目的研究白術(shù)中雙白術(shù)內(nèi)酯對Aβ淀粉樣蛋白(Aβ_25-35)神經(jīng)損傷的大鼠腎上腺嗜鉻細胞瘤細胞（PC12）及人骨髓神經(jīng)母細胞瘤細胞系（SH-SY5Y）誘導其線粒體膜通透性轉(zhuǎn)換孔（mPTP）的開放和細胞凋亡的的保護作用及其機制研究,并進一步提供基于藥代動力學和藥理學數(shù)據(jù)的系統(tǒng)網(wǎng)絡藥理學模型,以確定中藥白術(shù)的有效化合物和潛在靶點,為白術(shù)對阿爾茨海默病具有神經(jīng)保護作用提供理論基礎(chǔ)。方法體外培養(yǎng)PC12和SH-SY5Y細胞,用20μMAβ_25-35處理細胞構(gòu)建細胞損傷模型,對照組用不同濃度雙白術(shù)內(nèi)酯預處理2小時。實驗組如下:（1）空白對照組,（2）Aβ_25-35損傷組,（3）藥物對照組（低,中,高）。通過MTT法檢測細胞增殖;用羅丹明123染色檢測線粒體膜電位;ROS試劑盒檢測細胞內(nèi)ROS;蛋白質(zhì)印跡分析檢測線粒體相關(guān)蛋白CypD和CytC的變化。網(wǎng)絡藥理學能構(gòu)建生物網(wǎng)絡并分析網(wǎng)絡中成分、靶點和疾病之間的聯(lián)系。我們通過使用Cytoscape 3.2.1軟件構(gòu)建了以下網(wǎng)絡圖:（1）C-T（成分-靶點圖）,在TCMSP和SWISS數(shù)據(jù)... 

【文章來源】：湖南師范大學湖南省 211工程院校

【文章頁數(shù)】：84 頁

【學位級別】：碩士

【文章目錄】：
Abstract
摘要
Chapter1.Introduction
Chapter2.Experimental Materials
    2.1 Experimental cell
    2.2 Main Experimental instruments
    2.3 Main reagents and Kits
    2.4 Main reagent preparation
        2.4.1.Incubation of Aβ25-
        2.4.2.Preparation of BD
        2.4.3.Preparation of Rhodamine123 stain
        2.4.4.Western-blot main reagent preparation
            2.4.4.1 10%SDS
            2.4.4.2 10%Ammonium persulfate(APS)
            2.4.4.3 TBS-T buffer
            2.4.4.4 5%BSA(blocking solution)
            2.4.4.5 1×transfer buffer
            2.4.4.6 1×running buffer
        2.4.5.0.25%Trypsin working solution
        2.4.6.Cell Cryopreservation solution
        2.4.7.Preparation of MTT
Chapter3.Methods
    3.1 Network pharmacology
        3.1.1 Data set construction
        3.1.2 Active compound screening
        3.1.3 Network construction
        3.1.4 Pathway analysis
    3.2 Experimental Investigation
        3.2.1 Materials and Methods
        3.2.2 Cell culture
        3.2.3 Experimental grouping and drug treatment
        3.2.4 Cell viability assay(MTT assay)
        3.2.5 Detection of MMP by rhodamine123 staining
        3.2.6 Measurement of intracellular ROS
        3.2.7 Western blot analysis(CYPD,Cyt C)
    3.3 Statistical analysis
Chapter4.Results
    4.1 Experimental Results
        4.1.1 MTT assay
        4.1.2 Detection of MMP by rhodamine123 staining
        4.1.3 Measurement of intracellular ROS
        4.1.4 Protein characterization
    4.2 Networks Results
        4.2.1 Active compound screening
        4.2.2 Network construction and analysis:
            4.2.2.1 C-T network
            4.2.2.2 T-D network
            4.2.2.3 Target-pathway(T-P)network
Chapter5.Discussion
Chapter6.Conclusion
Table-1
Table-2
References
Review article
    References
Published papers
Acknowledgement


【參考文獻】：
期刊論文
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[2]中藥群體藥代動力學專家共識(征求意見稿)[J]. 謝雁鳴,王建農(nóng),賀福元,姜俊杰,熊昕.  中國中藥雜志. 2013(18)
[3]降香化學成分、藥理作用及藥代特征的研究進展[J]. 楊志宏,梅超,何雪輝,孫曉波.  中國中藥雜志. 2013(11)
[4]白術(shù)的化學成分研究[J]. 李偉,文紅梅,崔小兵,張發(fā)成,董潔.  中草藥. 2007(10)



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