烟草废弃物止血和抑菌有效部位的筛选
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摘要
目的:研究烟草废弃物不同提取部位的止血及抑菌作用,筛选止血和抑菌有效部位。方法:甲醇回流提取烟草废弃物,所得浸膏依次用石油醚,酸性乙酸乙酯,碱性乙酸乙酯和正丁醇萃取得各部位浸膏。首先通过体外凝血法(凝血板法和试管法)初步筛选有效止血部位,体内凝血法进一步考察初筛有效部位的不同剂量组对小鼠凝血时间的影响。将80只小鼠随机分为空白对照组,云南白药组(2 g·kg~(-1)),烟草废弃物石油醚部位高、中和低剂量组,酸性乙酸乙酯部位高、中和低剂量组。采用玻片法和毛细管法测量小鼠凝血时间,计算缩短率;采用微量稀释法测定烟草废弃物各部位对10种临床常见病原菌的最低抑菌浓度(MIC)。结果:体外凝血法表明:石油醚部位和酸性乙酸乙酯部位具有止血作用。体内凝血实验表明:与空白对照组比较,酸性乙酸乙酯部位的高、中剂量组可显著缩短小鼠的体内凝血时间,缩短率均高于60%(P<0.01),低剂量组可明显缩短小鼠体内凝血时间,缩短率高于39%(P<0.05)。酸性乙酸乙酯部位对痢疾杆菌,链球菌,白色念珠菌,犬小孢子菌,红色毛癣菌和石膏样小孢子菌有明显抑制作用,MIC值均小于6.25 g·L~(-1),对黑曲霉菌有一定的抑制作用,MIC值为50 g·L~(-1)。石油醚部位仅对白色念珠菌、链球菌和痢疾杆菌有较强的抑制作用,MIC值均小于50 g·L~(-1)。结论:酸性乙酸乙酯部位既有止血作用,又有抑菌作用,且抗菌范围广,故将酸性乙酸乙酯部位作为止血抑菌有效部位,为烟草废弃物的进一步开发利用提供依据。
Objective: To screen the effective parts of Tobacco Waste through hemostatic and Anti-bacterial activities. Method: Tobacco waste was firstly extracted by methanol and followed by petroleum ether, acidic ethyl acetate, basic ethyl acetate, and n-butanol to obtain the different parts of extraction. Vitro hemostatic method(clotting plate method and the test tube method) was utilized to screen the effective hemostatic parts. Effects of different dosages of the effective part on coagulation time in Mice were furtherly verificated through vivo hemostatic method. Eighty mices were randomly divided into blank group, Yunnanbaiyao group(2 g·kg~(-1)), and high-, middle- and low-dose of petroleum ether and acid ethyl acetate parts groups. The clotting time was measured by the slide method and capillary tube method, and the shortened rate was calculated. Micro dilution method was chosen to determine the minimum inhibitory concentration(MIC) of 10 different clinical pathogens. Result: The vitro hemostasis is initially indicated petroleum ether part and acidic ethyl acetate part have a hemostatic effect. the high and middle dose groups of acidic ethyl acetate part could significantly shorten the clotting time with a shortened rate that greater than 60%(P<0.01) and the low-dose group could obviously shorten the blood coagulation time with a shortened rate that higher than 39%(P<0.05) from vivo clotting experiments. Acidic ethyl acetate part has a strong inhibitory effect on Shigella Castellani, Streptococcus, Candida albicans, Microsporum canis, Trichophyton rubidium(MIC is less than 6.25 g·L~(-1)) and a certain inhibitory effect on Aspergillus niger(MIC is 50 g·L~(-1)).Petroleum ether part has a definite inhibitory effect on Candida albicans, Streptococcus and Shigella Castellani(MIC is lower than 50 g·L~(-1)). Conclusion: Acidic ethyl acetate part has obvious hemostatic and bacteriostatic effect, and the anti-bacterial range is wide. Therefore, acidic ethyl acetate part is the effective part of hemostasis and bacteriostasis, which provides the evidence for the further development and utilization of tobacco waste.
Objective: To screen the effective parts of Tobacco Waste through hemostatic and Anti-bacterial activities. Method: Tobacco waste was firstly extracted by methanol and followed by petroleum ether, acidic ethyl acetate, basic ethyl acetate, and n-butanol to obtain the different parts of extraction. Vitro hemostatic method(clotting plate method and the test tube method) was utilized to screen the effective hemostatic parts. Effects of different dosages of the effective part on coagulation time in Mice were furtherly verificated through vivo hemostatic method. Eighty mices were randomly divided into blank group, Yunnanbaiyao group(2 g·kg~(-1)), and high-, middle- and low-dose of petroleum ether and acid ethyl acetate parts groups. The clotting time was measured by the slide method and capillary tube method, and the shortened rate was calculated. Micro dilution method was chosen to determine the minimum inhibitory concentration(MIC) of 10 different clinical pathogens. Result: The vitro hemostasis is initially indicated petroleum ether part and acidic ethyl acetate part have a hemostatic effect. the high and middle dose groups of acidic ethyl acetate part could significantly shorten the clotting time with a shortened rate that greater than 60%(P<0.01) and the low-dose group could obviously shorten the blood coagulation time with a shortened rate that higher than 39%(P<0.05) from vivo clotting experiments. Acidic ethyl acetate part has a strong inhibitory effect on Shigella Castellani, Streptococcus, Candida albicans, Microsporum canis, Trichophyton rubidium(MIC is less than 6.25 g·L~(-1)) and a certain inhibitory effect on Aspergillus niger(MIC is 50 g·L~(-1)).Petroleum ether part has a definite inhibitory effect on Candida albicans, Streptococcus and Shigella Castellani(MIC is lower than 50 g·L~(-1)). Conclusion: Acidic ethyl acetate part has obvious hemostatic and bacteriostatic effect, and the anti-bacterial range is wide. Therefore, acidic ethyl acetate part is the effective part of hemostasis and bacteriostasis, which provides the evidence for the further development and utilization of tobacco waste.
引文
[1]马杰.印度梨形孢诱导烟草促生、抗病、抗逆作用及其机理的初步研究[D].浙江大学,2012.
[2]魏克强,杨俊仙,魏治中.新型烟草化学成分及医药成分的初步分析[J].天然产物研究与开发,2008,20(4):652-656.
[3]李莹,李钟,郭培国.烟草中主要生物活性成分的研究进展[J].天然产物研究与开发,2015(12):2157-2163.
[4]许晓风,郁锋,杨启银,等.烟草工业下脚料综合利用研究(I)--抗菌成分的提取与抑菌效果测定[J].安徽农业科学,2003,31(2):209-209.
[5]隋婧.青蒿体外止血活性部位筛选及其应用研究[D].重庆大学,2009.
[6]徐叔云,卞如濂,陈修.药理实验方法学[M].北京:人民卫生出版社,1991.1016-1018.
[7]黄艳,宁小清,原鲜玲,等.壮药白花九里明水煎液止血作用机制探讨[J].世界科学技术-中医药现代化,2013(7):1552-1556.
[8]海丽娜,张志伟,王金华,等.复方苦参注射液对小鼠的镇痛止血和抗应激作用[J].中国实验方剂学杂志,2012,18(2):199-202.
[9]王秋红,刘玉婕,吕邵娃,等.线叶菊抗菌作用研究[J].中国实验方剂学杂志,2011,17(22):141-145.
[10]Noake T,Kuriyama T,White P L,et al.Antifungal susceptibility of Candida species using the Clinical and Laboratory Standards Institute disk diffusion and broth microdilution methods.[J].Journal of Chemotherapy,2007,19(3):283-7.
[11]朱庆玲,冯翀.五爪龙不同提取物体外抗菌活性的研究[J].中国实验方剂学杂志,2011,17(11):232-233
[12]张伟芳,侯喜琴.NCCLSM27-A微量法测定白色念珠菌对伊曲康唑的药物敏感性[J].福建医药杂志,2013,35(4):75-77.
[13]李昕,赵月,苏昕.32味中药水提物对4种真菌抑菌实验研究[J].辽宁中医药大学学报,2015(3):36-39.
[14]陈秋荣.黄酮类化合物药理作用的分析[J].中国实用医药,2012,07(21):254-255.
[15]林塬,陈静.常用民族药植物中酚酸类化合物的提取、纯化及生物活性研究进展[J].中国民族医药杂志,2014,20(9):39-41.
[16]Sharififar F,Moshafi M H,Mansouri S H,et al.In vitro evaluation of antibacterial and antioxidant activities of the essential oil and methanol extract of endemic Zataria multiflora,Boiss[J].Food Control,2007,18(7):800-805.
[17]Cushnie T P,Lamb A J.Assessment of the antibacterial activity of galangin against 4-quinolone resistant strains of Staphylococcus aureus.[J].Phytomedicine,2006,13(3):187-191.
[18]孙雷雷,陈毓,张丽,等.小蓟及小蓟炭止血作用机理的研究概况[J].时珍国医国药,2005,16(12):1304-1305.
[2]魏克强,杨俊仙,魏治中.新型烟草化学成分及医药成分的初步分析[J].天然产物研究与开发,2008,20(4):652-656.
[3]李莹,李钟,郭培国.烟草中主要生物活性成分的研究进展[J].天然产物研究与开发,2015(12):2157-2163.
[4]许晓风,郁锋,杨启银,等.烟草工业下脚料综合利用研究(I)--抗菌成分的提取与抑菌效果测定[J].安徽农业科学,2003,31(2):209-209.
[5]隋婧.青蒿体外止血活性部位筛选及其应用研究[D].重庆大学,2009.
[6]徐叔云,卞如濂,陈修.药理实验方法学[M].北京:人民卫生出版社,1991.1016-1018.
[7]黄艳,宁小清,原鲜玲,等.壮药白花九里明水煎液止血作用机制探讨[J].世界科学技术-中医药现代化,2013(7):1552-1556.
[8]海丽娜,张志伟,王金华,等.复方苦参注射液对小鼠的镇痛止血和抗应激作用[J].中国实验方剂学杂志,2012,18(2):199-202.
[9]王秋红,刘玉婕,吕邵娃,等.线叶菊抗菌作用研究[J].中国实验方剂学杂志,2011,17(22):141-145.
[10]Noake T,Kuriyama T,White P L,et al.Antifungal susceptibility of Candida species using the Clinical and Laboratory Standards Institute disk diffusion and broth microdilution methods.[J].Journal of Chemotherapy,2007,19(3):283-7.
[11]朱庆玲,冯翀.五爪龙不同提取物体外抗菌活性的研究[J].中国实验方剂学杂志,2011,17(11):232-233
[12]张伟芳,侯喜琴.NCCLSM27-A微量法测定白色念珠菌对伊曲康唑的药物敏感性[J].福建医药杂志,2013,35(4):75-77.
[13]李昕,赵月,苏昕.32味中药水提物对4种真菌抑菌实验研究[J].辽宁中医药大学学报,2015(3):36-39.
[14]陈秋荣.黄酮类化合物药理作用的分析[J].中国实用医药,2012,07(21):254-255.
[15]林塬,陈静.常用民族药植物中酚酸类化合物的提取、纯化及生物活性研究进展[J].中国民族医药杂志,2014,20(9):39-41.
[16]Sharififar F,Moshafi M H,Mansouri S H,et al.In vitro evaluation of antibacterial and antioxidant activities of the essential oil and methanol extract of endemic Zataria multiflora,Boiss[J].Food Control,2007,18(7):800-805.
[17]Cushnie T P,Lamb A J.Assessment of the antibacterial activity of galangin against 4-quinolone resistant strains of Staphylococcus aureus.[J].Phytomedicine,2006,13(3):187-191.
[18]孙雷雷,陈毓,张丽,等.小蓟及小蓟炭止血作用机理的研究概况[J].时珍国医国药,2005,16(12):1304-1305.