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题名 胸腺五肽抗肿瘤和抗氧化作用的研究
姓名 常惠
院系 生命科学学院
专业 生物化学与分子生物学
学位名称 理学硕士
外文题名 Studies on Anti-tumor and Anti-oxidative Effects of Thymopentin (TP5)
第一导师姓名 陈强
关键词 胸腺五肽;白血病细胞;联合作用;细胞增殖;细胞凋亡;细胞分化;心肌;羟自由基;过氧化氢;PC12细胞
外文关键词 TP5;leukemia cells;coordinate effect;apoptosis;cell differentiation;myocardial injury;hydroxyl free radical;H2O2;PC12 cells
学科 理学
摘要 胸腺五肽(Thymopentin , TP5,Arg-Lys-Asp-Val-Tyr)是人工合成的在人体内具有免疫调节活性的五肽。本文主要研究TP5与化疗药物联合使用时在体外对人白血病细胞株HL-60细胞和K562细胞增殖与分化的影响。应用生长曲线测定法检测TP5与DMSO(二甲基亚砜)联合作用对人早幼粒白血病细胞株HL-60细胞增殖的影响以及TP5与ADM(阿霉素)联合作用对人慢性髓系白血病细胞株K562细胞增殖的影响;根据硝基蓝四氮唑(NBT)还原能力来测定白血病细胞的分化;用流式细胞仪检测细胞凋亡率的变化以及凋亡相关基因Bcl-2和CD95的表达。结果表明,TP5与DMSO联合应用对HL60细胞增殖有协同抑制作用,TP5能显著增强DMSO对HL60细胞的NBT还原能力,流式细胞仪检测发现TP5与DMSO联合应用诱导HL-60细胞发生凋亡;此外,TP5与ADM联合应用对K562细胞增殖有协同抑制作用,TP5能显著增强ADM对K562细胞的NBT还原能力,瑞氏-姬姆萨染色细胞结果显示TP5与 ADM联合应用使K562细胞形态发生显著变化,形态学结果表明K562细胞的分化增强。这些结果表明,TP5不但可以作为肿瘤化疗过程中的免疫调节剂,还可以作为一种潜在的白血病化疗药物。 本实验中我们还研究了TP5对外源羟自由基致大鼠心肌细胞损伤的作用。采用由Fenton体系产生的羟自由基损伤心肌细胞,MTT法检测细胞活力。结果显示,TP5对外源羟自由基致大鼠心肌细胞损伤具有显著的保护作用,这种保护作用可能是通过TP5对自由基的清除实现的。 我们还研究了TP5对H2O2致PC12细胞损伤的作用。结果显示TP5可以浓度依赖的显著削弱H2O2 (0.5mmol/L)对PC12细胞的损伤作用。推测TP5在神经系统损伤修复中发挥重要作用。
外文摘要 Thymopentin (Arg-Lys-Asp-Val-Tyr, TP5) has shown immuno-regulatory activities in human beings. In the present study, we investigated the effects of TP5 combined with chemotherapeutic agents (DMSO or ADM) on the proliferation and differentiation of two human leukemia cell lines (HL-60 and K562). The anti-proliferation and differentiating effects induced by TP5 and DMSO (or ADM) were determined by cell viability, Wright''s-Giemsa staining, and NBT-reduction activity. The percentage of apoptosis cells and the expression of Bcl-2 and CD95 were analysed by flow cytometry (FCM). The results showed that TP5 and DMSO displayed concentration-dependent inhibitory effects on the proliferation of HL-60 cells. TP5 could induce differentiation of HL-60 cells and enhance the effect when combined with DMSO. TP5 (500μmol/L) and DMSO (2%) induced apoptosis in HL-60 cells. In addition, TP5 could inhibit proliferation and induce differentiation of K562 cells and enhance the effects when combined with ADM. The significant morphology changes were observed when K562 cells were exposed to TP5 and ADM. All the results indicated that TP5 not only act as an immunomodulatory factor in cancer chemotherapy, but also could be a potential chemotherapeutic agent in the human leukemia therapy. We also investigated the protective effect of TP5 on myocardial injury induced by hydroxyl free radical in rats. The hydroxyl free radical was generated by the Fenton system established by FeSO4 / H2O2. The cell viability of cardiac myocyte was determined by MTT assay. The result indicated that TP5 significantly attenuated myocardial injury induced by the Fenton system. The protection effect of TP5 was possibly via scavenging hydroxyl free radical. We studied the protective effect of TP5 on cultured PC12 cells injured by hydrogen peroxide. PC12 cells were pretreated with different concentrations of TP5 for 2 hours, then, treated with H2O2 (0.5mmol/L) for 48h. The cell survival rate (MTT assay) was measured. The data indicated that TP5 can strikingly and dose-dependently attenuated cell injury induced by H2O2. Our results suggest that TP5 may play an important role in neuron system repairing.
研究领域 多肽生化与药化
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