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1.
目的探讨人参皂甙Rg3对耐顺铂(CDDP)人肺腺癌细胞系A549DDP的逆转作用及其机理.方法以A549DDP及其亲本细胞A549为研究对象,MTT法观察Rg3对A549DDP耐药的逆转作用,采用免疫组化及RT-PCR方法分别在蛋白质和mRNA水平检测耐药相关蛋白MDR1、MRP、LRP表达.结果 MTT法显示低细胞毒浓度Rg3(10 μmol)有效逆转A549DDP细胞耐药7.3倍,而20、30 μmol Rg3分别逆转耐药1.3、1.2倍;10 μmol Rg3预处理A549DDP细胞12、24、36及48 h后分别逆转耐药1.0、1.6、7.6及10.4倍,表明Rg3逆转耐药呈时间依赖性.免疫组化和RT-PCR显示:A549DDP细胞MDR1、MRP、LRP呈过量表达,以Rg3(10 μmol)预处理A549DDP 12、24、36及48 h后,MDR1、MRP表达减弱,呈时间依赖性,而LRP表达无明显时间依赖性.结论 Rg3具有中度逆转肿瘤耐药作用,并呈时间依赖性.  相似文献   

2.
目的:探讨染料木素(genistein)对非小细胞肺癌A549/DDP细胞的耐药逆转及对肺耐药相关蛋白(LRP)和多药耐药相关蛋白(MRP)表达的影响。方法:采用细胞计数法绘制A549/DDP及A549细胞生长曲线;MTT法测定12.5μg/ml染料木素预处理对A549/DDP细胞耐药的逆转作用,并在荧光显微镜下观察细胞形态变化;用RT-PCR和Western blotting分别检测染料木素对肿瘤细胞LRP和MRP的mRNA及蛋白表达的影响。结果:A549/DDP和A549细胞生长曲线相似,倍增时间分别为(27.38±0.25)h和(18.15±0.36)h;染料木素预处理后耐药倍数从2.28倍下降至1.57倍;镜下观察细胞形态出现体积过大、伪足过度伸展、细胞边缘模糊等变化;LRP及MRP mRNA表达均出现明显下调(P<0.05),但二者蛋白表达无明显改变。结论:A549/DDP细胞基本保留了亲代细胞的生长特性;染料木素可以逆转A549/DDP细胞的耐药性,作用机制可能与下调LRP及MRP mRNA表达有关。  相似文献   

3.
BSO逆转人肺腺癌细胞株多药耐药性的实验研究   总被引:3,自引:0,他引:3  
目的研究谷胱甘肽(GSH)合成酶抑制剂——BSO对人肺腺癌多药耐药细胞株A549DDP细胞内GSH含量影响;探讨BSO逆转多药耐药(MDR)作用机制及逆转效果。方法GSH还原酶循环法测定BSO对细胞内GSH含量的影响。MTT比色法测定经BSO预处理后顺氯氨铂(DDP)、阿霉素(ADM)对细胞50%抑制浓度(IC50)的影响。流式细胞仪检测BSO对MDR细胞内柔红霉素(DNR)荧光强度的影响。结果耐药细胞A549DDP细胞内GSH含量较人肺腺癌A549细胞内GSH含量明显增高。BSO在一定浓度范围内(50~200μmol·L-1)对A549细胞和耐药细胞A549DDP无明显细胞毒性作用(抑制率均小于10%)。BSO呈剂量依赖性非线性抑制细胞内GSH的合成,其对MDR细胞内GSH合成影响较为显著,而对A549细胞内GSH合成影响较小。BSO在一定浓度范围内能降低DDP、ADM对A549DDP细胞的IC50,而对A549细胞的IC50无明显影响。A549DDP细胞内DNR荧光强度较A549细胞显著降低,能不同程度提高A549DDP细胞内柔红霉素荧光强度,均较未处理组显著提高;与未经BSO处理的A549细胞比较,细胞内荧光强度轻度增高,但统计学上无显著性差异。结论BSO能有效逆转A549DDP细胞的MDR,其机制与降低MDR细胞内GSH含量有关。  相似文献   

4.
目的研究异钩藤碱脂质体(Isorhynchophylline Liposomes,IR-L)在体外对人肺腺癌细胞系A549/DDP对顺铂(DDP)的逆转作用。方法采用MTT法检测药物细胞毒性作用及耐药细胞逆转倍数,电感耦合等离子体质谱(ICP-MS)法测定细胞内顺铂浓度。结果IR-L无细胞毒浓度组(2.0μg/mL)使A549/DDP细胞对DDP的IC50由16.81 mg/L降至3.36 mg/L;低细胞毒浓度组(7.0μg/mL)的IC50降至2.34 mg/L;两组均明显提高DDP在A549/DDP细胞内的浓度,从0.92μg/L分别提高到4.62μg/L和6.89μg/L。结论IR能部分逆转A549/DDP细胞的MDR,其作用机制可能与增加细胞内药物浓度有关。  相似文献   

5.
目的:从逆转多药耐药的角度,研究金复康口服液对耐药人肺腺癌细胞A549/DDP的化疗增效的作用和分子机制。方法:应用血清药理学方法,选择A549、A549/DDP细胞,实验分为实验组和空白对照组,从细胞水平、分子水平进行研究。结果:金复康口服液低剂量能够逆转A549/DDP对DDP的耐药,逆转耐药倍数为3.45倍,与DDP(320μmol/L)合用,使DDP(320μmol/L)对A549/DDP细胞的抑制率由47.55%升高为73.32%,且q>1.15,表明金复康口服液与DDP合用有明显的协同增效作用,增加了DDP对A549/DDP细胞的敏感度;金复康口服液低剂量能够降低A549/DDP膜转运蛋白LRP、MRP的mRNA表达。结论:(1)低剂量金复康口服液与DDP合用能够增加DDP对A549/DDP的增殖抑制作用,二者具有显著的协同增效作用。(2)金复康口服液能够降低A549/DDP膜转运蛋白LRP、MRP的mRNA表达。金复康口服液可能通过降低耐药肿瘤细胞膜转运蛋白的表达而逆转肿瘤多药耐药,为扶正方药对化疗增效机理提供了新的实验依据。  相似文献   

6.
目的:背景与目的从中药功劳木中提取、分离出来的异喹啉类生物碱组分(Fraction 6,F6),具有抗病毒、抗炎、降血压等生理活性。近年有文献报道异喹啉类生物碱具有一定的逆转肿瘤细胞耐药的作用。本文以白血病多药耐药细胞(K562/ADM)为对象来研究F6逆转肿瘤多药耐药性(Mu ltidrug resistance,MDR)的效果及作用机制,以寻找具有多药耐药逆转活性的新型中药。方法:采用MTT法检测F6及阿霉素(Adriamyc in,ADM)对K562/S和K562/ADM细胞增殖的抑制作用;RT-PCR法检测F6对耐药肿瘤细胞MDR1基因mRNA表达的影响;流式细胞仪分析细胞内罗丹明123(Rhodam ine123,Rh123)浓度,以检测F6对肿瘤细胞膜P糖蛋白(P-glycoprote in,P-gp)泵功能的影响;免疫组化方法检测F6对肿瘤细胞膜P-gp表达水平的影响;流式细胞仪检测F6对肿瘤细胞凋亡的作用。结果:10 mg/L为F6的无毒剂量;ADM对K562/S和K562/ADM细胞的IC50分别为(1.68±0.08)mg/L和(80.25±1.06)mg/L;无毒剂量F6与ADM联合应用后对K562/S和K562/ADM细胞的IC50分别为(1.09±0.07)mg/L和(16.68±0.72)mg/L,此剂量F6使K562/ADM细胞的IC50下降4.81倍;无毒剂量的F6应用前后,K562/ADM细胞MDR1基因表达水平和细胞膜P-gp表达水平无明显差异;Rh123蓄积试验中无毒剂量的F6应用后使K562/ADM细胞内Rh123浓度由F6应用前的29.21%升高到85.35%,Rh123外排试验中无毒剂量的F6应用后使K562/ADM细胞内Rh123浓度由F6应用前的27.19%升高到59.22%;凋亡检测结果显示无毒剂量的F6使K562/ADM细胞凋亡率升高3.82倍。结论:F6能有效逆转白血病细胞的MDR;F6逆转白血病MDR的机制为通过抑制肿瘤细胞膜上P-gp的药泵功能,增加耐药细胞内化疗药物浓度逆转耐药性,而不是抑制MDR1基因和P-gp表达。  相似文献   

7.
目的 探讨和厚朴酚(HNK)对白血病细胞U937阿霉素(ADR)耐药细胞系U937/ADR多药耐药逆转作用及其机制.方法 以大剂量(IC50)ADR短时间诱导方法 ,构建U937/ADR细胞系.以低浓度HNK(IC20)与不同化疗药物联合作用于U937/ADR细胞系,检测其对不同化疗药物耐药逆转倍数.罗丹明123检测药物外排功能;荧光定量PCR(FQ-PCR)和Western blotting检测不同浓度HNK对U937/ADR细胞系核转录因子-κB(NF-κB)和耐药相关基因MDR1及其蛋白P-糖蛋白(P-gp)表达的影响;ELISA法检测NF-κB亚单位p65(NF-κB p65)的DNA结合活性.结果 成功构建了白血病多药耐药细胞系U937/ADR,对ADR耐药指数为亲代U937细胞的11倍.6.5 μg/mL HNK可以逆转U937/ADR对ADR的耐药,逆转倍数为2.20倍.HNK能够浓度依赖性地下调U937/ADR细胞NF-κB、MDR1 mRNA和P-gp表达,抑制NF-κB p65的活性.结论 HNK能有效逆转U937/ADR多药耐药,其机制可能与抑制NF-κB p65活性、下调MDR1和P-gp表达有关.  相似文献   

8.
目的探讨Src酪氨酸激酶抑制剂逆转人胃癌顺铂耐药细胞SGC7901/DDP的作用及机制。方法以Src酪氨酸激酶抑制剂逆转前后的人胃癌顺铂耐药细胞SGC7901/DDP为研究对象,应用Western blot观察细胞内p-Src表达的变化,MTS法检测细胞的药物敏感性,流式细胞仪检测细胞Rh123外排及P-gp表达的水平。结果 Src酪氨酸激酶抑制剂可下调SGC7901/DDP细胞中p-Src表达,在5μmol/L及10μmol/L Src酪氨酸激酶抑制剂作用下,SGC7901/DDP细胞对顺铂的药物敏感性分别提高了1.52倍和3.96倍,细胞中Rh123含量分别提高了1.24倍和2.64倍,P-gp表达水平分别降低了65.8%和47.4%。结论 Src酪氨酸激酶抑制剂可逆转SGC7901/DDP细胞多药耐药性,其耐药表型的逆转可能与降低细胞Rh123外排及P-gp表达水平有关。  相似文献   

9.
MDR1/MRP反义寡核苷酸逆转胃癌细胞耐药的研究   总被引:1,自引:0,他引:1  
目的:探讨MDR1/MRP反义寡脱氧核苷酸(Antisense oligodooxynucleotide,ASODN)片段逆转人胃腺癌耐药细胞SGC7901/ADM多药耐药(Multidrugresistance,MDR)的作用.方法:经MDR1或多药耐药相关蛋白(Multidrugresistance-asscociated protein,MRP)ASODN分别或联合转染SGC7901/ADM细胞后,采用RT-PCR法检测MDR1mRNA和MRPmRNA的表达.免疫细胞化学检测SGC7901/ADM细胞内p-糖蛋白(P-glycoprotein,P-gp)、MRP的表达,流式细胞术检测瘤细胞内Rh123的潴留状况,MTT法检测瘤细胞对阿霉素、卡铂等抗癌药的敏感性.结果:SGC7901/ADM细胞经ASODN转染12h后,MDR1和MRP mRNA的表达均降低,转染24h后下降最明显,48h后恢复至转染前水平.经转染ASODN48h后,SGC7901/ADM细胞P-gp、MRP的表达显著下降,瘤细胞内Rh123的潴留量显著高于转染前,且联合转染MDR1和MRP ASODN后瘤细胞内Rh123的潴留量显著高于单纯转染组.SGC7901/ADM细胞在联合转染MDR1和MRP ASODN后对阿霉素、卡铂等化疗药物的敏感性明显高于单纯转染ASODN.结论:分别转染MDR1或MRP ASODN可部分逆转人胃腺癌耐药细胞SGC7901/ADM多药耐药,而联合转染2种ASODN则可显著提高瘤细胞耐药逆转效率.  相似文献   

10.
目的探讨去甲斑蝥酸钠(SNCTD)对耐顺铂人肺腺癌细胞系A549/DDP的逆转作用及可能分子机制。方法(1)采用CCK法筛选出SNCTD对A549/DDP的无毒浓度(即对细胞抑制率〈10%的药物浓度),并检测出顺铂及与无毒浓度SNCTD联合对耐药细胞株的半数抑制浓度(IC50);(2)采用流式细胞仪分别检测1、2倍无毒浓度SNCTD对细胞内罗丹明123的蓄积情况;(3)采用逆转录-多聚酶链反应(RT—PCR)检测1、2倍无毒浓度SNCTD处理细胞48h后耐药相关基因mdr1mRNA、MRP1mRNA、GST—PimRNA表达,并检测药物处理72h后GST—PimRNA的表达。结果(1)SNCTD对A549/DDP的无毒浓度为5μg/ml,无毒浓度SNCTD与顺铂联合应用可使A549/DDP细胞对顺铂的IC50值下降为4.32μg/ml,逆转倍数为1.97倍;(2)无毒浓度SNCTD处理耐药细胞48h后细胞内罗丹明123荧光强度明显增强(P〈0.05);(3)5、10μg/ml SNCTD处理耐药细胞株48h后mdr1 mRNA、MRP1 mRNA表达均明显减弱(P〈0.05),而且10μg/ml SNCTD处理组减弱强度亦明显大干5μg/mlSNCTD处理组(P〈0.05),呈现浓度依赖性;而GST—PimRNA表达并无明显变化(P〉0.05),72h后GST—PimRNA表达也未出现下降(P〉0.05)。结论SNCTD可以逆转A549/DDP细胞的耐药作用。其作用机制可能与下调耐药相关基因mdr1、MRP1的表达,影响膜蛋白外排泵功能有关。  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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