粉防己碱逆转肿瘤多药抗药性细胞的凋亡抗性作用

目的探讨粉防己碱逆转ＭＤＲ细胞凋亡抗性的作用及其机制。方法ＭＤＲ细胞株ＭＣＦ－７／Ａｄｒ与其相应的敏感株ＭＣＦ－７进行对比研究。比较粉防己碱对阿霉素（Ｄｏｘ）诱导ＭＤＲ细胞及其相应的敏感株的凋亡作用。并比较粉防己碱对ＭＤＲ细胞及其相应的敏感株的细胞ｂｃｌ－２蛋白的表达的影响。细胞凋亡及ｂｃｌ－２蛋白表达的测定以流式细胞仪法。结果加入Ｄｏｘ共同培养２４ｈ可诱导７４．６％ＭＣＦ－７细胞凋亡，而只引起１４．３％的ＭＣＦ－７／Ａｄｒ细胞凋亡。只加入粉防己碱共同培养２４ｈ，未见明显增加ＭＤＲ细胞及其相应敏感细胞的凋亡百分比。Ｄｏｘ＋粉防己碱能显著地使ＭＤＲ细胞的凋亡增至４７．０％，与单独用Ｄｏｘ组相比较，差异有显著性（Ｐ＜０．０１），而敏感细胞株为７７．８％，与单独用Ｄｏｘ组相比较，差异无显著性（Ｐ＞０．０５）。ＭＤＲ细胞株与其相应的敏感株ｂｃｌ－２蛋白表达水平均较低且差异无显著性。加入粉防己碱对ＭＤＲ细胞株与其相应的敏感株ｂｃｌ－２蛋白表达水平均未见明显影响。结论粉防己碱能逆转ＭＤＲ细胞对Ｄｏｘ的凋亡抗性。其逆转机制可能与ｂｃｌ－２蛋白表达无关。粉防己碱逆转ＭＤＲ细胞ＭＣＲ－７／ＡＤＲ对Ｄｏｘ的凋亡抗性的机制有待进?     

复方黄连素注射液的HPLC测定

复方黄连素注射液的ＨＰＬＣ测定杜兴，阚学瑾（甘肃省药品检验所，兰州７３００００）ＤＥＴＥＲＭＩＮＡＴＩＯＮＯＦＣＯＭＰＯＵＮＤＢＥＲＢＲＩＮＥＩＮＪＥＣＴＩＯＮＢＹＨＰＬＣ￥ＤＵＸｉｎｇ；ＫＡＮＸｕｅ－Ｊｉｎ（ＧａｎｓｕＰｒｏｖｉｎｃｉａｌＩｎｓｔｉ...     

抗人CD3单抗在体外诱导LAK和TIL增殖的实验研究

目的 观察抗人ＣＤ３单抗（ａＣＤ３ＭｃＡｂ）对单核细胞（ＰＢＭＣ）和肿瘤组织中提取的浸润淋巴细胞（ＴＩＬ）增殖的影响。方法 将外周血中分离出的ＰＢＭＣ和ＴＩＬ进行扩增培养,同时测定其活性。结果 ａＣＤ３ＭｃＡｂ和白细胞介素－２（ＩＬ－２）的浓度是包被法制备ＣＤ３单克隆抗体激活的杀伤细胞（ＣＤ３ＡＫ）的主要影响因素。经选择ａＣＤ３ＭｃＡｂ的最适蛋白浓度为０．１ｕｇ／ｍｌ,ＩＬ－２为２０Ｕ／ｍｌ。结论     

番荔枝内酯克服肿瘤多药抗药性作用及机制

目的：探讨阿蒂莫耶番荔枝总内酯（ＡＡ）及其单体ｂｕｌｌａｔａｃｉｎ和ａｔｅｎｏｙａｃｉｎ－Ａ克服肿瘤多药抗药性（ＭＤＲ）的作用及其机制。方法：用两对ＭＤＲ细胞株及其相应的敏感株进行对比研究。结果：ＡＡ及其单体具有极强的细胞毒作用,且对ＭＤＲ细胞与其相应的敏感株的ＩＣ５０相近。加入ＡＡ,ｂｕｌｌａｔａｃｉｎ和ａｔｅｍｏｙａｃｉｎ－Ａ能使细胞内Ｆｕｒａ－２分别增加２．３９,３．１４和３．２４倍,但不能     

Modulation of multidrug resistance by three bisbenzyl-isoquinolines in comparison with verapamil

比较轮环藤碱（Ｃｙｃ）、海岛轮环藤碱（Ｉｎｓｒ）和海岛轮环藤酚碱（Ｉｎｓｎ）与维拉帕米（Ｖｅｒ）体外调节多药耐药性（ＭＤＲ）的作用．方法：细胞毒试验采用ＭＴＴ法，细胞内阿霉素（Ｄｏｘ）积累采用荧光分光光度法测定．结果：Ｃｙｃ，Ｉｎｓｒ，Ｉｎｓｎ和Ｖｅｒ在ＭＤＲ细胞系ＭＣＦ７／Ａｄｒ和ＫＢｖ２００能显著调节Ｄｏｘ和长春新碱的耐药性，且其作用呈剂量依赖性．Ｃｙｃ，Ｉｎｓｒ，Ｉｎｓｎ和Ｖｅｒ均能增加ＭＣＦ７／Ａｄｒ细胞内Ｄｏｘ的积累．Ｃｙｃ和Ｉｎｓｒ调节ＭＤＲ作用明显优于Ｖｅｒ，而Ｉｎｓｎ的作用类似于Ｖｅｒ．结论：Ｃｙｃ，Ｉｎｓｒ和Ｉｎｓｎ能通过增加ＭＤＲ细胞内Ｄｏｘ的积累而调节ＭＤＲ．     

Inhibitory effects of α-methyl-4-(3-oxo-2H-1,2-benzoisoselenazol-2-yl)benzeneacetic acid on Cu~(2+) and Fe~(2+) induced oxidative modification of human low density lipoprotein

为观察α－甲基－４－（３－氧－２Ｈ－１，２－苯并异硒唑－２－基）苯乙酸（ＭＢＢＡ）对Ｃｕ２＋及Ｆｅ２＋氧化修饰低密度脂蛋白（ＬＤＬ）的保护作用及其作用机理，采用分光光度法测定ＬＤＬ中丙二醛（ＭＤＡ）和共轭双烯（ＣＤ）的产生量．ＭＢＢＡ（０．２－２μｍｏｌ·Ｌ－１）能以剂量依赖性抑制Ｃｕ２＋及Ｆｅ２＋诱导的ＭＤＡ和ＣＤ生成．２μｍｏｌ·Ｌ－１的ＭＢＢＡ对Ｃｕ２＋诱导ＬＤＬ产生ＭＤＡ和ＣＤ的抑制率分别为８９．７％和６０．３％．０．５ｍｍｏｌ·Ｌ－１ＧＳＨ对ＬＤＬ产生ＭＤＡ无影响，但能显著增强ＭＢＢＡ对ＭＤＡ生成的抑制作用．上述结果表明ＭＢＢＡ对ＬＤＬ氧化修饰的抑制作用可能依赖于其ＧＳＨ－Ｐｘ样活性的作用和（或）直接还原脂质氢过氧化物的作用．     

氧氟沙星片剂的HPLC测定

氧氟沙星片剂的ＨＰＬＣ测定徐子硕，苗家生（沈阳克达制药厂，辽宁１１００４１）ＨＰＬＣＤＥＴＥＲＭＩＮＡＴＩＯＮＯＦＯＦＬＯＸＡＣＩＮＴＡＢＬＥＴＳ￥ＸＵＺｉ－Ｓｈｕｏ；ＭＩＡＯＪｉａ－Ｓｈｅｎｇ（ＳｈｅｎｙａｎｇＫｅｄａＰｈａｒｍａｃｅｕｔｉｃａｌＦ...     

康乐霉素C对T-和B-淋巴细胞活化的抑制作用(英文)

阐明康乐霉素Ｃ（Ｋａｎ）对脾细胞增殖和Ｔ－细胞亚型的作用。方法：氚掺入法或噻唑蓝（ＭＴＴ）比色法测定细胞增殖;用荧光激活细胞分选仪（ＦＡＣＳ）测定细胞亚型;曲利苯蓝排斥法测定细胞存活率．结果： Ｋａｎ ８, ４０, ８０和 ４００ ｎｍｏｌ· Ｌ－１,除抑制丝裂原（Ｃｏｎ Ａ, ＰＨＡ和 ＴＰＡ＋ＩＭ）和同种异型抗原刺激的小鼠脾细胞增殖外;与Ｃｉｃ不同,抑制ＬＰＳ （１０ｍｇ·Ｌ－１）刺激的脾细胞增殖;使Ｌ３Ｔ４＋／Ｌｙｔ２＋ Ｔ－细胞亚型比值倒置; Ｋａｎ于 Ｃｏｎ Ａ（ ５ｍｇ· Ｌ－１）刺激后 ２４ ｈ内加入,仍抑制脾细胞增殖． Ｋａｎ Ｂ－４００ ｎｍｏｌ· Ｌ－１不影响脾细胞存活率。结论：与Ｃｉｃ的作用方式不同,Ｋａｎ抑制Ｔ－和Ｂ－细胞活化的早期和晚期时相,抑制细胞增殖,对Ｔｎ－细胞有选择性。     

马杜拉放线菌NK73-NF4产生的新抗生素──decatromicin A和B

Ｍｏｍｏｓｅ等从马杜拉放线菌（Ａｃｌｉｎｏｍａｄｕｒａ　ｓｐ．）ＭＫ７３－ＮＦ４的培养液中分离获得含４－羟乙酰乙酸内酯的大环内酯类新抗生素ｄｅｃｔｒｏｍｉｃｉｎＡ、Ｂ，两种新抗生素对革兰氏阳性菌有抗菌活性，而对革兰氏阴性菌无活性。首先对产生菌ＮＫ７３－ＮＦ４进行了鉴定。用扫描电镜观察孢子和菌丝形态；ＨＰＬＣ和质谱分析甲基萘酿类物质，主要是ＭＫ－９（Ｈ６），其次是ＭＫ－９（Ｈ８）。细胞壁水解分析显示存在内消旋－二氨基庚二酸。全细胞水解分析显示存在马杜拉糖。因此将菌株ＭＫ７３－ＮＦ４归属马杜拉放线菌…     

三种双苄基异喹啉生物碱调节多药耐药性的作用及与维拉帕米的比较

目的：比较轮环藤碱（Ｃｙｃ）、海岛轮环藤碱（Ｉｎｓｒ）和海岛轮环藤酚碱（Ｉｎｓｎ）与维拉帕米（Ｖｅｒ）体外调节多药耐药性（ＭＤＲ）的作用。方法：细胞毒试验采用ＭＴＴ法，细胞内阿霉素（Ｄｏｘ）积累采用荧光分光光度法测定。结果：Ｃｙｃ，Ｉｎｓｒ，Ｉｎｓｎ和Ｖｅｒ在分光光度法测定。结果：Ｃｙｃ，Ｉｎｓｒ，Ｉｎｓｎ和Ｖｅｒ在ＭＤＲ细胞系ＭＣＦ－７／Ａｄｒ和ＫＢｖ２００能显著调节Ｄｏｘ和长春新碱的耐药性具有     

番荔枝内酯单体89-2实验治疗KBv200及KB细胞移植瘤的作用

目的番荔枝内酯单体89-2是自阿蒂莫耶番荔枝植物分离获得。本研究旨在探讨89-2对MDR肿瘤的实验治疗作用。方法以MTT法测定细胞毒;KBv200及KB细胞裸鼠移植瘤模型研究89-2对MDR肿瘤的实验治疗作用;以Fura-2-AM法测定P-糖蛋白(P-gp)功能。结果89-2对KBv200及KB细胞的IC₅₀分别为48.7和64.6 nmol·L^-1(P>0.05)。89-2对KB及KBv200细胞裸鼠移植瘤具有相似的剂量依赖性抑制作用,而毒性可以耐受。89-2剂量依赖性地增加KBv200细胞的Fura-2积累。结论89-2体内外均抑制KB和KBv200细胞生长,具有开发前景。     

Reversal of P-gp mediated multidrug resistance in-vitro and in-vivo by FG020318

Overexpression of P-glycoprotein (P-gp) by tumours results in multidrug resistance (MDR) to structurally and functionally unrelated chemotherapeutic drugs. Combined therapy with MDR-related cytotoxins and MDR modulators is a promising strategy to overcome clinical MDR. This study was performed to explore the MDR reversal activity of a novel compound 2-[4-(2-pyridin-2-yl-vinyl) phenyl]-4,5-bis-(4-N,N-diethylaminophenyl)-1(H)-imidazole (FG020318) in-vitro and in-vivo. Tetrazolium (MTT) assay was used to evaluate the ability of FG020318 to reverse drug resistance in two P-gp-expressing tumour cell lines, KBv200 and MCF-7/adr. Intracellular doxorubicin accumulation was determined by fluorescence spectrophotometry in MCF-7/adr cell line. The effect of FG020318 on P-gp function was demonstrated by rhodamine 123 (Rh123) accumulation in KBv200 cells. KBv200 cell xenograft models were established to study the in-vivo effect of FG020318 on reversing MDR. FG020318 was not cytotoxic by itself against P-gp expressing KBv200 cells and MCF-7/adr cells and their parental drug-sensitive KB cells and MCF-7 cells. FG020318 could significantly increase the sensitivity of MDR cells to antitumour drugs including doxorubicin and vincristine in MCF-7/adr cells and KBv200 cells, respectively. It was much stronger than the positive control verapamil in reversal of MDR. FG020318 also increased the intracellular accumulation of doxorubicin in a concentration-dependent manner in MCF-7/adr cells, but did not affect the accumulation of doxorubicin in drug-sensitive MCF-7 cells. The Rh123 accumulation in resistant KBv200 cells was also increased by the addition of FG020318, but Rh123 accumulation was not affected by FG020318 in drug-sensitive KB cells. FG020318 potentiated the antitumour activity of vincristine to KBv200 xenografts and was an efficacious modulator in-vivo. Our results suggested that FG020318 was a highly potent, efficacious MDR modulator not only in-vitro but also in-vivo. The reversal of drug resistance by FG020318 was probably related to the increased anticancer drug accumulation and its inhibition of P-gp function of MDR tumour cells.     

The anti-hepatitis drug DDB chemosensitizes multidrug resistant cancer cells <Emphasis Type="Italic">in vitro</Emphasis> and <Emphasis Type="Italic">in vivo</Emphasis> by inhibiting P-gp and enhancing apoptosis

Summary Purpose: DDB (dimethyl-4,4′-dimethoxy-5,6,5′6′-dimethylene dioxybiphenyl-2,2′-dicarboxylate) is a synthetic hepatoprotectant which has been widely used to treat chronic viral hepatitis B patients in China for more than 20 years. In this study, we evaluated DDB as a multidrug resistance (MDR) chemosensitizing agent. Methods: A panel of sensitive and resistant cancer cell lines were treated with various concentration of DDB, and the effect on chemosensitivity and accumulation of anticancer drugs; promotion of apoptosis and P-glycoprotein (P-gp) expression were determined by MTT (Dimethyl thiazolyl-2,5-diphenyltetrazolium bromide) assay, fluorospectrometry and flow cytometry respectively. Drug resistance reversal activity of DDB was also examined in BALB/c nude mice bearing both acquired MDR human nasopharyngeal carcinoma KBv200 and parental KB xenografts. The effect of DDB on the pharmacokinetics of Dox and hematological toxicity induced by Dox was measured in ICR and C₅₇/BL mice, respectively. Results: DDB at nontoxic concentrations of 12.5, 25 and 50 μM partly reversed the resistance to vincristine, doxorubicin, paclitaxel in acquired MDR breast carcinoma MCF-7/Adr cells, KBv200 and intrinsic MDR human hepatocarcinoma Bel₇₄₀₂ cells, whereas no chemosensitizing effect of DDB was observed in sensitive KB and MCF-7 cells. DDB increased the intracellular accumulation of doxorubicin and inhibited surface P-gp expression in MCF-7/Adr cells. Furthermore, it was found that DDB promoted doxorubicin-induced apoptosis of Bel₇₄₀₂ cells through enhanced caspase-3 activation. Co-administration of DDB at 300 and 500 mg/kg orally to nude mice increased the antitumor activity of vincristine to KBv200 xenografts without a significant increase in toxicity. In contrast, Co-administration of DDB did not inhibit the growth of KB xenografts. DDB also markedly reduced the decrease of leukocytes in doxorubicin-treated C₅₇/BL mice. Co-administration of DDB increased Dox concentration in ICR mice bearing S180 sarcoma, but no pharmacokinetical interaction with Dox was observed. Conclusion: These results indicate that DDB has MDR reversal activity by inhibiting P-gp and when used in combination with anti-cancer drugs, it could potentially be used as a clinical treatment for P-gp-mediated MDR cancers.     

Reversal of P-glycoprotein-mediated multidrug resistance by the novel tetrandrine derivative W6

Overexpression of ATP-dependent efflux pump P-glycoprotein (P-gp) is the main cause of multidrug resistance (MDR) and chemotherapy failure in cancer treatment. Inhibition of P-gp-mediated drug efflux is an effective way to overcome cancer drug resistance. The present study investigated the reversal effect of the novel tetrandrine derivative W6 on P-gp-mediated MDR. KBv200, MCF-7/adr and their parental sensitive cell lines KB, MCF-7 were used for reversal study. The intracellular accumulation with P-gp substrates of doxorubicin was determined by flow cytometry. The expression of P-gp and ERK1/2 was investigated by western blot and real-time-PCR (RT-PCR) analysis. ATPase activity of P-gp was performed by P-gp-Glo^TM assay systems. In comparison with P-gp-negative parental cells, W6 produced a favorable reversal effect in the MDR cells, as determined using the MTT assay. W6 significantly and dose-dependently increased intracellular accumulation of P-gp substrate doxorubicin (DOX) in P-gp overexpressing KBv200 cells, and also inhibited the ATPase activity of P-gp. W6 inhibited P-gp expression in KBv200 cells in a time-dependent manner, but it had no effect on MDR1 expression. In addition, W6 significantly decreased the ERK1/2 activation in KBv200 cells. Our results showed that W6 effectively reversed P-gp-mediated MDR by inhibiting the transport function and expression of P-gp, demonstrating the potential clinical utility of W6.     

Bullatacin克服肿瘤多药抗药性作用及其机理

目的：探讨bullatacin克服肿瘤多药抗药性(MDR)的作用及其机制。方法：以两对MDR细胞株及其相应的敏感株进行对比,比较两种细胞株的细胞毒、Fura-2及阿霉素细胞内积累。结果：bullatacin不仅对敏感细胞株具有很强的细胞毒活性,而且对MDR细胞株也同样具有很强的细胞毒活性,不受抗药性的影响。bullatacin能使MDR细胞内Fura-2的积累增加;也能增加MDR细胞内阿霉素的积累。结论：bullatacin具有克服MDR的作用,其作用机理与bullatacin影响MDR细胞P-gp的功能,使MDR细胞内抗癌药物积累增加有关。     

基因mdr1高表达多药耐药肿瘤细胞对力达霉素的药物敏感性

目的利用经药物诱导获得的mdr1基因高表达细胞株以及通过mdr1基因转染建立的稳定高表达细胞株，研究多药耐药肿瘤细胞对力达霉素(C-1027)的药物敏感性。方法构建mdr1重组真核表达质粒pcDNA3.1/mdr1，利用脂质体转染技术，获得mdr1高表达HepG2肝癌细胞。经RT-PCR、细胞荧光免疫化学及罗丹明外排实验，鉴定了细胞的mdr1表达水平和药物外排活性。MTT方法测定敏感细胞及相对应的多药耐药细胞对力达霉素等多种抗肿瘤药物的药物敏感性。结果mdr1稳定转染细胞株HepG2/mdr1、多药耐药KBv200细胞和MCF-7/ADR细胞对力达霉素的IC₅₀值分别为(0.020±0.011) nmol·L^-1，(0.24±0.20) nmol·L^-1和(0.028±0.011) nmol·L^-1。相对于各自的敏感细胞，多药耐药细胞HepG2/mdr1，KBv200和MCF-7/ADR对力达霉素的抗药倍数分别是1.3，6.8和1.6倍，对阿霉素的抗药倍数分别是8.8， 37.2和181.3倍，对紫杉醇的抗药倍数分别是40.3， 336.8和49.2倍。结论 mdr1高表达的多药耐药肿瘤细胞对力达霉素仍高度敏感，未表现出抗药性。     

脂质体阿霉素逆转肿瘤多药耐药的活性和机制研究

目的 探讨脂质体阿霉素( PLD)逆转肿瘤多药耐药(MDR)的活性及其逆转机制.方法 以噻唑蓝(MTT)方法检测PLD对多药耐药肿瘤细胞MCF-7/ADR及KBv200的耐药逆转活性.结果 PLD在体内具有较强的逆转活性,逆转活性大于公认的强逆转剂维拉帕米的活性;在5.0μmol/L浓度下使多药耐药细胞KBv200对长春新碱的敏感性增加了45倍.PLD浓度依赖性增加(0、2.5、5.0、10 μmol/L)KBv200细胞内的罗丹明蓄积.PLD的心脏毒性、骨髓抑制以及脱发等不良反应显著降低.结论 脂质体阿霉素具有较强的逆转MDR的活性,主要通过持续向肿瘤组织聚集,肿瘤局部药物浓度升高,抗肿瘤的活性增强.     

R—dl—型维拉帕米调低KBv200肿瘤细胞对长春新碱和多柔比星的多？…

目的：研究Ｒ－型维拉帕米对多药耐药的降低和及其急性动物毒性,并与消旋维闰帕米的相应结果作比较。方法：细胞毒性的测定用ＭＴＴ法;细胞内多柔比星积累的测定用萤光分光光度计法。急性毒性试验用ＢＡＬＢ－ｃ小鼠腹腔注射法。结果：Ｒ－型维拉帕米部分调低ＫＢｖ２００细胞对长春新碱和多柔比星的耐区性,其调低效应与作用肖度和作用时间有关。     

双苄基异喹啉类生物碱粉防己碱与小檗胺逆转多药抗药性的比较研究

比较了2种结构相近的双苄基异喹啉(BBI)生物碱粉防己碱(TTD)、小檗胺(BBM)与维拉帕米(VRP)逆转多药抗药性的作用。结果,TTD,BBM和VRP在多药抗药的MCF-7/Adr和KBv₂₀₀细胞对ADR和VCR均有明显增敏作用,且作用呈剂量依赖性。其中10μmol·L^-1TTD能完全逆转MCF-7/Adr细胞对ADR的抗药性。TTD,BBM和VRP均有增加MCF-7/Adr细胞内阿霉素积累的作用。TTD和BBM在结构上仅有微小差别,但TTD的逆转MDR作用优于VRP10倍,而BBM的作用与VRP相仿。TTD在裸鼠体内MCF-7/Adr实体瘤模型上也证实有明显逆转ADR抗药性的作用。