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1.
目的:研究纯种发酵淡豆豉异黄酮提取物在体外对人乳腺癌细胞MCF-7的生长影响。方法:用不同剂量组对体外培养的MCF-7细胞分别干预12、24、36、48、72h后,用MTT法观察其对MCF-7细胞的生长影响。结果:1纯种发酵淡豆豉异黄酮提取物作用MCF-7细胞12、24、36h,其浓度5、10、20、40mg/L(除24h 10mg/L外)均表现出对MCF-7细胞稳定的抑制作用,与对照组相比差异具有统计学意义(P0.05);2纯种发酵淡豆豉异黄酮对人乳腺癌MCF-7细胞抑制作用,在一定浓度、作用时间有依赖关系,与大豆异黄酮抑制作用相比差异无统计学意义(P0.05)。结论:纯种发酵淡豆豉有明确的抗乳腺癌的作用,其24h IC50为(7.58±1.57)mg/L。  相似文献   

2.
目的 研究天花粉多糖促人外周血单个核细胞(peripheral blood mononuclear cells,PBMC)增殖作用和对人乳腺癌细胞(MCF-7细胞)、人子宫颈癌细胞(HeLa细胞)生长抑制作用.方法 分离健康人PBMC,采用MTT法测定不同浓度天花粉多糖(0.5,1.0,2.0,5.0,10.0,20.0 mmol·L-1)作用于人PBMC 72h后,对人PBMC的促增殖作用;不同浓度天花粉多糖对MCF-7细胞和HeLa细胞生长的抑制作用.结果 2.0~20.0 mmol·L-1天花粉多糖具有显著促进人PBMC增殖(P<0.05),且呈一定的浓度依赖关系;5.0,10.0,20.0 mmol·L-1天花粉多糖均能有效抑制MCF-7细胞和HeLa细胞生长(P<0.05).结论 天花粉多糖具有促人PBMC的增殖作用,并可显著抑制肿瘤细胞MCF-7和HeLa细胞的生长.  相似文献   

3.
目的:研究陈皮多甲氧基黄酮类成分(CM)对人乳腺癌、肝癌细胞株的增殖抑制作用并对其敏感性进行比较.方法:采用MTT法观察CM对人乳腺癌MCF-7、肝癌HepG2细胞株的增殖抑制作用.结果:不同剂量CM(10mg/L、20mg/L、40mg/L)作用于MCF-7细胞株,24h细胞抑制率分别为18.75%、19.79%、34.38%,48h细胞抑制率分别为17.39%、36.96%、40.22%,72h细胞抑制率分别为35.57%、47.08%、50.69%;作用于HepG2细胞株,24h细胞抑制率分别为5.51%、11.81%、29.92%,48h细胞抑制率分别为13.54%、38.54%、51.04%,72h细胞抑制率分别为13.21%、43.40%、55.67%.结论:CM可以抑制人乳腺癌MCF-7、肝癌HepG2细胞株的生长并对两细胞株的增殖抑制呈现时间、浓度依赖性.同等条件下,在小剂量、短时间作用时,MCF-7对于CM作用的敏感性较高;在大剂量、长时间作用时,HepG2表现出比MCF-7更高的敏感性.  相似文献   

4.
[目的]探讨野西瓜总皂苷(CsS)对人肝癌HepG-2细胞的杀伤作用和诱导凋亡的作用.[方法]四甲基偶氮唑蓝(MTr)法研究CSS对HepG一2细胞的杀伤作用;荧光显微镜观察细胞凋亡形态;碘化吡啶(PI)单染经流式细胞仪检测CSS对HepG-2细胞周期及凋亡的影响.[结果]CSS对HepC-2细胞的生长增殖具有抑制作用;经CSS作用后.HepG-2细胞在出现特征性凋亡形态特征,并出现了G1期阻滞,G2期比例下降,高剂量组出现了G2期消失的现象,50 mg/L剂量作用48 h,凋亡细胞比率高达66.652%.[结论]CSS对人肝癌HepG-2有明显的杀伤和诱导凋亡作用,并出现细胞周期的改变.  相似文献   

5.
红花多糖对人PBMC增殖活性及NK、LAK细胞杀伤活性的影响   总被引:2,自引:0,他引:2  
目的:探讨红花多糖(Safflower polysaccharide,SPS)体外对人外周血单个核细胞(PBMC)的增殖作用及对NK细胞、LAK细胞杀伤活性的影响。方法:采集健康成人外周血,常规分离PBMC,体外与不同浓度的SPS共同培养,用3H-TdR法检测PBMC的增殖活性;MTT法检测NK、LAK细胞的杀伤活性。结果:SPS对PBMC增殖有促进作用,其促增殖作用呈现量效依赖性,以1.25mg.mL-1和0.625mg.mL-1剂量组促增殖作用明显(P0.05);SPS对NK细胞与LAK细胞的杀伤活性都有明显的提升作用(P0.05)。结论:SPS可促进PBMC的增殖,增强NK细胞、LAK细胞的杀伤活性,增强免疫功能。  相似文献   

6.
目的:研究柴胡皂苷D对紫杉醇(paclitaxel, PTX)耐药的乳腺癌细胞(MCF-7/PTX)耐药性的逆转作用及其机制。方法:采用噻唑蓝溴化四唑法(MTT法)测定不同质量分数(0、12.5、25、50、100、200 mg/L)PTX处理MCF-7/PTX细胞和MCF-7细胞48 h的细胞存活率,确定MCF-7/PTX细胞对PTX的耐药程度。采用MTT法测定不同质量分数(0.125、0.25、0.50、1.00 mg/L)柴胡皂苷D处理MCF-7/PTX细胞48 h的细胞增殖抑制率,确定柴胡皂苷D对MCF-7/PTX细胞的安全性。采用MTT法测定确定0.125、0.25和0.5 mg/L柴胡皂苷D能否调节MCF-7/PTX细胞对PTX的敏感性。采用蛋白免疫印迹法(Western Blot)和流式细胞术分别检测不同质量分数的柴胡皂苷D对Yes相关蛋白(YAP)和P糖蛋白(P-gp)表达的影响。进行罗丹明123(Rh123)积累和外流测定。结果:与0 mg/L PTX对比,MCF-7和MCF-7/PTX细胞存活率随PTX质量分数增加而降低,且呈现依赖性(P<0.05);确立10...  相似文献   

7.
赵明智  李雪 《天津中医药》2021,38(1):119-123
[目的]探索大蒜素对人乳腺癌MCF-7细胞顺铂耐药性的逆转作用及调控机制.[方法]以对数生长期人乳腺癌耐顺铂MCF-7/DDP细胞为受试细胞,设空白对照组(DMSO)、大蒜素(20μg/mL)单药组、顺铂(6μg/mL)单药组、联合组(大蒜素20μg/mL+顺铂6μg/mL).药物干预48 h后,噻唑蓝(MTT)染色法...  相似文献   

8.
目的观察复方中药紫龙金对人乳腺癌细胞系MCF-7增殖的抑制作用及对其凋亡的诱导作用,探讨紫龙金治疗乳腺癌的作用机制。方法依据培养液紫龙金浓度的差异,实验共设4组:(1)对照组:不加紫龙金干预的1640培养液;(2)紫龙金低浓度组:1.5 mg生药/mL紫龙金培养液;(3)紫龙金中浓度组:3 mg生药/mL紫龙金培养液;(4)紫龙金高浓度组:6 mg生药/mL紫龙金培养液。采用细胞计数法绘制生长曲线及四甲基偶氮唑蓝(MTT)比色法检测不同浓度紫龙金对MCF-7细胞增殖能力的影响,并应用流式细胞术、Hoechst 33342核染色法及DNA梯度形成法测定紫龙金诱导凋亡的作用。结果(1)与对照组比较,紫龙金各剂量组的细胞计数在各时间点(24、48、72、96、120、144 h)明显减少,其差异均有统计学意义(P0.05);紫龙金各剂量组间的生长抑制率差异除低、中浓度组间在24、72 h无统计学意义外(P0.05),各剂量组间两两比较在不同时间点(24、48、72、96、120、144 h),其差异均有统计学意义(P0.05)。紫龙金对MCF-7细胞的增殖抑制作用呈时间依赖性和剂量依赖性;(2)紫龙金使细胞阻滞在G_0/G_1期;(3)中、高浓度紫龙金诱导MCF-7细胞凋亡,并形成DNA ladder。结论紫龙金能抑制人乳腺癌细胞MCF-7细胞增殖,并诱导细胞凋亡,从而起到抗肿瘤的作用。  相似文献   

9.
目的研究黄荆子乙酸乙酯提取物(EVn-50)对人乳腺癌MCF-7细胞Mcl-1、Bcl-2、Bax蛋白的作用。方法应用Wersten Bloting分析EVn-50对人乳腺癌MCF-7细胞Mcl-1、Bax和Bcl-2蛋白表达的影响。结果 Evn-50使Mcl-1及Bcl-2值下调,Bax值上调,促进细胞凋亡。且EVn-50中浓度组相对灰度面积比值与低浓度组比较有显著性差异(P均0.05),EVn-50组48 h相对灰度面积比值与24 h比较有显著性差异(P均0.05)。结论 EVn-50具有诱导人乳腺癌MCF-7细胞凋亡的作用,最佳浓度为3.0 mg/L,最佳作用时间为48 h。  相似文献   

10.
目的 研究沙苑子总黄酮(FAC)对人乳腺癌细胞MCF-7的增殖抑制作用、诱导凋亡作用以及其对NF-кB表达作用影响.方法 以不同浓度(0.5,0.25,0.125,0.062 5 mg/ ml) 处理MCF-7细胞6,12,24,48,72 h后,SRB法测定细胞增殖抑制;FITC-AnnexinⅤ/PI 双染法检测凋亡率变化;用Western blot法检测NF-кB蛋白表达.结果 SRB法显示, 0.5,0.25 mg/ml浓度沙苑子黄酮作用24,48 72 h以及0.125 mg/ml作用48,72 h后MCF-7细胞增殖抑制作用与对照组相比差异具有显著性(P<0.01);0.062 5 mg/ml作用48,72 h后以及0.125 mg/ml作用24 h后对MCF-7的增殖抑制作用与对照组相比也有差异性(P<0.05).FITC-AnnexinⅤ/PI 双染法FCM(flow cytometry)检测显示,不同浓度沙苑子黄酮(0.5,0.25,0.125,0.062 5 mg/ ml)作用72 h后,MCF-7细胞早期凋亡率分别为71.46%,60.34%,51.27%,46.83%, 晚期凋亡率分别为4.12%,3.27%,2.59%,1.26%.与对照组比较0.5,0.25,0.125 mg/ml组早、晚期凋亡率差异具有显著性(P﹤0.01),0.062 5 mg/ml组的早、晚期凋亡率也有差异性(P﹤0.05),0.5 mg/ml剂量组的早、晚期凋亡率与5-Fu组比较差异无显著性(P﹥0.05).Western blot 检测NF-κB表达,再用 Band Leader软件分析.结果 0.125,0.25,0.5mg/ml浓度组与对照组相比NF-кB表达减少差异具有显著性(P<0.01);0.062 5 mg/ml组与对照组相比NF-кB表达也有差异性(P<0.05).结论 沙苑子黄酮对MCF-7具有增殖抑制和诱导凋亡作用;其作用可能与其下调NF-кB表达有关.  相似文献   

11.
姜黄素对Raji细胞体外抗癌作用的实验研究   总被引:6,自引:0,他引:6       下载免费PDF全文
目的研究姜黄素对Raji细胞体外抗癌作用及其机制,对比研究其对RaIi细胞和人单个核细胞的细胞毒性。方法用MTT法检测姜黄素对Raji细胞及人单个核细胞增殖的影响,Annexin—V/PI双标流式术、缺口末端标记法检测姜黄素对Raji细胞及人单个核细胞凋亡的影响,PI单标流式细胞术测定姜黄素对Raji细胞DNA含量分布的影响。结果(1)姜黄素对Raji细胞具有明显的增殖抑制作用。(2)姜黄素可以时间和剂量依赖性方式诱导Raji细胞凋亡。(3)姜黄素组Raji细胞周期发生变化,细胞周期被阻滞于G0/G1和G2/M期,S期比例减少。(4)姜黄素对人单个核细胞无明显抑制增殖和诱导凋亡作用。结论姜黄素能够调控Raji细胞的细胞周期并诱导其凋亡,从而抑制Raji细胞增殖;姜黄素对人单个核细胞无明显细胞毒作用,而选择性作用于肿瘤细胞。  相似文献   

12.
 目的表达、纯化重组葡萄球菌肠毒素B突变体蛋白D9N(rSEB-D9N),了解表达产物rSEB-D9N的细胞毒性、促淋巴细胞增殖和肿瘤生长抑制作用。方法不同浓度IPTG诱导SEB-D9N突变体原核表达系统pET32a-SEB-D9N-E.coliBL21DE3表达rSEB-D9N,经Ni-NTA亲和层析法纯化rSEB-D9N,10%SDS-PAGE和高效液相色谱检测其纯度。以Vero细胞为靶细胞,进行rSEB-D9N的细胞毒性作用,并计算TCID50值。采用MTT法分别检测不同浓度的rSEB-D9N促人淋巴细胞增殖及抑制KB及MCF-7癌细胞株生长的作用。流式细胞术检测rSEB-D9N刺激人淋巴细胞后CD3、CD4、CD8的表达。结果rSEB-D9N表达产量约占细菌总蛋白30%,纯化后获得纯度为90.48%的rSEB-D9N。rSEB和rSEB-D9N对Vero细胞的TCID50值分别为3.12,8.85μg,10.0,20.0mg·L-1的rSEB-D9N具有显著的促PBMC增殖的作用(P<0.05),且主要上调CD3,CD4和CD8的表达。5.0~20.0mg·L-1rSEB-D9N作用的PBMC上清对KB和MCF-7细胞生长均可产生明显抑制作用(P<0.05)。结论获得并纯化了重组葡萄球菌肠毒素B突变体蛋白(rSEB-D9N),其细胞毒性作用有所降低,但促人淋巴细胞增殖和抑制肿瘤细胞生长作用仍较强,可作为研制升白细胞、抗肿瘤相关药物的候选突变体。  相似文献   

13.
The cytotoxicity of naphthazarin derivatives isolated from the roots of Onosma arenaria on human cervix adenocarcinoma cells (HeLa) and leukaemia K562 cells, as well on non-malignant peripheral blood mononuclear cells (PBMC) was studied. The results show that beta-hydroxyisovalerylalkannin, acetylalkannin and the pigment fraction exhibited high cytotoxicity in vitro against the tested cell lines, as well the healthy PBMC before or after activation with phytohaemagglutinin.  相似文献   

14.

Aim of the study

Various parts of Carica papaya Linn. (CP) have been traditionally used as ethnomedicine for a number of disorders, including cancer. There have been anecdotes of patients with advanced cancers achieving remission following consumption of tea extract made from CP leaves. However, the precise cellular mechanism of action of CP tea extracts remains unclear. The aim of the present study is to examine the effect of aqueous-extracted CP leaf fraction on the growth of various tumor cell lines and on the anti-tumor effect of human lymphocytes. In addition, we attempted to identify the functional molecular weight fraction in the CP leaf extract.

Materials and methods

The effect of CP extract on the proliferative responses of tumor cell lines and human peripheral blood mononuclear cells (PBMC), and cytotoxic activities of PBMC were assessed by [3H]-thymidine incorporation. Flow cytometric analysis and measurement of caspase-3/7 activities were performed to confirm the induction of apoptosis on tumor cells. Cytokine productions by PBMC were measured by ELISA. Gene profiling of the effect of CP extract treatment was performed by microarray analysis and real-time RT-PCR.

Results

We observed significant growth inhibitory activity of the CP extract on tumor cell lines. In PBMC, the production of IL-2 and IL-4 was reduced following the addition of CP extract, whereas that of IL-12p40, IL-12p70, IFN-γ and TNF-α was enhanced without growth inhibition. In addition, cytotoxicity of activated PBMC against K562 was enhanced by the addition of CP extract. Moreover, microarray analyses showed that the expression of 23 immunomodulatory genes, classified by gene ontology analysis, was enhanced by the addition of CP extract. In this regard, CCL2, CCL7, CCL8 and SERPINB2 were representative of these upregulated genes, and thus may serve as index markers of the immunomodulatory effects of CP extract. Finally, we identified the active components of CP extract, which inhibits tumor cell growth and stimulates anti-tumor effects, to be the fraction with M.W. less than 1000.

Conclusion

Since Carica papaya leaf extract can mediate a Th1 type shift in human immune system, our results suggest that the CP leaf extract may potentially provide the means for the treatment and prevention of selected human diseases such as cancer, various allergic disorders, and may also serve as immunoadjuvant for vaccine therapy.  相似文献   

15.
评价中药寒热药性的实验方法研究   总被引:3,自引:0,他引:3  
目的通过考察寒凉药与温热药对人乳腺癌细胞MCF-7体外生长增殖的影响来建立评价中药寒热药性的实验方法。方法用噻唑蓝比色法(MTT法)考察4种寒凉药和5种温热药对人乳腺癌细胞MCF-7体外生长增殖的影响,倒置显微镜观察寒凉药、温热药对细胞形态特征的影响,台盼蓝染色法分析所选寒凉药和温热药的细胞毒性作用。结果寒凉药(50~800μg/mL黄连、虎杖、竹叶、夏枯草)抑制MCF-7生长增殖,随质量浓度增大抑制作用增强。温热药干姜和白胡椒(50~800μg/mL)促进MCF-7生长增殖,干姜的促进作用随质量浓度增大而增强,白胡椒的促进作用先增大后减小。其余温热药低质量浓度(肉桂与花椒50~200μg/mL、仙茅50~400μg/mL)促进MCF-7生长增殖,促进作用随质量浓度增大而增强;高质量浓度(肉桂与花椒400~800μg/mL、仙茅600~800μg/mL)抑制MCF-7生长增殖,抑制作用随质量浓度增大而增强。形态学观察表明寒凉药(黄连、虎杖、竹叶、夏枯草)在所选质量浓度范围内使MCF-7密度减小,细胞固缩变圆;温热药(干姜、白胡椒、肉桂、仙茅、花椒)在各自起促进作用的质量浓度范围内使MCF-7密度增大,生长旺盛。台盼蓝染色表明各寒凉药与温热药对MCF-7没有细胞毒性作用。结论本方法有可能用于中药寒热药性的实验评价。  相似文献   

16.
A neem leaf preparation (NLP) was investigated for its role in the induction of tumor cell apoptosis to elucidate the mechanism of NLP mediated immunoprophylaxis in tumor growth restriction. As NLP did not induce direct apoptosis of human tumor cell lines KB, MCF7 and K562, it was used instead to stimulate human peripheral blood mononuclear cells (PBMC) for 72 h. The PBMC derived culture supernatant (NLP-CS) was observed to induce the restriction of tumor cell proliferation as well as apoptosis. An enzyme linked immunosorbant assay revealed the presence of cytotoxic cytokines, IFN-gamma and TNF-alpha, in the NLP-CS. The inhibition of secretion of IFN-gamma and TNF-alpha in NLP-CS caused a significant decrease in tumor cell apoptosis. Furthermore, stimulation of these tumor cells with NLP-CS resulted in upregulation of the caspase 3 and downregulation of the Bcl 2 and cyclin D1. These observations suggested that NLP could induce tumor cellular apoptosis by releasing cytotoxic cytokines from human PBMC.  相似文献   

17.
黄芪提取物免疫调节活性的体外实验研究   总被引:85,自引:5,他引:85       下载免费PDF全文
目的:研究黄芪提取物(Astragalus membranaceus extract,AME)对人的外周血免疫细胞(PBIC)免疫功能的调节作用。方法:采用^3H-TdR掺入法分析AME对外周血单个核细胞(PBMC)的增殖活性及对外周血粘附单核细胞(PBAM)吞噬肿瘤细胞的影响;采用^51Cr释放法测定AME对杀伤性T细胞(CTL)杀伤肿瘤细胞的影响;用ELISA法和生物学法研究AME对外周血B细胞(PBBC)产生IgG及对PBAM产生细胞因子的影响;采用SDS-PAGE分析AME的蛋白组成。结果:AME能促进PBMC的增殖;提高CTL对肿瘤细胞的杀伤活性;增强PBAM对肿瘤细胞的吞噬和产生细胞因子的功能;促进了PBBC产生IgG;AME含有多种蛋白成分。结论:AME对人免疫功能有增强作用,提高了抗肿瘤免疫效应,可应用于临床调节免疫功能和治疗肿瘤等疾病。  相似文献   

18.
 目的研究松口蘑发酵菌丝体糖蛋白MTS03的理化性质及抗肿瘤作用。方法采用SDS-PAGE鉴定MTS03的纯度及相对分子质量,氨基酸自动分析仪分析其中的氨基酸组成;MTT法研究糖蛋白MTS03体外对人乳腺癌细胞MCF-7的增殖抑制率,同时考察MTS03对人正常肝细胞L-02的杀伤率,评价其安全性。结果SDS-PAGE电泳鉴定为单一组分,测定其相对分子质量约为2.37×104,经氨基酸自动分析仪分析含有17种氨基酸;MTS03对MCF-7具有直接的细胞毒作用,在一定的浓度范围内呈现量效和时效关系,MTS03对MCF-7作用24,48,72h的IC50分别为25.81,12.46,8.86mg·L-1,而对正常肝细胞的IC50分别为1366.49,190.60,95.58mg·L-1,对MCF-7的治疗指数TI分别为54.16,18.74,12.25。结论MTS03对于体外培养的乳腺癌细胞具有良好的直接杀伤作用,而对正常肝细胞的毒性较小,可能是一种高效低毒的生物活性物质。  相似文献   

19.
Natural killer (NK) activity of peripheral blood mononuclear cells (PBMC) from 28 patients with systemic lupus erythematosus (SLE) was measured using enzyme-release assay. The SLE patients had significantly decreased NK activity in comparing with normal controls. The levels of NK activity correlated with disease activity. Pre-incubation of PBMC separately with Astragalus membranaceus and Tripterygium hypoglaucum or with their mixture considerably stimulated NK cytotoxicity both in SLE patients and healthy donors. The extent of enhancement was dose-dependent and relevant to pre-incubation periods. The release of a soluble natural killer cytotoxic factor (NKCF) by peripheral blood mononuclear cells was tested by cytotoxicity assay induced in K562 cells. Natural killer cytotoxic factor release was significantly lower in SLE patients than in controls. The levels of natural killer cytotoxic factor were correlated well with NK activities, but correlated negatively with clinical activity. Pre-incubated supernatants from peripheral blood mononuclear cells with above-mentioned agents caused much higher percentage of lysis on K562 targets than that of without pre-incubation.  相似文献   

20.
冠心病血瘀证单核细胞凝血与纤溶活性研究   总被引:2,自引:1,他引:1  
采用发色底物法检测冠心病血瘀证和非血瘀证患者外周血单核细胞 (PBMC) 促凝活性(PCA) ,组织型纤溶酶原激活物 (t- PA) 及其抑制物 (PAI- 1) 活性 ,研究冠心病血瘀证患者外周血单核细胞凝血 /纤溶活性改变及其在发病中的意义。 80例冠心病患者作为病例组 ,其中血瘀证组 3 0例 ,非血瘀证组 50例 (包括痰浊证 2 7例 ,气滞证 2 3例 ) ,健康对照组 2 0例。结果表明 ,冠心病血瘀证患者 PBMC PCA显著高于非血瘀证组和正常对照组 (P<0 .0 1) ;但血瘀证和非血瘀证患者 PBMC的 PAI-1活性无显著差异 (P>0 .0 5 ) ,均高于正常对照组 (P<0 .0 1) ;血瘀证患者 PBMC的 t-PA活性显著低于非血瘀证组 (P<0 .0 1) 。故得出冠心病血瘀证患者单核细胞促凝活性增高 ,纤溶活性下降 ,上述凝血 /纤溶改变可能是血瘀证形成的重要因素之一 ,提示冠心病血瘀证患者单核细胞已处于血栓前状态  相似文献   

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