Scientia Agricultura Sinica ›› 2014, Vol. 47 ›› Issue (1): 170-181.doi: 10.3864/j.issn.0578-1752.2014.01.018

• ANIMAL SCIENCE·VETERINARY SCIENCERE·SOURCE INSECT • Previous Articles     Next Articles

Study on Inhibitory Effect of the Swainsonine from Alkaloid of Astragalus strictus Grah.Ex Bend on Bovine Viral Diarrhea Virus

 HAO  Bao-Cheng-1, 2 , WU  Fan-Lin-2, XING  Xiao-Yong-2, XIANG  Hai-Tao-2, WEN  Feng-Qin-2, WANG  Xue-Hong-1, QUAN  Xiao-Di-1, 2 , HU  Yong-Hao-2, LIANG  Jian-Ping-1, 2Ping-1 , 2   

  1. 1.Key Laboratory of New Animal Drug Project,Gansu Province/Key Laboratory of Veterinary Pharmaceutics Discovery, Ministry of Agriculture /Lanzhou Institute of Husbandry and Pharmaceutical Sciences of Chinese Academy of Agricultural Sciences ,    Lanzhou 730050;
    2.College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070
  • Received:2013-06-17 Online:2014-01-01 Published:2013-09-30

Abstract: 【Objective】 Locoweed is one of the poisonous weeds in western China that influences the healthy development of grassland and animal husbandry. Locoweeds mainly belong to Astragalus and Oxytropis genera. As a serious lack of forage in winter season, livestock such as cattle and sheep are forced to feed them. Locoweed can decrease the production performance, toxicity and death of animals, and the direct economic loss of the grassland ecological and animal husbandry is about billions of dollars, which is the most serious poisonous weed. At present, the economic losses of animal death caused by locoweed poisoning continues to soar. Swainsonine is the main toxic component and believed to be the cause of animal locoweed poisoning. Locoweed is widely distributed and is rich in resources, and how to improve the grassland ecology and use of locoweed is becoming the topic of research subject and direction. In recent years, with the study and expansion of locoweed, people have found that swainsonine not only has a good anti-tumor effect, also it can be used as an immunomodulator, anti HIV and anti proliferation inhibitor, anti viral and cell protective agent and other drugs. At present, the domestic research on swainsonine anti-tumor and regulation of immunity is hot, However, no research reports are found in antiviral activity and mechanism of action of swainsonine up to the present. This experiment was designed to investigate the anti-virus mechanism of the swainsonine from alkaloid of Astragalus on bovine viral diarrhea virus (BVDV), and to make clear of the antiviral effect in vitro, it will provide references for the screening of anti BVDV drugs. 【Method】 Using cell culture techniques, and combined with CPE observation method and MTT colorimetric method, different concentrations SW toxicity with bovine kidney primary cells (Madin-Darby Bovine Kidney Cells, MDBK) were detected, the concentration of the drug’s safety and TD50 was determined. They were measured by four administration routes of virus before swainsonine, virus after swainsonine, virus-and-swainsonine after 2 hours, and virus-and-swainsonine after swainsonine, which are the effect of different concentrations of SW against blocking effect on BVDV invasion, replicatting inhibition, directly killing effect and combining effects, respectively. The treatment indexes under different interaction modes were also calculated.【Result】It was found that the TCID50 of BVDV-MDV is 10-4.7. SW was not toxic to MDBK when its concentration was below 0.256 μg•mL-1, and TD50 is 2.512 μg•mL-1. Under the way of virus after SW, IC50 is 2.399 μg•mL-1, TI is 1.05. Under the way of SW after virus, IC50 is 1.018 μg•mL-1, TI is 2.47. Under the way of 2 h after mixed virus with SW, IC50 is 1.205 μg•mL-1, TI is 2.08. Under the way of synthetic action, IC50 is 0.782 μg•mL-1,TI is 3.21. After the TI of different modes of action was compared, it showed that the comprehensive effect on BVDV of SW (65.29%,P<0.01) and the inhibition effect of SW on the reproduction of BVDV (65.05%,P<0.01) were better, and SW had a certain direct killing effect on BVDV, but the blocking effect of SW on the invasion of BVDV was poor.【Conclusion】The effect of swainsonine (SW)-anti-BVDV activity in vitro was better, and the authors infered that the antiviral mechanism of SW might be that it could enter into the cells and inhibit the reproduction of BVDV, or it could play a role by killing the uncombined BVDV.

Key words: Astragalus strictus Grah. Ex Bend , swainsonine (SW) , bovine viral diarrhea virus (BVDV) , anti-viral

[1]赵月兰, 左玉柱, 范京惠, 张宁, 秦建华, 刘占民, 杨汉春. 牛病毒性腹泻/黏膜病病毒河北分离株的生物学特性. 中国农学通报, 2006, 22(12): 1-4.

Zhao Y L, Zuo Y Z, Fan J H, Zhang N, Qin J M, Liu Z M, Yang H C. Biological characteristics of bovine viral diarrhoea-mucosal disease virus HB strain. Chinese Agricultural Science Bulletin, 2006, 22(12): 1-4. (in Chinese)

[2]王丽荣, 董永军, 梁美兰. 牛病毒性腹泻病毒的病原学与诊治. 安徽农业科学, 2005, 33(12): 2348-2349.

Wang L R, Dong Y J, Liang M L. Etiology, diagnosis and treatment of bovine viral diarrhea virus. Journal of Anhui Agricultural Sciences, 2005, 33(12): 2348-2349. (in Chinese)

[3]王镇, 丁明孝. 牛病毒性腹泻病毒的分子生物学研究进展. 中国病毒学, 1997, 12(2): 112-118.

Wang Z, Ding M X. Advance in molecular biology of bovine viral diarrhea virus. Virologica Sinica, 1997, 12(2): 112-118. (in Chinese)

[4]冯彦君. 猪瘟兔化弱毒苗对牛病毒性腹泻-粘膜病的防治作用. 中国奶牛, 1995(2): 39-40.

Feng Y J. The preventive effect of Hog Cholera Lapinized Virus vaccine with bovine viral diarrhea-Mucosal disease. China Dairy Cattle, 1995(2): 39-40. (in Chinese)

[5]郝宝成, 梁剑平, 王学红, 郭文柱, 郭志廷, 杨贤鹏. 牛病毒性腹泻病研究进展及防控建议. 中国奶牛, 2013(2): 45-48.

Hao B C, Liang J P, Wang X H, Guo W Z, Guo Z T, Yang X P. Analysis of bovine viral diarrhea prevalence and comprehensive prevention and control in china. China Dairy Cattle, 2013(2): 45-48. (in Chinese)

[6]王新平, 任文陟, 朱维正, 李佑民, 宣华, 常国权, 丁壮. 双抗体夹心ELISA检测牛病毒性腹泻病毒的研究. 中国兽医学报, 1995, 15(3): 246-249.

Wang X P, Ren W Z, Zhu W Z, Li Y M, Xuan H, Chang G Q, Ding Z. Study on double-antibody sandwich ELISA for detection of bovine viral diarrhea virus. Chinese Journal of Veterinary Science, 1995, 15(3): 246-249. (in Chinese)

[7]李佑民, 刘振润, 武银莲. 牛病毒学腹泻粘膜病病毒株(长春184)的分离与鉴定. 中国人民解放军兽医大学学报, 1983, 3(2): 113-121.

Li Y M, Liu Z R, Wu Y L. Isolation and identification the Bovine viral diarrhea and mucosal disease virus strains (Changchun 184). Journal of PLA Veterinary Medicine University, 1983, 3(2): 113-121. (in Chinese)

[8]郑志刚, 刘佩兰, 郑增忍, 王永玲, 杜元钊. 关于牛病毒性腹泻粘膜病血清中和抗体的调查报告. 动物检疫, 1991(5): 40-42.

Zheng Z G, Liu P L, Zheng Z R, Wang Y L, Du Y Z. The investigation report of the bovine viral diarrhea mucosal disease antibodies in serum. Journal of Animal Quarantine, 1991(5): 40-42. (in Chinese)

[9]韩鹏, 刘亚刚, 华莎, 索化夷. 我国牛病毒性腹泻-粘膜病的流行及防制状况. 畜禽业, 2003, 11: 8-10.

Han P, Liu Y G, Hua S, Suo H Y. The situation of prevalence and control of bovine virral diarrhoea-mucosal disease in our country. Livestock and Poultry Industry, 2003, 11: 8-10. (in Chinese)

[10]张宝宁. 牛病毒性腹泻-粘膜病病毒的分离、RT-PCR鉴定、NS2-3区基因克隆及变异分析[D]. 保定:河北农业大学硕士论文, 2006.

Zhang B N. Isolation and identification on BVDV and cloning, analysis of genetic variation on NS2-3[D]. Baoding: Hebei Agricultural University, 2006. (in Chinese)

[11]陈备娟, 高全新, 朱毅兴, 陆仁初, 李媛媛. 牛病毒性腹泻病的流行病学调查. 上海畜牧兽医通讯, 2010(5): 43-44.

Chen B J, Gao Q X, Zhu Y X, Lu R C, Li Y Y. Epidemiological investigation of bovine viral diarrhea disease. Shanghai Journal of Animal Husbandry and Veterinary Medicine, 2010(5): 43-44. (in Chinese)

[12]侯炜, 杨占秋, 陈科力, 杨继江, 王卫华, 肖红, 程丽. 大黄有效成分抗单纯疱疹病毒作用的实验研究. 药物分析杂志, 2003, 4(23): 259.

Hou W, Yang Z Q, Chen K L, Yang J J, Wang W H, Xiao H, Cheng L. Extrinsic experimental study on Rheum officinale Baill solution antivirotic in Herpes Simplex Virus. Journal of Pharmaceutical Analysis, 2003, 4(23): 259. (in Chinese)

[13]王志洁, 黄铁牛, 刘焱文, 鲁遂荣, 杨占秋, 方学韫. 黄芪多种成分抗人疱疹病毒的初步实验研究. 中国现代应用药学杂志, 2002, 19(5): 356.

Wang Z J, Huang T N, Liu Z W, Lu S R, Yang Z Q, Fang X W. The experimental research of antiherpes simplex virus effects of various components in the root of Astragalus membranaceus. Chinese Journal of Modern Applied Pharmacy, 2002, 19(5): 356. (in Chinese)

[14]刘媛媛, 杨占秋, 何静, 石丽桥, 肖红. MTT分析法、空斑减数法及CPE观察法评估药物体外抗病毒效果的比较与分析. 武汉大学学报, 2005, 26(2): 199-202.

Liu Y Y, Yang Z Q, He J, Shi L Q, Xiao H. Comparison and analysis of CPE assay, MTT assay and plaque reductive assay for evaluating medicines in antiviral effects in vitro. Journal of Hubei Medical University, 2005, 26(2): 199-202. (in Chinese)

[15]四朗玉珍, 吴玉江, 益西多吉, 吴金措姆, 赵宝玉. 西藏疯草研究现状. 西藏科技, 2013, 3: 72-75.

Silang Y Z, Wu Y J, Yixi D J, Wujin C M, Zhao B Y. Locoweed research in Tibet. Tibet Science and Technology, 2013, 3: 72-75. (in Chinese)

[16]陈基萍, 赵宝玉, 路浩, 徐创明, 马尧, 周启武. 产苦马豆素菌株Aspergillus ustus发酵工艺研究. 中国兽医学报, 2013, 33(3): 423-427.

Chen J P, Zhao B Y, Lu H, Xu C M, Ma Y, Zhou Q W. The fermentation condition optimization of swainsonine-producing endophyte Aspergillus ustus. China Journal of Veterinary Sciences, 2013, 33(3): 423-427. (in Chinese)

[17]赵清梅, 余永涛, 何生虎, 王建华. 苦马豆素的来源及其生物合成研究进展. 农业科学研究, 2012, 33(4): 82-88.

Zhao Q M, Yu Y T, He S H, Wang J H. Research advances in sources and biosynthesis of swainsonine. Journal of Agricultural Science, 2012, 33(4): 82-88. (in Chinese)

[18]宋岩岩, 赵宝玉, 路浩, 徐向军, 拓晓玲, 王金屿, 李蓉. 急弯棘豆生物碱成分薄层色谱分析及苦马豆素分离. 西北农业学报, 2012, 21(7): 25-29.

Song Y Y, Zhao B Y, Lu H, Xu X J, Tuo X L, Wang J Y, Li R. The alkaloids analysis of oxytropis deflexa by the thinlayer chromatography and separation of swainsonine. Acta Agricultural Boreali-occidentalis Sinica, 2012, 21(7): 25-29. (in Chinese)

[19]张淑英, 敖云格日勒, 王丹丹. 吉兰太地区家畜疯草中毒的调查及防治. 中国畜牧兽医文摘, 2012, 28(1): 81.

Zhang S Y, Aoyungerile, Wang D D. Investigation and prevention locoweed poisoning in livestock Jilantai region. China Animal Husbandry and Veterinary Digest, 2012, 28(1): 81. (in Chinese)

[20]赵宝玉, 刘忠艳, 万学攀, 霍星华, 郭玺, 王建军, 刘忠艳, 孙莉莎, 石志诚. 中国西部草地毒草危害及治理对策. 中国农业科学, 2008, 41(10): 3094-3103.

Zhao B Y, Liu Z Y, Wan X P, Huo X H, Guo X, Wang J J, Liu Z Y, Sun L S, Shi Z C. Damage and control of poisonous-weeds in Chinese western grassland. Scientia Agricultura Sinica, 2008, 41(10): 3094-3103. (in Chinese)

[21]周启武, 赵宝玉, 路浩, 王姗姗, 张梁, 温伟利, 杨晓雯. 中国西部天然草地疯草生态及动物疯草中毒研究与防控现状. 中国农业科学, 2013, 46(6): 1280-1296.

Zhou Q W, Zhao B Y, Lu H, Wang S S, Zhang L , Wen L W, Yang X W. The research and control situation of ecology and animal poisoning of locoweed in western natural grassland of China. Scientia Agricultura Sinica, 2013, 46(6): 1280-1296. (in Chinese)

[22]汪兰芳, 方优妮, 王平, 黄婕.正交试验优选失眠贴中黄连乙醇回流提取工艺.湖北中医学院学报, 2010, 12(5): 31-33.

Wang L F, Fang Y N, Wang P, Huang J. Orthogonal test for optimizing ethanol reflux extraction process of Rhizoma Coptidis in Insomnia Patch. Journal of Hubei University of Chinese Medicine, 2010, 12(5): 31-33. (in Chinese)

[23]邹恒琴, 徐峰, 张忠义, 孙逊. 一种具有前景的抗癌药苦马豆素的研究进展. 中草药. 1997, 28(7): 437-439.

Zou H Q, Xu F, Zhang Z Y, Sun X. Research Progress a kind of promising anti-cancer drugs —Swainsonine. Chinese Traditional and Herbal Drugs, 1997, 28(7): 437-439. (in Chinese)

[24]刘文明, 童德文, 李 立. 苦马豆素对鸡马立克氏病血液学指标的影响. 西北农林科技大学学报: 自然科学版, 2008, 36(3): 85-89.

Liu W M, Tong D W, Li L. Effects of swainsonine on hematology of Marek′s disease. Journal of Northwest A & F University: Nat. Sci. Ed.. 2008, 36(3): 85-89. (in Chinese)

[25]刘文明. 苦马豆素对鸡马立克氏病的作用研究[D]. 杨凌:西北农林科技大学, 2007.

Liu W M. The effects of swainsonine to Marek’s disease[D]. Yangling: Northwest Agriculture and Forestry University, 2007. (in Chinese)

[26]Humphries M J, Mastumoto K, White S L, Molyneux R J, Olden K . Augmentation of murine natural Killer cell activity by swainsonine, a new antimetastatic immunomodulator. Cancer Research, 1988, 48: 1410-1415.

[27]崔建国. 白毛藤生物碱抗菌抗病毒及免疫药理学研究[D]. 长沙:湖南农业大学, 2005.

Cui J G. Studies on the antibacterial, antiviral effect and immunopharmacology of solanum lyratum thunb alkaloids[D]. Changsha: Hunan Agricultural University, 2006. (in Chinese)

[28]郝宝成, 梁剑平, 杨贤鹏, 王学红, 陶蕾, 王保海, 刘建枝, 刘宇, 郭文柱, 尚若锋, 郭志廷, 华兰英. 一种苦马豆素的酶法提取工艺. CN201210176457. 9, 2012.

Hao B C, Liang J P, Yang X P, Wang X H, Tao L, Wang B H, Liu J Z, Liu Y, Guo W Z, Shang R F, Guo Z T, Hua L Y. One kind of cellulase extraction of swainsonine. CN201210176457. 9, 2012. (in Chinese)

[29]郝宝成, 杨贤鹏, 王学红, 陈慧, 郭文柱, 郭志廷, 刘宇, 尚若锋, 胡永浩, 梁剑平. 茎直黄芪中苦马豆素的提取工艺研究. 湖北农业科学, 2013, 52(8): 1886-1888.

Hao B C, Yang X P, Wang X H, Chen H, Guo W Z, Guo Z T, Liu Y, Shang R F, Hu Y H, Liang J P. Study on the extraction technology of swainsonine from astragalus strictus. Hubei Agricultural Science, 2013, 52(8): 1886-1888. (in Chinese)

[30]刘晶星, 陈福祥, 陆德源, 陈曙霞, 钱富荣. 苦参总碱抗柯萨奇B病毒作用机理的研究. 上海第二医科大学学报, 1996, 10(3): 183-185.

Liu J X, Chen F X, Lu D Y, Chen S X, Qian F R. Preliminary study on the mechanism of anti-coxsackievirus B of sophora flavescens alkaloids. Acta University Medicine Second Shanghai, 1996, 10(3): 183-185. (in Chinese)

[31]李智. 中药疫毒清制剂抗流感病毒作用研究[D]. 武汉:湖北中医学院, 2009.

Li Z. Research on the traditional Chinese medicine depestia with the effect of anti-influenza virus[D]. Wuhan: Hubei Chinese Medicine College, 2009. (in Chinese)

[32]殷震, 刘景华. 动物病毒学. 北京: 科学出版社, 1997.

Yin Z, Liu J H. Animal Virology. Beijing:Science Press, 1997. (in Chinese)
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