CAJ | 학술논문

背景:阿尔茨海默病(Alzheimer disease,AD)的病理病因虽然还没有完全清楚,但目前认为β-淀粉样肽(amyloid β-peptide,Aβ)是引起AD的胆碱能神经功能紊乱的主要成分.因此,Aβ的产生及其对神经细胞的影响,以及寻找该病的发病机制、治疗方法及其有效的治疗药物成了医药界的研究热点.目的:观察三七总皂甙(PNS)对NG108-15细胞老年性痴呆模型的影响,为寻找该病的发病机制提供佐证,为研究和开发中药的新成分提供理论依据.设计:体外平行对照实验.单位:广西中医学院新药开发中心,广西中医学院生物化学教研室.对象:实验在广西中医学院新药开发中心完成.NG108-15细胞:中国科学院上海细胞研究所提供.PNS:云南玉溪维和制药厂生产.Aβ25～35片段:德国Sigma公司产品,批号:042K49501.MTT(四甲基噻唑蓝),德国Sigma公司产品,批号:020609.cAMP(环一磷酸腺苷)日本ヤマサ酱油株式会社产品,批号:00207.胚胎牛血清(FBS):由美国Hyclone公司提供,批号:0405.DMEM培养基:美国GIBCO公司产品,批号:0311.干预:NG108-15细胞培养法:含50 mL/L胚牛血清、10 g/L HAT、10 g/L青霉素和链霉素(1:1)的DMEM培养液,置100 mL/L CO2、37℃用的培养箱内培养殖,每隔一天换一次液,4 d传代一次.主要观察指标:利用MTT法、细胞计数法和细胞形态学检测法观察PNS对NG108-15细胞存活率和细胞突起的改变.结果:PNS能增加Aβ25～35片段神经毒性诱导的NG108-15细胞存活率[增殖状态:(311±21)%,分化状态:(113±31)%,P＜0.05,F=5.567]、细胞突起率[增殖状态:(25.66±3.65)%,分化状态:(67.97±2.47)%,P＜0.05,F=6.471]和细胞突起平均长度[增殖状态:(25.66±3.09)μm,分化状态:(25.66±3.65)μm,P＜0.001,F=12.641].结论:PNS具有减轻细胞对Aβ的神经毒性反应和促进细胞突起生长的作用,表明PNS能对老年性痴呆的病理发展有拮抗作用.
배경:아이자해묵병(Alzheimer disease,AD)적병리병인수연환몰유완전청초,단목전인위β-정분양태(amyloid β-peptide,Aβ)시인기AD적담감능신경공능문란적주요성분.인차,Aβ적산생급기대신경세포적영향,이급심조해병적발병궤제、치료방법급기유효적치료약물성료의약계적연구열점.목적:관찰삼칠총조대(PNS)대NG108-15세포노년성치태모형적영향,위심조해병적발병궤제제공좌증,위연구화개발중약적신성분제공이론의거.설계:체외평행대조실험.단위:엄서중의학원신약개발중심,엄서중의학원생물화학교연실.대상:실험재엄서중의학원신약개발중심완성.NG108-15세포:중국과학원상해세포연구소제공.PNS:운남옥계유화제약엄생산.Aβ25～35편단:덕국Sigma공사산품,비호:042K49501.MTT(사갑기새서람),덕국Sigma공사산품,비호:020609.cAMP(배일린산선감)일본ヤマサ장유주식회사산품,비호:00207.배태우혈청(FBS):유미국Hyclone공사제공,비호:0405.DMEM배양기:미국GIBCO공사산품,비호:0311.간예:NG108-15세포배양법:함50 mL/L배우혈청、10 g/L HAT、10 g/L청매소화련매소(1:1)적DMEM배양액,치100 mL/L CO2、37℃용적배양상내배양식,매격일천환일차액,4 d전대일차.주요관찰지표:이용MTT법、세포계수법화세포형태학검측법관찰PNS대NG108-15세포존활솔화세포돌기적개변.결과:PNS능증가Aβ25～35편단신경독성유도적NG108-15세포존활솔[증식상태:(311±21)%,분화상태:(113±31)%,P＜0.05,F=5.567]、세포돌기솔[증식상태:(25.66±3.65)%,분화상태:(67.97±2.47)%,P＜0.05,F=6.471]화세포돌기평균장도[증식상태:(25.66±3.09)μm,분화상태:(25.66±3.65)μm,P＜0.001,F=12.641].결론:PNS구유감경세포대Aβ적신경독성반응화촉진세포돌기생장적작용,표명PNS능대노년성치태적병리발전유길항작용.
BACKGROUND: Although the pathology and etiology of Alzheimer disease(AD) have not been clear completely yet, amyloid β-peptide(Aβ) is known as the main component induced cholinergic neural functional disturbance in AD. Therefore, the hot-topics on researches in the medical field are focused on Aβ production and its influence on neural cells and searching for the mechanism on the onset of disease, therapeutic methods and the effective therapeutic medications.OBJECTIVE: To observe the influence of panaxnotoginsengsaponins(PNS) on NG108-15 cells in senile dementia model, to provide evidences for searching for the mechanism on onset of disease and provide theoretic basis for researching and developing the new components of Chinese herbs.DESIGN: Parallel controlled trial in vitro.SETTING: Developing Center of New Drugs of Guangxi University of Chinese Medicine, Teaching-Research Room of Biochemistry in Guangxi University of Chinese Medicine.PARTICIPANTS: The experiment was accomplished in Developing Center of New Drugs of Guangxi University of Chinese Medicine. NG108-15 cells:provided by Shanghai Cellular Institute of China Scientific Academy. PNS:produced by Yunnan Yuxi Wethe Pharmaceutical Factory. Aβ25-35 segments: the product from German Sigma Company, No. 042K49501. MTT:the product from German Signa Company, No. 020609. cAMP: the product American Hyclone Company, No. 0405. DMEM medium: the product from American GIBCO Company, No. 0311.INTERVENTIONS: NG108-15 cellular cultivation: DMEM culture fluid contained 50 mL/L FBS, 10 g/L HAT, and 10 g/L penicillin and streptomycin(1: 1) was placed in a culture box of 100 mL/L CO2, at 37 ℃ for cultivated proliferation. The solution changed once every other day and the generations passed down once every 4 days.MAIN OUTCOME MEASURES: To observe the changes of PNS on survival rates and process growth of NG108-15 cells determined by MTT method, cell counting methods and cytomorphology.RESULTS: PNS can increase , by the neural toxicity of Aβ25-35 segments, the survival rate of NG108-15 cells [proliferation: (311±21)%,differentiation: ( 113 ± 31 ) %, P ＜ 0. 05, F = 5. 567 ], the process growth rate [ proliferation: ( 25.66 ± 3.65 ) %, differentiation: (67.97 ± 2.47 ) %,P ＜ 0.05, F=6.471] and the average length of cell process growth[proliferation: (25.66 ± 3.09) μm, differentiation: (25.66 ± 3.65) μm,P ＜0.001, F=12.641].CONCLUSION: PNS can minimize the neural toxic reaction of Aβ to cells and promote the growth of cellular process, indicating the antagonism of PNS to the pathological development of senile dementia.