大蒜油抗肿瘤作用的流式细胞光度术分析

用流式细胞光度术（ｆｌｏｗｅｙｔｏｍｅｔｒｙ,ＦＣＭ）+Ｓ０．２ｍｌ／日。吐温对照组ｌＰ吐温８００．２ｍｌ／日,以排除大蒜油制剂中溶剂（吐温８０）的影响。实验组ｌＰＧＯ１００ｍｇ／ｋｇ／．日。于连续４次注射后的６ｈ,２、３、５、１０天及１０次注射ＧＯ后７天取材,行ＦＣＭ样品制备。三、ＦＣＭ样品制备与测定中国中医研究院基础所腹水癌细胞用ＰＢＳ洗涤,７０％酒精固定,ＲＮａｓｅ消化细胞中的ＲＮＡ,ＰＩ染色。用腹腔内淋巴细胞作标准二倍体细胞。用４２０型荧光激活细胞分选仪（美国Ｂｅｃｔｏ－Ｄｉｃｋｓｏｎ公司产品）测单个肿瘤细胞的ＤＮＡ含量,用组方图显示肿瘤细胞ＤＮＡ倍体性质。图中第一个是ＤＮＡ二倍体（ＺＣ）峰,处于该峰的细胞为Ｇ;／Ｇ。期细胞,第二个峰为４ＣＤＮＡ峰,该处细胞为Ｇ。－Ｉ－－Ｍ期细胞,从２。到４。之间的细胞为Ｓ期细胞。大于４。ＤＮＡ峰细胞为高倍体细胞‘’：。根据不同峰值大小判定用药后肿瘤细胞倍体性质的变化。结果ＮＳ与吐温对照组肿瘤细胞都为非整倍体性,且呈多倍体性（图１,２）。连续４次给药后６ｈ,绝大部分多倍体肿瘤细胞被杀伤,残存肿瘤细胞以二倍体为主,多倍体肿瘤细胞峰值显著下降,甚至在组方图上几乎显不出多倍     

中国大蒜(Allium sativum L.)18个品种的酯酶同工酶多态性分析

采用聚丙烯酰胺凝胶电泳(PAGE)分离了中国大蒜3个生态型(低温反应敏感型、低温反应中间型和低温反应迟钝型)中18个较典型的品种的酯酶同工酶,并用排序分析法对18个品种的亲缘关系进行了分析,将18个品种分为3个变种群：1．“苏联”蒜变种群(var．Russia),2．吉木萨尔白皮蒜变种群(var．：Jimusaer。),3．中国内陆大蒜变种群(var．China)。其中中国内陆大蒜变种群又可分为5个品种群：①关中蒜品种群,②西北蒜品种群,③西南蒜品种群,④云贵蒜品种群,⑤华东蒜品种群。实验结果初步证实,大蒜生态型不能完全等同于基因型,酯酶同工酶的变化可能更能说明大蒜的亲缘进化关系。     

Cytotoxic effects of two extracts from garlic (Allium sativum L.) cultivars on the human squamous carcinoma cell line SCC-15

Garlic (Allium sativum L., Alliaceae) has acquired a reputation as a therapeutic agent and herbal remedy to prevent and treat several pathologies. The aim of the present study was to determine the effect of two Allium sativum L. cultivars, Harna? and Morado, on reactive oxygen species (ROS) production, viability and apoptotic process in human squamous carcinoma cell line SCC-15. The experiments were conducted on SCC-15 cell line exposed to increasing concentrations of garlic extracts of 0.062, 0.125, 0.250, 0.500 and 1.000 mg/mL. After the experiments, ROS formation, caspase-3 activity and neutral red uptake were measured in the cells, and in a collected medium lactate dehydrogenase (LDH) release was measured. The Spanish cultivar Morado has demonstrated higher potential to stimulate ROS production in SCC-15 cells after a short time period (6 h) than the Polish cultivar Harna?. However, the Polish cultivar Harna? manifested more prolonged potential to stimulate ROS production in SCC-15 cells. Both studied garlic extracts induced cytotoxicity on SCC-15 cell line which was probably ROS-dependent. We also determined that in SCC-15 cells high concentrations of studied extracts did not cause activation of caspase-3 which suggested caspase-independent or necrotic cell death.     

Garlic Oil Attenuated Nitrosodiethylamine-Induced Hepatocarcinogenesis by Modulating the Metabolic Activation and Detoxification Enzymes

Nitrosodiethylamine (NDEA) is a potent carcinogen widely existing in the environment. Our previous study has demonstrated that garlic oil (GO) could prevent NDEA-induced hepatocarcinogenesis in rats, but the underlying mechanisms are not fully understood. It has been well documented that the metabolic activation may play important roles in NDEA-induced hepatocarcinogenesis. Therefore, we designed the current study to explore the potential mechanisms by investigating the changes of hepatic phase Ⅰ enzymes (including cytochrome P450 enzyme (CYP) 2E1, CYP1A2 and CYP1A1) and phase Ⅱ enzymes (including glutathione S transferases (GSTs) and UDP- Glucuronosyltransferases (UGTs)) by using enzymatic methods, real-time PCR, and western blotting analysis. We found that NDEA treatment resulted in significant decreases of the activities of CYP2E1, CYP1A2, GST alpha, GST mu, UGTs and increases of the activities of CYP1A1 and GST pi. Furthermore, the mRNA and protein levels of CYP2E1, CYP1A2, GST alpha, GST mu and UGT1A6 in the liver of NDEA-treated rats were significantly decreased compared with those of the control group rats, while the mRNA and protein levels of CYP1A1 and GST pi were dramatically increased. Interestingly, all these adverse effects induced by NDEA were simultaneously and significantly suppressed by GO co-treatment. These data suggest that the protective effects of GO against NDEA-induced hepatocarcinogenesis might be, at least partially, attributed to the modulation of phase I and phase II enzymes.     

Effect of garlic and allium-derived products on the growth and metabolism of Spironucleus vortens

Spironucleus is a genus of small, flagellated parasites, many of which can infect a wide range of vertebrates and are a significant problem in aquaculture. Following the ban on the use of metronidazole in food fish due to toxicity problems, no satisfactory chemotherapies for the treatment of spironucleosis are currently available. Using membrane inlet mass spectrometry and automated optical density monitoring of growth, we investigated in vitro the effect of Allium sativum (garlic), a herbal remedy known for its antimicrobial properties, on the growth and metabolism of Spironucleus vortens, a parasite of tropical fish and putative agent of hole-in-the-head disease. The allium-derived thiosulfinate compounds allicin and ajoene, as well as an ajoene-free mixture of thiosulfinates and vinyl-dithiins were also tested. Whole, freeze-dried garlic and allium-derived compounds had an inhibitory effect on gas metabolism, exponential growth rate and final growth yield of S. vortens in Keister’s modified, TY-I-S33 culture medium. Of all the allium-derived compounds tested, the ajoene-free mixture of dithiins and thiosulfinates was the most effective with a minimum inhibitory concentration (MIC) of 107 μg ml⁻¹ and an inhibitory concentration at 50% (IC_50%) of 58 μg ml⁻¹. It was followed by ajoene (MIC = 83 μg ml⁻¹, IC_50% = 56 μg ml⁻¹) and raw garlic (MIC >20 mg ml⁻¹, IC_50% = 7.9 mg ml⁻¹); allicin being significantly less potent with an MIC and IC_50% above 160 μg ml⁻¹. All these concentrations are much higher than those reported to be required for the inhibition of most bacteria, protozoa and fungi previously investigated, indicating an unusual level of tolerance for allium-derived products in S. vortens. However, chemically synthesized derivatives of garlic constituents might prove a useful avenue for future research.     

硫对大蒜生理生化指标及营养品质的影响

采用盆栽试验探讨了硫对大蒜生理生化指标及营养品质的影响．结果表明,土壤硫含量8．41mg·kg^-1时,大蒜叶片硝酸还原酶、POD活性、光合色素含量、光合速率、气孔导度和蒸腾速率较高;可溶性蛋白质含量及CAT活性却是以土壤硫含量为7．92mg·kg^-1时最高;SOD活性以较低土壤硫含量6．45mg·kg^-1时较高．同时,土壤硫含量为8．41mg·kg^-1时,鳞茎中大蒜素、可溶性糖和Vc及蒜薹中大蒜素、可溶性糖和可溶性蛋白质含量分别比对照增加33．67％、23．75％、68．82％、40．95％、3．45％和7．45％;较高土壤硫含量9．48mg·kg^-1有利于蒜薹中Vc和游离氨基酸及鳞茎中游离氨基酸和可溶性蛋白质含量的提高,分别比对照高出15．17％、20．93％、6．49％和8．07％．     

Generation of chlorsulfuron-resistant transgenic garlic plants (Allium sativum L.) by particle bombardment

We established an effective biolistic transformation procedure fortransferring foreign genes into garlic (Allium sativumL.),which we demonstrated by generating transgenic plants resistant tochlorsulfuron, a sulfonylurea herbicide. We subcultured callus tissue from theapical meristem of garlic cloves and repeatedly selected calli with brittle,non-mucilaginous surfaces for over six months, to increase transformationefficiency. We then constructed recombinant DNA that contained the acetolactatesynthase (ALS) gene from a chlorsulfuron-resistantArabidopsis mutant, the cauliflower mosaic virus 35Spromoter, the -glucuronidase (GUS) reporter gene, and the hygromycinphosphotransferase (HPT) selectable marker gene. The garlic calli werebombarded twice with tungsten particles coated with the DNA constructs. Transformed calliwere efficiently selected by embedding them in solid agar medium containing 50mg l^–1 hygromycin B. Selected propagules wereregenerated into 12 independent plants. We confirmed that the transgenes wereintegrated and expressed in the plants using PCR-Southern and Northern blotanalyses and by -glucuronidase expression assay forGUS. The regenerated plants survived in the presence of 3mg l^–1 chlorsulfuron, demonstrating that theirALS was insensitive to this herbicide. These results illustrate the successfultransformation of foreign genes into garlic plants. The set of proceduresdeveloped in this study is applicable to the generation of transgenic garlicplants with other agronomically beneficial traits. These authors contributed equally to this work     

云南蒜油化学成分的研究

本文对云南曲靖产大蒜(Allium sativum L.)蒜油用气相色谱和色谱-质谱法进行了分析,鉴定了20个化合物。其中:6-甲基-1-硫杂-2,4-环己二烯(6-methyl-1-thi-2,4-cyclohexadiene),5-甲基-1,2-二硫杂-3-环戊烯(5-methyl-1,2-dithi-3-cyclopentene),4-甲基-1,2-二硫杂-3-环戊烯(4-methyl-1,2-dithi-3-cyclopentene),4-乙烯基-1,2,3-三硫杂-5-环己烯(4-vinyl-1,2,3-trithi-5-cyclohexene)及甲基烯丙基五硫醚(allyl methyl pentasulfide)等为蒜油中首次报道的化合物。     

富硒大蒜含硒蛋白的分离、鉴定和生物活性研究

本文报道了富硒大蒜含硒蛋白的分离、鉴定和生物活性研究。富硒大蒜水溶性蛋白多为酸性蛋白。通过Sephadex G—200柱层析和聚丙烯酰胺梯度凝胶电泳分离出两种分子量约为30,000和33,000的酸性含硒蛋白。大蒜含硒蛋白和亚硒酸钠一样,具有提高含硒谷胱甘肽过氧化物酶(SeGSHpx)活性,促进软骨细胞生长的生理效应。实验结果提示,富硒大蒜有可能成为良好的补硒来源。