环孢霉素A引起癫痫发作二例

环孢霉素Ａ引起癫痫发作二例王秀婷贺海涛郭永亮冯筱林我们应用环孢霉素Ａ（ＣｓＡ）治疗１例系统性红斑狼疮（ＳＬＥ）及１例重型再生障碍性贫血（ＳＡＡ）患者，治疗过程中出现了癫痫大发作，报道如下。例１，女，３０岁。自１９９３年９月始间断发热，渐乏力、颜面及下...     

环孢霉素A滴眼液治疗干眼症的临床观察

目的：观察环孢霉素A滴眼液治疗干眼症的疗效。方法：对诊断为干眼症的37例(74眼)患者，给予0.5％环孢霉素A滴眼液滴眼，3～4次／d，1个月后，改为2次／d，观察3～6个月。结果：所有患者症状减轻，体征好转71眼(95．9％)，除有轻度刺痛感外，无不良反应。结论：环孢霉素A滴眼液可有效治疗干眼症。     

环孢霉素A逆转人白血病耐药细胞系HL-60／ADM后氧自由基水平的变化

本研究旨在探讨将抗阿霉素的人急性早幼粒细胞白血病细胞系HL-60／ADM的耐药性逆转后细胞内氧自由基水平的变化特点。选择环孢霉素A（CsA）作为耐药逆转剂，分别采用MTT法、流式细胞术和免疫组织化学法分析CsA对人白血病耐药细胞系的毒性及逆转效果；用化学比色法检测逆转前后细胞内丙二醛（MDA）、超氧化岐化酶（SOD）和谷胱甘肽（GSH）的含量。结果表明：当CsA在4μg／ml以下时对HL-60／ADM无明显毒性作用，超过此浓度，其毒性呈剂量效应（P〈0．001）。当CsA浓度为0．5μg／ml时就有明显逆转作用，随着CsA剂量增加，逆转作用逐渐增强（P〈0．001），当CsA剂量达8μg／ml以上时对细胞存活产生明显的影响。流式细胞仪检测细胞内药物浓度发现，逆转耐药细胞系HL-60／ADM＋CsA细胞内阿霉素的浓度明显高于耐药细胞系HL-60／ADM。免疫组织化学检测结果显示，HL-60／ADM细胞膜上P—gP高表达，经CsA作用后P—gP表达下降。氧自由基测定结果显示：HL-60／ADM细胞经4μg／ml CsA作用72小时后细胞内SOD的活性与对照组相比显著降低（P〈0．001），细胞内的脂质过氧化产物MDA的含量与对照组相比有所升高（P〈0．05），抗氧化剂GSH的含量较对照组明显降低（p〈0．001）。结论：CsA能有效逆转HL-60／ADM的耐药性；逆转后细胞内氧自由基的含量升高，抗氧化剂的活性明显减低，过多的氧自由基可影响细胞的功能状态，导致细胞死亡。     

中药复方君子汤联合环孢霉素A逆转白血病细胞諯562/VCR耐药性的实验研究

本研究观察中药复方君子汤(FFJZ)联合环孢霉素A(cyclosporine A,CsA)逆转白血病耐药细胞系K562/VCR细胞耐药性的效果,以便寻找有效的联合逆转剂.采用MTT法、流式细胞术研究FFJZ联合CsA逆转白血病耐药细胞系K562/VCR耐药性的作用.结果表明FFJZ联合CsA在体外对K562/VCR细胞的耐药性有明显的逆转作用(p＜0.01),能提高K562/VCR细胞对阿霉素(ADM)的敏感性,且在药物有效浓度范围内对细胞本身无毒性作用.FFJZ联合CsA对K562/VCR细胞的P-gp表达的阳性率无明显影响.结论复方君子汤联合环孢霉素A可能成为治疗白血病多药耐药的安全有效的耐药逆转药物.     

脑缺血时环孢霉素A对无机离子的影响

脑血管病严重危害人类健康 ,其发病趋势已向青壮年扩展 ,复发率高 ,已成为世界三大疑难病之一。环孢霉素Ａ(ＣＳＡ)是由真菌产生的环状十一肽活性物质[1] ,作为线粒体渗透性迁移通道的特异阻滞剂 ,对脑缺血再灌损伤的保护作用 ,已引起人们的注意。1　对象与方法1.1　动物模型的     

五味子提取物逆转K562／DOX细胞阿霉素耐药的实验研究

目的探讨五味子提取物对阿霉素耐药细胞株K562／DOX的耐药逆转作用。方法MTT法检测单独应用阿霉素或阿霉素与五味子提取物联合作用的K562／DOX细胞的生存率;RT-PCR检测MDR1、MRP1 mRNA表达;Western blotting检测MDR1蛋白的表达。结果与对照组相比,单独应用阿霉素组细胞的生存率未见明显变化;而阿霉素与五味子提取物联合作用组细胞存活率显著降低,MDR1、MRP mRNA表达降低;MDR1蛋白表达下降。结论五味子提取物可能通过下调MDR1、MRP水平进而逆转K562／DOX细胞对阿霉素的耐药性,增加肿瘤耐药细胞对化疗药物的敏感性。     

五种逆转剂对耐药细胞系K562／VCR,K562／HHT逆转作用的研究

五种逆转剂对耐药细胞系Ｋ５６２／ＶＣＲ、Ｋ５６２／ＨＨＴ逆转作用的研究罗梅华，卞寿庚，薛艳萍，孟庆祥，冯敏，秘营昌肿瘤细胞对治疗药物的耐药性是急性白血病治疗未能取得很大进展的重要原因。克服耐药的方法之一是使用逆转剂，但多数由于副作用大，致体内难以达到...     

Cyclosporine diminishes multidrug resistance in K562/ADM cells and improves complete remission in patients with acute myeloid leukemia

This study was designed to investigate the effects of cyclosporine A (CsA) on a multidrug resistance cultured cell line, and its effect on complete remission in patients with acute myeloid leukemia (AML). A multidrug resistant K562/ADM cell line and drug-sensitive K562 cell line was used. The intracellular concentration of daunorubicin and the accumulation of Rhodamine 123 (Rh123) in the K562/ADM and K562 cells were evaluated. Clinical effects of CsA were also studied in 65 patients with AML. In the K562/ADM cells, the 50% of inhibition concentration (IC50) of daunorubicin only group was 23.0 ± 5.2 μmol/L, which was greater than in other groups co-administered with CsA (1.2 ± 4.8 μmol/L), verapamil (1.5 ± 5.4 μmol/L) or CsA + verapamil (1.4 ± 4.3 μmol/L) (all P < 0.01). The relative fluorescence intensity of Rh123 in the K562/ADM cells treated with CsA and daunorubicin was increased from 48.9% to 69.8% (P < 0.05). CsA also improved the complete remission rate in the AML patients (72.7% vs 21.9%, P < 0.01). We conclude that CsA can significantly diminish the multidrug resistance in K562/ADM cells. It also enhances the complete remission rates in patients with AML. CsA may be used as an integral part of the chemotherapy for AML.     

环孢霉素A逆转复发难治性白血病多药耐药的临床研究

目的：探讨环孢霉素Ａ（ＣｓＡ）逆转复发难治性急性白血病的临床意义及推广价值。方法：采用免疫组织化学ＡＢＣ法，ｐ１７０单抗ＪＳＢ－１检测复发难治性白血病的ｐ１７０表达，并对阳性患者随机分组，用ＣｓＡ加联合化疗进行逆转多药耐药研究，同时采用高效液相色谱法检测ＣｓＡ血药浓度，探讨ＣｓＡ血药浓度和逆转疗效间的关系。结果：ｐ１７０在复发难治性白血病中有高表达；ＣｓＡ有较好逆转复发难治性白血病多药耐药的作用（Ｐ＜０．０５）；ＣｓＡ血药浓度和逆转疗效呈正相关。结论：ＣｓＡ可能成为安全有效的逆转药物，在治疗复发难治性ＡＬ中有较大临床意义及推广价值。     

K562细胞及其耐药细胞株K562/A02细胞白血病耐药相关microRNA筛选

目的 通过检测慢性粒细胞白血病急变细胞系K562及其阿霉素耐药株K562/A02的微小RNA(microRNA、miR)表达差异,探讨microRNA与白血病化疗耐药的关系.方法 MTT法检测K562/A02及其亲本细胞系K562的耐药性能;流式细胞术检测K562与K562/A02细胞的P-gp表达;运用microRNA芯片技术筛查K562与K562/A02细胞之间差异表达的microRNA,随后用实时荧光定量RT-PCR方法进一步证实.结果 阿霉素耐药株K562/A02相对于其亲本细胞系K562对阿霉素的耐药倍数为180倍;K562细胞P-gp的表达率为0.2%,K562/A02细胞P-gp的表达率为86%;microRNA芯片结果显示K562/A02与K562细胞之间有22种microRNA表达存在显著的差异(P<0.01),表达差异在2倍以上的有9种,其中miR-221、miR-155、miR-451在K562/A02细胞表达上调,而miR-98、miR-181a、let-7f、miR-424、let-7g和miR-563则表达下调.实时荧光定量RT-PCR进一步证实了上述结果,并显示miR-451、miR-155、miR-221、let-7f、miR-424在两种细胞中表达差异显著.结论 K562/A02与K562细胞存在microRNA表达差异,其中miR-451、miR-155和miR-221在K562/A02中表达显著上调,而let-7f、miR-424则显著下调,提示microRNA可能参与白血病耐药形成,差异表达的microRNA可能为逆转白血病耐药提供新的作用靶点.     

K562细胞及其耐药细胞株K562/A02细胞白血病耐药相关microRNA筛选

目的 通过检测慢性粒细胞白血病急变细胞系K562及其阿霉素耐药株K562/A02的微小RNA(microRNA、miR)表达差异,探讨microRNA与白血病化疗耐药的关系.方法 MTT法检测K562/A02及其亲本细胞系K562的耐药性能;流式细胞术检测K562与K562/A02细胞的P-gp表达;运用microRNA芯片技术筛查K562与K562/A02细胞之间差异表达的microRNA,随后用实时荧光定量RT-PCR方法进一步证实.结果 阿霉素耐药株K562/A02相对于其亲本细胞系K562对阿霉素的耐药倍数为180倍;K562细胞P-gp的表达率为0.2%,K562/A02细胞P-gp的表达率为86%;microRNA芯片结果显示K562/A02与K562细胞之间有22种microRNA表达存在显著的差异(P<0.01),表达差异在2倍以上的有9种,其中miR-221、miR-155、miR-451在K562/A02细胞表达上调,而miR-98、miR-181a、let-7f、miR-424、let-7g和miR-563则表达下调.实时荧光定量RT-PCR进一步证实了上述结果,并显示miR-451、miR-155、miR-221、let-7f、miR-424在两种细胞中表达差异显著.结论 K562/A02与K562细胞存在microRNA表达差异,其中miR-451、miR-155和miR-221在K562/A02中表达显著上调,而let-7f、miR-424则显著下调,提示microRNA可能参与白血病耐药形成,差异表达的microRNA可能为逆转白血病耐药提供新的作用靶点.     

S3核糖体蛋白基因的过表达与K562/DOX细胞耐药性的关系

目的 观察S3核糖体蛋白（S3rp)基因表达的改变对白血病耐药性的影响。通过RT－PCR方法获得含S3rp基因完整开放阅读框架（ORF）的cRNA，并将其克隆到pGEM-T Easy载体里，然后通过亚克隆方法将pGEM-T Easy重组质粒里的S3rp cDNA片段克隆到真核表达载体pcDNA3.1( )中，分别构建正向插入和反向插入的S3rp cDNA重组质粒，进行基因转染。将正义S3rp cDNA真核表达质粒转染K562细胞，使其S3rp基因表达增加，观察其对化疗药物敏感性的改变；将反义S3rp cDNA真核表达质粒转染具有多药耐药（MDR）表型的K562／DOX细胞，阻碍其S3rp基因的表达，观察其对化疗药物耐药性的改变。结果 转染正义S3rp cDNA真核表达质粒后，K562细胞对阿霉素的耐药性比未转染的K562细胞增强5．8倍；转染反义S3rp cDNA真核表达质粒后，K562／DOX细胞对阿霉素的耐药性比未转染的K562／DOX细胞降低69％。结论 S3rp基因过表达在K562／DOX细胞耐药性的形成中起重要作用。     

Reversal effect of substituted 1,3-dimethyl-1H-quinoxalin-2-ones on multidrug resistance in adriamycin-resistant K562/A02 cells

QA1 and QA3 are the derivatives of substituted 1,3-dimethyl-1H-quinoxalin-2-ones that may selectively antagonize P-glycoprotein (P-gp) in multidrug resistance (MDR) cancer cells. Herein, we examined the reversal effect of two compounds on MDR in adriamycin (Adr)-induced resistant K562/A02 cells. MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) assay showed that QA1 and QA3 weakly inhibited the growth of tumor cells. However, the compounds increased Adr-induced cytotoxicity toward K562/A02 cells. The IC₅₀ values of Adr toward K562/A02 were decreased in the presence of QA1 or QA3. The maximal reversal fold (RF) of QA1 and QA3 was reached 6.9 and 9.0, respectively. The action of QA1 and QA3 was also confirmed by the increase of intracellular Adr accumulation in K562/A02 cells. In mechanism study, the intracellular accumulation and efflux of Rh123 were measured using multilabel counter with excitation/emission wavelengths of 485/535 nm. An increase of intracellular Rh123 and the decrease of efflux were observed in K562/A02 cells incubation with QA1 or QA3, indicating that the activity of P-gp was blocked. These results suggested that the derivatives of substituted 1,3-dimethyl-1H-quinoxalin-2-ones might reverse MDR in K562/A02 cells via inhibition activity of P-gp. QA1 and QA3 might be the candidate agents for reversing MDR of cancer.     

K562细胞及其耐药细胞株K562/A02细胞白血病耐药相关microRNA筛选

目的 通过检测慢性粒细胞白血病急变细胞系K562及其阿霉素耐药株K562/A02的微小RNA(microRNA、miR)表达差异,探讨microRNA与白血病化疗耐药的关系.方法 MTT法检测K562/A02及其亲本细胞系K562的耐药性能;流式细胞术检测K562与K562/A02细胞的P-gp表达;运用microRNA芯片技术筛查K562与K562/A02细胞之间差异表达的microRNA,随后用实时荧光定量RT-PCR方法进一步证实.结果 阿霉素耐药株K562/A02相对于其亲本细胞系K562对阿霉素的耐药倍数为180倍;K562细胞P-gp的表达率为0.2%,K562/A02细胞P-gp的表达率为86%;microRNA芯片结果显示K562/A02与K562细胞之间有22种microRNA表达存在显著的差异(P<0.01),表达差异在2倍以上的有9种,其中miR-221、miR-155、miR-451在K562/A02细胞表达上调,而miR-98、miR-181a、let-7f、miR-424、let-7g和miR-563则表达下调.实时荧光定量RT-PCR进一步证实了上述结果,并显示miR-451、miR-155、miR-221、let-7f、miR-424在两种细胞中表达差异显著.结论 K562/A02与K562细胞存在microRNA表达差异,其中miR-451、miR-155和miR-221在K562/A02中表达显著上调,而let-7f、miR-424则显著下调,提示microRNA可能参与白血病耐药形成,差异表达的microRNA可能为逆转白血病耐药提供新的作用靶点.     

环孢菌素D衍生物PSC 833逆转K562/A02细胞多药耐药的研究

目的 证实PSC833逆转剂的高效性，探讨PSC833逆转肿瘤细胞多药耐药的机制。方法 以人红白血病细胞系K562及其耐药细胞系K562／A02（耐阿霉素）为实验研究对象，采用MTT法检测细胞毒性；直接免疫荧光测定法检测P糖蛋白（P-gp）表达水平；RT－PCR法检测mdr1 mRNA水平，以流式细胞术测定两种细胞系内柔红霉素（DNR）的潴留来的反映P-gp的外排功能。结果 与K562细胞系相比，K562／A02耐药细胞系mdr1 mRNA及P－gp高表达，DNR潴留减少。1μmol/L的PSC833对K562／A02铁mdr1 mRNA及P－gp表达水平无明显影响（P＞0.05），PSC833对K562／A02细胞的DNR细胞毒性有剂量依赖性增敏作用，其增敏作用至少是环孢菌素A（CsA）、维拉帕米（Ver）的3倍。PSC833能增加K562／A02耐药细胞系的DNR潴留。1μmol/L的PSC833能使K562／A02细胞内DNR潴留量恢复至K562细胞的100.9%，而10μmol/L CsA只恢复至K562细胞的86.9%，PSC833对K562细胞系的DNR细胞毒性及DNR潴留均无明显影响（P＞0.05）。结论 PSC833较CsA、Ver逆转活性至少高3－10倍，其逆转K562／A02多药耐药的机制可能是通过抑制P－gp功能，而非直接下调mdr1 mRNA及P-gp水平。     

多药耐药基因反义寡核苷酸逆转肿瘤细胞耐药的初步研究

目的：克服肿瘤细胞的多药耐药（ＭＤＲ）。方法：用人工合成互补于ｍｄｒ１基因５′端转录起始部位的反义寡核苷酸（ＯＤＮ），直接转染耐药细胞株ＫＢ－８－５细胞，或以脂质体ｌｉｐｏｆｅｃｔｉｎ为载体进行基因转染实验，通过ＭＴＴ法检测细胞对柔红霉素（ＤＮＲ）的敏感性，流式细胞仪分析细胞内ＤＮＲ含量及免疫组化方法确定细胞表面糖蛋白（Ｐｇｐ）的表达水平。结果：ＯＤＮ可增加ＫＢ－８－５细胞内的ＤＮＲ浓度从而提高耐药细胞对ＤＮＲ的敏感性，ｌｉｐｏｆｅｃｔｉｎ进一步加强上述作用。在ＤＮＲ浓度为３．０ｍｇ／Ｌ组中，约有７４．４３％的被转染细胞对ＤＮＲ敏感而致死，基本达到药物敏感细胞株ＫＢ－３－１的水平。ＯＤＮ转染的ＫＢ－８－５细胞的Ｐｇｐ为弱阳性表达，低于阳性对照ＫＢ－８－５细胞的Ｐｇｐ表达水平。结论：ＯＤＮ的逆转作用可能与其互补结合的ｍｄｒ１ｍＲＮＡ降解或直接阻滞了Ｐｇｐ合成使其表达降低有关。     

Reversal effect of Tween-20 on multidrug resistance in tumor cells in vitro

Multidrug resistance (MDR) is a major barrier for chemotherapy of many cancers. Non-ionic surfactants have great potential to reverse the MDR by preventing onset or delay progression of the carcinogenic process. However, the role of Tween-20 in the development of MDR remains unknown. The aim of this study was to explore the reversal effect and potential mechanism of Tween-20 on tumor cells in vitro. Alamar Blue assay was used to examine the reversal index of Tween-20 to vincristine (VCR), doxorubicin (DOX) and 5-fluorouracil (5-FU) in KBv200, HepG2/R and Bel-7402/5-FU, respectively. Morphological change was determined by Gimsa and Hoechst 33258 staining. The acumulation of DOX was confirmed by spectrofluorimetric assay. Cell cycle analysis was performed using flow cytometry. The mRNA and protein expression levels of MDR were assessed by semiquantitative RT-PCR and dot blot, respectively. The results showed that Tween-20 at concentrations of 0.0025%, 0.005%, 0.01% had little cytotoxicity. When combined with the cancer drugs, it significantly promoted the sensitivity of MDR cells. Fluorescence staining confirmed that the percentage of apoptotic cell increased when combined with Tween-20. This notion was further supported by the observation that Tween-20 treatment potentiated VIN-induced G2/M arrest of the cell cycle. Furthermore, Tween-20 treatment increased significantly intracellular accumulation of DOX. RT-PCR and dot blot revealed that Tween-20 could downregulate the expression of MDR and P-glycoprotein. Low concentrations of Tween-20 can efficiently reverse the multidrug resistance phenotype by enhancing accumulation of the anticancer drugs. The potential mechanism may be via inhibiting the multidrug-resistant gene expression.