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1.
目的 探讨黄芩素联合U0126促进人膀胱癌细胞系T24凋亡的分子机制.方法 用黄芩素和U0126处理人膀胱癌T24细胞株,(1)CCK8检测细胞增殖;(2)流式细胞仪检测T24细胞周期的变化;(3)显微镜观察细胞凋亡;(4)流式细胞仪检测早期细胞凋亡和晚期细胞凋亡;(5)Real Time PCR检测细胞增殖和凋亡相关蛋白的表达水平;(6)Western blot检测细胞增殖和凋亡相关蛋白的表达水平.结果 黄芩素或U0126浓度越高,T24细胞增殖能力逐渐下降(P<0.05);不同浓度的黄芩素刺激人膀胱癌细胞株T24,T24细胞数量随浓度上升而逐渐减少(P<0.05);黄芩素与U0126联合处理组的T24细胞株S期的细胞数量显著低于黄芩素、U0126单独处理组(P<0.05);黄芩素与U0126联合处理组的T24细胞株早期凋亡和晚期凋亡均显著高于黄芩素、U0126单独处理组(P<0.05);黄芩素与U0126联合处理组的T24细胞株ERK1/2、Bcl-2和P38的mRNA表达水平显著低于黄芩素或U0126单独处理组(P<0.05);黄芩素与U0126联合处理组的T24细胞ERK1/2、P38的磷酸化水平以及Bcl-2的蛋白表达量均显著低于黄芩素或U0126单独处理组(P<0.05).结论 黄芩素和U0126均可显著抑制人膀胱癌细胞增殖,诱导T24早期凋亡和晚期凋亡且具有浓度依赖性,且两者具有协同效应.因此,黄芩素和U0126可用于治疗膀胱癌.  相似文献   

2.
黄芩苷联合黄芩素诱导乳腺癌细胞凋亡的机制研究   总被引:1,自引:0,他引:1  
目的:探讨黄芩苷和黄芩素联合应用对乳腺癌细胞凋亡的诱导作用及其相关机制。方法将不同浓度的黄芩苷、黄芩素以及两者联合作用于人乳腺癌(Michigan Cancer Foundation 7,MCF-7)细胞。检测上述化合物对乳腺癌细胞的活力、凋亡情况以及凋亡相关蛋白表达的影响。结果黄芩苷或黄芩素作用于乳腺癌细胞后,与作用前相比细胞生长抑制率显著升高(P〈0.05);两者联合应用,与两者单独作用相比抗增殖作用明显增强(P〈0.05)。流式细胞术检测发现两者作用后尤其是两者联用后细胞凋亡显著增加。蛋白印迹分析显示黄芩苷和黄芩素联合作用后Caspase-3、Caspase-9、Bax和p53的表达增加而Bcl-2表达下降,同时p-38活化亦增加。结论黄芩苷和黄芩素均能诱导乳腺癌细胞的凋亡,两者联合应用后作用明显增强,其机制可能与其促进促凋亡相关蛋白Caspase-3、Caspase-9、Bax和p53等的表达,并抑制抗凋亡蛋白Bcl-2的表达有关。  相似文献   

3.
目的探讨Que促人乳腺癌MCF-7细胞凋亡的分子机制。方法用不同浓度的槲皮素处理人乳腺癌MCF-7细胞,用MTT法测定槲皮素对MCF-7细胞增殖的抑制作用,用流式细胞仪检测MCF-7细胞的凋亡水平,用免疫印迹法检测Que处理人乳腺癌MCF-7细胞的Bcl-2和Bax蛋白表达水平的变化。结果 Que能够明显的抑制MCF-7细胞增殖,促进其凋亡;Que处理人乳腺癌MCF-7细胞的Bcl-2蛋白表达水平明显下调,而Bax蛋白表达水平明显上调。结论 Que通过调节Bcl-2和Bax蛋白表达水平促使人乳腺癌MCF-7细胞发生凋亡。  相似文献   

4.
姜黄素对人乳腺癌MCF-7细胞增殖抑制作用及机制研究   总被引:4,自引:0,他引:4  
目的:探讨姜黄素对人乳腺癌MCF-7细胞增殖抑制、诱导凋亡作用及其分子作用机制。方法:MTT法检测姜黄素对MCF-7细胞的增殖抑制作用;流式细胞术PI单染法检测细胞周期;Annexin V/PI双染法检测细胞凋亡;RT-PCR法检测凋亡相关基因Bcl-2和Bax的mRNA表达水平。结果:姜黄素对MCF-7细胞生长有明显抑制作用,并呈剂量、时间依赖性;FCM结果显示姜黄素能使MCF-7细胞阻滞在G1/S期;Annexin V/PI双染法验证姜黄素可以诱导细胞凋亡;RT-PCR结果显示Bax mRNA水平明显上调,而Bcl-2的mRNA表达水平降低。结论:姜黄素对人乳腺癌MCF-7细胞增殖具有显著的抑制作用并可诱导细胞凋亡,其作用机制可能与其上调Bax基因表达水平的同时,下调Bcl-2基因表达水平,从而诱导细胞凋亡有关。  相似文献   

5.
目的:研究ERK/MAP激酶抑制剂U0126是否增强索拉菲尼对原代胃癌细胞的抑制效应,并探索其可能机制。方法:选取病理确诊胃癌患者外科手术标本培养原代胃癌细胞。使用索拉菲尼和U0126分别或同时处理原代细胞。MTT试验检测处理后细胞增殖情况;流式细胞术检测细胞凋亡情况;Q-PCR和Western Blotting分别检测转录水平和翻译水平增殖和凋亡相关分子的表达情况。结果:使用索拉菲尼和U0126同时处理的原代胃癌细胞,其增殖明显受到抑制,凋亡增加,并均强于单独处理组。同时处理组中p-ERK和p-Raf表达水平降低,Bim、Bax和Bad水平升高,水平变化均强于单独处理组,P<0.01。结论:U0126能够增强索拉菲尼抑制胃癌细胞增殖和促进细胞凋亡的效应,其机制可能是二者在抑制ERK/MAPK通路及诱导Bcl-2家族中促凋亡分子表达上有协同效应。  相似文献   

6.
目的 探究丙泊酚联合卡铂对乳腺癌细胞增殖、凋亡的影响及其与酪氨酸激酶JAK/转录因子STAT(JAK/STAT)信号通路的关系,以期为疾病的临床治疗提供新思路。方法 在含有100?μmol丙泊酚、20?μmol/L 卡铂、100?μmol丙泊酚+20?μmol/L卡铂培养基中分别培养MCF-7细胞,CCK-8法检测细胞抑制情况;Hoechst33342染色法观察各组细胞形态变化;平板细胞克隆实验检测菌落形成情况;PI/Annexin V-FITC双染法检测细胞凋亡情况;Western blotting检测p-JAK2、p-STAT3、Bcl-2、Bax、Cleaved Caspase-3蛋白表达情况。结果 与对照组比较,丙泊酚组、卡铂组细胞抑制率升高,菌落数量均降低,细胞凋亡率升高(P?<0.05);与丙泊酚组、卡铂组比较,联合组细胞抑制率升高,菌落数量均降低,细胞凋亡率升高(P?<0.05)。与正常乳腺上皮细胞MCF-10A比较,MCF-7细胞p-JAK2、p-STAT3蛋白表达均升高(P?<0.05)。与对照组比较,丙泊酚组、卡铂组Cleaved Caspase-3、Bax蛋白表达均升高(P?<0.05),p-JAK2、p-STAT3、Bcl-2蛋白表达均降低(P?<0.05)。与丙泊酚组、卡铂组比较,联合组Cleaved Caspase-3、Bax蛋白表达均升高,p-JAK2、p-STAT3、Bcl-2蛋白表达均降低(P?<0.05)。结论 丙泊酚联合卡铂可抑制乳腺癌细胞增殖,诱导其凋亡,其机制可能与抑制JAK/STAT活性,上调凋亡蛋白表达有关。  相似文献   

7.
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。  相似文献   

8.
目的 探究补骨脂乙素(IBC)对人乳腺癌MCF-7细胞死亡的作用及其可能机制。方法 使用不同浓度IBC处理MCF-7细 胞24、48、72 h后,用MTT法检测IBC对MCF-7细胞增殖的影响;将MCF-7细胞设置10、20、40 μmol/L IBC处理组,用AnnexinV-FITC/PI双染结合流式细胞仪、荧光显微镜检测IBC对MCF-7细胞凋亡的影响;免疫印迹法检测凋亡、自噬相关蛋白(Bax、Bcl-2、Akt、p-Akt、p62、LC3)表达的变化;透射电镜观察40 μmol/L IBC处理MCF-7后的细胞亚显微结构;利用JC-1试剂盒、ATP试剂盒和活性氧试剂盒检测IBC对细胞线粒体功能的影响。结果 MTT实验结果显示,IBC具有抑制MCF-7细胞增殖的作用,并呈现浓度和时间依赖性,其作用24、48、72 h的IC50值分别为38.46、31.31、28.26 μmol/L。IBC诱导MCF-7细胞发生凋亡,呈现浓度依赖性。预处理凋亡抑制剂z-VAD-fmk后,其细胞存活率显著高于单独处理IBC组(P<0.05),而程序性坏死抑制剂Nec-1没有保护作用。免疫印迹结果显示IBC增加促凋亡蛋白Bax(P<0.05)表达,下调Bcl-2(P<0.05)、Akt(P<0.05)及其Ser-473位的磷酸化水平(P<0.05)而诱导凋亡。同时,随着IBC浓度的增加,自噬相关蛋白p62的表达逐渐增加,LC3-II/I比值升高,透射电镜下也观察到IBC处理的MCF-7细胞胞浆内有自噬泡以及自噬小体。试剂盒检测到IBC导致线粒体膜电位降低、细胞内ATP水平下降、产生和积聚活性氧(ROS)(P<0.05)。结论 IBC诱导人乳腺癌MCF-7细胞发生凋亡和自噬等多种死亡形式,可成为今后抗乳腺癌创新药物研发的先导化合物。  相似文献   

9.
[目的] 探讨毛酸浆活性成分毛酸浆内酯(PPB)诱导人乳腺癌MCF-7细胞凋亡的机制。[方法] 采用噻唑蓝法(MTT法)检测PPB对MCF-7细胞及人外周血单核细胞(hPBMC)增殖的影响;克隆形成实验考察PPB对MCF-7细胞克隆形成的影响;MTT法考察泛半胱天冬氨酸蛋白酶(Caspase)抑制剂Z-VAD-fmk对PPB诱导MCF-7细胞死亡的作用;Western Blot法考察PPB处理后凋亡相关蛋白PARP、Caspase-7/8/9、Cytochrome c、Bax、Bcl-2、Fas及FADD的表达水平。[结果] PPB时间、剂量依赖性抑制MCF-7细胞生长,且对hPBMC无明显细胞毒性;PPB呈剂量依赖性地抑制MCF-7细胞克隆形成;PPB处理后,凋亡特征性蛋白PARP、原型Caspase-7蛋白表达量显著减少,而Cleaved PARP、Cleaved Caspase-7显著增加;同时内源性凋亡途径相关蛋白Bax、Cleaved Caspase-9、Cytochrome c表达量显著上调,而Bcl-2及Caspase-9原型形式显著下调。外源性凋亡相关蛋白Fas、Cleaved Caspase-8及FADD表达量上调,而Caspase-8原型形式显著下调;加入Z-VAD-fmk可显著逆转PPB诱导的MCF-7细胞死亡。[结论] 毛酸浆活性成分PPB可以诱导MCF-7细胞发生内源性和外源性凋亡。  相似文献   

10.
目的?探讨三黄煎剂对乳腺癌MCF-7、MDA-MB-231细胞凋亡的影响及Aurora激酶A(Aurora A)蛋白表达及功能的影响,并探讨其内在机制。方法?采用CCK-8法检测三黄煎剂对乳腺癌细胞MCF-7、MDA-MB-231增殖的影响。AnnexinV-FITC/PI法检测MCF-7、MDA-MB-231细胞凋亡率。q-PCR法检测MCF-7、MDA-MB-231细胞Aurora A、p53的mRNA表达水平。Western Blot法检测MCF-7、MDA-MB-231细胞凋亡相关蛋白的表达及Aurora A蛋白的表达。结果?三黄煎剂对MCF-7、MDA-MB-231细胞增殖抑制率呈浓度梯度依赖增长(P<0.05),给药48?h疗效好于24?h(P<0.05),与给药72?h无统计学差异(P>0.05)。三黄煎剂能够诱导MCF-7、MDA-MB-231细胞凋亡,并上调c-PARP、c-Caspase 3、Bax蛋白的表达,下调Bcl-2蛋白的表达,呈浓度梯度依赖。三黄煎剂能够下调Aurora A蛋白及mRNA的表达、上调p53蛋白及mRNA的表达。结论?三黄煎剂能够通过下调Aurora A蛋白及mRNA的表达,抑制Aurora A的生物活性,抑制MCF-7、MDA-MB-231细胞增殖,诱导其凋亡。   相似文献   

11.
Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P < 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (>0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P <0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P < 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.  相似文献   

12.
Objective: To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods: SD rats served as both donors and recipients. 4℃ sodium lactate Ringer's was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava (IVC) inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results: Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion: This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.  相似文献   

13.
Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P< 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P< 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.  相似文献   

14.
目的:评价使用安心颗粒对急诊经皮冠状动脉介入术(PPCI)术后生活质量的影响.方法:将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组(术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次)和对照组(仅接受基础药物治疗).所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表(MIDAS)、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果:入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异(P>0.05);出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低(P<0.05);组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低(P<0.05).两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血(P>0.05).两组发生轻度血小板减少症的患者数比较无差异(P>0.05).结论:PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.  相似文献   

15.
Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P<0.01);the MAP decreased(P<0.01)and the HR increased drastically(P<0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients.  相似文献   

16.
Objective:To investigate the gene expression of osteoprotegerin(OPG) and osteoclast differentiation factor(ODF) in the bone tissue of patients with hip fracture due to osteoporosis. Methods:OPGmRNA and ODFmRNA in the bone tissue in 50 cases of osteoporosis sufferers(over 50 years old) with hip fracture(Observer Group) and 30 cases of hip facture sufferers with no osteoporosis(Control group) were analyzed with the Semi-Quantitative RT-PCR method. Results:The mRNA expressed of ODF, OPG were both high in the patients with hip fracture. In the control group, the expression of OPG mRNA was observed, while the expression of ODF mRNA was very slight. Conclusion:Aged patients contained all signals including OPG, ODF that are essential for inducing osteoclastogenesis and promoting bone resorption.  相似文献   

17.
Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P.  相似文献   

18.
Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine.  相似文献   

19.
Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.  相似文献   

20.
A clinical guideline for the therapeutic interventions of integrative medicine may be defined as a written document which states a series of recommendations on therapeutic interventions of integrative medicine for a special disease or condition. The guideline may provide assistance to medical professionals in making clinical decisions aimed at improving the clinical outcome of patients and reducing the costs of medical care(~'4~. Recommendations issued by a guideline should be based on the best available evidence in both Western and Chinese medicine. For fulfilling this purpose, the development of clinical guidelines for therapeutic interventions in the field of integrative medicine should follow scientific principles and undergo a rigorous processes.  相似文献   

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