本文關(guān)鍵詞：茴香霉素對(duì)小鼠淋巴細(xì)胞行為及其同種異基因皮膚移植的影響　出處：《暨南大學(xué)》2007年碩士論文　論文類(lèi)型：學(xué)位論文

  更多相關(guān)文章： 茴香霉素 淋巴細(xì)胞 增殖 周期 活化 細(xì)胞毒作用 混合淋巴細(xì)胞發(fā)應(yīng) 遲發(fā)性超敏反應(yīng) 皮膚移植 小鼠

【摘要】： 目的：研究茴香霉素對(duì)小鼠淋巴細(xì)胞行為及其同種異基因皮膚移植的影響。方法：以活體染料羧基熒光素乙酰乙酸琥珀酰亞胺酯染色，建立在多克隆刺激劑刀豆蛋白A(ConA)刺激下評(píng)價(jià)小鼠淋巴細(xì)胞增殖的模型，通過(guò)流式細(xì)胞術(shù)和MTT法分析茴香霉素在不同劑量下對(duì)淋巴細(xì)胞增殖的作用；采用碘化丙錠染色分析茴香霉素對(duì)ConA或佛波醇酯(PDB)加離子霉素(Ion)刺激的小鼠淋巴細(xì)胞周期變化的作用；利用熒光標(biāo)記的單克隆抗體雙染技術(shù)和流式細(xì)胞術(shù)觀(guān)察茴香霉素對(duì)小鼠CD3~+T細(xì)胞早期及中期活化標(biāo)志分子CD69和CD25表達(dá)的影響。用MTT法檢測(cè)茴香霉素作用下淋巴細(xì)胞在單向或雙向混合淋巴細(xì)胞反應(yīng)(MLR)中的反應(yīng)性、對(duì)大鼠肝癌細(xì)胞(7919)的殺傷效應(yīng)以及茴香霉素對(duì)原代淋巴細(xì)胞和7919細(xì)胞系的直接細(xì)胞毒作用。建立小鼠同種異基因皮膚移植模型，觀(guān)察不同劑量茴香霉素對(duì)皮膚移植存活時(shí)間、遲發(fā)性超敏反應(yīng)(DTH)及移植后小鼠淋巴細(xì)胞反應(yīng)性的影響。結(jié)果：茴香霉素濃度在0.01～0.5ng／ml時(shí)，能夠增強(qiáng)ConA對(duì)淋巴細(xì)胞的促增殖作用，以0.5ng／ml茴香霉素促進(jìn)作用最明顯；1.0～25.0ng／ml茴香霉素則能抑制ConA對(duì)淋巴細(xì)胞的促增殖作用。進(jìn)一步發(fā)現(xiàn)，0.01～0.5ng／ml茴香霉素能夠促進(jìn)ConA誘導(dǎo)的淋巴細(xì)胞周期進(jìn)入G2／M期，促進(jìn)PDB加Ion刺激的淋巴細(xì)胞周期進(jìn)入S期：1.0～25.0ng／ml茴香霉素能使ConA或PDB加Ion刺激的淋巴細(xì)胞周期停滯于G0／G1期，阻止其進(jìn)入S期，以10.0ng／ml和25.0ng／ml茴香霉素的抑制作用最為明顯。據(jù)此選用最佳劑量10.0ng／ml，茴香霉素能夠明顯抑制CD3~+T細(xì)胞表面分子CD69和CD25的表達(dá)(P均＜0.01)，，且能抑制單向或雙向MLR中淋巴細(xì)胞的反應(yīng)性(P均＜0.01)，其抑制作用與地塞米松相比無(wú)明顯差異(P＞0.05)；茴香霉素劑量從0.1ng／ml逐漸增至100.0ng／ml，淋巴細(xì)胞對(duì)大鼠肝癌細(xì)胞的殺傷效應(yīng)逐漸減弱，以10.0ng／ml和100.0ng／ml劑量組抑制作用最為明顯(P＜0.01)；在上述劑量下，茴香霉素對(duì)小鼠原代淋巴細(xì)胞和大鼠肝癌7919細(xì)胞系無(wú)直接細(xì)胞毒作用。體內(nèi)研究進(jìn)一步發(fā)現(xiàn)，15.0mg／kg茴香霉素能夠顯著延長(zhǎng)小鼠皮膚移植存活時(shí)間、抑制DTH發(fā)展及體內(nèi)小鼠淋巴細(xì)胞的反應(yīng)性，且上述作用明顯強(qiáng)于環(huán)孢素A(CsA) (P＜0.01或P＜0.05)。結(jié)論：茴香霉素能明顯抑制小鼠淋巴細(xì)胞的增殖、周期、活化、抗原反應(yīng)性和殺傷效應(yīng)等，抑制小鼠同種異基因皮膚移植的排斥反應(yīng)，且作用優(yōu)于CsA，這些發(fā)現(xiàn)可能為其作為新的免疫抑制劑的研發(fā)提供理論和實(shí)驗(yàn)依據(jù)。
[Abstract]:Objective: to study the effect of anisomycin on lymphocyte behavior and allogeneic skin transplantation in mice. Methods: in vivo dye carboxyl fluorescein acetoacetate succinimide ester staining was used. A model was established to evaluate lymphocyte proliferation in mice stimulated by concanavalin Cona, a polyclonal stimulator. The effect of anisomycin on lymphocyte proliferation was analyzed by flow cytometry and MTT. Using iodized propyl ingot staining, the effect of anisomycin on lymphocyte cycle changes induced by ConA or phorbol ester PDBs plus ionomycin Ion (Ion) in mice was analyzed. Observation of the effect of anisomycin on mouse CD _ 3 ~ ~ by fluorescence labeled monoclonal antibody double staining technique and flow cytometry. The expression of CD69 and CD25 in the early and middle stage of T cell activation. The lymphocyte reaction in unidirectional or bidirectional mixed lymphocyte was detected by MTT method. The reactivity in MLR. The cytotoxicity of anisomycin on primary lymphocytes and 7919 cell line was studied. The allogeneic skin transplantation model of mice was established. The survival time of different doses of anisomycin on skin transplantation was observed. Delayed hypersensitivity reaction (DTH) and lymphocyte reactivity after transplantation. Results: the concentration of anisome was 0. 01 ~ 0. 5 ng / ml. The effect of ConA on lymphocyte proliferation was enhanced, especially by 0.5 ng / ml anisamycin. Anisomycin at a dose of 25.0ng / ml inhibited the proliferation of lymphocytes induced by ConA. 0. 01 ~ 0. 5 ng / ml anisomycin could promote lymphocyte cycle induced by ConA into G 2 / M phase. To promote the lymphocyte cycle stimulated by PDB and Ion into S phase:. 1.0 ng / ml anisomycin could arrest the lymphocyte cycle induced by ConA or PDB plus Ion in G _ 0 / G _ 1 phase. The inhibitory effect of anisomycin was the most obvious in 10.0ng / ml and 25.0ng / ml of anisomycin, and the best dose was 10.0ng / ml. Anisamycin could significantly inhibit the expression of CD69 and CD25 on CD3T cells (P < 0.01). The reactivity of lymphocytes in unidirectional or bidirectional MLR was inhibited (P < 0.01), and there was no significant difference compared with dexamethasone (P > 0.05). The dose of anisomycin increased from 0.1 ng / ml to 100.0 ng / ml, and the killing effect of lymphocytes on hepatoma cells decreased gradually. The inhibitory effect of 10.0ng / ml and 100.0ng / ml groups was the most obvious (P < 0.01). Under the above dosage, anisomycin had no direct cytotoxic effect on primary lymphocytes of mice and rat hepatoma 7919 cell line. Anisomycin 15.0mg / kg could significantly prolong the survival time of mouse skin graft and inhibit the development of DTH and the reactivity of lymphocytes in vivo. The above effects were significantly stronger than that of cyclosporine (P < 0.01 or P < 0.05). Conclusion: anisomycin can significantly inhibit the proliferation, cycle and activation of lymphocytes in mice. The antigenic reactivity and killing effect could inhibit the rejection of allogeneic skin transplantation in mice and its effect was superior to that of CSA. These findings may provide a theoretical and experimental basis for the development of new immunosuppressants.
【學(xué)位授予單位】：暨南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類(lèi)號(hào)】：R392

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