锦绣杜鹃花抗菌、抗炎活性及其有效部位研究
|
摘要
为了研究锦绣杜鹃花的抗菌及抗炎活性,并比较不同提取方法对其抗菌活性物质的提取效率.本实验采用了甲醇冷浸法、乙醇回流法和微波萃取法分别对锦绣杜鹃花的有效成分进行提取,并采用微孔板稀释法测试提取物对多种阳性菌、阴性菌和耐药菌的抑制活性.基于其较好的体外抗菌活性,还采用RAW264.7细胞炎症模型评价了粗提物对炎症因子TNF-α的影响,并测定了各萃取物的总黄酮含量.此外,为了获取锦绣杜鹃花抗菌作用的有效部位,本实验将提取物分别用石油醚、乙酸乙酯和水萃取得到不同极性的萃取物组分,比较其抑菌活性.甲醇冷浸法获得的粗提物的抗菌活性最佳;各萃取组分中,乙酸乙酯萃取组分的抗菌效果最好,其对金黄色葡萄球菌26003的MIC值达到0.512mg/mL;提取物能明显抑制LPS诱导的炎症因子TNF-α的增加.本研究证实了锦绣杜鹃花具有较好的的抗菌抗炎活性,其抗菌作用的有效部位为乙酸乙酯萃取部位,为锦绣杜鹃花在抗菌、抗炎药物和产品研究开发方面提供了实验依据.
In this study,the antibacterial activity of the extract of Rhododendron pulchrum flower and its different extraction methods for antibacterial active substances were investigated.Extraction methods including methanol soaking,ethanol refluxing and microwave extracting were carried out.The antibacterial activities of the extracted components were evaluated by microporous plate dilution test using various positive bacteria,negative bacteria and antibiotic resistant bacteria as the tested bacteria.Based on its potent antibacterial activity,the effect of the extract on the inflammatory factor TNF-αin the RAW264.7 macrophage was also investigated.Additionally,the extractions with petroleum ether and ethyl acetate fromthe water suspension of crude extract were also carried out to got three different polar components,which were also subjected to the antibacterial test to obtained the effective part of the antibacterial activity of Rhododendron pulchrum flower.The results indicated thatthe methanol soaking was superior to the other extract methods in the aspect of antibacterial activity.Furthermore,the ethyl acetate phase,holding the greatest antibacterial effects,was the optimal active fractionof Rhododendron pulchrum flower.It displayed outstanding antibacterial activity againstStaphylococcus aureus1.2155 with MIC value of 0.512 mg/mL.The extract could significantly inhibit the increase of the TNF-αinduced by LPS.It can be concluded thatRhododendron pulchrum flower exhibited antibacterial and antiinflammatory activities,and the ethyl acetate extracting after a methanol soaking of the medicinal plant was a proper method to obtain its antibacterial active ingredients.
In this study,the antibacterial activity of the extract of Rhododendron pulchrum flower and its different extraction methods for antibacterial active substances were investigated.Extraction methods including methanol soaking,ethanol refluxing and microwave extracting were carried out.The antibacterial activities of the extracted components were evaluated by microporous plate dilution test using various positive bacteria,negative bacteria and antibiotic resistant bacteria as the tested bacteria.Based on its potent antibacterial activity,the effect of the extract on the inflammatory factor TNF-αin the RAW264.7 macrophage was also investigated.Additionally,the extractions with petroleum ether and ethyl acetate fromthe water suspension of crude extract were also carried out to got three different polar components,which were also subjected to the antibacterial test to obtained the effective part of the antibacterial activity of Rhododendron pulchrum flower.The results indicated thatthe methanol soaking was superior to the other extract methods in the aspect of antibacterial activity.Furthermore,the ethyl acetate phase,holding the greatest antibacterial effects,was the optimal active fractionof Rhododendron pulchrum flower.It displayed outstanding antibacterial activity againstStaphylococcus aureus1.2155 with MIC value of 0.512 mg/mL.The extract could significantly inhibit the increase of the TNF-αinduced by LPS.It can be concluded thatRhododendron pulchrum flower exhibited antibacterial and antiinflammatory activities,and the ethyl acetate extracting after a methanol soaking of the medicinal plant was a proper method to obtain its antibacterial active ingredients.
引文
[1]张梅,潘大仁.杜鹃属植物黄酮类化合物的研究进展[J].海峡科学,2012,65(5):3-14.
[2]陈国联,张德山,王桂山,等.秦岭10种药用杜鹃的抗菌作用的实验研究[J].中医药学报,1990,23(5):51-52.
[3]任茜,李强,李万波,等.十种秦岭杜鹃的抗菌作用研究[J].陕西林业科技,1990,17(3):31-32.
[4]张德山,王桂山,李凌夫,等.汶川杜鹃的抗菌实验及镇咳作用的临床观察[J].中医药学报,1998,22(4):36-38.
[5]任茜,陈国联,李万波.30种杜鹃属植物抗菌作用的试验研究[J].中国园艺文摘,2012,28(3):3-4.
[6]李少泓,孙欣.杜鹃属植物的化学成分及药理作用研究进展[J].中华中医药学刊,2010,28(11):2435-2437.
[7]赵宝琴,高陆.满山红化学成分和药理学研究进展[J].人参研究,2015,14(1):42-44.
[8]李雪萍,高亚曼,孔令米,等.蜂蜜的药用价值:对人类健康特别是对血糖调节影响[J].聊城大学学报(自然科学版),2016,29(4):31-38.
[9]张梅,潘大仁.HPLC同时测定锦绣杜鹃四种黄酮类成分的含量[J].西北林学院学报,2013,28(1):119-122.
[10]吴丹慧.锦绣杜鹃提取物制备工艺及质量标准研究[D].湖北中医药大学,2011.
[11]赵玺,车文实.满山红的化学成分和提取方法研究进展[J].安徽医科大学学报,2013,48(5):580-582.
[12]蔡秋华.千层金总黄酮的提取研究[J].中国农业信息,2015,34(19):159-160.
[13]黎璐,吕昱,汤洪敏.长裙竹荪3部分石油醚提取物的对比[J].食品科学,2013,35(6):73-77.
[14]仇燕,王明珠,赵紫华.菜芙蓉总黄酮提取物抑菌作用及其稳定性研究[J].北方园艺,2016,41(8):150-153.
[15]傅力明,李华峰,姚杰,等.中药蒲公英花提取物的抑菌性能研究[J].山西医药杂志,2015,44(8):947-949.
[16]张朝绅,安晔,张振秋.大孔吸附树脂纯化满山红总黄酮工艺研究[J].中国药师,2010,13(7):969-973.
[17]徐炳祥,韩公羽,刘明珠.映山红有效成份的研究-Ⅲ、乌索酸的分离和鉴定[J].第二军医大学学报,1982,8(4):298-299.
[18]马微微,王军,穆立蔷.紫椴花提取物体外抗菌活性[J].东北林业大学学报,2013,41(9):127-132.
[19]张彩霞,戎敢,张志刚.罗汉果花中黄酮的提取及结构表征[J].光谱实验室,2013,30(3):1389-1394.
[20]孟辉,邹向宇,刘国良,等.两种微波辅助萃取法萃取满山红叶中总黄酮[J].湖南农业大学学报(自然科学报),2006,32(2):210-213.
[21]彭姣阳,林梦婷,谭雨琪,等.含3-芳基吡唑的绕丹宁衍生物的合成及抗菌活性评价[J].井冈山大学学报,2017,38(5):96-103.
[22] Clinical and Laboratory Standards Institute.Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically,Approved Standard M7-A6[S].Wayne,PA,USA,2003.
[23]曹婷婷.丹参对原代246.7细胞炎性因子作用机制的研究[D].华中科技大学,2005.
[24]陈滴,陈星,刘昕.分光光度法测定玉米须总黄酮含量研究[J].玉米科学,2007,15(2):147-152.
[25]梁军.银杏叶黄酮类化合物提取工艺的优化研究[J].辽宁中医杂志,2011,38(4):706-709.
[26]徐璐,夏国华,徐黎明,等.水芹中总黄酮类化合物提取工艺的优化及其含量测定[J].抗感染药学,2017,14(9):1654-1657.
[27]谢天艳,何开泽,赵海,等.4种浮萍提取物的抗菌活性和黄酮含量[J].应用与环境生物学报,2014,20(2):238-244.
[28]黄晓冬.4种龙眼核提取物的总黄酮含量、体外抗菌活性与抗氧化活性[J].食品科学,2011,32(11):43-47.
[29]黄晓冬.赤楠叶醇提物抗菌活性及成分总黄酮的研究[J].泉州师范学院学报,2007(4):98-102.
[2]陈国联,张德山,王桂山,等.秦岭10种药用杜鹃的抗菌作用的实验研究[J].中医药学报,1990,23(5):51-52.
[3]任茜,李强,李万波,等.十种秦岭杜鹃的抗菌作用研究[J].陕西林业科技,1990,17(3):31-32.
[4]张德山,王桂山,李凌夫,等.汶川杜鹃的抗菌实验及镇咳作用的临床观察[J].中医药学报,1998,22(4):36-38.
[5]任茜,陈国联,李万波.30种杜鹃属植物抗菌作用的试验研究[J].中国园艺文摘,2012,28(3):3-4.
[6]李少泓,孙欣.杜鹃属植物的化学成分及药理作用研究进展[J].中华中医药学刊,2010,28(11):2435-2437.
[7]赵宝琴,高陆.满山红化学成分和药理学研究进展[J].人参研究,2015,14(1):42-44.
[8]李雪萍,高亚曼,孔令米,等.蜂蜜的药用价值:对人类健康特别是对血糖调节影响[J].聊城大学学报(自然科学版),2016,29(4):31-38.
[9]张梅,潘大仁.HPLC同时测定锦绣杜鹃四种黄酮类成分的含量[J].西北林学院学报,2013,28(1):119-122.
[10]吴丹慧.锦绣杜鹃提取物制备工艺及质量标准研究[D].湖北中医药大学,2011.
[11]赵玺,车文实.满山红的化学成分和提取方法研究进展[J].安徽医科大学学报,2013,48(5):580-582.
[12]蔡秋华.千层金总黄酮的提取研究[J].中国农业信息,2015,34(19):159-160.
[13]黎璐,吕昱,汤洪敏.长裙竹荪3部分石油醚提取物的对比[J].食品科学,2013,35(6):73-77.
[14]仇燕,王明珠,赵紫华.菜芙蓉总黄酮提取物抑菌作用及其稳定性研究[J].北方园艺,2016,41(8):150-153.
[15]傅力明,李华峰,姚杰,等.中药蒲公英花提取物的抑菌性能研究[J].山西医药杂志,2015,44(8):947-949.
[16]张朝绅,安晔,张振秋.大孔吸附树脂纯化满山红总黄酮工艺研究[J].中国药师,2010,13(7):969-973.
[17]徐炳祥,韩公羽,刘明珠.映山红有效成份的研究-Ⅲ、乌索酸的分离和鉴定[J].第二军医大学学报,1982,8(4):298-299.
[18]马微微,王军,穆立蔷.紫椴花提取物体外抗菌活性[J].东北林业大学学报,2013,41(9):127-132.
[19]张彩霞,戎敢,张志刚.罗汉果花中黄酮的提取及结构表征[J].光谱实验室,2013,30(3):1389-1394.
[20]孟辉,邹向宇,刘国良,等.两种微波辅助萃取法萃取满山红叶中总黄酮[J].湖南农业大学学报(自然科学报),2006,32(2):210-213.
[21]彭姣阳,林梦婷,谭雨琪,等.含3-芳基吡唑的绕丹宁衍生物的合成及抗菌活性评价[J].井冈山大学学报,2017,38(5):96-103.
[22] Clinical and Laboratory Standards Institute.Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically,Approved Standard M7-A6[S].Wayne,PA,USA,2003.
[23]曹婷婷.丹参对原代246.7细胞炎性因子作用机制的研究[D].华中科技大学,2005.
[24]陈滴,陈星,刘昕.分光光度法测定玉米须总黄酮含量研究[J].玉米科学,2007,15(2):147-152.
[25]梁军.银杏叶黄酮类化合物提取工艺的优化研究[J].辽宁中医杂志,2011,38(4):706-709.
[26]徐璐,夏国华,徐黎明,等.水芹中总黄酮类化合物提取工艺的优化及其含量测定[J].抗感染药学,2017,14(9):1654-1657.
[27]谢天艳,何开泽,赵海,等.4种浮萍提取物的抗菌活性和黄酮含量[J].应用与环境生物学报,2014,20(2):238-244.
[28]黄晓冬.4种龙眼核提取物的总黄酮含量、体外抗菌活性与抗氧化活性[J].食品科学,2011,32(11):43-47.
[29]黄晓冬.赤楠叶醇提物抗菌活性及成分总黄酮的研究[J].泉州师范学院学报,2007(4):98-102.