不同中药与盐酸恩诺沙星联用对大肠杆菌耐药菌株体外抑菌作用研究
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摘要
本试验旨在研究不同中药与盐酸恩诺沙星联用对大肠杆菌耐药菌株的体外抑菌作用。采用微量肉汤法测定临床上一种常用药物盐酸恩诺沙星和不同中药对大肠杆菌耐药菌株及其联合使用的最小抑菌浓度(minimum inhibitory concentration,MIC)。结果发现,单味中药水提物中侧柏叶、金樱子和五味子的抑菌效果较好,其MIC值分别为3.91、3.91和7.81 mg/mL;单味中药的醇提物中,侧柏叶、马齿苋抑菌效果较好,其MIC值分别为7.81和3.91 mg/mL。盐酸恩诺沙星分别与五味子和金樱子水提物联合作用后,对大肠杆菌耐药菌株的MIC值分别降低到0.0078和0.0039 mg/mL;与马齿苋、赤芍、侧柏叶和金樱子的醇提物联合后,对大肠杆菌耐药菌株MIC值均明显降低。盐酸恩诺沙星分别与鱼腥草水提物、赤芍水提物、椿皮水提物、马齿苋醇提物、苍术醇提物、三颗针醇提物、赤芍醇提物、薤白醇提物、拳参醇提物、侧柏叶醇提物、金樱子醇提物和败酱草醇提物联合作用后,对大肠耐药菌株呈现相加或协同抑菌作用,其FIC值分别为0.75、0.62、0.62、0.27、0.75、0.75、0.31、0.62、0.50、0.53、0.28和0.62。初步认为不同中药中含有能有效降低大肠杆菌耐药性的中药,也含有与盐酸恩诺沙星组合,具有协同或相加作用的中药,为寻找有效防治大肠杆菌病的药物组合来对抗多重耐药菌提供了可能。
The purpose of this experiment was to study the in vitro bacteriostasis of different kinds of traditional Chinese herbal medicines combined with enrofloxacin hydrochloride against drug-resistant strains of Escherichia coli(E.coli).The minimum inhibitory concentration(MIC) of enrofloxacin hydrochloride(a commonly used drug) and different Chinese herbal medicines against E.coli resistant strains and their combined use was determinated by micro-broth method.The results showed that the antimicrobial effects of Cacumen Biotae,Rosa Laevigata and Schisandra Chinensis Fructus were better in water extracts of single traditional Chinese herbal medicine,and the MIC value were 3.91,3.91 and 7.81 mg/mL,respectively;The antimicrobial effects of Cacumen Biotae and Portulaca Oleracea were better in ethanol extracts of single traditional Chinese herbal medicine,and the MIC value were 7.81 and 3.91 mg/mL,respectively.When enrofloxacin hydrochloride was combined with water extracts of Schisandra Chinensis and Rosa Laevigata,the MIC value of drug-resistant strains of E.coli reduced to 0.0078 and 0.0039 mg/mL,respectively.When enrofloxacin hydrochloride was combined with ethanol extracts of Portulaca Oleracea,Radix Paeoniae Rubra,Cacumen Biotae and Rosa Laevigata,the MIC value of drug-resistant strains significantly decreased.When the enrofloxacin hydrochloride was combined with water extract of Houttuynia Herba,Radix Paeoniae Rubra and Ailanthi Cortex,and alcohol extract of Portulaca Herba,Atractylodis Rhizoma,Berberidis Radix,Paeoniae Radix Rubra,Allii Macrostemi Bulbus,Allii Macrostemonis Bulbus,Platycladi Cacumen,Rosa Laevigatae and Ixeris Denticulata,it showed adding or synergistic bacteriostatic effect on the E.coli resistant strains,the FIC values were 0.75,0.62,0.62,0.27,0.75,0.75,0.31,0.62,0.50,0.53,0.28 and 0.62,respectively.It was preliminarily believed that there were traditional Chinese herbal medicines that could effectively reduce the drug resistance of E.coli among the different kinds of traditional Chinese herbal medicines,there were also traditional Chinese herbal medicines combined with enrofloxacin hydrochloride,which had synergistic or additive effects.It was possible to find effective drug combinations for the prevention and treatment of E.coli disease to against multi-drug-resistant bacteria.
The purpose of this experiment was to study the in vitro bacteriostasis of different kinds of traditional Chinese herbal medicines combined with enrofloxacin hydrochloride against drug-resistant strains of Escherichia coli(E.coli).The minimum inhibitory concentration(MIC) of enrofloxacin hydrochloride(a commonly used drug) and different Chinese herbal medicines against E.coli resistant strains and their combined use was determinated by micro-broth method.The results showed that the antimicrobial effects of Cacumen Biotae,Rosa Laevigata and Schisandra Chinensis Fructus were better in water extracts of single traditional Chinese herbal medicine,and the MIC value were 3.91,3.91 and 7.81 mg/mL,respectively;The antimicrobial effects of Cacumen Biotae and Portulaca Oleracea were better in ethanol extracts of single traditional Chinese herbal medicine,and the MIC value were 7.81 and 3.91 mg/mL,respectively.When enrofloxacin hydrochloride was combined with water extracts of Schisandra Chinensis and Rosa Laevigata,the MIC value of drug-resistant strains of E.coli reduced to 0.0078 and 0.0039 mg/mL,respectively.When enrofloxacin hydrochloride was combined with ethanol extracts of Portulaca Oleracea,Radix Paeoniae Rubra,Cacumen Biotae and Rosa Laevigata,the MIC value of drug-resistant strains significantly decreased.When the enrofloxacin hydrochloride was combined with water extract of Houttuynia Herba,Radix Paeoniae Rubra and Ailanthi Cortex,and alcohol extract of Portulaca Herba,Atractylodis Rhizoma,Berberidis Radix,Paeoniae Radix Rubra,Allii Macrostemi Bulbus,Allii Macrostemonis Bulbus,Platycladi Cacumen,Rosa Laevigatae and Ixeris Denticulata,it showed adding or synergistic bacteriostatic effect on the E.coli resistant strains,the FIC values were 0.75,0.62,0.62,0.27,0.75,0.75,0.31,0.62,0.50,0.53,0.28 and 0.62,respectively.It was preliminarily believed that there were traditional Chinese herbal medicines that could effectively reduce the drug resistance of E.coli among the different kinds of traditional Chinese herbal medicines,there were also traditional Chinese herbal medicines combined with enrofloxacin hydrochloride,which had synergistic or additive effects.It was possible to find effective drug combinations for the prevention and treatment of E.coli disease to against multi-drug-resistant bacteria.
引文
[1] 高海涛,韩俊丽,关道明.大肠杆菌耐药现状的严峻性[J].生命科学,2017,29(5):514-520.GAO H T,HAN J L,GUAN D M.The grim situation of drug resistance in Escherichia coli[J].Life Sciences,2017,29(5):514-520.(in Chinese)
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[3] BLONDEAU J M,LASKOWSKI R,BORSOS S.In-vitro activity of cefepime and seven other antimicrobial agents against 1518 non-fermentative Gram-negative bacilli collected from 48 Canadian health care facilities.Canadian Afermenter Study Group[J].Journal of Antimicrobial Chemotherapy,1999,44(4):545-588.
[4] 李继霞,公衍文.多重耐药菌医院感染状况分析[J].检验医学,2013,28(9):784-788.LI J X,GONG Y W.Analysis of multiple-resistant strain hospital infections[J].Laboratory Medicine,2013,28(9):784-788.(in Chinese)
[5] NATIONAL COMMITTEE FOR CLINICAL AND LABORATORY STANDARDS.Performance standards for antimicrobial disk and dilution susceptibility tests for bacterial isolated from animals:Approved standard M31-A2[S].2nd Edition.2004.
[6] 解长银,魏红.细菌吲哚试验快速纸片法检测[J].现代检验医学杂志,2006,21(6):65-66.XIE C Y,WEI H.Bacterial indole test fast paper method detection[J].Journal of Modern Laboratory Medicine,2006,21(6):65-66.(in Chinese)
[7] 孙颖悟.临床微生物学[M].大连:大连出版社,1990.SUN Y W.Clinical Microbiology[M].Dalian:Dalian Publishing House,1990.(in Chinese)
[8] 林万明.医学细菌生化试验鉴定手册[M].北京:人民卫生出版社,1985.LIN W M.Manual for Identification of Medical Bacterial Biochemical Tests[M].Beijing:People’s Health Press,1985.(in Chinese)
[9] 黄衍强,黄宏思,张莲,等.6种中草药抗耐药性大肠杆菌的抑菌效果[J].右江民族医学院学报,2009,31(3):368-369.HUANG Y Q,HUANG H S,ZHANG L,et al.Bacterial effects of 6 Chinese herbal resistance to E.coli[J].Journal of Youjiang Institute of Ethnic Medicine,2009,31(3):368-369.(in Chinese)
[10] 岳立.九种中药煎液对猪大肠杆菌的体外抑菌作用[J].畜禽业,2014,1:50-51.YUE L.Effects of nine kinds of Chinese medicine decoction on the antibacterial effect of Escherichia coli in vitro[J].Livestock Industry,2014,1:50-51.(in Chinese)
[11] 胡建军,刘利林,黄金奎,等.植物提取物体外抑菌试验的研究[J].塔里木农垦大学学报,2004,2:5-8.HU J J,LIU L L,HUANG J K,et al.Study on in vitro antibacterial test of plant extracts[J].Journal of Tarim Agricultural Reclamation University,2004,2:5-8.(in Chinese)
[12] 相自来,单红志,柴方红,等.复方金樱子提取液对仔猪白痢的疗效研究[J].吉林畜牧兽医,2015,36(5):31-32.XIANG Z L,SHAN H Z,CHAI F H,et al.Study on the effect of compound extract of Rosa Laeviagta on piglet dysentery[J].Jilin Animal Husbandry and Veterinary Medicine,2015,36(5):31-32.(in Chinese)
[13] 张景皓,肖明霞,季萍.黄芩、黄连、五味子对5种多重耐药菌抑菌作用的实验研究[J].新疆医学,2012,42(2):4-7.ZHANG J H,XIAO M X,JI P.Experimental study on baicalensis of Scutellaria,Coptis and Schisandra on five kinds of multi-drug resistant bacteria[J].Xinjiang Medicine,2012,42(2):4-7.(in Chinese)
[14] 闫绍悦.抗鸡大肠杆菌病单味中药筛选及其成分的研究[D].泰安:山东农业大学,2014.YAN S Y.Screening of single Chinese herb and analysis of its compositions for curing chicken colibacillosis[D].Tai’an:Shandong Agricultural University,2014.(in Chinese)
[15] 孙宁宁.马齿苋治疗菌痢[J].中国民间疗法,2018,26(7):74.SUN N N.Purslane treatment of bacterial diarrhea[J].Chinese Folk Therapy,2018,26(7):74.(in Chinese)
[16] 陈国妮,孙飞龙,闫亚茹.马齿苋类黄酮提取工艺及抑菌效果的研究[J].包装与食品机械,2016,34(1):6-10.CHEN G N,SUN F L,YAN Y R.Study on extraction process of flavonoids from purslane and their antibacterial effect[J].Packaging and Food Machinery,2016,34(1):6-10.(in Chinese)
[17] 孙燕杰.马齿苋提取物对产ESBLs大肠杆菌体外抑菌作用及抑菌机理的研究[D].南宁:广西大学,2016.SUN Y J.Study on antibacterial effect and mechanism of Portulaca Oleracea L.against extended-spectrum-β-lactamase-producing Escherichia coli[D].Nanning:Guangxi University,2016.(in Chinese)
[18] 梁统,覃艳梅,梁念慈.侧柏叶醇提物抗炎作用的研究[J].中国药科大学学报,2001,32(3):224-226.LIANG T,TAN Y M,LIANG N C.Study on the anti-inflammatory effect of Biota Orientalis[J].Journal of the China Pharmaceutical University,2001,32(3):224-226.(in Chinese)
[19] 徐婷.大肠埃希菌对恩诺沙星及苯扎溴铵交叉耐药变迁的分子机制研究[D].重庆:西南大学,2016.XU T.Study of cross-resistance change molecular mechanism of E.coli to enrofloxacin and benzaz ammonium bromide[D].Chongqing:Southwestern University,2016.(in Chinese)
[20] 张庭廷,潘继红,聂刘旺,等.金樱子多糖的抑菌和抗炎作用研究[J].生物学杂志,2005,22(2):41-42.ZHANG T T,PAN J H,NIE L W,et al.Study on the anti-microbial and anti-inflammatory activities of Rosa Lacvigata Michx polysaccharides[J].Journal of Biology,2005,22(2):41-42.(in Chinese)
[21] 林树乾,秦炳燕,冯敏燕,等.五味子水提物和阿米卡星诱导细菌耐药性试验[J].中兽医医药杂志,2018,37(4):42-44.LIN S Q,QIN B Y,FENG M Y,et al.Experiment on water extract from Fructus Schisandrae and amikacin inducing bacterial resistance[J].Chinese Journal of Veterinary Medicine,2008,37(4):42-44.(in Chinese)
[22] 刘晓庆.MDR AB氨基糖苷类耐药机制及中草药抑菌探索[D].南昌:南昌大学,2012.LIU X Q.Resistance mechanism of MDR AB aminoglycoside and antibacterial exploration of Chinese herbal medicine[D].Nanchang:Nanchang University,2012.(in Chinese)
[23] 石磊,段博文,高进贤,等.大肠埃希菌对喹诺酮类药物的耐药机制分析[J].中华医院感染学杂志,2017,27(20):4568-4571.SHI L,DUAN B W,GAO J X,et al.Resistance mechanism of Escherichia coli to quinolones[J].Chinese Journal of Nosocomiology,2017,27(20):4568-4571.(in Chinese)
[24] 陈敬民,李友娣.金樱子醇提物的抗炎作用[J].中国民族民间医药,2011,20(5):33-35.CHEN J M,LI Y D.Experimental study on antiinflammation effects of Jinyingzi[J].Chinese Journal of Ethnomedicine and Ethnopharmacy,2011,20(5):33-35.(in Chinese)
[25] 邬苏晓,肖正中,周建超.中西药联用对猪大肠杆菌体外抗菌活性分析[J].甘肃畜牧兽医,2009,39(1):9-11.WU S X,XIAO Z Z,ZHOU J C.Analysis of in vitro antibacterial activity of Chinese and Western medicine combined with pig Escherichia coli[J].Gansu Animal Husbandry Veterinarian,2009,39(1):9-11.(in Chinese)
[26] 潘婕,刘增援,梁政,等.赤芍水提物与抗菌药联合对耐药大肠杆菌抑菌作用的研究[J].中国畜牧兽医,2015,42(11):3080-3086.PAN J,LIU Z Y,LIANG Z,et al.Study on the anti-bacterial effect of aqueous extract of Radix Paeoniac Rubra combined with antibiotics against drug-resistant E.coli[J].China Animal Husbandry & Veterinary Medicine,2015,42(11):3080-3086.(in Chinese)
[2] 植婵萍,郭小婷,姚旭,等.有机农场鸡、猪源大肠杆菌耐药性研究[J].中国畜牧兽医,2015,42(12):3351-3357.ZHI C P,GUO X T,YAO X,et al.Study on drug resistance of Escherichia coli isolated form chicken and swine in organic farm[J].China Animal Husbandry & Veterinary Medicine,2015,42(12):3351-3357.(in Chinese)
[3] BLONDEAU J M,LASKOWSKI R,BORSOS S.In-vitro activity of cefepime and seven other antimicrobial agents against 1518 non-fermentative Gram-negative bacilli collected from 48 Canadian health care facilities.Canadian Afermenter Study Group[J].Journal of Antimicrobial Chemotherapy,1999,44(4):545-588.
[4] 李继霞,公衍文.多重耐药菌医院感染状况分析[J].检验医学,2013,28(9):784-788.LI J X,GONG Y W.Analysis of multiple-resistant strain hospital infections[J].Laboratory Medicine,2013,28(9):784-788.(in Chinese)
[5] NATIONAL COMMITTEE FOR CLINICAL AND LABORATORY STANDARDS.Performance standards for antimicrobial disk and dilution susceptibility tests for bacterial isolated from animals:Approved standard M31-A2[S].2nd Edition.2004.
[6] 解长银,魏红.细菌吲哚试验快速纸片法检测[J].现代检验医学杂志,2006,21(6):65-66.XIE C Y,WEI H.Bacterial indole test fast paper method detection[J].Journal of Modern Laboratory Medicine,2006,21(6):65-66.(in Chinese)
[7] 孙颖悟.临床微生物学[M].大连:大连出版社,1990.SUN Y W.Clinical Microbiology[M].Dalian:Dalian Publishing House,1990.(in Chinese)
[8] 林万明.医学细菌生化试验鉴定手册[M].北京:人民卫生出版社,1985.LIN W M.Manual for Identification of Medical Bacterial Biochemical Tests[M].Beijing:People’s Health Press,1985.(in Chinese)
[9] 黄衍强,黄宏思,张莲,等.6种中草药抗耐药性大肠杆菌的抑菌效果[J].右江民族医学院学报,2009,31(3):368-369.HUANG Y Q,HUANG H S,ZHANG L,et al.Bacterial effects of 6 Chinese herbal resistance to E.coli[J].Journal of Youjiang Institute of Ethnic Medicine,2009,31(3):368-369.(in Chinese)
[10] 岳立.九种中药煎液对猪大肠杆菌的体外抑菌作用[J].畜禽业,2014,1:50-51.YUE L.Effects of nine kinds of Chinese medicine decoction on the antibacterial effect of Escherichia coli in vitro[J].Livestock Industry,2014,1:50-51.(in Chinese)
[11] 胡建军,刘利林,黄金奎,等.植物提取物体外抑菌试验的研究[J].塔里木农垦大学学报,2004,2:5-8.HU J J,LIU L L,HUANG J K,et al.Study on in vitro antibacterial test of plant extracts[J].Journal of Tarim Agricultural Reclamation University,2004,2:5-8.(in Chinese)
[12] 相自来,单红志,柴方红,等.复方金樱子提取液对仔猪白痢的疗效研究[J].吉林畜牧兽医,2015,36(5):31-32.XIANG Z L,SHAN H Z,CHAI F H,et al.Study on the effect of compound extract of Rosa Laeviagta on piglet dysentery[J].Jilin Animal Husbandry and Veterinary Medicine,2015,36(5):31-32.(in Chinese)
[13] 张景皓,肖明霞,季萍.黄芩、黄连、五味子对5种多重耐药菌抑菌作用的实验研究[J].新疆医学,2012,42(2):4-7.ZHANG J H,XIAO M X,JI P.Experimental study on baicalensis of Scutellaria,Coptis and Schisandra on five kinds of multi-drug resistant bacteria[J].Xinjiang Medicine,2012,42(2):4-7.(in Chinese)
[14] 闫绍悦.抗鸡大肠杆菌病单味中药筛选及其成分的研究[D].泰安:山东农业大学,2014.YAN S Y.Screening of single Chinese herb and analysis of its compositions for curing chicken colibacillosis[D].Tai’an:Shandong Agricultural University,2014.(in Chinese)
[15] 孙宁宁.马齿苋治疗菌痢[J].中国民间疗法,2018,26(7):74.SUN N N.Purslane treatment of bacterial diarrhea[J].Chinese Folk Therapy,2018,26(7):74.(in Chinese)
[16] 陈国妮,孙飞龙,闫亚茹.马齿苋类黄酮提取工艺及抑菌效果的研究[J].包装与食品机械,2016,34(1):6-10.CHEN G N,SUN F L,YAN Y R.Study on extraction process of flavonoids from purslane and their antibacterial effect[J].Packaging and Food Machinery,2016,34(1):6-10.(in Chinese)
[17] 孙燕杰.马齿苋提取物对产ESBLs大肠杆菌体外抑菌作用及抑菌机理的研究[D].南宁:广西大学,2016.SUN Y J.Study on antibacterial effect and mechanism of Portulaca Oleracea L.against extended-spectrum-β-lactamase-producing Escherichia coli[D].Nanning:Guangxi University,2016.(in Chinese)
[18] 梁统,覃艳梅,梁念慈.侧柏叶醇提物抗炎作用的研究[J].中国药科大学学报,2001,32(3):224-226.LIANG T,TAN Y M,LIANG N C.Study on the anti-inflammatory effect of Biota Orientalis[J].Journal of the China Pharmaceutical University,2001,32(3):224-226.(in Chinese)
[19] 徐婷.大肠埃希菌对恩诺沙星及苯扎溴铵交叉耐药变迁的分子机制研究[D].重庆:西南大学,2016.XU T.Study of cross-resistance change molecular mechanism of E.coli to enrofloxacin and benzaz ammonium bromide[D].Chongqing:Southwestern University,2016.(in Chinese)
[20] 张庭廷,潘继红,聂刘旺,等.金樱子多糖的抑菌和抗炎作用研究[J].生物学杂志,2005,22(2):41-42.ZHANG T T,PAN J H,NIE L W,et al.Study on the anti-microbial and anti-inflammatory activities of Rosa Lacvigata Michx polysaccharides[J].Journal of Biology,2005,22(2):41-42.(in Chinese)
[21] 林树乾,秦炳燕,冯敏燕,等.五味子水提物和阿米卡星诱导细菌耐药性试验[J].中兽医医药杂志,2018,37(4):42-44.LIN S Q,QIN B Y,FENG M Y,et al.Experiment on water extract from Fructus Schisandrae and amikacin inducing bacterial resistance[J].Chinese Journal of Veterinary Medicine,2008,37(4):42-44.(in Chinese)
[22] 刘晓庆.MDR AB氨基糖苷类耐药机制及中草药抑菌探索[D].南昌:南昌大学,2012.LIU X Q.Resistance mechanism of MDR AB aminoglycoside and antibacterial exploration of Chinese herbal medicine[D].Nanchang:Nanchang University,2012.(in Chinese)
[23] 石磊,段博文,高进贤,等.大肠埃希菌对喹诺酮类药物的耐药机制分析[J].中华医院感染学杂志,2017,27(20):4568-4571.SHI L,DUAN B W,GAO J X,et al.Resistance mechanism of Escherichia coli to quinolones[J].Chinese Journal of Nosocomiology,2017,27(20):4568-4571.(in Chinese)
[24] 陈敬民,李友娣.金樱子醇提物的抗炎作用[J].中国民族民间医药,2011,20(5):33-35.CHEN J M,LI Y D.Experimental study on antiinflammation effects of Jinyingzi[J].Chinese Journal of Ethnomedicine and Ethnopharmacy,2011,20(5):33-35.(in Chinese)
[25] 邬苏晓,肖正中,周建超.中西药联用对猪大肠杆菌体外抗菌活性分析[J].甘肃畜牧兽医,2009,39(1):9-11.WU S X,XIAO Z Z,ZHOU J C.Analysis of in vitro antibacterial activity of Chinese and Western medicine combined with pig Escherichia coli[J].Gansu Animal Husbandry Veterinarian,2009,39(1):9-11.(in Chinese)
[26] 潘婕,刘增援,梁政,等.赤芍水提物与抗菌药联合对耐药大肠杆菌抑菌作用的研究[J].中国畜牧兽医,2015,42(11):3080-3086.PAN J,LIU Z Y,LIANG Z,et al.Study on the anti-bacterial effect of aqueous extract of Radix Paeoniac Rubra combined with antibiotics against drug-resistant E.coli[J].China Animal Husbandry & Veterinary Medicine,2015,42(11):3080-3086.(in Chinese)