双黄连可溶性粉不同组分抗菌抗病毒活性研究

doi:10.16431/j.cnki.1671-7236.2021.08.042

摘要/Abstract

摘要： 为了深度开发双黄连可溶性粉（金银花、连翘、黄芩，1：1：2），并为家禽、家畜合理选择用药提供科学依据，本研究以双黄连口服液生产工艺生产的双黄连可溶性粉及其醇沉物（多糖组分）、滤液（黄酮组分）作为研究材料，以哺乳动物牦牛源轮状病毒、鸡新城疫病毒，牦牛源致病性大肠杆菌、沙门氏菌、金黄色葡萄球菌为研究对象，采用牛津杯结合平板二倍稀释法，探讨其体外抗菌活性；以牦牛MA-104细胞株、鸡胚成纤维细胞株UMNSAH/DF-1为宿主细胞，采用噻唑蓝比色法（MTT）和细胞病变（CPE）检测法，探讨其体外抗病毒活性。研究结果表明，双黄连可溶性粉对革兰氏阴性菌（牦牛源大肠杆菌、牦牛源沙门氏菌、大肠杆菌质控菌株）的抑菌效果优于对革兰氏阳性菌（牦牛源金黄色葡萄球菌、金黄色葡萄球质控菌株）的抑菌效果，从双黄连可溶性粉中分离的黄酮组分可能是双黄连可溶性粉抑菌的主要成分，而多糖组分则无抑菌活性；黄酮组分对革兰氏阳性菌的抑菌效果优于对革兰氏阴性菌的抑菌效果；双黄连可溶性粉及其黄酮组分和多糖组分对鸡胚胎成纤维细胞UMNSAH/DF-1的最小无毒浓度（TC₀）分别为：2.58、1.17和1.56 μg/mL，对牦牛细胞MA-104的TC₀分别为：1.290、0.585和0.780 μg/mL，对牦牛轮状病毒均具有良好的抗病毒效果，双黄连可溶性粉对鸡新城疫病毒的抗病毒效果优于黄酮和多糖组分。研究结果为双黄连可溶性粉深度开发和对畜禽合理选择用药提供了科学依据。

关键词: 双黄连可溶性粉; 黄酮组分; 多糖组分; 抗菌活性; 抗病毒活性

Abstract: In order to provide scientific basis for the in-depth development of Shuanghuanglian soluble powder(honeysuckle, forsythia, Scutellaria baicalensis, 1:1:2) and the rational selection of medicine for poultry and livestock.In this study, Shuanghuanglian soluble powder produced by production technology of Shuanghuanglian oral liquid and its alcohol precipitates (polysaccharide components) and filtrate (flavonoids components) were used as research materials.The mammals Yak source rotavirus, Newcastle disease virus, yak source pathogenic Escherichia coli, Salmonella, Staphylococcus aureus as the research object, Oxford cup combined with two-fold dilution method was used to investigate the antibacterial activity in vitro.Using yak MA-104 cell line and chicken embryo fibroblast cell line UMNSAH/DF-1 as host cells, Thiazolium blue colorimetry (MTT) and cytopathic detection (CPE) were used to investigate the antiviral activity in vitro.The results showed that the antibacterial effect of the Shuanghuanglian soluble powder on Gram-negative bacteria(Yak Escherichia coli, Yak Salmonella, Escherichia coli quality control strain) was better than that on Gram-positive bacteria (Staphylococcus aureus, Staphylococcus aureus quality control strain from yaks).The flavonoids isolated from the Shuanghuanglian soluble powder might be the main antibacterial component of the Shuanghuanglian soluble powder, while the polysaccharide component had no antibacterial activity.The antibacterial effect of soluble powder flavone components of Shuanghuanglian on Gram-positive bacteria was better than that on Gram-negative bacteria.The minimum nontoxic concentration (TC₀) of Shuanghuanglian soluble powder and its flavonoids and polysaccharides on UMNSAH/DF-1 cells were 2.58, 1.17 and 1.56 μg/mL, respectively.The TC₀ of MA-104 cells were 1.290, 0.585 and 0.780 μg/mL, respectively.They all had good antiviral effect for the Yak rotavirus.The antiviral effect of Shuanghuanglian soluble powder against Newcastle disease virus was better than that of flavonoids and polysaccharides components.The results provided scientific basis for the in-depth development for the Shuanghuanglian soluble powder and rational selection of medicine for livestock and poultry.

Key words: Shuanghuanglian soluble powder; flavonoid components; polysaccharide components; antibacterial activity; antiviral activity

中图分类号:

R285.5

杨彦文, 李梓密, 刘群. 双黄连可溶性粉不同组分抗菌抗病毒活性研究[J]. 中国畜牧兽医, 2021, 48(8): 3087-3096.

YANG Yanwen, LI Zimi, LIU Qun. Study on Antibacterial and Antiviral Activity of Different Components of Shuanghuanglian Soluble Powder[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(8): 3087-3096.

参考文献

[1] 朱国福.中药学[M].北京:清华大学出版社,2012. ZHU G F.Traditional Chinese Medicine[M].Beijing:Tsinghua University Press,2012.(in Chinese)
[2] 宋光熠.中药药理学[M].北京:人民卫生出版社,2009. SONG G Y.Pharmacology of Chinese Medicine[M].Beijing:People's Medical Publishing House,2009.(in Chinese)
[3] 国家药典委员会.中华人民共和国药典(一部)[M].2010版.北京:中国医药科技出版社,2010. NATIONAL PHARMACOPOEIA COMMISSION.Pharmacopoeia of the People's Republic of China (I)[M].2010 edition.Beijing:China Medical Science Press,2010.(in Chinese)
[4] 道列提汗.双黄连制剂在兽医临床上的应用[J].中国畜牧兽医文摘,2014,30(5):143. DAO L T H.Application of Shuanhuanglian preparation in veterinary clinic[J].Chinese Animal Husbandry and Veterinary Abstract,2014,30(5):143.(in Chinese)
[5] 易文龙,方峰,宁琴,等.双黄连、鱼腥草注射液抗鼠肝炎病毒3型的体外实验研究[J].东南大学学报(医学版),2006,4:228-231. YI W L,FANG F,NING Q,et al.In vitro study on the effect of Shuanghuanglian and Houttuynia injection on Rat hepatitis virus 3[J].Journal of Southeast University (Medical Edition),2006,4:228-231.(in Chinese)
[6] 王莉莉,胡凯.双黄连对内毒素诱导小鼠肝脏组织结构的影响[J].中国畜禽种业,2019,15(1):73-74. WANG L L,HU K.Effect of Shuanghuanglian on liver tissue structure induced by endotoxin in mice[J].China Livestock and Poultry Seed Industry,2019,15(1):73-74.(in Chinese)
[7] 贾雷,高梅,程志杰,等.双黄连对小鼠细胞免疫功能及IL-2产生的影响[J].吉林中医药,2000,1:62. JIA L,GAO M,CHENG Z J,et al.Effects of Shuanghuanglian on cellular immune function and IL-2 in mice[J].Jilin Chinese Medicine,2000,1:62.(in Chinese)
[8] 杨志,曾耀英,黄秀艳,等.双黄连粉针剂对小鼠T淋巴细胞体外活化与增殖的影响[J].细胞与分子免疫学杂志,2008,8:774-777. YANG Z,ZENG Y Y,HUANG X Y,et al.Effects of Shuanghuanglian powder on the activation and proliferation of T lymphocytes in mice in vitro[J].Journal of Cellular and Molecular Immunology,2008,8:774-777.(in Chinese)
[9] 孙翠翠,程江,唐棣,等.藏药绿萝花水溶性多糖提取工艺研究[J].黑龙江畜牧兽医,2017,4:208-213. SUN C C,CHENG J,TANG D,et al.A study on the extraction technology of water-soluble polysaccharide from Chloranthus chinensis[J].Heilongjiang Animal Science and Veterinary Medicine,2017,4:208-213.(in Chinese)
[10] 刘芮,唐棣,刘群.藏药波棱瓜花水溶性多糖提取工艺实验研究[J].西南民族大学学报(自然科学版),2019,45(6):570-578. LIU R,TANG D,LIU Q.Experimental study on the extraction process of water-soluble polysaccharide from Tibetan medicine Herpetospermum pedunculosum flower[J].Journal of Southwest University for Nationalities (Natural Science Edition),2019,45(6):570-578.(in Chinese)
[11] 魏雄辉,张建斌.天然药物化学[M].北京:北京大学出版社,2013. WEI X H,ZHANG J B.Natural Medicine Chemistry[M].Beijing:Peking University Press,2013.(in Chinese)
[12] 邱园园,韩金潭,孙翠翠,等."香连"溶液干膏制剂体外抑菌试验研究[J].中国畜牧兽医,2015,42(3):708-713. QIU Y Y,HAN J T,SUN C C,et al.Experimental study on antibacterial activity of "Xianglian" solution dry paste preparation in vitro[J].China Animal Husbandry & Veterinary Medicine,2015,42(3):708-713.(in Chinese)
[13] 江洁芳.花椒中抑菌活性成分提取工艺的研究[J].中国调味品,2011,36(3):30-32. JIANG J F.Study on the extraction technology of antibacterial active ingredients from prickly ash[J].Chinese Condiment,2011,36(3):30-32.(in Chinese)
[14] CAVAR S,MILKAM M,VIDIC D,et al.Chemical composition and antioxidant activity of essential oil of Artenisia annua L.from Bosnia[J].Crops and Products,2011,10(1):2-7.
[15] 马建凤,刘华钢,朱丹.中药体外抑菌研究的方法学进展[J].药物评价研究,2010,33(1):42-45. MA J F,LIU H G,ZHU D.Advances in the study of traditional Chinese medicine in vitro bacteriostasis[J].Drug Evaluation Research,2010,33(1):42-45.(in Chinese)
[16] 颜娓娓,徐佳馨,王继锋,等.白头翁与蛴螬不同配伍比例体外抗病毒作用研究[J].长春中医药大学学报,2018,34(3):453-456. YAN W W,XU J X,WANG J F,et al.Study on the antiviral effect of Grubs in vitro with different compatibility ratios of radix holothuriae and crubs[J].Journal of Changchun University of Chinese Medicine,2012,34(3):453-456.(in Chinese)
[17] 赵昕.人参皂苷Rb1体外抗PRRSV的作用及其机制[D].太谷:山西农业大学,2013. ZHAO X.The anti-PRRSV effect of ginsenoside Rb1 and its mechanism in vitro[D].Taigu:Shanxi Agricultural University,2013.(in Chinese)
[18] 张盼盼,焦方文,周建国,等.发酵全蝎粉酶解前后水提液抗病毒活性比较[J].中南学报,2016,14(3):267-269. ZHANG P P,JIAO F W,ZHOU J G,et al.Comparison of antiviral activity of water extract before and after enzymatic hydrolysis of fermented scorpion powder[J].Acta Zhongnan Sinica,2016,14(3):267-269.(in Chinese)
[19] 刘云,朱善元,龚祝南,等.6种中药体外抗猪繁殖与呼吸综合征病毒的作用[J].河南农业科学,2019,48(10):147-154. LIU Y,ZHU S Y,GONG Z N,et al.Anti-porcine reproductive and respiratory syndrome virus effect of 6 traditional Chinese medicines in vitro[J].Journal of Henan Agricultural Sciences,2019,48(10):147-154.(in Chinese)
[20] 黄培成,杨强,程小耕,等.金银花-连翘药对抗肠道病毒71型的初步探究[J].实用药物与临床,2019,22(6):589-592. HUANG P C,YANG Q,CHENG X G,et al.A preliminary study of honeysuckle forsythia capsule against enterovirus-71[J].Practical Medicine and Clinical Practice,2019,22(6):589-592.(in Chinese)
[21] 王成虎,李延萍,李群堂,等.潜阳封髓丹加减治疗多重耐药菌感染反复发烧3例[J].世界中西医结合杂志,2017,12(3):422-425. WANG C H,LI Y P,LI Q T,et al.Qianyang Fengsui Dan in the treatment of recurrent fever caused by multiple drug-resistant bacteria infection in 3 cases[J].World Journal of Integrated Traditional Chinese and Western Medicine, 2017,12(3):422-425.(in Chinese)
[22] SI W,QIAO Y L,LIU Z,et al.Combination of multi-model statistical analysis and quantitative fingerprinting in quality evaluation of Shuang-huang-lian oral liquid[J].Analysis and Bioanalytical Chemistry, 2020,412(26):412-426.
[23] 梁传军,郭秋萍,李建香.药加味二仙汤配合抗菌药物治疗女性患者慢性尿路感染的临床疗效评价[J].抗感染药学,2016,13(6):1433-1435. LIANG C J,GUO Q P,LI J X.Clinical evaluation of Jiawei Erxian decoction combined with antibiotics in the treatment of female patients with chronic urinary tract infection[J].Anti-infective Pharmacy,2016,13(6):1433-1435.(in Chinese)
[24] 蒋锦琴,黄卫平,张小贤.中药结核丸对结核分枝杆菌体内外抗菌效果的观察[J].江西医学检验,2006,23(6):521-522. JIANG J Q,HUANG W P,ZHANG X X.Observation on the antibacterial effect of traditional Chinese medicine tuberculosis pill on Mycobacterium tuberculosis in vivo and in vitro[J].Jiangxi Medical Laboratory,2006,23(6):521-522.(in Chinese)
[25] 曾春晖,杨柯,徐明光,等.广西藤茶总黄酮对金黄色葡萄球菌抗菌机制研究[J].中国实验方剂学杂志,2013,19(10):249-252. ZENG C H,YANG K,XU M G,et al.Study on the antibacterial mechanism of Guangxi Tengcha total flavonoids against Staphylococcus aureus[J].Chinese Journal of Experimental Formulas, 2013,19(10):249-252.(in Chinese)
[26] 李海华,郭蔚冰,陈志强,等.45味中药对多重耐药大肠杆菌的抑菌效果[J].中国现代中药,2019,21(6):791-796. LI H H,GUO W B,CHEN Z Q,et al.Antibacterial effect of 45 traditional Chinese medicines on multi-drug resistant Escherichia coli[J].Modern Chinese Medicines,2019,21(6):791-796.(in Chinese)
[27] 冯沙沙,宋鹏琰,翟福展,等.42种常见中药对猪链球菌的体外抑菌试验[J].黑龙江畜牧兽医,2017,24:131-133. FENG S S,SONG P Y,ZHAI F Z,et al.In vitro antibacterial test of 42 kinds of common Chinese medicines on Streptococcus suis[J].Heilongjiang Animal Science and Veterinary Medicine,2017,24:131-133.(in Chinese)
[28] 胡欢,左国营,张泽萍.36种中药材体外抗菌活性筛选研究[J].广西植物,2018,38(4):22-34. HU H,ZUO G Y,ZANG Z P.Screening of in vitro antibacterial activity of 36 kinds of traditional Chinese medicine[J].Plants in Guangxi,2018,38(4):22-34.(in Chinese)
[29] 邱志宏,张彩虹.蒲公英水提物对临床耐药菌体外抑菌作用的研究[J].临床合理用药杂志,2018,11(35):131-132. QIU Z H,ZHANG C H.Study on the bacteriostatic effect of aqueous extract of dandelion on clinical drug-resistant bacteria in vitro[J].Journal of Clinical Rational Drug Use,2018,11(35):131-132.(in Chinese)
[30] 张昊.山丹鳞茎总黄酮提取分离和抑菌活性的研究[D].延安:延安大学,2016. ZHANG H.Study on the extraction and isolation of total flavonoids from the bulbs of Shandan and their antimicrobial activity[D].Yan'an:Yan'an University,2016.(in Chinese)
[31] 苏建桥,赵雅婵,常云玲,等.苍耳子中黄酮的提取工艺优化及抑菌活性研究[J].饲料研究,2017,3(8):28-31. SU J Q,ZHAO Y C,CHANG Y L,et al.Study on optimization of extraction technology and antibacterial activity of Xanthium flavone[J].Feed Research,2017,3(8):28-31.(in Chinese)
[32] 徐林.藤椒水提取液中黄酮含量测定及抑菌、抗氧化活性的研究[D].成都:四川农业大学,2016. XU L.Determination of flavonoid content in water extract of Rattan pepper and study on antibacterial and antioxidant activities[D].Chengdu:Sichuan Agricultural University,2016.(in Chinese)
[33] MA Q,LIANG D,SONG S,et al.Comparative study on the antivirus activity of Shuang-Huang-Lian injectable powder and its bioactive compound mixture against human adenovirus Ⅲ in vitro[J].Viruses.2017,9(4):79.
[34] 王剑,侯林,陈亚乔,等.黄芩提取物体外抗病毒药效学研究[J].中成药,2017,39(9):1924-1927. WANG J,HOU L,CHEN Y Q,et al.Study on antiviral pharmacodynamics of Scutellaria baicalensis extract in vitro[J].Proprietary Chinese Medicine,2017,39(9):1924-1927.(in Chinese)
[35] 邵文爱,张丽媛,李康生.抗病毒药物筛选中两种方法的比较[J].汕头大学医学院学报,2006,1:52-54. SHAO W A,ZHANG L Y,LI K S.Comparison of two methods in antiviral drug screening[J].Journal of Medical College of Shantou University,2006,1:52-54.(in Chinese)