雙白術(shù)內(nèi)酯對(duì)Aβ 25-35 損傷神經(jīng)細(xì)胞的保護(hù)及其與mPTP開放的研究
發(fā)布時(shí)間：2021-09-17 19:37
目的研究白術(shù)中雙白術(shù)內(nèi)酯對(duì)Aβ淀粉樣蛋白(Aβ25-35)神經(jīng)損傷的大鼠腎上腺嗜鉻細(xì)胞瘤細(xì)胞（PC12）及人骨髓神經(jīng)母細(xì)胞瘤細(xì)胞系（SH-SY5Y）誘導(dǎo)其線粒體膜通透性轉(zhuǎn)換孔（mPTP）的開放和細(xì)胞凋亡的的保護(hù)作用及其機(jī)制研究,并進(jìn)一步提供基于藥代動(dòng)力學(xué)和藥理學(xué)數(shù)據(jù)的系統(tǒng)網(wǎng)絡(luò)藥理學(xué)模型,以確定中藥白術(shù)的有效化合物和潛在靶點(diǎn),為白術(shù)對(duì)阿爾茨海默病具有神經(jīng)保護(hù)作用提供理論基礎(chǔ)。方法體外培養(yǎng)PC12和SH-SY5Y細(xì)胞,用20μMAβ25-35處理細(xì)胞構(gòu)建細(xì)胞損傷模型,對(duì)照組用不同濃度雙白術(shù)內(nèi)酯預(yù)處理2小時(shí)。實(shí)驗(yàn)組如下:（1）空白對(duì)照組,（2）Aβ25-35損傷組,（3）藥物對(duì)照組（低,中,高）。通過MTT法檢測(cè)細(xì)胞增殖;用羅丹明123染色檢測(cè)線粒體膜電位;ROS試劑盒檢測(cè)細(xì)胞內(nèi)ROS;蛋白質(zhì)印跡分析檢測(cè)線粒體相關(guān)蛋白CypD和CytC的變化。網(wǎng)絡(luò)藥理學(xué)能構(gòu)建生物網(wǎng)絡(luò)并分析網(wǎng)絡(luò)中成分、靶點(diǎn)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(luò)圖:（1）C-T（成分-靶點(diǎn)圖）,在TCMSP和SWISS數(shù)據(jù)...
【文章來源】：湖南師范大學(xué)湖南省 211工程院校
【文章頁數(shù)】：84 頁
【學(xué)位級(jí)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(xiàn)】：
期刊論文
[1]網(wǎng)絡(luò)藥理學(xué)研究相關(guān)技術(shù)與應(yīng)用[J]. 彭曉飛,王麗,邊育紅. 天津中醫(yī)藥大學(xué)學(xué)報(bào). 2015(02)
[2]中藥群體藥代動(dòng)力學(xué)專家共識(shí)(征求意見稿)[J]. 謝雁鳴,王建農(nóng),賀福元,姜俊杰,熊昕. 中國(guó)中藥雜志. 2013(18)
[3]降香化學(xué)成分、藥理作用及藥代特征的研究進(jìn)展[J]. 楊志宏,梅超,何雪輝,孫曉波. 中國(guó)中藥雜志. 2013(11)
[4]白術(shù)的化學(xué)成分研究[J]. 李偉,文紅梅,崔小兵,張發(fā)成,董潔. 中草藥. 2007(10)
本文編號(hào)：3399369
【文章來源】：湖南師范大學(xué)湖南省 211工程院校
【文章頁數(shù)】：84 頁
【學(xué)位級(jí)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(xiàn)】：
期刊論文
[1]網(wǎng)絡(luò)藥理學(xué)研究相關(guān)技術(shù)與應(yīng)用[J]. 彭曉飛,王麗,邊育紅. 天津中醫(yī)藥大學(xué)學(xué)報(bào). 2015(02)
[2]中藥群體藥代動(dòng)力學(xué)專家共識(shí)(征求意見稿)[J]. 謝雁鳴,王建農(nóng),賀福元,姜俊杰,熊昕. 中國(guó)中藥雜志. 2013(18)
[3]降香化學(xué)成分、藥理作用及藥代特征的研究進(jìn)展[J]. 楊志宏,梅超,何雪輝,孫曉波. 中國(guó)中藥雜志. 2013(11)
[4]白術(shù)的化學(xué)成分研究[J]. 李偉,文紅梅,崔小兵,張發(fā)成,董潔. 中草藥. 2007(10)
本文編號(hào)：3399369
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