小鼠各种组织细胞对N-甲酰溶肉瘤素敏感性的比较——病理形态和细胞计数的研究

本实验使用形态学方法在切片上计算细胞数以比较N-甲酰溶肉瘤素对小鼠各种器官所引起病变的发展过程,并从数量上去衡量各种细胞对药物的敏感性。结果发现小鼠腹腔內注射N-甲酰溶肉瘤素150毫克/公斤后,病变主要见于骨髓、胸腺、淋巴结、脾脏、小肠、大肠和睾丸。各种组织的病变发展过程不同,一般在给药后2—4天最为严重。在敏感的器官中,不同类型组织细胞对N-甲的敏感性有很大差別,最敏感的是睾丸的精原细胞和骨髓的嗜酸性粒细胞。     

N-甲酰溶肉瘤素对小鼠精原细胞的损害作用

用计算细胞数的方法,观察了N-甲酰溶肉瘤素对小鼠造精细胞的损害作用。结果表明,在造精细胞系列中,精原细胞对N-甲酰溶肉瘤素最为敏感,表现为细胞变性、核分裂消失、细胞数极度减少,在各型精原细胞中,又以B型更为敏感。考虑到N-甲酰溶肉瘤素是一种在临床对精原细胞瘤有显著疗效的药物,本文讨论了药物对精原细胞的损害作用和对精原细胞瘤的治疗作用的相关性.     

关于抗肿瘤药物的研究 Ⅰ.207种合成化合物对动物移植性肿瘤的影响

本文报告几年来从合成化合物中筛选抗肿瘤药的结果,在207种合成化合物中,有效药物主要都是氮芥衍生物,计有N-甲酰溶肉瘤素(M₂),N-乙酰溶肉瘤素(M₃),N-甲酰溶肉瘤素酰苯丙氨酸乙酯(M₄),邻苯二甲谷氨酰溶肉瘤秦(M₅)及α-硝基-β-双(2-氯乙基)氨基己酸乙酯(M₉),α-硝基-β-双(2-氯乙基)氨基-β-(p-二甲苯基氨基)丙酸乙酯(M₁₀)。在其他类型化合物中,只丙烷异硫脲酒石酸锑对吉田腹水肉瘤有一定疗效。有效化合物中以N-甲酰溶肉瘤素(M₂),N-乙酰溶肉瘤素(M₃),N-甲酰溶肉瘤素酰苯丙氨酸乙酯(M₄)及邻苯二甲谷氨酰溶肉瘤素(M5)的疗效比较突出。已经在临床前药理研究的基础上,将N-甲酰溶肉瘤素及N-甲酰溶肉瘤素酰苯丙氨酸乙酯推荐给临床试用。     

几种氮芥类抗肿瘤药与大鼠血浆的结合

本文用超过滤法比较了几种苯丙氨酸氮芥与大鼠血浆的结合,并以N-甲酰溶肉瘤素(以下简称N-甲)为例,对影响结合的因素进行了探讨。N-甲等氨基被酰化的衍生物与大鼠血浆结合的程度均大于60%,而溶肉瘤素等氨基未被酰化的衍生物结合程度均低于24%。分子中的氯乙基可能是此种结合所必要的基团,而游离羧基并非必要。但此结合不受溫度的影响。N-甲与血浆结合的最适pH为3.6;在一定范围內,结合程度随血浆稀释度增加而减少。所试各种组织匀浆与N-甲结合程度以下列次序递减:肝,81.5%;血浆,71.9%;肾,69.7%;梭形细胞肉瘤B₂₂(N-甲不敏感瘤),60.9%;吉田肉瘤(N-甲敏感瘤)57.9%。     

几种氮芥类抗肿瘤药与大鼠血浆的结合

本文用超过滤法比较了几种苯丙氨酸氮芥与大鼠血浆的结合,并以N-甲酰溶肉瘤素(以下简称N-甲)为例,对影响结合的因素进行了探讨。 N-甲等氨基被酰化的衍生物与大鼠血浆结合的程度均大于60％,而溶肉瘤素等氨基未被酰化的衍生物结合程度均低于24％。分子中的氯乙基可能是此种结合所必要的基团,而游离羧基并非必要。但此结合不受溫度的影响。N-甲与血浆结合的最适pH为3.6;在一定范围內,结合程度随血浆稀释度增加而减少。所试各种组织匀浆与N-甲结合程度以下列次序递减:肝,81.5％;血浆,71.9％;肾,69.7％;梭形细胞肉瘤B_(22)(N-甲不敏感瘤),60.9％;吉田肉瘤(N-甲敏感瘤)57.9％。     

关于抗肿瘤药物的研究 Ⅶ.N-甲酰溶肉瘤素的抗瘤谱及毒性

N-甲酰溶肉瘤素是一种有效的抗肿瘤药,抗瘤譜广,在12种动物肿瘤中,对9种具有明显的抑制作用。对大鼠吉田肉瘤腹水型及实体型,Walker癌-肉瘤256等抑制作用最明显,抑制率为96—100%。对小鼠网織細胞肉瘤L₂抑制率为60—70%;对腹水瘤L₁及Krebs-2腹水癌亦有明显抑制作用,分別延长寿命5天及7天;对Ehrlich癌腹水型小鼠,可延长寿命2.2—2.6天;对Ehrlich癌实体型及梭形細胞肉瘤B₂₂抑制率分別为30—41%及36—43%。在不致中毒的剂量下,对小鼠肉瘤180,肉瘤AK,黑色素瘤Me則无明显抑制作用。口服N-甲酰溶肉瘤素对小鼠及大鼠的半数致死量(LD₅₀±S.E.)分別为730±79毫克/公斤及700±79毫克/公斤;腹腔內給药則分別为152±7毫克/公斤及80±10毫克/公斤。靜脉注射对小鼠的半数致死量为155±10毫克/公斤。     

关于抗肿瘤药物的研究——Ⅳ.N-甲酰溶肉瘤素与溶肉瘤素的效价比较

对药物的评价不仅决定于治疗剂量的大小,还取决于药物治疗剂量与中毒剂量差距的大小(即治疗指数及安全限).我们在寻找抗肿瘤新药的研究中,发现N-甲酸溶肉瘤素(以下简称N-甲)有明显的抗肿瘤作用.从化学结构看来,N-甲与溶肉瘤素非常相似,但临床观察表明,N-甲对溶肉瘤素不敏感的淋巴肉瘤、何杰金氏病却有明显疗效.因此,我们对N-甲及溶肉瘤素在三种动物肿瘤作用的效价进行了比较.本文即研究初步结果的简报.材料及方法药物系由我所药物化学系合成室供给.N-甲批号是6103,熔点为148-151℃,实验前将药物混悬于2％淀粉糊中.溶肉瘤素批号是S615,熔点为178-181℃,实验前将药物用70-80℃蒸馏水溶解,现用现配.     

关于抗肿瘤药物的研究Ⅸ.N-甲酰溶肉瘤素的一些药理作用

抗肿瘤药N-甲酰溶肉瘤素(简称N-甲)的毒性低.小鼠于每天口服25-150毫克/公斤连续10天,游泳耐力不减,肝脏破坏戊巴比妥钠功能正常.小鼠于腹腔注射N-甲10-20毫克/公斤/天,连续10天,或150毫克/公斤一次,对因戊巴比妥钠产生的睡眠时间无改变.大鼠于每天腹腔注射N-甲10毫克/公斤,连续10天,其利尿功能,肾上腺内维生素C含量及微血管渗进性均无明显改变.麻醉兔于静脉注射N-甲10-40毫克/公斤后,血压亦无明显改变.当N-甲20毫克/公斤/天,连续10天时,于注射后5天,大鼠利尿功能受到抑制,但于停药后3天逐渐恢复正常.大鼠于腹腔注射N-甲80毫克/公斤后2小时,肾上腺内维生素C含量明显下降,微血管渗透性显著增加.     

从姑息性治疗转向根治性治疗，时空几何？

1肿瘤化疗能否划上句号 1.1肿瘤化疗的现状和水平 我国现代肿瘤内科治疗始于20世纪50年代.自放线菌素K研制成功以来,北京、上海各自合成了几种烷化剂,如N-甲酰溶肉瘤素、消瘤芥、甲氧芳芥、甘磷酰芥等,对常见肿瘤均有一定疗效.     

肉苁蓉总苷对照射损伤小鼠敏感器官超微结构的保护作用

观察肉苁蓉总苷 ( GCs)对小鼠经60 Coγ射线照射后胸腺、脾、睾丸超微结构变化的影响 ;结果表明辐射损伤后小鼠胸腺、脾、睾丸生物膜及细胞器受到不同程度的损伤 ,GCs可促进其恢复 ;推论 GSs对受照小鼠敏感器官超微结构有保护作用 ,其机制可能与抗氧化、促进 DNA、RNA合成有关。     

五种抗肿瘤药物对小鼠各种组织损害作用的比较

用计算细胞数的方法,比较了五种抗肿瘤药物对小鼠各种正常组织的损害作用.结果表明,不同的抗肿瘤药物,对正常组织的损害作用是有差别的.这种差别,不仅存在于烷化剂与抗代谢物两类药物之间,也存在于各种烷化剂之间.研究这些差别,将对寻找新药和临床应用方面提供参考资料.     

Alterations in the morphology and functional activity of bone marrow phagocytes following benzene treatment of mice.

Benzene is a well-established hematotoxin that affects developing leukocytes and erythrocytes as well as bone marrow stromal cells. In the present studies we analyzed the effects of benzene on the morphology and functional activity of bone marrow phagocytes. Male Balb/c mice were treated with benzene (660 mg/kg) once per day for 3 days. Bone marrow cells were then isolated and fractionated by density gradient centrifugation. Using highly sensitive techniques in flow cytometry/cell sorting, we found that we could separate three distinct populations of bone marrow cells that differed with respect to size and density. Monoclonal antibody binding and cell sorting revealed a large, dense population that consisted predominantly of granulocytes, a smaller, less dense population of lymphocytes, and a population of intermediate size and density consisting of mononuclear phagocytes and precursor cells. Differential staining of sorted mononuclear phagocytes revealed that benzene treatment of mice caused a marked increase in the number of mature, morphologically activated macrophages in the bone marrow. Benzene treatment of mice also resulted in enhanced chemotaxis and production of hydrogen peroxide by bone marrow granulocytes and mononuclear phagocytes. In contrast, treatment of mice with the combination of hydroquinone and phenol (50 mg/kg each, 1 x/day, 3 days), two metabolites of benzene, resulted in a significant (p < or = 0.02) depression of granulocyte chemotaxis and had no effect on hydrogen peroxide production by bone marrow phagocytes compared to cells from control animals. Taken together these results demonstrate that benzene causes increased differentiation and/or activation of phagocytes in the bone marrow.     

Immunotoxicity of the anticancer drug CI-994 in rats: effects on lymphoid tissue

CI-994 (acetyldinaline) is an investigative oral anticancer drug currently in clinical trials. To characterize the effects of CI-994 on lymphoid tissue, male rats were administered single oral doses at 0 (vehicle control), 10, 23, and 45 mg/kg and killed up to 7 days after dosing for evaluation of white blood cell differentials, bone marrow differentials, lymphoid tissue weights, and selected histopathology of lymphoid tissue. Statistically significant dose-related reductions in blood lymphocytes (CD-3⁺, CD-4⁺, CD-8⁺, CD-20⁺), monocytes, neutrophils, and bone marrow lymphoid cells were observed in all drug-treated groups on days 1 and/or 3. Statistically significant reductions in bone marrow myeloid cells were also observed on days 1 and 3 at 23 and 45 mg/kg. Complete or partial reversal of most parameters was evident on day 7. Spleen and/or thymus weights were significantly decreased in the groups administered 23 and 45 mg/kg on days 1, 3, and/or 7. Minor reductions in lymphoid organ weights were noted at 10 mg/kg. Minimal to moderate lymphoid depletion of the spleen and thymus was noted on day 3 in animals dosed at 23 mg/kg. In vitro, CI-994 inhibited mitogen-stimulated blood lymphocyte proliferation with a 50% inhibitory concentration (IC₅₀) value of 3 μM. The results demonstrate that CI-994 can effect lymphoid tissue in rats within 1 day of a single oral dose, that effects are generally reversible within 7 days, and that inhibition of lymphocyte proliferation is a sensitive indicator of CI-994 immunotoxicity in vitro. Received: 23 September 1998 / Accepted: 12 January 1999     

In Vivo Micronucleus Assay and GST Activity in Assessing Genotoxicity of Plumbagin in Swiss Albino Mice

Information available on the mutagenicity of a large number of indigenous drugs commonly employed in the Siddha and Ayurveda systems of medicine is scanty. In this context, the current investigation on plumbagin, 5-hydroxy-2methyl-1,4-napthoquinone, an active principle in the roots of Plumbago zeylanica used in Siddha and Ayurveda for various ailments, was carried out; 16 mg/kg b.w. (LD₅₀) was fixed as the maximum dose. Subsequent dose levels were fixed as 50% and 25% of LD₅₀ amounting to 8 mg and 4 mg/kg b.w., respectively, and given orally for 5 consecutive days in 1% Carboxyl Methyl Cellulose (CMC) to Swiss albino mice weighing 25–30 g. The micronucleus assay was done in mouse bone marrow. Plumbagin was found to induce micronuclei at all the doses studied (4 mg/kg, 8 mg/kg, 16 mg/kg b.w.), and it proves to be toxic to bone marrow cells of Swiss albino mice. Animal treated with cyclophosphamide (40 mg/kg b.w.) served as positive control. In addition, glutathione S-transferase (GST) activity was observed in control, plumbagin (4 mg, 8 mg, 16 mg/kg b.w., respectively), and genotoxin-treated experimental group of animals. No significant change in GST activity was observed with plumbagin dose of 4 mg/kg b.w., whereas GST activity was significantly inhibited by higher doses of plumbagin (8 mg and 16 mg/kg b.w.) and cyclophosphamide.     

Protective effects of ginsenoside Rg<Subscript>3</Subscript> against cyclophosphamide-induced DNA damage and cell apoptosis in mice

Despite the significant anti-tumor activities, cyclophosphamide (CP) also shows cytotoxicity to normal cells. In order to explore the protective effects of drugs against CP-induced adverse effects, 20(S)-ginsenoside Rg₃ was tested for its possibly protective activities on CP-induced DNA damage and cell apoptosis in mouse bone marrow cells or peripheral lymphocyte cells. In the current study, the alkaline single cell gel electrophoresis (comet assay), flow cytometry assay with annexin V-FITC/PI and AO/EB staining assay were employed to measure DNA strand breakage and cell apoptosis, respectively. The activities of SOD and GPx and the contents of MDA were also tested by the various colormetric methods. The results showed that CP at a dose of 100 mg/kg, i.p. significantly caused DNA damages in both mouse bone marrow cells and peripheral lymphocyte cells, and markedly inhibited the activities of GPx and SOD and increased MDA contents in mouse blood. Moreover, CP at a dose of 200 mg/kg, i.p. triggered apoptosis in mouse bone marrow cells. On the other hand, 20(S)-ginsenoside Rg₃ orally administered at a dose of 20 mg/kg to the animals once a day for 2 days significantly inhibited CP-induced DNA damages in mouse bone marrow cells and peripheral lymphocyte cells, decrease the apoptotic numbers of bone marrow cells, antagonized the reduction of the activities of SOD and GPx, and the increase in MDA contents. In conclusion, 20(S)-ginsenoside Rg₃ showed the significant protective effects on CP-induced cell DNA damage and apoptosis. These effects might be partially attributed to its protective actions against CP-induced oxidative stress.     

Effects of cyclophosphamide on hemic precursor cells in mouse bone marrow and spleen

Cyclophosphamide severely inhibits the function of various hemic precursor cells in mice. In these studies we compared the effects of Cy on early B-lymphocytic precursor cells in mouse bone marrow to those effects on bone marrow myeloid precursor cells and splenic lymphoid precursor cells. These studies demonstrated that: 1) Pre-ARC in the bone marrow were extremely sensitive to even low doses of Cy (50 mg/kg) with virtually no renewal of this precursor population 11 days after injection of 200 mg/kg. 2) The sensitivity of pre-ARC to Cy along with the lack of their renewal was exemplified by the virtual absence of functional ARC in the spleen when assessed 11 days after injection of 200 mg/kg of Cy. 3) A rapid multiphasic renewal pattern was observed for myelocytic progenitor cells. 4) In general, LPS responsive cells were suppressed to a greater extent than were Con A responsive cells, especially with lower doses of Cy.     

Behavioral effects of apomorphine isomers in the rat: Selective locomotor-inhibitory effects of S(+)N-n-propylnorapomorphine

The optical isomers of apomorphine (APO) and N-n-propylnorapomorphine (NPA) were evaluated behaviorally in the rat. Both R(-) isomers induced motor-excitatory effects and strong stereotyped sniffing, licking, and gnawing, as has been reported previously. The S(+) isomers selectively inhibited locomotor activity and did not induce stereotypy or catalepsy. These actions of the S(+) aporphines were selective against locomotor activity stimulated by low doses of R(-) isomers. (+)NPA (ID₅₀=0.2 mg/kg) was 20 times more potent than (+)APO (ID₅₀=4 mg/kg) in antagonizing the locomotor arousal-inducing effects of (-)APO (at ED₅₀=0.3 mg/kg). (+)NPA also inhibited spontaneous locomotor activity much more potently (ID₅₀=3.0 mg/kg) than did (+)APO (ID₅₀>50 mg/kg). Neither S(+) aporphine had a significant effect against stereotypy induced by the R(-) isomers, even at high doses (up to 30 mg/kg). Inhibition of the effects of (-)APO by (+)NPA appeared not to be due to altered uptake of (-)APO into brain. These results suggest that S(+)NPA or its congeners and analogs may have selective antidopaminergic actions in limbic rather than striatal areas of mammalian brain.     

Prolonged antidopaminergic actions of single doses of butyrophenones in the rat

Rats were treated once with doses of haloperidol or of droperidol below and above the acute ID₅₀ vs the dopamine agonist apomorphine; they were later challenged with an acute dose of apomorphine (0.3mg/kg, SC) and rated for stereotyped behavioral responses. The two neuroleptics were similar in acute anti-apomorphine potency (ID₅₀=0.12 and 0.18mg/kg for haloperidol and droperidol, respectively). The antidopaminergic effects of droperidol persisted for nearly 1 week and those of haloperidol lasted for 20–40 days, depending on the dose given. The computed half-time of disappearance of their antidopaminergic effects was 7.6±1.0 days and 0.59±0.17 days for haloperidol and droperidol, respectively, following a dose of 0.3 mg/kg, and these indices of duration of action did not vary significantly at doses between 0.1 and 1.0mg/kg. Haloperidol reduced the acute entry of ³H-apomorphine into brain by 21.5% 1 week later. Treatment with apomorphine alone just prior to haloperidol (both at 0.3 mg/kg) prevented the prolonged antidopaminergic effects of the neuroleptic evaluated 1 week later. These results indicate that some neuroleptics may have very prolonged activity or retention in tissue at sites of action, even after moderate, single doses. Caution is recommended in the interpretation of studies which assume neuroleptic-free conditions of subjects previously exposed to a neuroleptic agent.