發(fā)布時(shí)間：2018-06-17 19:39

  本文選題：金屬蛋白酶抑制劑 + 膜型TRAIL�。� 參考：《第四軍醫(yī)大學(xué)》2005年博士論文

【摘要】：目的 研究蛋白酶抑制劑對(duì)膜型TRAIL(Membrane TRAIL,mTRAIL)和可溶型TRAIL(Soluble TRAIL,sTRAIL)表達(dá)與功能的調(diào)節(jié)。方法:(1)采用辛酸法純化FMU-TRAIL3單克隆抗體(Monoclonal antibody,mAb)并用SDS-PAGE鑒定。采用間接免疫熒光染色及流式細(xì)胞術(shù)檢測(cè)mTRAIL的分布;用免疫沉淀及免疫印跡方法鑒定mAb對(duì)重組TRAIL(Recombinant TRAIL,rTRAIL)和細(xì)胞裂解液中TRAIL分子的識(shí)別。應(yīng)用凋亡檢測(cè)、~(51)Cr釋放實(shí)驗(yàn)鑒定FMU-TRAIL3 mAb的中和活性。(2)以不同蛋白酶抑制劑處理Jurkat、Daudi和Colo205細(xì)胞,采用間接免疫熒光染色及流式細(xì)胞術(shù)分析蛋白酶抑制劑對(duì)mTRAIL表達(dá)的影響。用RT-PCR法分析Jurkat、Daudi和Colo205細(xì)胞內(nèi)ADAM10(Adisintegrin and a metollaprotease, ADAM10)和ADAM17基因的表達(dá)。(3)分別用PMA和PHA刺激Jurkat細(xì)胞不同時(shí)間,有的實(shí)驗(yàn)加入金屬蛋白酶抑制劑1,10-phenanthroline或ADAM抑制劑TAPI-1(TNF-αprotease inhibitor-1),用間接免疫熒光染色及流式細(xì)胞術(shù)分析mTRAIL表達(dá);用ELISA檢測(cè)細(xì)胞培養(yǎng)上清中sTRAIL水平。并觀察IFN-α和金
[Abstract]:Objective to investigate the regulation of protease inhibitor on the expression and function of membrane TRAILMembrane trail and soluble TRAILLE Soluble TRAILSTRAIL. Methods FMU-TRAIL3 monoclonal antibody (FMU-TRAIL3) was purified by octanoic acid method and identified by SDS-PAGE. The distribution of mTRAIL was detected by indirect immunofluorescence staining and flow cytometry, and the recognition of recombinant trail Recombinant trail rTRAIL and trail in cell lysate was identified by immunoprecipitation and immunoblotting methods. The neutralization activity of FMU-TRAIL3 mAb was assayed by apoptosis assay. The neutralization activity of FMU-TRAIL3 mAb was assayed with different protease inhibitors. Jurkatron Daudi and Colo205 cells were treated with different protease inhibitors. The effects of protease inhibitors on the expression of mTRAIL were analyzed by indirect immunofluorescence staining and flow cytometry. The expression of ADAM10Adisintegrin and a metollaprotease, (ADAM10) and ADAM17 gene in Jurkatine Daudi and Colo205 cells were analyzed by RT-PCR. The expression of ADAM17 gene in Jurkat cells was stimulated by PMA and PHA at different time points, respectively. In some experiments, the expression of mTRAIL was detected by indirect immunofluorescence staining and flow cytometry, and the level of sTRAIL in the supernatant of cell culture was detected by Elisa, and the expression of mTRAIL was detected by indirect immunofluorescence staining and flow cytometry, by adding metalloproteinase inhibitor 1-phenanthroline or Adam inhibitor TAPI-1TNF- 偽 protease inhibitor. IFN- 偽 and gold were observed.
【學(xué)位授予單位】：第四軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2005
【分類號(hào)】：R392

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 林承曦;毛慶;高見(jiàn)書;康麗娜;徐標(biāo);;ILK轉(zhuǎn)染的BM-MSCs通過(guò)旁分泌作用抑制CFB增殖和膠原合成[J];江蘇醫(yī)藥;2011年14期

2 ;[J];;年期

3 ;[J];;年期

4 ;[J];;年期

5 ;[J];;年期

6 ;[J];;年期

7 ;[J];;年期

8 ;[J];;年期

9 ;[J];;年期

10 ;[J];;年期

相關(guān)會(huì)議論文 前1條

1 陳燁;王繼德;周殿元;;幽門螺桿菌CagA及其它疫苗候選抗原對(duì)T細(xì)胞FasLigand介導(dǎo)凋亡的調(diào)節(jié)作用[A];中國(guó)中西醫(yī)結(jié)合學(xué)會(huì)第十二次全國(guó)消化系統(tǒng)疾病學(xué)術(shù)研討會(huì)論文匯編[C];2000年

相關(guān)重要報(bào)紙文章 前7條

1 翟義;未來(lái)5年全球肝癌用藥市場(chǎng)潛力大[N];中國(guó)高新技術(shù)產(chǎn)業(yè)導(dǎo)報(bào);2002年

2 付東紅;腫瘤轉(zhuǎn)移有重要標(biāo)記物[N];健康報(bào);2002年

3 ;輝瑞公司長(zhǎng)江后浪推前浪[N];中國(guó)高新技術(shù)產(chǎn)業(yè)導(dǎo)報(bào);2001年

4 鄭莉麗 王雪飛 符壯才;介入術(shù)后能防冠脈再狹窄嗎[N];健康報(bào);2002年

5 付東紅;北大基礎(chǔ)醫(yī)學(xué)院基質(zhì)金屬蛋白酶作用機(jī)理初步揭示[N];人民日?qǐng)?bào)海外版;2003年

6 陳茂梁 余寧寧;浙醫(yī)二院解決腦癌治療難題[N];中國(guó)醫(yī)藥報(bào);2001年

7 香港麥迪信醫(yī)學(xué)出版有限公司供稿;最佳時(shí)機(jī) 最佳劑量 最佳方案[N];醫(yī)藥經(jīng)濟(jì)報(bào);2002年

相關(guān)博士學(xué)位論文 前1條

1 李妍;金屬蛋白酶抑制劑參與膜型和可溶型凋亡誘導(dǎo)配體TRAIL表達(dá)與功能的調(diào)節(jié)[D];第四軍醫(yī)大學(xué);2005年

相關(guān)碩士學(xué)位論文 前2條

1 劉鳳潔;基質(zhì)金屬蛋白酶-2及其抑制劑-2在妊高征患者胎盤中的表達(dá)[D];吉林大學(xué);2005年

2 于爽;局灶性腦缺血大鼠腦組織EAATs、mGluR、TIMP的表達(dá)[D];延邊大學(xué);2008年

，

本文編號(hào)：2032225