宽筋藤有效部位对D-半乳糖联合Aβ2535所致AD大鼠海马蛋白组学的影响

Effects of Effective Part of Tinospora sinensis on Hippocampus Proteomics of AD Rats Induced by Beta-amyloid Protein and D-galactose

目的探讨宽筋藤大孔树脂提取物大孔树脂提取物体积分数80%,100%乙醇洗脱部位对D-半乳糖联合Aβ2535,所致阿尔茨海默病(AD)模型大鼠海马蛋白组表达的影响。方法采用D-半乳糖联合Aβ2535复制AD大鼠模型,随机分成假手术组、模型组、多奈哌齐组(多奈哌齐,6.0 mg·kg^-1)、宽筋藤80%提取部位组(生药量6 g·kg^-1)。多奈哌齐组灌胃多奈哌齐0.1mL·10 g^-1,给药宽筋藤有效部位组灌胃0.1 mL·10 g^-1,模型组与假手术组灌胃等容量的生理盐水,每日1次,连续给药15 d后分离大鼠海马提取蛋白,Nanol-ESI液相-质谱联用系统检测,Protein Discovery软件进行蛋白鉴定,SIEVE软件对海马蛋白进行相对定量定性分析PANTHER Classification System软件对差异蛋白进行GO分析,运用IPAD软件进行信号通路富集。结果后得结果与模型组相比,给药组大鼠共有66个差异蛋白包含微管蛋白、热休克蛋白、能量代谢相关蛋白、囊泡生成/转运相关蛋白和脑保护相关蛋白等与AD密切相关的蛋白以上差异蛋白共涉及21条信号通路。结论宽筋藤可能通过上调网格蛋白等与囊泡生成转运及神经递质释放相关蛋白,促进了神经递质的合成与释放,改善了脑内胆碱能功能来达到干预AD病理进程的作用。

OBJECTIVE To investigate the effect of the 80%ethanol elution part of Tinospora sinensis macroporous resin extract on the expression of hippocampus proteome in Alzheimer’s disease(AD)model rats induced by D-galactose combined with Aβ2535METHODS The AD rats model replicated by D-galactose combined with Aβ2535,The AD rat model was replicated by D-galactose combined with Aβ2535,and randomly divided into sham operation group,model group,Donepezil group(donepezil,6.0 mg·kg^-1)and 80%extraction of Tinospora sinensi group(crude drug 6 g·kg^-1).Donepezil group:donepezil 0.1 mL·10 g^-1 ig.80%extraction of Tinospora sinensi group:Tinospora sinensis effective part extraction 0.1 mL·10 g^-1 g.Model group and sham-operation group:physiological saline 0.1 mL·10 g^-1 ig.Once a day,continuous administration for 15 d.Separating the hippocampus and extracting the protein,take the system test with nanol-ESI liquid-mass spectrometry,protein discovery software was used for identification,and qualitative analysis different groups of hippocampal proteins by SIEVE software.Take the GO analysis on differential protein with the ANTHER classification system and use IPAD to enrich the pathway.RESULTS Compared with the model group,the drug-administered group had 66 differential proteins,including tubulin,heat shock proteins,energy metabolism-related proteins,vesicle production/transport related proteins,and brain protectior-related proteins,which are closely related to AD.The above differential proteins involve a total of 21 signaling pathways.CONCLUSION Tinospora sinensis may promote the synthesis and release of neurotransmitters by up-regulating clathrin and vesicle-forming transport and neurotransmitter release,and improve the function of cholinergic function in the brain to achieve the pathological process of AD.