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国家自然科学基金(81360501)

作品数:8 被引量:6H指数:2
相关作者:马丹方琴王季石王萍孙嘉更多>>
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发文基金:国家自然科学基金贵州省科学技术基金贵州省省长基金更多>>
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Upregulated heme oxygenase-1 expression of mouse mesenchymal stem cells resists to chemotherapy-induced bone marrow suppression
2014年
Background Bone marrow hematopoietic function suppression is one of the most common side effects of chemotherapy.After chemotherapy,the bone marrow structure gets destroyed and the cells died,which might cause the hematopoietic function suppression.Heme oxygenase-1 (HO-1) is a key enzyme of antioxidative metabolism that associates with cell proliferation and resistance to apoptosis.The aim of this study was to restore or resist the bone marrow from the damage of chemotherapy by the HO-1 expression of mouse mesenchymal stem cells (mMSCs) homing to the mice which had the chemotherapy-induced bone marrow suppression.Methods One hundred and sixty female Balb/c mice (6-8-weeks old) were randomly divided into four groups.Each group was performed in 40 mice.The control group was intraperitoneally injected for 5 days and tail intravenously injected on the 6th day with normal saline.The chemotherapy-induced bone marrow suppression was established by intraperitoneally injecting cyclophosphamide (CTX) into the mice which performed as the chemotherapy group.The mMSCs were tail intravenously injected into 40 chemotherapically damaged mice which served as the mMSCs group.The difference between the HO-1 group and the mMSCs group was the injected cells.The HO-1 group was tail intravenously injected into the mMSCs that highly expressed HO-1 which was stimulated by hemin.The expression of HO-1 was analyzed by Western blotting and RT-PCR.Cell proliferation was measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay.Histopathologic examinations were performed 1 week after injection.Results Compared with the control group,the expression levels of HO-1 mRNA and protein were significantly higher in the HO-1 group (all P <0.05),even obviously than the mMSCs group.CTX treatment induced apoptosis and inhibited proliferation.After injected,the white blood cell (WBC),red blood cell (RBC) and platelet (PLT) declined fast and down to the bottom at the 7th day.The bone marrow
Chen ShuyaWang JishiFang QinGao RuiShi QianyingZhang HuiZhao Jiangyuan
高表达ALDH2基因改善血管内皮细胞功能的初步研究被引量:1
2016年
目的:探讨乙醛脱氢酶2(ALDH2)基因导入人脐静脉内皮细胞(HUVECs)后对其增殖和抗氧化损伤的影响。方法:将培养的HUVECs分为转染目的基因组(转基因组)、转染空载体组(空质粒对照组)和未转染质粒组(阴性对照组),转基因组采用脂质体介导的方法将含人ALDH2基因的真核表达载体导入HUVECs中,采用RT-PCR和Western blot检测各组ALDH2基因和蛋白的表达;采用不同浓度过氧化氢(H_2O_2)(0、50、75、100和125μmol/L)孵育HUVECs 48 h后,检测各组细胞增殖水平及氧化损伤程度,检测各组细胞中血红素氧合酶(HO-1)、ROS及一氧化氮(NO)含量的变化。结果:转基因组荧光强度和数量比阴性对照组显著提高,提示导入的ALDH2基因在HUVECs细胞得到高表达;与空质粒对照组和阴性对照组比较,转染72 h后,转基因组细胞ALDH2 mRNA和蛋白表达增加(P<0.01);不同浓度H_2O_2孵育48 h后,转基因组HUVECs存活率明显升高(P<0.05);转基因组可抑制一定浓度H_2O_2诱导的ROS水平的升高、降低HO-1表达及增加NO的产生,差异有统计学意义(P<0.05)。结论:ALDH2基因高表达可降低低氧自由基引起的HUVECs细胞的损伤,保护内皮细胞的生物学功能。
胡秀英方琴王季石马丹蒋丽
关键词:乙醛脱氢酶2转染内皮细胞ALDEHYDEDEHYDROGENASE
17-丙烯胺基-17-去甲氧基格尔德霉素增强慢性粒细胞白血病细胞对伊马替尼敏感性的机制研究
2014年
目的探索伊马替尼(IM)联合使用17-丙烯胺基-17-去甲氧基格尔德霉素(17-AAG)增加慢性粒细胞白血病(CML)细胞株K562和K562A02凋亡率的机制。方法采用四甲基偶氮唑蓝法观察细胞增殖抑制率,Annexin V/PI双染色法检测细胞凋亡率,逆转录-聚合酶链反应法检测Bcr-abl、MDR1、Bcl-2 mRNA的表达及Western blot法检测Bcr-abl、p-gp、Bcl-2蛋白的表达。结果0.31μg·mL-1伊马替尼联合1.25μg·mL-117-丙烯胺基-17-去甲氧基格尔德霉素分别作用于K562和K562A02细胞48 h,增殖抑制率分别为(70.78±1.01)%和(39.27±1.52)%,凋亡率为(48.74±2.51)%和(45.0±0.4)%。2.5μg·mL-1伊马替尼联合10μg·mL-117-丙烯胺基-17-去甲氧基格尔德霉素作用于K562A02细胞48 h,增殖抑制率达(65.63±0.93)%。逆转录-多聚酶链反应和Western blot结果显示,联合用药后K562细胞的Bcr-abl和Bcl-2基因显著降低,MDR1呈弱表达,无明显变化。而K562A02细胞中,Bcr-abl、Bcl-2和MDR1基因的表达均有明显下调(P<0.05)。结论伊马替尼联合17-丙烯胺基-17-去甲氧基格尔德霉素可有效抑制K562和K562A02细胞增殖和诱导凋亡,两者联合应用具有克服伊马替尼耐药的作用。
方琴谢建琼王季石胡秀英柴柏胜
关键词:17-丙烯胺基-17-去甲氧基格尔德霉素伊马替尼慢性粒细胞白血病耐药
血红素氧合酶-1在减轻硝酸甘油耐受中的作用研究
2017年
目的探讨血红素氧合酶-1(HO-1)在减轻硝酸甘油耐受中的作用。方法建立人脐静脉内皮细胞(HUVECs)的硝酸甘油耐受模型;采用HO-1诱导剂Hemin和抑制剂锌原卟啉(ZnPP)分别进行干预处理后,利用RT-PCR以及Western blot检测HO-1的表达情况。分别采用硝酸还原酶法和荧光分光光度法检测药物干预后各组细胞NO及活性氧ROS的表达水平。结果与对照组相比,Hemin诱导组细胞增殖水平增高(P<0.05),并且HO-1蛋白表达上调可以增加NO的含量(P<0.01),促进硝酸甘油的生物转化,该过程伴随着ROS的减少(P<0.01)。结论通过Hemin诱导HO-1过表达可以一定程度上减轻硝酸甘油的耐受,该过程与氧化应激水平的降低有关。
胡秀英方琴王季石马丹蒋丽
关键词:耐受硝酸甘油血管内皮细胞氧化应激HO-1
白桦脂酸联合硼替佐米诱导U266细胞凋亡及其机制研究被引量:2
2015年
目的:探讨白桦脂酸(BA)联合硼替佐米诱导多发性骨髓瘤(MM)U266细胞凋亡及其机制。方法:用不同浓度的BA(20、40、60及80 mg/L)、硼替佐米(10、80及320 nmol/L)、40 mg/L BA分别联合10、80及320 nmol/L硼替佐米(联合1~3组)处理U266细胞,MTT法检测上述各组U266细胞增殖抑制率;Real-time PCR和蛋白免疫印迹法(Western blot)分别检测40 mg/L BA、80 nmol/L硼替佐米及40 mg/L BA联合80 nmol/L硼替佐米(联合4组)的U266细胞Survivin、Cyto-C、BCL-2及Bax的mRNA和蛋白表达水平。结果:联合1~3组对U266细胞增殖抑制率显著高于BA组和硼替佐米组(P〈0.05);与硼替佐米组和BA组相比,联合4组U266细胞的cyto-C,Bax mRNA和蛋白表达水平显著升高(P〈0.05),而Survivin、Bcl-2 mRNA和蛋白表达水平显著降低(P〈0.05)。结论:BA联合硼替佐米可以促进MM肿瘤细胞凋亡,机制可能与下调Survivin、Bcl-2表达,上调Cyto-c、Bax表达有关。
孙嘉马丹王萍方琴
关键词:白桦脂酸硼替佐米U266细胞凋亡基因表达
Over-expression of heme oxygenase-1 in peripheral blood predicts the progression and relapse risk of chronic myeloid leukemia
2014年
Background There are limited eligible clinical markers at present to monitor the progress of chronic myeloid leukemia (CML).Heme oxygenase-1 (HO-1),as one of the most important oxidation-regulating enzymes in vivo,suggests the onset and progression of cancer when highly expressed.Furthermore,HO-1 level is related with the occurrence and development of hematological diseases.But the relationship between HO-1 expression and progression/relapse of CML has seldom been studied hitherto.This study aimed to investigate the relationship between them to find out a new molecular marker for prediction.Methods A total of 60 peripheral blood and bone marrow (BM) samples from 25 CML patients in different phases were collected respectively to detect the expressions of HO-1 and bcr/abl using real-time PCR.Routine blood test was performed to detect the changes of leukocyte and platelet counts.The proportion of primitive cells in BM was detected by flow cytometry.The relationship between high HO-1 expression and CML progression and relapse was explored by the analysis of variance by Wilcoxon test and linear regression analysis.The diagnostic accuracy and cutoff values were determined by receiver operating characteristic curve.Results Relative expression of HO-1 mRNA in CML patients peripheral blood was significantly higher than that of donors (P 〈0.0001),which were 0.57±3.78 and (1.417±1.125)×10-6,respectively.HO-1 expression level in CML patients was 0.061 5±0.062 4,which decreased to 0.009 4±0.006 7 upon CMoR,and remained remarkably higher 0.016 3±0.017 5 than that of normal donors (1.417±1.125)× 10-6,P 〈0.001.When relapse occurred,HO-1 expression significantly increased from 0.020 6±0.021 0 to 3.852±10.285 in CMoR stage and undergoing relapse.According to progression of CML,HO-1 expression level in CML patients increased from CP (0.009 5±0.017 6) to AP (0.028 0±0.055 7) and then to BP (0.276 7± 0.447 0).And there was a linear correlation between HO-1 expression and proportion
Wei SixiWang YatingChai QixiangFang QinZhang YamingLu YinghaoWang Jishi
关键词:PROGRESSIONRELAPSE
白桦脂酸联合沙利度胺诱导U266细胞凋亡机制研究被引量:1
2015年
目的:探讨白桦脂酸联合沙利度胺诱导多发性骨髓瘤(MM)U266细胞凋亡的机制。方法:分别用白桦脂酸(20、40、60和80 mg/L,白桦脂酸组)、沙利度胺(10、50和100 mg/L,沙利度胺组)及白桦脂酸(40 mg/L)联合沙利度胺(联合组,白桦脂酸40 mg/L,沙利度胺10、50和100 mg/L)分别处理U266细胞,同期设对照组;MTT法和流式细胞术分别检测不同浓度白桦脂酸组、沙利度胺组及联合组U266细胞增殖抑制率和凋亡率;Real-time PCR检测4组U266细胞中Survivin、Cyto-C、Bcl-2、Bax mRNA的表达,蛋白免疫印迹法检测4组U266细胞中Survivin、Cyto-C、Bcl-2、Bax蛋白表达水平。结果:随着白桦脂酸浓度的增加,U266细胞增殖抑制率增加,差异有统计学意义(P<0.05),最适浓度为40 mg/L;与沙利度胺组相比,联合组U266细胞的增殖抑制率和凋亡率明显升高,差异具有统计学意义(P<0.05);与沙利度胺组或白桦脂酸组相比,联合组U266细胞中Survivin、Bcl-2 mRNA的表达明显降低,Cyto-C和Bax mRNA表达明显升高,差异具有统计学意义(P<0.05);与沙利度胺、白桦脂酸组相比,联合组U266细胞中Survivin、Bcl-2蛋白表达明显降低,Cyto-C和Bax蛋白表达明显升高,差异具有统计学意义(P<0.05)。结论:白桦脂酸联合沙利度胺可以促进多发性骨髓瘤U266细胞凋亡,其机制可能与凋亡分子Survivin、Bcl-2、Cyto-c和Bax相关。
孙嘉马丹王萍方琴
关键词:白桦脂酸沙利度胺多发性骨髓瘤U266细胞增殖凋亡
调控血红素加氧酶-1诱导K562A02细胞增殖、凋亡及耐药机制研究被引量:2
2016年
目的通过血红素加氧酶-1(HO-1)诱导剂Hemin及抑制剂ZNPP IX调控HO-1,并联合阿霉素逆转K562A02细胞化疗耐药机制的研究,为慢性髓系白血病(CML)的逆转耐药提供新的策略。方法培养K562及K562A02细胞,采用荧光原位杂交(FISH)法检测K562A02细胞中bcr-abl融合基因表达。分别用HO-1诱导剂Hemin及抑制剂ZNPP IX调控HO-1基因表达联合阿霉素处理K562A02细胞后;流式细胞术检测药物诱导细胞凋亡情况。Western blot检测耐药相关基因及凋亡基因蛋白表达水平。结果 K562A02细胞中bcr-abl融合基因阳性细胞占94%。阿霉素处理细胞后,随着阿霉素浓度的增加,HO-1表达下降,耐药相关基因MDR1、NF-κB(P65)、MRP1、TopoⅡα、ABCD2表达亦降低;用HO-1诱导剂Hemin、抑制剂ZNPP IX、阿霉素单药分别及联合处理K562A02细胞后,显示HO-1高表达后耐药相关基因表达升高,细胞凋亡率下降。而降低HO-1表达,耐药相关基因表达下降,细胞凋亡率增加。结论 HO-1可作为逆转耐药的靶基因,可以使K562A02对阿霉素重新敏感,起到增敏效应。
柴其翔韦四喜王娅婷方琴王季石
关键词:BCR-ABL融合基因阿霉素K562血红素加氧酶-1
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