中西医结合学报
中西醫結閤學報
중서의결합학보
JOURNAL OF CHINESE INTEGRATIVE MDEICINE
2009年
3期
242-248
,共7页
战丽彬%牛新萍%隋华%宫晓洋
戰麗彬%牛新萍%隋華%宮曉洋
전려빈%우신평%수화%궁효양
阿尔茨海默病%滋补脾阴方药%淀粉样p蛋白%血清诱导激酶%树突棘相关RaP特异性GTP酶活化蛋白%受体,N-甲基-D-天冬氨酸%突触
阿爾茨海默病%滋補脾陰方藥%澱粉樣p蛋白%血清誘導激酶%樹突棘相關RaP特異性GTP酶活化蛋白%受體,N-甲基-D-天鼕氨痠%突觸
아이자해묵병%자보비음방약%정분양p단백%혈청유도격매%수돌극상관RaP특이성GTP매활화단백%수체,N-갑기-D-천동안산%돌촉
Alzheimer disease%Zibu Piyin Recipe%amyloid beta-peptide%serum-inducible kinase%spine-associated Rap guanosine triphosphatase activating protein:receptors,N-methyl-D-aspartate%synapses
目的:观察β淀粉样蛋白(amyloid β-peptide,Aβ)神经毒性、血清诱导激酶(serum-inducible kinase,SNK)-树突棘相关Rap特异性GTP酶活化蛋白(spine-associated Rap guanosine triphosphatase activating protein,SPAR)途径、N-甲基-D-门冬氨酸受体(N-methyl-D-aspartate receptor,NMDAR)三者的关系,探讨滋补脾阴方药保护Aβ损伤原代培养大鼠海马神经元的作用机制.方法:将干粉Aβ1-40配制成溶液,在37℃恒温箱中孵育72 h,获得聚集态纤维状Aβ1-40.原代培养大鼠海马神经元,以血清药理学方法制备滋补脾阴方药含药血清,建立Aβ1-40(5 μmoi/L)损伤神经元模型,滋补脾阴方药含药血清为干预组,采用逆转录聚合酶链式反应方法观察SNK、SPAR及NMDAR亚基NRl、NR2A、NR2B mRNA表达.结果:与对照组相比,神经元暴露于Aβ1-40(5 μmol/L)2 h后,SNK mRNA表达上调,SPAR、NRl、NR2A和NR2B mRNA表达下调,差异有统计学意义(P<0.01,P<0.05).与Aβ1-40(5 μmol/L)组比较,各浓度滋补脾阴方药含药血清组SNK mRNA表达下调,SPAR、NRl、NR2A和NR2B mRNA表达上调.以2%滋补脾阴方药含药血清效果最显著(P<0.05).结论:Aβ引起神经元损伤的过程与SNK-SPAR途径和NMDAR相关;滋补脾阴方药对Aβ损伤神经元有保护作用,其保护机制可能与调节NMDAR表达,阻断SNK-SPAR途径有关.
目的:觀察β澱粉樣蛋白(amyloid β-peptide,Aβ)神經毒性、血清誘導激酶(serum-inducible kinase,SNK)-樹突棘相關Rap特異性GTP酶活化蛋白(spine-associated Rap guanosine triphosphatase activating protein,SPAR)途徑、N-甲基-D-門鼕氨痠受體(N-methyl-D-aspartate receptor,NMDAR)三者的關繫,探討滋補脾陰方藥保護Aβ損傷原代培養大鼠海馬神經元的作用機製.方法:將榦粉Aβ1-40配製成溶液,在37℃恆溫箱中孵育72 h,穫得聚集態纖維狀Aβ1-40.原代培養大鼠海馬神經元,以血清藥理學方法製備滋補脾陰方藥含藥血清,建立Aβ1-40(5 μmoi/L)損傷神經元模型,滋補脾陰方藥含藥血清為榦預組,採用逆轉錄聚閤酶鏈式反應方法觀察SNK、SPAR及NMDAR亞基NRl、NR2A、NR2B mRNA錶達.結果:與對照組相比,神經元暴露于Aβ1-40(5 μmol/L)2 h後,SNK mRNA錶達上調,SPAR、NRl、NR2A和NR2B mRNA錶達下調,差異有統計學意義(P<0.01,P<0.05).與Aβ1-40(5 μmol/L)組比較,各濃度滋補脾陰方藥含藥血清組SNK mRNA錶達下調,SPAR、NRl、NR2A和NR2B mRNA錶達上調.以2%滋補脾陰方藥含藥血清效果最顯著(P<0.05).結論:Aβ引起神經元損傷的過程與SNK-SPAR途徑和NMDAR相關;滋補脾陰方藥對Aβ損傷神經元有保護作用,其保護機製可能與調節NMDAR錶達,阻斷SNK-SPAR途徑有關.
목적:관찰β정분양단백(amyloid β-peptide,Aβ)신경독성、혈청유도격매(serum-inducible kinase,SNK)-수돌극상관Rap특이성GTP매활화단백(spine-associated Rap guanosine triphosphatase activating protein,SPAR)도경、N-갑기-D-문동안산수체(N-methyl-D-aspartate receptor,NMDAR)삼자적관계,탐토자보비음방약보호Aβ손상원대배양대서해마신경원적작용궤제.방법:장간분Aβ1-40배제성용액,재37℃항온상중부육72 h,획득취집태섬유상Aβ1-40.원대배양대서해마신경원,이혈청약이학방법제비자보비음방약함약혈청,건립Aβ1-40(5 μmoi/L)손상신경원모형,자보비음방약함약혈청위간예조,채용역전록취합매련식반응방법관찰SNK、SPAR급NMDAR아기NRl、NR2A、NR2B mRNA표체.결과:여대조조상비,신경원폭로우Aβ1-40(5 μmol/L)2 h후,SNK mRNA표체상조,SPAR、NRl、NR2A화NR2B mRNA표체하조,차이유통계학의의(P<0.01,P<0.05).여Aβ1-40(5 μmol/L)조비교,각농도자보비음방약함약혈청조SNK mRNA표체하조,SPAR、NRl、NR2A화NR2B mRNA표체상조.이2%자보비음방약함약혈청효과최현저(P<0.05).결론:Aβ인기신경원손상적과정여SNK-SPAR도경화NMDAR상관;자보비음방약대Aβ손상신경원유보호작용,기보호궤제가능여조절NMDAR표체,조단SNK-SPAR도경유관.
Obiective:To observe the relationship among amyloid-peptide(Aβ)-induced neurotoxicity,serum-inducible kinase(SNK).spine.associated Rap guanosine triphosphatase activating protein(SPAR)pathway and N-methyl-D-aspartate receptor(NMDAR),and to explore the mechanism of the protective effect of spleen'yln nourishing recipe(Zibu Piyin Recipe,ZBPYR)in hippocampal neu rons against Aβ-induced neu rotoxicity.Methods:The Aβ1-40 powder was dissolved in 1×PBS and incubated at 37℃,and then agg regated fibrillar Aβ1-40 was obtained 72 h later.We used rat primary hippocampal neu rons as cell model.ZBPYR-containing serum was gained by the method of serum pharmacology.ZBPYR-containing serum was added to the culture 1 h before Aβ1-40(5 μmol/L)exposure.Cells were harvested 2 h after Aβ1-10 exposu re for total RNA extracting.Then the mRNA expression levels of SNK,SPAR and NMDAR subunits NR1,NR2A and NR2B were detected by reverse transcription-polymerase chain reaction(RT-PCR).ResuIts:After 2-hour Aβl-10 exposure,we found that the expression level of SNK mRNA was up-regulated and the expression Ievels of SPAR,NR1,NR2A and NR2B mRNAs were down-regulated in hippocampal neu rons as compared with control group(P<0.01,P<0.05).While with ZBPYR-containing serum pretreatment,the expression level of SNK mRNA was down-regulated and the levels of SPAR,NRt,NR2A and NR2B were up-regulated as compared with Aβ1-40 exposure,and 2%ZBPYR-containing serum showed the best effect(P<0.05).Conclusion:Aβ-induced neurotoxicity was related to SNK.SPAR pathway and NMDAR;ZBPYR-containing serum can protect neurons from Aβ-induced neu rotoxicity,and this protective effect may be performed by regulating the expression of NMDAR and blocking of the SNK-SPAR pathway.
