中药小蓟不同药用部位体外抗炎、促凝血的作用研究
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摘要
目的:研究小蓟不同部位化学成分的差异与抗炎、促凝血病症疗效关系,研究生药不同部位的精准应用。方法:使用HPLC色谱法建立不同部位的指纹图谱,MTT法测定不同部位提取物对炎症细胞RAW264.7增值的抑制作用,ELISA法测定不同部位提取物对小鼠凝血酶原(PT)指标的影响,并使用灰色关联软件,将色谱峰的峰面积与抗炎、凝血酶原指标相关联,得到各个成分的贡献率。结果:小蓟根、茎、叶、花中的成分种类与含量均有不同。在体外抗炎药效研究中,小蓟花的效果最好,对炎症细胞RAW264.7的抑制率达到63.25%。小蓟根次之,小蓟叶与茎对炎症细胞的抑制作用不大。用ELISA法测定小鼠血浆中凝血酶原指标,小蓟花对指标影响最大,叶次之,茎与根对指标变化影响不大。灰色关联度计算软件结果显示,绿原酸、橙皮苷、咖啡酸、蒙花苷对抑制炎症细胞增值(抗炎)的相关程度分别为0.7250,0.5249,0.5105,0.5096。与升高凝血酶原指标(止血)的相关程度分别为0.5982,0.5088,0.5037,0.5034。其关联程度与其他成分相比,差距较大。结论:小蓟不同部位和抗炎、促凝血病症的药效存在差异,明确了生药小蓟不同部位的精准应用。
Objective: To study the difference of chemical constituents and pesticide effects of anti-inflammation and promote coagulation in different parts of Cirsium setosum, and set up the HPLC fingerprint of different parts of Cirsium setosum. Methods: The MTT assay was used to test the inhibitory action to the RAW264.7 cells of different parts of Cirsium setosum and use the ELISA assay to measure the effects of extracts from different parts on prothrombin(PT) in mice. Then use the GreyModeling V3.0 software to calculate the correlation coefficients of peak area of chromatographic peaks and the pesticide effects. Results: Different species and contents of components in the roots, stems, leaves, and flowers are different in Cirsium setosum. The flowers of Cirsium setosum had the best effects on inhibition of RAW 264.7 cells which the inhibition rate was 63.25%. The leaves of Cirsium setosum ranked second. The prothrombin in mice was determined by ELASA assay. The flowers of Cirsium setosum had the greatest impact on the indicator. The roots of Cirsium setosum ranked second. The results of GreyModeling software shows that the chlorogenic acid, hesperidin, caffeic acid, linarin have greater degrees of contribution on anti-inflammation and promote coagulation which compared with others. The correlation coefficients of four components on anti-inflammatory were 0.7250, 0.5249, 0.5105 and 0.5096, on coagulant effects were 0.5982, 0.5088, 0.5037, and 0.5034. Conclusion: Different groups of components in different parts of Cirsium setosum may prove to be the major factor to the different pesticide effects.
Objective: To study the difference of chemical constituents and pesticide effects of anti-inflammation and promote coagulation in different parts of Cirsium setosum, and set up the HPLC fingerprint of different parts of Cirsium setosum. Methods: The MTT assay was used to test the inhibitory action to the RAW264.7 cells of different parts of Cirsium setosum and use the ELISA assay to measure the effects of extracts from different parts on prothrombin(PT) in mice. Then use the GreyModeling V3.0 software to calculate the correlation coefficients of peak area of chromatographic peaks and the pesticide effects. Results: Different species and contents of components in the roots, stems, leaves, and flowers are different in Cirsium setosum. The flowers of Cirsium setosum had the best effects on inhibition of RAW 264.7 cells which the inhibition rate was 63.25%. The leaves of Cirsium setosum ranked second. The prothrombin in mice was determined by ELASA assay. The flowers of Cirsium setosum had the greatest impact on the indicator. The roots of Cirsium setosum ranked second. The results of GreyModeling software shows that the chlorogenic acid, hesperidin, caffeic acid, linarin have greater degrees of contribution on anti-inflammation and promote coagulation which compared with others. The correlation coefficients of four components on anti-inflammatory were 0.7250, 0.5249, 0.5105 and 0.5096, on coagulant effects were 0.5982, 0.5088, 0.5037, and 0.5034. Conclusion: Different groups of components in different parts of Cirsium setosum may prove to be the major factor to the different pesticide effects.
引文
1中国药典编写委员会.中华人民共和国药典一部.北京:中国医药科技出版社, 2015.
2 宋·苏颂编撰,尚志钩辑校.本草图经.合肥:安徽科学技术出版社,1994.
3 宋·赵佶.圣济总录.北京:人民卫生出版社, 2013.
4 杨柄友,杨春丽,刘艳,等.小蓟的研究进展.中草药. 2017, 48(23):5039-5048.
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6 柯睿,朱恩圆,侴桂新.小蓟中一个新的苯丙素苷类化合物.药学学报, 2010, 45(7):879-882.
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9 李璐,罗建光,孔令义.小蓟炒炭前后蒙花苷的含量测定.中国现代中药, 2010, 12(5):23-25.
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11 许浚,张铁军,龚苏晓,等.小蓟止血活性部位的化学成分研究.中草药, 2010, 41(4):542-544.
12 冯子明,杨桠楠,姜建双,等.小蓟的化学成分研究.中国实验方剂学杂志, 2012, 18(6):87-89.
13 王鹤辰,樊佳新,孟宪生,等.基于灰色关联法研究胆矾中元素对A549细胞的体外抑制作用.中国现代应用药学, 2017, 34(11):1522-1525.
14 刘昌孝,陈士林,肖小河,等.中药质量标志物(Q-Marker):中药产品质量控制的新概念.中草药, 2016, 47(09):1443-1457.
15 Wang H, Li T, Bao Y, et al. A multifunctional integrated simultaneously online screening microfluidic biochip for the examination of“efficacytoxicity”and compatibility of medicine. Chinese Chemical Letters, 2019,30(2):403-405.
16 Coussens L M, Werb Z. Inflammation and cancer. Nature, 2002, 420(6917):860.
17 杨晓丽,张君利,王京峰,等.绿原酸抗肿瘤作用及机制研究进展.中国实验方剂学杂志, 2018, 24(19):229-234
18 夏荣木,陆红玲.橙皮苷抗肿瘤活性研究进展.重庆医学, 2018, 47(21):2846-2849.
19 包伊凡,沈新春,汪芳.咖啡酸及其主要衍生物的研究进展及开发前景.天然产物研究与开发, 2018:1-15.
20 吴亚军,苏洁,黄浦俊,等.蒙花苷对TNF-α诱导的血管内皮细胞炎症损伤及TLR4/IκBα/NF-κB信号通路的影响.中国现代应用药学,2017, 34(5):637-643.
21 刘昌孝.精准药学:从转化医学到精准医学探讨新药发展.药物评价研究, 2016, 39(1):1-18.
22 罗国安,谢媛媛,梁琼麟,等.中医药整合医学—三论创建新医药学.世界科学技术-中医药现代化, 2015, 17(1):7-15.
23 罗国安,谢媛媛,王义明,等.精准医学与中医药现代化研究——五论创建新医药学.世界科学技术-中医药现代化, 2017, 19(1):19-29.
2 宋·苏颂编撰,尚志钩辑校.本草图经.合肥:安徽科学技术出版社,1994.
3 宋·赵佶.圣济总录.北京:人民卫生出版社, 2013.
4 杨柄友,杨春丽,刘艳,等.小蓟的研究进展.中草药. 2017, 48(23):5039-5048.
5 顾玉诚,屠呦呦.小蓟化学成分研究.中国中药杂志. 1992, 17(9):547-548.
6 柯睿,朱恩圆,侴桂新.小蓟中一个新的苯丙素苷类化合物.药学学报, 2010, 45(7):879-882.
7 孟永海,王秋红,姜海,等.小蓟化学成分研究.中药材, 2009, 32(1):58-61.
8 Sun Q, Chang L, Ren Y, et al. Simultaneous analysis of 11 main active components in Cirsium setosum based on HPLC-ESI-MS/MS and combined with statistical methods. J Sep Sci. 2012, 35(6):2897-2907.
9 李璐,罗建光,孔令义.小蓟炒炭前后蒙花苷的含量测定.中国现代中药, 2010, 12(5):23-25.
10 樊佳新,王帅,孟宪生,等.基于微流控芯片技术的荆芥诱导肺肿瘤细胞凋亡谱效关系研究.药学学报, 2017, 52(1):126-131.
11 许浚,张铁军,龚苏晓,等.小蓟止血活性部位的化学成分研究.中草药, 2010, 41(4):542-544.
12 冯子明,杨桠楠,姜建双,等.小蓟的化学成分研究.中国实验方剂学杂志, 2012, 18(6):87-89.
13 王鹤辰,樊佳新,孟宪生,等.基于灰色关联法研究胆矾中元素对A549细胞的体外抑制作用.中国现代应用药学, 2017, 34(11):1522-1525.
14 刘昌孝,陈士林,肖小河,等.中药质量标志物(Q-Marker):中药产品质量控制的新概念.中草药, 2016, 47(09):1443-1457.
15 Wang H, Li T, Bao Y, et al. A multifunctional integrated simultaneously online screening microfluidic biochip for the examination of“efficacytoxicity”and compatibility of medicine. Chinese Chemical Letters, 2019,30(2):403-405.
16 Coussens L M, Werb Z. Inflammation and cancer. Nature, 2002, 420(6917):860.
17 杨晓丽,张君利,王京峰,等.绿原酸抗肿瘤作用及机制研究进展.中国实验方剂学杂志, 2018, 24(19):229-234
18 夏荣木,陆红玲.橙皮苷抗肿瘤活性研究进展.重庆医学, 2018, 47(21):2846-2849.
19 包伊凡,沈新春,汪芳.咖啡酸及其主要衍生物的研究进展及开发前景.天然产物研究与开发, 2018:1-15.
20 吴亚军,苏洁,黄浦俊,等.蒙花苷对TNF-α诱导的血管内皮细胞炎症损伤及TLR4/IκBα/NF-κB信号通路的影响.中国现代应用药学,2017, 34(5):637-643.
21 刘昌孝.精准药学:从转化医学到精准医学探讨新药发展.药物评价研究, 2016, 39(1):1-18.
22 罗国安,谢媛媛,梁琼麟,等.中医药整合医学—三论创建新医药学.世界科学技术-中医药现代化, 2015, 17(1):7-15.
23 罗国安,谢媛媛,王义明,等.精准医学与中医药现代化研究——五论创建新医药学.世界科学技术-中医药现代化, 2017, 19(1):19-29.