苦参碱对白血病细胞HL-60黏附、运动和侵袭的抑制作用

目的:研究不同浓度苦参碱对白血病细胞HL-60黏附、运动和侵袭的影响.方法:HL-60细胞体外培养,经0、0.1、0.15、0.2 g/L苦参碱分别处理细胞,细胞黏附实验检测细胞黏附力,细胞迁移实验检测细胞运动力,细胞侵袭实验检测细胞侵袭力,RT-PCR检测HL-60细胞MMP-2和MMP-9 mRNA的表达.结果:与对照组比较,0.1、0.15、0.2 g/L苦参碱不仅能有效抑制HL-60细胞的黏附力、运动力和侵袭力(P＜0.01),而且还能下调HL-60细胞MMP-2和MMP-9 mRNA的表达(P＜0.01).结论:苦参碱能抑制HL-60细胞的浸润转移能力,其机制可能与下调MMP-2和MMP-9 mRNA的表达使HL-60细胞的黏附力、运动力和侵袭力下降有关.     

青藤碱对体外人胆管癌细胞的抑制作用

目的研究青藤碱对体外人胆管癌细胞的抑制作用。方法 CCK-8和平板克隆实验检测青藤碱对胆管癌细胞存活率及增殖率的影响,流式细胞术检测青藤碱对胆管癌细胞凋亡作用的影响,Transwell检测青藤碱对胆管癌细胞侵袭能力的影响,细胞划痕检测青藤碱对胆癌细胞迁移能力的影响,Western blot检测青藤碱对胆管癌细胞中凋亡相关蛋白caspase-3和caspase-9和侵袭相关蛋白E-Cadherin、Vimentin表达的影响。结果随着青藤碱浓度的提高以及时间的延长,胆管癌细胞的存活率逐渐下降(P0.05,P0.01),细胞集落形成数量逐渐减少(P0.05,P0.01)。青藤碱可呈剂量依赖性地促进胆管癌细胞凋亡(P0.01),抑制胆管癌细胞的侵袭迁移(P0.01),并且可以使cleaved caspase-3、cleaved caspase-9蛋白以及E-cadherin蛋白表达逐渐升高,vimentin蛋白表达逐渐下降(P0.05,P0.01)。结论青藤碱可以有效地抑制体外胆管癌细胞的增殖,侵袭和迁移能力,诱导细胞凋亡,这可能与其上调cleaved caspase-3、cleaved caspase-9和E-cadherin蛋白表达,以及下调vimentin蛋白表达有关。     

Inhibitory effects of the root cortex of Paeonia suffruticosa on interleukin-8 and macrophage chemoattractant protein-1 secretions in U937 cells

In an effort to elucidate the mechanism of the anti-inflammatory effect of mudanpi, the root cortex of Paeonia suffruticosa Andrews (Ranunculaceae), we determined the effects of the methanolic extract of mudanpi (MEM) on the secretions of interleukin (IL)-8, a major mediator of acute neutrophil-mediated inflammation, and macrophage chemoattractant protein (MCP)-1, a major mediator of chronic macrophage-mediated inflammation, in human monocytic U937 cells stimulated with phorbol myristate acetate (PMA). MEM significantly inhibited PMA-induced secretions of IL-8 and MCP-1 proteins in a dose-dependent manner. The inhibition of these chemokines by MEM was due to its suppression of IL-8 and MCP-1 genes. In addition, 1,2,3,4,6-penta-O-galloyl-beta-D-glucose, one of major constituents isolated from MEM, inhibited PMA-induced secretions of IL-8 and MCP-1 proteins by its suppression of IL-8 and MCP-1 genes. Thus, one possible anti-inflammatory mechanism of mudanpi, an anti-inflammatory Chinese crude drug, may be to inhibit the secretions of inflammatory chemokines.     

蟾酥总蟾蜍甾烯对H22荷瘤小鼠的抗肿瘤作用及组织中代谢物的初步分析

目的:评价蟾酥总蟾蜍甾烯(total bufadienolides from toad venom,TBFs)对H22荷瘤小鼠的肿瘤抑制作用,同步对体内组织中的代谢物进行初步分析.方法:采用HPLC和LC-MS对制备的受试药TBFs的化学组成进行表征;采用灌胃和腹腔注射2种方式给予H22荷瘤小鼠高、中、低3个剂量的受试药,给药后第13天处死,剥取瘤组织,称重,计算抑瘤率;同时取心、肝、脾、肺、肾各脏器组织,采用HPLC和LC-MS对其中的药物进行初步分析.结果:在对照品对照,在线紫外吸收光谱和质谱数据的基础上,明确了受试药TBFs的化学组成,以日蟾毒它灵、沙蟾毒精、蟾毒它灵、酯蟾毒精、华蟾毒它灵、蟾毒灵、华蟾酥毒基和酯蟾毒配基为主.与空白对照组比,灌胃给予TBFs对H22荷瘤小鼠的体重无影响,低(5 mg· kg-1),中(10 mg·kg-1),高剂量组(20 mg·kg-1)的抑瘤率分别为14.76％,16.38％,10.32％;腹腔注射给予TBFs,与空白对照组比,中、高剂量组在给药第5天时体重增加较慢,第13天时恢复正常;低(1.5mg· kg-1),中(3 mg·kg-1),高剂量组(6 mg·kg-1)的抑瘤率分别为17.30％,19.80％,40.95％,呈一定的剂量依赖性.小鼠心、肝、脾、肺、肾脏器组织HPLC分析表明,肝组织中存在蟾蜍甾烯类成分,LC-DAD-MS初步推测其中的11个化合物.结论:灌胃给予TBFs对H22荷瘤小鼠肿瘤生长有一定的抑制趋势,但作用不明显;腹腔注射则展示了显著性抑瘤作用,同时给药初期也表现了一定的毒性反应;肝组织中推测的蟾蜍甾烯类化合物为进一步研究其体内代谢途径奠定基础.     

毛蕊花糖苷抑制脂多糖诱导的BV-2小胶质细胞炎症反应及机制研究

探究毛蕊花糖苷(acteoside,ACT)对细菌脂多糖(lipopolysaccharide,LPS)诱导的BV-2小胶质细胞神经炎症反应的抑制作用及潜在机制。该实验以LPS诱导BV-2细胞作为炎症模型,将BV-2细胞分为正常对照组、模型组、给药组,ACT按指定浓度(12.5,25,50μmol·L-1)作用于细胞。采用NO试剂盒检测NO炎症因子的释放量,ELISA法检测TNF-α和IL-6的表达量,Western blot法检测炎症相关蛋白(i NOS,COX-2,p-IKKβ,IKKβ,p-IκB,IκB)的表达。此外,通过检测细胞核/浆中NF-κB p65的表达以及借助免疫荧光技术,阐明ACT对炎症转录因子NF-κB p65核转位的作用。结果显示,ACT可显著降低LPS诱导的BV-2小胶质细胞神经炎症,且呈剂量依赖性。具体表现在:显著降低炎症因子NO,IL-6和TNF-α的释放量,抑制炎症相关蛋白i NOS和COX-2的表达。同时,ACT可明显抑制NF-κB信号通路中关键蛋白IKKβ和IκB的磷酸化,且明显抑制NF-κB p65的细胞核转位。以上结果表明,毛蕊花糖苷可显著降低LPS诱导的小胶质细胞神经炎症反应,抑制炎症相关蛋白的表达,其潜在机制是通过抑制NF-κB信号通路实现的。     

Inhibitory effects of Agaricus blazei extracts on human myeloid leukemia cells

Ethnopharmacological relevance

Agaricus blazei has been used as an adjuvant in cancer chemotherapy and is found to inhibit the growth of various types of tumor cells.

Aim of the study

Our study has adopted a systematic and bioassay-guided approach to optimize the extraction of Agaricus blazei for anti-leukemic bioactive components. The tumor-selective growth inhibitory activity of the extracts on leukemic cell lines was evaluated in vitro and in vivo using tumor-bearing nude mice.

Materials and methods

Agaricus blazei extracts were prepared using different methods. MTT and tritiated thymidine incorporation assays were used to evaluate the in vitro anti-leukemic effects. The most potent extract was further investigated using NB-4 cells-bearing nude mice and mechanistic studies using DNA fragmentation assay and cell death detection ELISA.

Results

The JAB80E70 extract showed the most potent tumor-selective growth inhibitory activity against human leukemia NB-4 and K-562 cells. This is the first report of anti-leukemic activity of JAB80E70 in athymic nude mice bearing NB-4 cells. Using DNA fragmentation assays and cell death detection ELISA, JAB80E70 was found to induce apoptosis in NB-4 cells. However, the polysaccharide enriched fractions failed to show significant cytotoxicity on NB-4 cells in vitro.

Conclusions

The JAB80E70 extract exhibited potent anti-leukemic effect in vitro and in vivo. The effect can be attributed, at least in part, to the induction of apoptosis. Besides, polysaccharides in Agaricus blazei may not possess direct anti-leukemic activity in vitro.     

艾叶挥发油对A549细胞的抑制作用

目的研究艾叶挥发油对A549细胞的抑制作用。方法 MTT法检测挥发油对A549细胞的抗增殖活性,流式细胞仪检测细胞凋亡率及细胞周期。皮下注射A549细胞建立肺癌模型后,裸鼠随机分为阴性对照组、5-氟尿嘧啶组(10 mg/kg)及艾叶挥发油低、中、高剂量组(57.5、115、230 mg/kg),考察挥发油对肿瘤生长及体质量的影响。HE染色观察病理组织。结果挥发油明显抑制细胞增殖,并呈浓度依赖性。424.8μg/mL挥发油处理细胞48 h后,细胞凋亡率最高(67.1%),G0/G1期细胞比例降低,G2/M期细胞比例无明显变化,S期细胞比例增加。与阴性对照组比较,挥发油组肿瘤质量和体积显著降低(P<0.05,P<0.01),裸鼠体质量无明显变化(P>0.05)。结论艾叶挥发油可阻滞A549细胞于S期,从而抑制细胞增殖,诱导细胞凋亡,而且安全性高。     

Inhibitory effects of Vietnamese medicinal plants on tube-like formation of human umbilical venous cells

Seven of 58 plant materials from Vietnamese medicinal plants showed strong to moderate inhibitory activity on the tube-like formation induced by human umbilical venous endothelial cells in the in vitro angiogenesis assay. These plant materials include the herb of Ephedra sinica, leaves and stem of Ceiba pentandra, seed of Coix lachryma-jobi, rhizome of Drynaria fortunei, fruits and stem of Illicium verum and stem of Bombax ceiba. Of these, the methanol extracts of the herb of Ephedra sinica and stem of -Ceiba pentandra exhibited the strongest activities with inhibition percentages of 89.3% and 87.5% at 30 and 100 microgram/mL, respectively.     

Inhibitory effects of Keishi-bukuryo-gan on free radical induced lysis of rat red blood cells

Keishi-bukuryo-gan (KBG) prevents the progression of atherosclerosis in cholesterol-fed rabbits by its antioxidative effect. The present study was to test our hypothesis that ingestion of KBG would protect red blood cell (RBC) membranes from free radical induced oxidation if polyphenolic antioxidants in KBG could be absorbed and circulated in the blood. When incubated with a RBC suspension, KBG and four of five herb medicines constituting KBG provided strong protection for RBC membranes against haemolysis induced by 2,2-azo-bis (2-amidinopropane) dihydrochloride (AAPH), an azo free radical initiator. The inhibitory effect was dose dependent at concentrations of 100-1000 microg/mL. Furthermore, the ingestion of 200 mg of KBG was associated with a significant decrease in susceptibility of RBC to haemolysis in rats.     

Inhibitory effects of diterpenoid alkaloids on the growth of A172 human malignant cells

The cytotoxicity against A172 human malignant glioma cells was examined for 14 alkaloids from the roots of Aconitum yesoense var. macroyesoense and of Aconitum japonicum and from the seeds of Delphinium elatum as well as for 25 semisynthetic derivatives. The major alkaloid constituents of A. yesoense var. macroyesoense, kobusine (2) and pseudokobusine (3), a minor alkaloid constituent of A. japonicum, aljesaconitine A (5), and six alkaloid derivatives, N-deethyldelcosine (10), N-deethyldelsoline (11), 12-benzoylluciculine (18), 12-anisoylluciculine (19), 6,11-dibenzoylpseudokobusine (28), and 6-veratroylpseudokobusine (29), had only very weak activity. Four acylated alkaloid derivatives, 12-acetylluciculine (23), 11-veratroylpseudokobusine (30), 11-(m-trifluoromethylbenzoyl)pseudokobusine (32), and 11-(m-trifluoromethylbenzoyl)kobusine (39), exhibited more potent activity, while pseudokobusine 11-cinnamoate (31), 11-anisoate (33), and 11-p-nitrobenzoate (34) were found to be the most potent cytotoxic agents.     

Inhibitory effects of Okbyungpoong-Gamhmi on anaphylactic responses

We investigated the effect of a herbal formulation Okbyungpoong-Gamhmi (OG) on mast cell-dependent anaphylactic reactions by intra-rectal administration. OG concentration dependently inhibited compound 48/80-induced anaphylaxis-like response and ear swelling response with doses of 0.01-1g/kg. OG also inhibited the passive cutaneous anaphylaxis at the same concentrations. The histamine release induced by compound 48/80 or IgE from the rat peritoneal mast cells was reduced by 64.2 and 63.6%, respectively, at 1g/l. These results provide evidence that intra-rectal therapy of OG may be beneficial in the treatment of anaphylactic response.     

蟾蜍二烯酸内酯类成分的生物转化和生物合成研究进展

蟾蜍二烯酸内酯类化合物是名贵中药蟾酥的主要活性成分,具有显著的抗肿瘤、强心、镇痛和抗炎等药理活性。目前,蟾蜍二烯酸内酯成分的主要来源方法是从蟾蜍类药材中直接提取,但此方法成本高、收益低,且药用资源十分短缺,相关新药开发与应用研究严重受阻。近年来,中药活性成分的生物合成在中药资源保护方面发挥起越来越重要的作用。基于合成生物学原理,设计和改造微生物菌株发酵生产天然产物的方法已被广泛使用。该方法的应用需要建立在目标产物生物合成途径完全解析的基础之上。基于此,总结近年来蟾蜍二烯酸内酯的生物转化以及生物合成途径解析方面的研究进展,并进一步推测蟾蜍二烯酸内酯类的生物合成途径,并进一步展望与其相关的生物合成发展方向。     

和厚朴酚抑制视网膜新生血管形成的实验研究

目的:观察和厚朴酚对视网膜新生血管形成的抑制作用。方法:氧诱导幼鼠建立视网膜NV模型。治疗组分别腹腔注射和厚朴酚100 mg/kg和50 mg/kg,每日2次共5 d;对照组注射等量BSS。视网膜铺片了解血管改变;组织切片计数突破视网膜内界膜血管内皮细胞核;免疫组化检测视网膜VEGF和MMP-2表达。结果:治疗组视网膜血管密度降低,新生血管数减少。治疗组突破内界膜胞核数较对照组明显减少;大剂量组减少更显著。治疗组视网膜VEGF和MMP-2表达较对照组明显减少;大剂量组更少,两者差异显著。结论:和厚朴酚可以有效抑制视网膜新生血管形成,呈剂量依赖关系。     

Inhibitory effects of triterpenes isolated from Hoelen on free radical-induced lysis of red blood cells

Hoelen, sclederma of Poria cocos Wolf, has long been used as a sedative and diuretic in traditional medicine. Formerly, we demonstrated that Hoelen in vitro protects red blood cells from AAPH-induced hemolysis. In this study, tests were carried out to identify the main ingredient of Hoelen that has the scavenging effect on free-radicals. Triterpene carboxylic acids isolated from the methanol extract of Hoelen, i.e. pachymic acid, polyporenic acid, 3-epidehydrotumulosic acid, 3beta-hydroxylanosta-7,9(11), 24-trien-21-oic acid and 3-o-acetyl-16 alpha -hydroxytrametenolic acid, were found to have inhibitory activities against AAPH-induced lysis of red blood cells.     

Rabdosin B对急性早幼粒白血病细胞HL-60分化的诱导作用

目的研究螺环内酯型对映-贝壳杉烷型二萜rabdosin B(RB)诱导HL-60向成熟粒细胞分化的特征及机制。方法采用台盼蓝染色、吉姆萨染色、硝基四氮唑蓝(NBT)还原力测定及流式细胞术检测RB对HL-60细胞增殖、细胞周期、NBT还原力、细胞吞噬及细胞表面抗原CD11b表达的影响。结果 RB在1.0、3.0、5.0μmol/L下抑制HL-60细胞增殖,引起HL-60细胞G0/G1期阻滞,增加HL-60细胞肾形核和分叶核细胞比率,并伴随细胞NBT还原力和吞噬能力显著增强以及CD11b阳性细胞率明显上升。进一步检测显示,活性氧(ROS)清除剂N-乙酰-L-半胱氨酸(NAC,300μmol/L)及NADPH氧化酶抑制剂夹竹桃麻素(APO,100μmol/L)和二联苯碘锚(DPI,0.1μmol/L)不仅可显著抑制RB诱导的细胞内ROS升高,并且可显著抑制RB诱导的HL-60细胞分化的各项特征指标的改变。结论 RB可通过上调HL-60细胞NADPH氧化酶活性,升高胞内ROS浓度,诱导HL-60细胞向成熟粒细胞分化。     

Inhibitory effects of Korean folk medicine `Hi-Chum' on histamine release from mast cells in vivo and in vitro

The effect of aqueous extract of Siegesbeckia glabrescence (Compositae) whole plants (SGWP) on systemic or local anaphylaxis was studied. SGWP inhibited compound 48/80-induced systemic anaphylaxis 100% with a dose of 1000 mg/kg. Oral administration of SGWP (100 mg/kg) showed a marked inhibition rate in local immunoglobulin E (IgE)-mediated passive cutaneous anaphylaxis reaction. When SGWP was pretreated at concentration ranging from 0.1 to 1000 mg/kg, the serum histamine levels were reduced in a dose-dependent manner. Moreover, SGWP dose-dependently inhibited the histamine release from peritoneal mast cells by compound 48/80. These results indicate that SGWP possess strong antianaphylactic activity by inhibition of histamine release from mast cells.     

薯蓣皂苷对动脉粥样硬化大鼠白介素-6的影响

目的 探讨薯蓣皂苷对动脉粥样硬化模型大鼠白介素6 (interleukin-6,IL-6)的影响.方法 采用随机数字表法将60只雄性SD大鼠随机分成:假手术组、模型组、阿司匹林组、薯蓣皂苷低剂量组及高剂量组.饲养7d后,除假手术组,余大鼠于左侧颈总动脉行主动脉内膜球囊拉伤术.3个月后,行颈总动脉HE染色,采用图象分析仪...     

六神丸中蟾蜍二烯内酯类成分的含量测定

目的：考察2家企业生产六神丸中蟾蜍二烯内酯类化合物的含量。方法：采用高效液相色谱法／DAD检测器同时测定4种蟾蜍二烯内酯类化合物。结果：该测定方法的回收率高于96％，线性范围在1μg／L-100μg／mL，方法重现性良好。六神丸标准方中的蟾蜍二烯内酯含量显著高于在其他配伍方式样品中含量。结论：六神丸配伍均匀程度可能对蟾蜍二烯内酯类化合物具有一定的增溶作用，促进了这种活性物质的释放。     

丹参酮ⅡA抑制大鼠血管平滑肌细胞增殖及其机制研究

目的：观察丹参酮ⅡA（tanshinone ⅡA，TA）对10％胎牛血清诱导的大鼠血管平滑肌细胞（RVSMC）增殖的抑制作用，并探讨其可能的作用机制。方法：体外培养大鼠主动脉平滑肌细胞A7r5细胞株，以终浓度为10％的胎牛血清（FBS）作为刺激因素，用细胞计数法、噻唑蓝（MTY）比色法和5-溴脱氧尿嘧啶（BrdU）掺入法测定TA对细胞增殖的影响，用流式细胞术（FCM）分析细胞周期分布特征，用Western blot实验测定细胞外信号调节激酶1／2（ERK1／2）磷酸化活性，用RT—PCR测定c—fos的表达水平。结果：细胞计数、MTY比色法和BrdU掺入实验表明丹参酮Ⅱ。能抑制10％FBS所诱导的VSMC增殖，作用强度呈剂量依赖性；细胞周期分析显示，TA处理组G0／G1期细胞百分比高于10％FBS组，而S期比例低于10％FBS组，表明TA可阻止10％FBS所诱导的细胞周期由G0／G1期向S期推进；Western blot结果显示与10％FBS组相比，TA处理组ERK1／2磷酸化活性降低；RT—PCR结果显示TA处理组c—fos表达水平降低。结论：丹参酮可抑制10％FBS诱导的体外培养大鼠动脉平滑肌细胞增殖，此作用可能与其阻止细胞周期由G0／G1期向S期推进，抑制MAPK信号转导通路激活，进而下调c-fos表达有关。