川芎醇提物抗氧化活性及抗食源性致病菌特性分析
|
摘要
为了研究川芎醇提物(Ligusticum chuanxiong alcohol extract,LCAE)抗氧化活性及对食源性致病菌的抑制特性。对LCAE的主要活性组分进行了初步分析,并测定其还原力、总抗氧化能力及对羟自由基、超氧阴离子的清除效果;采用打孔法和倍比稀释法研究了LCAE对5种常见食源性致病菌的抑制作用,并探究了各试验菌的生长趋势。结果表明:LCAE中主要的活性组分川芎嗪(tetramethylpyrazine,TMP)、阿魏酸、游离酚及结合酚的含量分别为55.7、31.45、177.38及42.86 g/kg。LCAE溶液稀释100倍后的还原力与14.79μg/m L VC相当,LCAE总抗氧化能力为4.46 U/mg。0.9 g/L LCAE对羟自由基清除率为超氧阴离子的1.12倍。LCAE对革兰氏阴性菌抑制效果强于阳性菌:沙门氏菌>大肠杆菌>假单胞菌>金黄色葡萄球菌>枯草芽孢杆菌,且存在明显的浓度依赖性,MIC介于6.25～25 g/L之间,MBC介于12.5～50 g/L之间。等价浓度下,纯品TMP和阿魏酸的抑菌效果比LCAE弱。药物处理下各种菌生长曲线特性显示,随着LCAE浓度的增加,其中革兰氏阴性试验菌在对数期生长受到明显抑制,阳性菌稳定期延长。
To study the antioxidant activity of Ligusticum chuanxiong alcohol extract( LCAE) and its inhibition to food-borne pathogens.The mainly active components of LCAE were analyzed and their reducing power,total antioxidant capacity even the scavenging effect on hydroxyl radical and superoxide anion were also measured. Adopting duple diluent method and stiletto method have been adopted to study the inhibition of LCAE on 5 common food-born pathogens,the growth trends of the tested bacteria were also studied. The results indicated that the content of TMP,ferulic acid,free phenolic and bound phenolic was55.7,31.45,177.38 and 42.86 g/kg,respectively. After LCAE diluted 100 times,the reducing power was equivalent to14.79 μg/m L VC.The total antioxidant of LCAE was 4.46 U/mg. The clearance rate of 0.9 g/L LCAE on hydroxy radical was1.12 times of that on superoxide anion. The inhibitory effect of LCAE on gram-negative bacteria was stronger than that of positive bacteria: Salmonella > Escherichia coli > Pseudomonas > Staphylococcus aureus > Bacillus subtilis,and the inhibition in a dose-dependent manner.The MIC were between 6.25～25 g/L while the MBC were lies between 12.5～50 g/L.Inhibitory effect of pure TMP and ferulic acid were lower compared with LCAE at equivalent concentration.Under drug treament,5 tests bacteria growth curve characteristic showed that with the concentration of LCAE increased,the growth of gram-negative test bacteria was significantly inhibited in the logarithmic phase,while the stable phase of positive bacteria was prolonged.
To study the antioxidant activity of Ligusticum chuanxiong alcohol extract( LCAE) and its inhibition to food-borne pathogens.The mainly active components of LCAE were analyzed and their reducing power,total antioxidant capacity even the scavenging effect on hydroxyl radical and superoxide anion were also measured. Adopting duple diluent method and stiletto method have been adopted to study the inhibition of LCAE on 5 common food-born pathogens,the growth trends of the tested bacteria were also studied. The results indicated that the content of TMP,ferulic acid,free phenolic and bound phenolic was55.7,31.45,177.38 and 42.86 g/kg,respectively. After LCAE diluted 100 times,the reducing power was equivalent to14.79 μg/m L VC.The total antioxidant of LCAE was 4.46 U/mg. The clearance rate of 0.9 g/L LCAE on hydroxy radical was1.12 times of that on superoxide anion. The inhibitory effect of LCAE on gram-negative bacteria was stronger than that of positive bacteria: Salmonella > Escherichia coli > Pseudomonas > Staphylococcus aureus > Bacillus subtilis,and the inhibition in a dose-dependent manner.The MIC were between 6.25～25 g/L while the MBC were lies between 12.5～50 g/L.Inhibitory effect of pure TMP and ferulic acid were lower compared with LCAE at equivalent concentration.Under drug treament,5 tests bacteria growth curve characteristic showed that with the concentration of LCAE increased,the growth of gram-negative test bacteria was significantly inhibited in the logarithmic phase,while the stable phase of positive bacteria was prolonged.
引文
[1]Maríaángeles Argudín. Food poisoning and Staphylococcus aureus enterotoxins[J].Toxins,2010,2(7):1751-1773.
[2]Dillen C A,Pinsker B L,Marusina A I,et al. Clonally expandedγδT cells protect against Staphylococcus aureus skin reinfection[J]. Journal of Clinical Investigation,2018,128(3):1026-1042.
[3]Nataro J P,Kaper J B. Diarrheagenic Escherichia coli[J].Clinical Microbiology Reviews,1998,11(1):142-201.
[4] Ganesan R,Hos N J,Gutierrez S,et al. Salmonella typhimurium disrupts Sirt1/AMPK checkpoint control of m TOR to impair autophagy[J].Plos Pathogens,2017,13(2):e1006227.
[5]郝士卿,杨浩宁,尤文文,等.372株铜绿假单胞菌的临床分布特点及耐药性分析[J].临床合理用药杂志,2017,10(1):154-155.
[6]Setlow P. Spores of Bacillus subtilis:Their resistance to and killing by radiation,heat and chemicals[J]. J Appl Microbiol,2010,101(3):514-525.
[7]赵冉冉,孙彬青,高晶晶,等.中草药提取物的研究现状及应用[J].广州化工,2017,45(12):5-7.
[8]孔秋莲,张明融,章丽丽.中草药提取物对两种果蔬致腐真菌的抑制作用[J].保鲜与加工,2002,2(3):17-18.
[9]Miao C,Wu S,Luo B,et al. A new sesquiterpenoid from Ligusticum chuanxiong Hort[J]. Fitoterapia,2010,81(8):1088-1090.
[10]Zhang X,Han B,Feng Z M,et al. Ferulic acid derivatives from Ligusticum chuanxiong[J].Fitoterapia,2018,125:147-154.
[11]Lu C,Xu W,Shao J,et al. Nrf2 activation is required for Ligustrazine to inhibit hepatic steatosis in alcohol-preferring mice and hepatocytes[J].Toxicological Sciences An Official Journal of the Society of Toxicology,2016,155(2):432-443.
[12]Hwang H,Hartman T,Rosen R,et al.Formation of pyrazines from the maillard reaction of glucose and Lysine-α-amine-15N[J].Journal of Agricultural and Food Chemistry,1994,42(4):1000-1004.
[13]Ge H F,Chen Y Z,Chen J C,et al.Evaluation of antioxidant activities of ethanol extract from Ligusticum subjected to in-vitro gastrointestinal digestion[J].Food&Chemical Toxicology,2018,119:417-424.
[14]刘晓丽,霍展样,张金莲.盐酸川芎嗪体外抗氧化作用的研究[J].中国航天医药杂志,2003,5(2):37-39.
[15]唐杰,冯甦,候若彤,等.川芎提取物的抑菌杀虫作用研究[J].时珍国医国药,2008,19(10):2437-2439.
[16]吴培源,俞晓红,谢建生.四物汤抗氧化活性及抗菌活性研究[J].光明中医,2017,32(16):2339-2340.
[17]许理恕.中药“十味败毒汤”效能及抗菌机制的初步报告[J].西安交通大学学报(医学版),1959(1).
[18] Sangeetha M,Mathammal R. Studies on the growth,characterization,physicochemical properties and anti-bacterial activity of ferulic acid crystals[J].Int J Pharm Pharm Sci,2016,8(1):121-126.
[19]Killiny N,Valim M F,Jones S E,et al. Metabolically speaking:Possible reasons behind the tolerance of‘Sugar Belle’mandarin hybrid to huanglongbing[J]. Plant Physiology&Biochemistry,2017,116:36-47.
[20]段靖,尹琼玉,李燕思,等.自拟当归活血丸再度提高质量标准的成分检测研究[J].实用医药杂志,2017,34(9):815-817.
[21]肖乃玉,卢曼萍,陈少君,等.阿魏酸-胶原蛋白抗菌膜在腊肠保鲜中的应用[J].食品与发酵工业,2014,40(4):210-215.
[22]王振斌,孙亚钊,郭强.超声波对无花果多糖抗氧化活性的影响[J].食品工业科技,2013,34(3):97-99,103.
[23]葛慧芳,陈继承,张孝林,等.黄芩抗金黄葡萄球菌提取工艺的优化[J].热带生物学报,2017,8(2):220-224.
[24]Liu G,Ren G,Zhao L,et al. Antibacterial activity and mechanism of bifidocin A against Listeria monocytogenes[J].Food Control,2016,73:854-861.
[25]Li Y,Song P,Zhu Q,et al.Liguzinediol improved the heart function and inhibited myocardial cell apoptosis in rats with heart failure[J].2014,35(10):1257-1264.
[26]Gu J,Su S,Guo J,et al. Anti-inflammatory and antiapoptotic effects of the combination of Ligusticum chuanxiong and Radix paeoniae against focal cerebral ischaemia via TLR4/MyD88/MAPK/NF-κB signalling pathway in MCAO rats[J].Journal of Pharmacy&Pharmacology,2018,70(2):268-277.
[27]刘亚杰.川芎嗪衍生物的合成及其抗菌活性的研究[D].广州:广东药科大学,2016.
[28]高慧娟,景凤,潘媛,等.灵芝发酵川芎产物的抑菌性试验[J].中成药,2016,38(2):434-437.
[29]Xu J,Song Z,Guo Q,et al.Synergistic effect and molecular mechanisms of traditional Chinese medicine on regulating tumor microenvironment and cancer cells[J]. Biomed Res Int,2016,2016(5):1490738.
[30]肖悦,刘天佳,黄正蔚,等.天然药物对粘性放线菌生长和产酸影响的体外研究[J].四川大学学报(医学版),2002,33(2):253-255.
[2]Dillen C A,Pinsker B L,Marusina A I,et al. Clonally expandedγδT cells protect against Staphylococcus aureus skin reinfection[J]. Journal of Clinical Investigation,2018,128(3):1026-1042.
[3]Nataro J P,Kaper J B. Diarrheagenic Escherichia coli[J].Clinical Microbiology Reviews,1998,11(1):142-201.
[4] Ganesan R,Hos N J,Gutierrez S,et al. Salmonella typhimurium disrupts Sirt1/AMPK checkpoint control of m TOR to impair autophagy[J].Plos Pathogens,2017,13(2):e1006227.
[5]郝士卿,杨浩宁,尤文文,等.372株铜绿假单胞菌的临床分布特点及耐药性分析[J].临床合理用药杂志,2017,10(1):154-155.
[6]Setlow P. Spores of Bacillus subtilis:Their resistance to and killing by radiation,heat and chemicals[J]. J Appl Microbiol,2010,101(3):514-525.
[7]赵冉冉,孙彬青,高晶晶,等.中草药提取物的研究现状及应用[J].广州化工,2017,45(12):5-7.
[8]孔秋莲,张明融,章丽丽.中草药提取物对两种果蔬致腐真菌的抑制作用[J].保鲜与加工,2002,2(3):17-18.
[9]Miao C,Wu S,Luo B,et al. A new sesquiterpenoid from Ligusticum chuanxiong Hort[J]. Fitoterapia,2010,81(8):1088-1090.
[10]Zhang X,Han B,Feng Z M,et al. Ferulic acid derivatives from Ligusticum chuanxiong[J].Fitoterapia,2018,125:147-154.
[11]Lu C,Xu W,Shao J,et al. Nrf2 activation is required for Ligustrazine to inhibit hepatic steatosis in alcohol-preferring mice and hepatocytes[J].Toxicological Sciences An Official Journal of the Society of Toxicology,2016,155(2):432-443.
[12]Hwang H,Hartman T,Rosen R,et al.Formation of pyrazines from the maillard reaction of glucose and Lysine-α-amine-15N[J].Journal of Agricultural and Food Chemistry,1994,42(4):1000-1004.
[13]Ge H F,Chen Y Z,Chen J C,et al.Evaluation of antioxidant activities of ethanol extract from Ligusticum subjected to in-vitro gastrointestinal digestion[J].Food&Chemical Toxicology,2018,119:417-424.
[14]刘晓丽,霍展样,张金莲.盐酸川芎嗪体外抗氧化作用的研究[J].中国航天医药杂志,2003,5(2):37-39.
[15]唐杰,冯甦,候若彤,等.川芎提取物的抑菌杀虫作用研究[J].时珍国医国药,2008,19(10):2437-2439.
[16]吴培源,俞晓红,谢建生.四物汤抗氧化活性及抗菌活性研究[J].光明中医,2017,32(16):2339-2340.
[17]许理恕.中药“十味败毒汤”效能及抗菌机制的初步报告[J].西安交通大学学报(医学版),1959(1).
[18] Sangeetha M,Mathammal R. Studies on the growth,characterization,physicochemical properties and anti-bacterial activity of ferulic acid crystals[J].Int J Pharm Pharm Sci,2016,8(1):121-126.
[19]Killiny N,Valim M F,Jones S E,et al. Metabolically speaking:Possible reasons behind the tolerance of‘Sugar Belle’mandarin hybrid to huanglongbing[J]. Plant Physiology&Biochemistry,2017,116:36-47.
[20]段靖,尹琼玉,李燕思,等.自拟当归活血丸再度提高质量标准的成分检测研究[J].实用医药杂志,2017,34(9):815-817.
[21]肖乃玉,卢曼萍,陈少君,等.阿魏酸-胶原蛋白抗菌膜在腊肠保鲜中的应用[J].食品与发酵工业,2014,40(4):210-215.
[22]王振斌,孙亚钊,郭强.超声波对无花果多糖抗氧化活性的影响[J].食品工业科技,2013,34(3):97-99,103.
[23]葛慧芳,陈继承,张孝林,等.黄芩抗金黄葡萄球菌提取工艺的优化[J].热带生物学报,2017,8(2):220-224.
[24]Liu G,Ren G,Zhao L,et al. Antibacterial activity and mechanism of bifidocin A against Listeria monocytogenes[J].Food Control,2016,73:854-861.
[25]Li Y,Song P,Zhu Q,et al.Liguzinediol improved the heart function and inhibited myocardial cell apoptosis in rats with heart failure[J].2014,35(10):1257-1264.
[26]Gu J,Su S,Guo J,et al. Anti-inflammatory and antiapoptotic effects of the combination of Ligusticum chuanxiong and Radix paeoniae against focal cerebral ischaemia via TLR4/MyD88/MAPK/NF-κB signalling pathway in MCAO rats[J].Journal of Pharmacy&Pharmacology,2018,70(2):268-277.
[27]刘亚杰.川芎嗪衍生物的合成及其抗菌活性的研究[D].广州:广东药科大学,2016.
[28]高慧娟,景凤,潘媛,等.灵芝发酵川芎产物的抑菌性试验[J].中成药,2016,38(2):434-437.
[29]Xu J,Song Z,Guo Q,et al.Synergistic effect and molecular mechanisms of traditional Chinese medicine on regulating tumor microenvironment and cancer cells[J]. Biomed Res Int,2016,2016(5):1490738.
[30]肖悦,刘天佳,黄正蔚,等.天然药物对粘性放线菌生长和产酸影响的体外研究[J].四川大学学报(医学版),2002,33(2):253-255.