-sin-1, SYN)染色,ELISA法检测神经元突触内突触后膜蛋白-95(PSD-95)和突触体素(synaptophysin,SYP)蛋白的浓度,Western blot法检测Aβ1-42、钙调蛋白激酶激酶2( calci-um/calmodulin-dependent protein kinase kinase 2, CAMKK2)、磷酸化单磷酸腺苷活化蛋白激酶α1(5’-adenosine mono-phosphate-activated protein kinaseα1, p-AMPKα1)、AMPKα1的蛋白表达。结果感染APP的神经元显现较强的APP阳性,并生成有神经毒性的Aβ1-42。与模型组比较,给药组明显增强神经细胞的存活能力,增加 synapsin-1的表达量,提高PSD-95和SYP的浓度,降低CAMKK2、p-AMPKα1蛋白的表达。结论蛇床子素对感染APP基因的神经元突触造成的损伤具有保护作用,此神经保护作用可能是与抑制CAMKK2/AMPK信号通路有关。
목적：탐토사상자소( osthole,Ost)대감염APP( amy-loid precursor protein)기인적신경원돌촉적보호작용급기궤제。방법장배양적신경원분위대조조(감염GFP조)、모형조(감염APP조)화급약조( Ost+APP조)。통과감염함유돌변위점적APP기인래모의아이자해묵병, CCK-8법검측신경원적존활능력,면역형광화학법대돌촉소-1( synap-sin-1, SYN)염색,ELISA법검측신경원돌촉내돌촉후막단백-95(PSD-95)화돌촉체소(synaptophysin,SYP)단백적농도,Western blot법검측Aβ1-42、개조단백격매격매2( calci-um/calmodulin-dependent protein kinase kinase 2, CAMKK2)、린산화단린산선감활화단백격매α1(5’-adenosine mono-phosphate-activated protein kinaseα1, p-AMPKα1)、AMPKα1적단백표체。결과감염APP적신경원현현교강적APP양성,병생성유신경독성적Aβ1-42。여모형조비교,급약조명현증강신경세포적존활능력,증가 synapsin-1적표체량,제고PSD-95화SYP적농도,강저CAMKK2、p-AMPKα1단백적표체。결론사상자소대감염APP기인적신경원돌촉조성적손상구유보호작용,차신경보호작용가능시여억제CAMKK2/AMPK신호통로유관。
Aim To investigate the effect of osthole on neuron synapses infected APP gene and its underlying mechanism. Methods The neurons were divided into three groups:GFP, APP, APP+Ost groups. The neu-rons were infected APP gene with containing mutational site in vitro for mimicking the characterstics of Alzhei-mer’ s disease ( AD) . The cell viability was assessed by CCK-8 , the expression of synapsin-1 was deter-mined by immunofluorescence, and the concentration of PSD-95 and SYP were detected by ELISA. The ex-pressions of Aβ1-42 , CAMKK2 , phoshorylated AMPKα1 , AMPKα1 protein were determined by West-ern blot. Results Strong APP staining was visible in neurons infected with APP and abundant expression of Aβ1-42 , a neurotoxic oligomer. Compared with APP group, APP+Ost group significantly increased cell vi-ability, promoted the expression of synapsin-1, up-reg-ulated the concentration of PSD-95 and SYP, and de-creased the expressions of CAMKK2 and p-AMPKα1 . Conclusions Ost can protect the neuron synapses a-gainst infected with APP gene. Its neuroprotective effect may be related to inhibiting the CAMKK2/AMPK signal pathway.