發(fā)布時(shí)間：2018-03-03 20:39

  本文選題：Suramin　切入點(diǎn)：淀粉樣纖維　出處：《南方醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：研究背景:艾滋病(Acquired immunodeficiency sydrom,AIDS)是由人類免疫缺陷病毒(Human immunodeficiency virus type 1,HIV-1)引起的一種免疫能力喪失的傳染性疾病,嚴(yán)重威脅著人類的生命和健康。全球范圍內(nèi),新感染病例數(shù)居高不下,其中80%新感染病例來自性傳播,性傳播已成為HIV-1最主要的傳播途徑。殺微生物劑是種有效預(yù)防HIV-1性傳播的重要手段。但迄今為止,仍未開發(fā)出有效的殺微生物劑。精液來源的病毒增強(qiáng)因子(SEVI)能削弱殺微生物劑的抗病毒活性同時(shí)顯著促進(jìn)病毒的感染。SEVI是前列腺酸性磷酸酶(PAP)裂解的多肽片段PAP248-286自我組裝形成的淀粉樣纖維。SEVI的陽離子特性能消除病毒和細(xì)胞的靜電排斥,增大病毒和靶細(xì)胞接觸幾率,從而增強(qiáng)HIV-1感染。因此,SEVI是殺微生物劑的一個(gè)重要靶點(diǎn)。我們前期研究表明,HIV-1進(jìn)入抑制劑ADS-J1既能拮抗SEVI,與抗病毒藥物合用具有協(xié)同抗病毒效果。但由于ADS-J1具有潛在的致癌性,限制了它的臨床應(yīng)用。我們?cè)贏DS-J1類似物中篩選到一個(gè)本身具有抗HIV-1活性的化合物Suramin,它是臨床治療錐蟲病的經(jīng)典老藥,具有強(qiáng)大的藥理活性:抑制蛋白酶和阻斷生長(zhǎng)因子等。前期研究表明,Suramin能抑制多肽PAP248-286形成SEVI,本文將深入研究Suramin抑制及拮抗SEVI的作用及機(jī)制;由于Suramin本身具有抗HIV-1活性,我們也將探討其與抗病毒藥物合用協(xié)同抗病毒效果,期望將Suramin開發(fā)為殺微生物劑或其輔助用藥,更好地預(yù)防HIV-1性傳播。研究目的從具備拮抗HIV-1性傳播過程作用多靶點(diǎn)能力的角度闡述Suramin用于預(yù)防HIV-1性傳播的新的可能及機(jī)制,從老藥新用的角度尋求擴(kuò)大Suramin臨床適應(yīng)癥,為預(yù)防HIV-1性傳播的殺微生物劑研發(fā)提供新的研究思路。研究方法1.剛果紅染色、透射電鏡、激光共聚焦顯微鏡、圓二色譜、Western blot和病毒感染實(shí)驗(yàn)方法檢測(cè)Suramin對(duì)SEVI形成及纖維本身的影響。2.病毒感染實(shí)驗(yàn)檢測(cè)Suramin合用抗逆轉(zhuǎn)錄病毒藥物(ARVs)的抗病毒效果。3.XTT法檢測(cè)Suramin對(duì)生殖道和淋巴細(xì)胞的毒性作用。通過家兔陰道粘膜刺激性實(shí)驗(yàn),Suramin凝膠劑進(jìn)行安全性初步評(píng)價(jià),其中包括組織病理學(xué)檢查、ELISA法檢測(cè)陰道灌洗液細(xì)胞因子表達(dá)水平和PCNA免疫組化評(píng)價(jià)家兔陰道組織的炎癥。4.多肽PAP248-286與Suramin進(jìn)行分子動(dòng)力學(xué)(MD)模擬,從分子動(dòng)力學(xué)角度證明Suramin抑制SEVI形成機(jī)制。研究結(jié)果1.Suramin能濃度依賴性抑制多肽PAP248-286纖維化;Suramin能結(jié)合SEVI,阻斷它和HIV-1的結(jié)合,降低SEVI促進(jìn)HIV-1的感染能力。2.Suramin在精液中聯(lián)合ARVs可以抑制CCR5受體和CXCR4受體嗜性HIV-1,50%抑制率時(shí)的聯(lián)合指數(shù)CI值范圍為0.233-0.739。3.Suramin在體外對(duì)生殖道細(xì)胞和淋巴細(xì)胞基本沒有毒性作用,它對(duì)各細(xì)胞系的CC50均大于1000 μM。組織病理學(xué)檢查結(jié)果顯示Suramin凝膠組的陰道粘膜上皮細(xì)胞均較完整,固有層組織僅輕度水腫;Suramin凝膠不會(huì)促進(jìn)炎癥因子的釋放;Suramin凝膠組的陰道上皮細(xì)胞和基質(zhì)細(xì)胞PCNA陽性表達(dá)率與生理鹽水組無顯著性差異。4.MD模擬表明分子結(jié)構(gòu)非常相似的3個(gè)化合物與PAP248-286結(jié)合模式不同,分子結(jié)構(gòu)趨向于立體的Suramin能把PAP248-286完全包裹住,另外其改變PAP248-286單體結(jié)構(gòu)的程度最大:Suramin幾乎與PAP248-286 上所有的氨基酸有氫鍵或疏水作用:氫鍵有19個(gè),疏水作用數(shù)為45。研究結(jié)論1.Suramin能緊密結(jié)合多肽PAP248-286,抑制其自我聚合形成淀粉樣纖維SEVI。2.Suramin阻斷SEVI和HIV-1結(jié)合,拮抗其增強(qiáng)病毒感染能力。3.Suramin在精液中分別聯(lián)合不同作用機(jī)制的ARVs均具有協(xié)同抗病毒作用,且對(duì)生殖道細(xì)胞和淋巴細(xì)胞基本無毒性作用,對(duì)家兔陰道無明顯致炎作用,有較高的安全性。
[Abstract]:Background: AIDS (Acquired immunodeficiency, Sydrom, AIDS) is by the human immunodeficiency virus (Human immunodeficiency virus type 1, HIV-1) caused a loss of immunity to infectious diseases, serious threat to human life and health. Globally, the number of new infections is high, of which 80% new infections from spreading. Sexual transmission has become the most important route of transmission of HIV-1. A microbicide is an important means for effective prevention of sexual transmission of HIV-1. But so far, has not yet developed effective microbial kill agent. Semen derived virus enhancement factor (SEVI) can weaken the microbicide antiviral activity also significantly promote.SEVI virus infection is prostatic acid phosphatase (PAP) characteristics of cationic polypeptide fragment PAP248-286 cleavage of the self-assembly of amyloid fibrils formed by the.SEVI can eliminate the virus and cell The electrostatic repulsion, the risk of viral and target cell contact increases, thereby enhancing the infection of HIV-1. Therefore, SEVI is an important target for microbicides. Our previous study showed that HIV-1 can antagonize SEVI ADS-J1 entry inhibitors, in combination with antiviral drugs has synergistic antiviral effect. But because ADS-J1 has potential carcinogenicity, limiting the clinical the application of it. We screened ADS-J1 analogues in a compound with anti HIV-1 activity of Suramin, it is a classic old drug treatment of trypanosomiasis, has strong pharmacological activity: inhibition of protease and blocking growth factor. Previous studies have shown that Suramin can inhibit the formation of peptide PAP248-286 SEVI, effect and mechanism this paper will deeply Research on Suramin inhibition and antagonism of SEVI; because Suramin itself has anti HIV-1 activity, we will also discuss the Chinese medicine combined with synergistic antiviral effect, Expect to exploit Suramin as a microbicide or auxiliary medicine for better prevention of sexual transmission of HIV-1. The purpose of the study on the Suramin from antagonism to HIV-1 communication process multi-target capability perspective for possible new mechanism and prevention of sexual transmission of HIV-1, seeking to expand Suramin clinical indications from the angle of the new old drugs and provide a new idea for the microbicide to prevent sexual transmission of HIV-1. Methods 1. Congo red staining, transmission electron microscopy, laser confocal microscope, round two chromatography, detection of Suramin on the formation of SEVI and blot test method of Western virus infection and the effect of fiber.2. virus infection assay Suramin combined antiretroviral drugs the detection of Suramin (ARVs) toxicity of the antiviral effect of.3.XTT method on genital and lymphocytes. The rabbit vaginal mucosa irritation test, Suramin gel into. Preliminary evaluation, including histopathological examination, detection of vaginal lavage cytokine ELISA expression and immunohistochemistry of PCNA.4. peptide PAP248-286 and Suramin in the assessment of inflammatory rabbit vaginal tissue by molecular dynamics (MD) simulations show that Suramin inhibits SEVI formation mechanism from the view of molecular dynamics. The research results of 1.Suramin concentration dependence inhibition of polypeptide PAP248-286 fibrosis; Suramin can bind to SEVI, blocking its combination with HIV-1, reduce HIV-1 infection combined with ARVs promote the ability of.2.Suramin in semen SEVI index can be combined inhibition of CCR5 receptor and CXCR4 receptor tropism HIV-1,50% inhibition rate when the CI value is in the range of 0.233-0.739.3.Suramin in vitro on reproductive tract cells and lymphocytes basically no toxic effects. It is on the cell line CC50 was greater than 1000 M. histopathological examination showed vaginal gel group Suramin Epithelial cells were relatively complete propria tissue only mild edema; Suramin gel does not promote the release of inflammatory factors; Suramin gel group PCNA vaginal epithelial cells and stromal cells and the positive expression rate of saline group had no significant difference between the.4.MD simulation shows that the molecular structure is similar to 3 compounds and PAP248-286 binding mode, molecular the three-dimensional structure tends to Suramin the PAP248-286 can completely wrap, also the change of PAP248-286 monomer structure maximum: Suramin and PAP248-286 almost all amino acids have hydrogen bonds and hydrophobic interaction of hydrogen bonds 19, hydrophobic interaction number 45. conclusions 1.Suramin can combine polypeptide PAP248-286, inhibiting its self polymerization to form starch kind of fiber SEVI.2.Suramin blocking SEVI and HIV-1 combination, enhance the ability of.3.Suramin antagonistic virus infection in semen respectively with different effect The mechanism of ARVs has synergistic antiviral effect, and has no toxic effect on reproductive tract cells and lymphocytes. It has no obvious inflammation effect on rabbit vagina, and has high safety.

【學(xué)位授予單位】：南方醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R512.91

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 張華群;曹潔;潘衛(wèi);;人精液源性病毒感染增強(qiáng)因子研究進(jìn)展[J];國(guó)際生物制品學(xué)雜志;2010年02期

相關(guān)博士學(xué)位論文 前1條

1 李琳;酸酐修飾卵清蛋白作為候選殺微生物劑預(yù)防HIV性傳播研究[D];南方醫(yī)科大學(xué);2010年

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本文編號(hào)：1562658