Article(id=1208491499709382865, tenantId=1146029695717560320, journalId=1189982191388893191, issueId=1208491462300385385, articleNumber=null, orderNo=null, doi=10.16438/j.0513-4870.2021-0676, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1620403200000, receivedDateStr=2021-05-08, revisedDate=1622044800000, revisedDateStr=2021-05-27, acceptedDate=null, acceptedDateStr=null, onlineDate=1766056426216, onlineDateStr=2025-12-18, pubDate=1628697600000, pubDateStr=2021-08-12, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1766056426216, onlineIssueDateStr=2025-12-18, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1766056426216, creator=13701087609, updateTime=1766056426216, updator=13701087609, issue=Issue{id=1208491462300385385, tenantId=1146029695717560320, journalId=1189982191388893191, year='2021', volume='56', issue='8', pageStart='2039', pageEnd='2324', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1766056417298, creator=13701087609, updateTime=1766137099178, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1208829866691130129, tenantId=1146029695717560320, journalId=1189982191388893191, issueId=1208491462300385385, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1208829866691130130, tenantId=1146029695717560320, journalId=1189982191388893191, issueId=1208491462300385385, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=2093, endPage=2101, ext={EN=ArticleExt(id=1208491501575848254, articleId=1208491499709382865, tenantId=1146029695717560320, journalId=1189982191388893191, language=EN, title=The effect of Moutan cortex carbonisata nano-components on the blood-cooling and hemostatic, columnId=1208491496576238458, journalTitle=Acta Pharmaceutica Sinica, columnName=Special Reports: Structural Chinese Medicine, runingTitle=null, highlight=null, articleAbstract=

At present, the research of Moutan cortex carbonisata (MCC) mainly focuses on the changes of chemical composition before and after charcoal production, and there is a lack of material basic research directly related to the efficacy at home and abroad. In this study, Moutan cortex, as a precursor, and was calcined to MCC at high temperature. The Moutan cortex carbonisata nano-components (MCC-NCs) were extracted and separated from MCC to explore its cooling-blood and hemostatic effects. In the experiment, the MCC was calcined at a high temperature in a muffle furnace (350℃, 1 h), and then MCC-NCs were extracted for MCC, and characterized by transmission electron microscopy and UV-vis absorption spectroscopy, fluorescence spectroscopy, Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy. In addition, the study evaluated the bloodcooling and hemostatic effects of MCC-NCs. The results showed that MCC-NCs have a size distribution of 0.80-2.8 nm, a lattice spacing of 0.26 nm. MCC-NCs are mainly composed of C, O and N elements and have abundant surface functional groups such as OH, C=O, C-N and C=C. The fluorescence yield of MCC-NCs was 7.18%. The experiments complied with the Animal Ethics Committee of Beijing University of Chinese Medicine. The result indicated that pretreatment MCC-NCs can significantly (P < 0.05) reduce the high, medium, and low viscosity of whole blood and plasma viscosity, and reduce hematocrit, red blood cell distribution width, hemoglobin and red blood cell level. In addition, MCC-NCs significantly reduced the levels of activated partial thromboplastin time, thrombin time and fibrinogen (P < 0.05). The pathological examination results showed that MCC-NCs can significantly reduce lung tissue damage, reduce bleeding and inflammatory cell infiltration. At the same time, it can also significantly reduce the symptoms of gastric mucosal bleeding. In conclusion, the results indicated that MCC-NCs has significantly the effect of blood cooling and hemostasis, and its hemostatic effect is mainly related to the activation of endogenous coagulation pathway or fibrinogen system, which provided a novel strategy for exploring the material basis of traditional Chinese medicine for hemostasis.

, correspAuthors=Hui KONG, Yan ZHAO, authorNote=null, correspAuthorsNote=null, copyrightStatement=Copyright ©2021 Acta Pharmaceutica Sinica. All rights reserved., copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, authorCompany=null, fund=null, authors=null, authorsList=Juan LUO, Jie HU, Yu-han LIU, Xue BAI, Yu-min CHEN, Yu-ru LI, Ya-fang ZHAO, Hui KONG, Yan ZHAO, Hui-hua QU), CN=ArticleExt(id=1208491503727526409, articleId=1208491499709382865, tenantId=1146029695717560320, journalId=1189982191388893191, language=CN, title=丹皮炭纳米类成分的凉血止血作用研究, columnId=1208491496861451165, journalTitle=药学学报, columnName=专题报道:结构中药学, runingTitle=null, highlight=null, articleAbstract=

目前国内外对于丹皮炭的研究主要集中在制炭前后化学成分的变化,缺乏与药效直接关联的物质基础研究。本研究以丹皮为前驱体,高温烧制成丹皮炭,从丹皮炭中提取分离出丹皮炭纳米类成分(Moutan cortex carbonisatanano-components,MCC-NCs),并探究其凉血止血作用。实验通过马弗炉(350℃,1 h)高温烧制丹皮炭,再从中提取分离出MCC-NCs,并利用透射电子显微镜、紫外-可见吸收光谱、荧光光谱、傅里叶变换红外光谱和X射线光电子能谱对其进行了表征;此外,该研究评估了MCC-NCs的凉血止血作用。结果显示,MCC-NCs的尺寸分布范围为0.80~2.8 nm,晶格间距为0.26 nm,主要由C、O和N元素组成,具有OH、C=O、C-N、C=C等丰富的表面官能团,荧光量子产率为7.18%。所有动物实验均符合北京中医药大学伦理委员会的规定。结果表明,预先给予MCC-NCs可显著降低全血高、中、低黏度和血浆黏度;降低红细胞压积和红细胞分布宽度,减少血红蛋白含量和红细胞计数(P < 0.05);此外,MCC-NCs可以显著降低活化部分凝血活酶时间、凝血酶时间和纤维蛋白原水平(P < 0.05)。病理学检查结果发现,MCC-NCs可以明显减轻肺组织损伤,减小出血和炎性细胞浸润;同时,也可以明显减轻胃黏膜出血症状。综上,实验结果显示MCC-NCs具有凉血止血的作用,其止血作用主要与内源性凝血途径或纤维蛋白原系统的激活有关,为探索中药止血的物质基础提供了新的策略。

, correspAuthors=孔慧, 赵琰, authorNote=null, correspAuthorsNote=
*孔慧, Tel: 86-10-64286705, E-mail: ;
赵琰, E-mail:
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J Ethnopharmacol, 2011, 135: 510-514, articleTitle=Dragon's Blood extract has antithrombotic properties, affecting platelet aggregation functions and anticoagulation activities, refAbstract=null)], funds=[Fund(id=1208491514821460238, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1208491499709382865, awardId=2019-JYB-TD-001, language=CN, fundingSource=中央高校基本科研业务费专项资金资助项目(2019-JYB-TD-001), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1208491504004350504, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1208491499709382865, xref=null, ext=[AuthorCompanyExt(id=1208491504008544809, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1208491499709382865, companyId=1208491504004350504, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1. 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Beijing Institute of Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing 100029, China), AuthorCompanyExt(id=1208491505560437335, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1208491499709382865, companyId=1208491505522688592, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=3.北京中医药大学中医药研究院, 北京 100029)])], figs=[ArticleFig(id=1208491512099356810, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1208491499709382865, language=EN, label=null, caption=null, figureFileSmall=01YEKhGzJOhOfmhCl73MBg==, figureFileBig=HOeJ+x4oWgFA7QIPyrT5iA==, tableContent=null), ArticleFig(id=1208491512195825811, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1208491499709382865, language=CN, label=Figure 1, caption= Illustration of the formation process of nano-components from <i>Moutan cortex</i> (MC) by high temperature calcinations , figureFileSmall=01YEKhGzJOhOfmhCl73MBg==, figureFileBig=HOeJ+x4oWgFA7QIPyrT5iA==, tableContent=null), ArticleFig(id=1208491512451678373, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1208491499709382865, language=EN, label=null, caption=null, figureFileSmall=4Iq97kmHDcTMKJimHopP8Q==, figureFileBig=djjHIYPWomWF7ZLdnZsUeg==, tableContent=null), ArticleFig(id=1208491512560730285, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1208491499709382865, language=CN, label=Figure 2, caption= Morphological characterization of <i>Moutan cortex</i> carbonisata nano-components (MCC-NCs). A: Transmission electron microscopy (TEM) images of MCC-NCs displaying ultra-small particles; B: TEM size distribution of MCC-NCs; C: High-resolution TEM image of MCC-NCs; D: UV-vis spectrum of MCC-NCs; E: Excitation and emission fluorescence spectra; F: Fourier transform infrared spectroscopy (FTIR) spectra of MCC-NCs , figureFileSmall=4Iq97kmHDcTMKJimHopP8Q==, figureFileBig=djjHIYPWomWF7ZLdnZsUeg==, tableContent=null), ArticleFig(id=1208491512699142329, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1208491499709382865, language=EN, label=null, caption=null, figureFileSmall=rv6OZC8Z7PMnyHxDR//ZgA==, figureFileBig=JXidsBDxUIlr4id++Nf0vA==, tableContent=null), ArticleFig(id=1208491512778834114, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1208491499709382865, language=CN, label=Figure 3, caption= X-ray photoelectron spectroscopy (XPS) survey spectrum of MCC-NCs (A). XPS spectrum of C1s region (B), O1s region (C), and N1s region (D) , figureFileSmall=rv6OZC8Z7PMnyHxDR//ZgA==, figureFileBig=JXidsBDxUIlr4id++Nf0vA==, tableContent=null), ArticleFig(id=1208491512913051853, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1208491499709382865, language=EN, label=null, caption=null, figureFileSmall=ie9WLTlXt2gvIcxN6/QG4Q==, figureFileBig=QZ1LiNO8jCOk3P8SW8jbYw==, tableContent=null), ArticleFig(id=1208491514133594322, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1208491499709382865, language=CN, label=Figure 4, caption= Whole blood viscosity (A). Blood plasma viscosity (B). 1, 5, 30, 200: Whole blood viscosity low cut (1, 5 s), medium cut (30 s) and high cut (200 s); Positive: Yunnan Baiyao powder; H: High dose of MCC-NCs groups (1.25 g·kg<sup>-1</sup>); M: Medium dose of MCC-NCs groups (0.63 k·kg<sup>-1</sup>); L: Low of MCC-NCs groups (0.31 g·kg<sup>-1</sup>). <i>n</i> = 10, <i><span class="mag-xml-overline" style="border-top:1px solid black">x</span></i>± <i>s</i>. <sup>#</sup><i>P</i> < 0.05, <sup>##</sup>P < 0.01 <i>vs</i> blank group; <sup>*</sup><i>P</i> < 0.05, <sup>**</sup><i>P</i> < 0.01 <i>vs</i> model group , figureFileSmall=ie9WLTlXt2gvIcxN6/QG4Q==, figureFileBig=QZ1LiNO8jCOk3P8SW8jbYw==, tableContent=null), ArticleFig(id=1208491514234257630, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1208491499709382865, language=EN, label=null, caption=null, figureFileSmall=+i3J0dt/u0LnFC6GwQeH/g==, figureFileBig=eb2FS322tVhuuFMGPKnc+g==, tableContent=null), ArticleFig(id=1208491514318143716, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1208491499709382865, language=CN, label=Figure 5, caption= Blood parameters. A: Hematocrit (HCT); B: Red blood cell distribution width (RDW); C: Hemoglobin (Hb); D: Red blood cell (RBC). <i>n</i> = 10, <i><span class="mag-xml-overline" style="border-top:1px solid black">x</span></i> ± <i>s</i>. <sup>#</sup><i>P</i> < 0.05, <sup>##</sup><i>P</i> < 0.01 <i>vs</i> blank group; <sup>*</sup><i>P</i> < 0.05 <i>vs</i> model group , figureFileSmall=+i3J0dt/u0LnFC6GwQeH/g==, figureFileBig=eb2FS322tVhuuFMGPKnc+g==, tableContent=null), ArticleFig(id=1208491514431389932, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1208491499709382865, language=EN, label=null, caption=null, figureFileSmall=vhXUtu67aYdcm8uc5eCDUw==, figureFileBig=qvDiJK6ST5PPgl3ez0i8mg==, tableContent=null), ArticleFig(id=1208491514527858934, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1208491499709382865, language=CN, label=Figure 6, caption= Effects on coagulation parameters. A: Activated partial thromboplastin time (APTT); B: Thrombin time (TT); C: Prothrombin time (PT); D: Fibrinogen (FIB). <i>n</i> = 10, <i><span class="mag-xml-overline" style="border-top:1px solid black">x</span></i> ± <i>s</i>. <sup>#</sup><i>P</i> < 0.05, <sup>##</sup><i>P</i> < 0.01 <i>vs</i> blank group; <sup>*</sup><i>P</i> < 0.05, <sup>**</sup><i>P</i> < 0.01 <i>vs</i> model group , figureFileSmall=vhXUtu67aYdcm8uc5eCDUw==, figureFileBig=qvDiJK6ST5PPgl3ez0i8mg==, tableContent=null), ArticleFig(id=1208491514603356415, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1208491499709382865, language=EN, label=null, caption=null, figureFileSmall=NffiRbY0zqahsPV/mIdxqg==, figureFileBig=Pawx0FeeBmDUpX7+U5Rbeg==, tableContent=null), ArticleFig(id=1208491514699825415, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1208491499709382865, language=CN, label=Figure 7, caption= The macrogram and histopathological examination of lung (HE, ×100) and stomach (HE, ×400) of rats. A: Blank group; B: Model group; C: Positive group; D: MCC-NCs high-dose group; E: MCC-NCs medium-dose group; F: MCC-NCs low-dose group (#: Mucosal hemorrhagic). 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丹皮炭纳米类成分的凉血止血作用研究
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罗娟 1 , 胡杰 1 , 刘育含 1 , 白雪 1 , 陈玉民 1 , 李郁茹 2 , 赵亚芳 1 , 孔慧 1, * , 赵琰 1, * , 屈会化 3
药学学报 | 专题报道:结构中药学 2021,56(8): 2093-2101
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药学学报 | 专题报道:结构中药学 2021, 56(8): 2093-2101
丹皮炭纳米类成分的凉血止血作用研究
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罗娟1, 胡杰1, 刘育含1, 白雪1, 陈玉民1, 李郁茹2, 赵亚芳1, 孔慧1, * , 赵琰1, * , 屈会化3
作者信息
  • 1.北京中医药大学中医学院, 北京 100029
  • 2.北京中医药大学中药学院, 北京 100029
  • 3.北京中医药大学中医药研究院, 北京 100029

通讯作者:

*孔慧, Tel: 86-10-64286705, E-mail: ;
赵琰, E-mail:
The effect of Moutan cortex carbonisata nano-components on the blood-cooling and hemostatic
Juan LUO1, Jie HU1, Yu-han LIU1, Xue BAI1, Yu-min CHEN1, Yu-ru LI2, Ya-fang ZHAO1, Hui KONG1, * , Yan ZHAO1, * , Hui-hua QU3
Affiliations
  • 1. School of Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing 100029, China
  • 2. School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 100029, China
  • 3. Beijing Institute of Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing 100029, China
出版时间: 2021-08-12 doi: 10.16438/j.0513-4870.2021-0676
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目前国内外对于丹皮炭的研究主要集中在制炭前后化学成分的变化,缺乏与药效直接关联的物质基础研究。本研究以丹皮为前驱体,高温烧制成丹皮炭,从丹皮炭中提取分离出丹皮炭纳米类成分(Moutan cortex carbonisatanano-components,MCC-NCs),并探究其凉血止血作用。实验通过马弗炉(350℃,1 h)高温烧制丹皮炭,再从中提取分离出MCC-NCs,并利用透射电子显微镜、紫外-可见吸收光谱、荧光光谱、傅里叶变换红外光谱和X射线光电子能谱对其进行了表征;此外,该研究评估了MCC-NCs的凉血止血作用。结果显示,MCC-NCs的尺寸分布范围为0.80~2.8 nm,晶格间距为0.26 nm,主要由C、O和N元素组成,具有OH、C=O、C-N、C=C等丰富的表面官能团,荧光量子产率为7.18%。所有动物实验均符合北京中医药大学伦理委员会的规定。结果表明,预先给予MCC-NCs可显著降低全血高、中、低黏度和血浆黏度;降低红细胞压积和红细胞分布宽度,减少血红蛋白含量和红细胞计数(P < 0.05);此外,MCC-NCs可以显著降低活化部分凝血活酶时间、凝血酶时间和纤维蛋白原水平(P < 0.05)。病理学检查结果发现,MCC-NCs可以明显减轻肺组织损伤,减小出血和炎性细胞浸润;同时,也可以明显减轻胃黏膜出血症状。综上,实验结果显示MCC-NCs具有凉血止血的作用,其止血作用主要与内源性凝血途径或纤维蛋白原系统的激活有关,为探索中药止血的物质基础提供了新的策略。

丹皮炭  /  纳米类成分  /  凉血止血  /  物质基础  /  止血机制

At present, the research of Moutan cortex carbonisata (MCC) mainly focuses on the changes of chemical composition before and after charcoal production, and there is a lack of material basic research directly related to the efficacy at home and abroad. In this study, Moutan cortex, as a precursor, and was calcined to MCC at high temperature. The Moutan cortex carbonisata nano-components (MCC-NCs) were extracted and separated from MCC to explore its cooling-blood and hemostatic effects. In the experiment, the MCC was calcined at a high temperature in a muffle furnace (350℃, 1 h), and then MCC-NCs were extracted for MCC, and characterized by transmission electron microscopy and UV-vis absorption spectroscopy, fluorescence spectroscopy, Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy. In addition, the study evaluated the bloodcooling and hemostatic effects of MCC-NCs. The results showed that MCC-NCs have a size distribution of 0.80-2.8 nm, a lattice spacing of 0.26 nm. MCC-NCs are mainly composed of C, O and N elements and have abundant surface functional groups such as OH, C=O, C-N and C=C. The fluorescence yield of MCC-NCs was 7.18%. The experiments complied with the Animal Ethics Committee of Beijing University of Chinese Medicine. The result indicated that pretreatment MCC-NCs can significantly (P < 0.05) reduce the high, medium, and low viscosity of whole blood and plasma viscosity, and reduce hematocrit, red blood cell distribution width, hemoglobin and red blood cell level. In addition, MCC-NCs significantly reduced the levels of activated partial thromboplastin time, thrombin time and fibrinogen (P < 0.05). The pathological examination results showed that MCC-NCs can significantly reduce lung tissue damage, reduce bleeding and inflammatory cell infiltration. At the same time, it can also significantly reduce the symptoms of gastric mucosal bleeding. In conclusion, the results indicated that MCC-NCs has significantly the effect of blood cooling and hemostasis, and its hemostatic effect is mainly related to the activation of endogenous coagulation pathway or fibrinogen system, which provided a novel strategy for exploring the material basis of traditional Chinese medicine for hemostasis.

Moutan cortex carbonisata  /  nano-component  /  blood-cooling and hemostatic effect  /  material basis  /  hemostatic mechanism
罗娟, 胡杰, 刘育含, 白雪, 陈玉民, 李郁茹, 赵亚芳, 孔慧, 赵琰, 屈会化. 丹皮炭纳米类成分的凉血止血作用研究. 药学学报, 2021 , 56 (8) : 2093 -2101 . DOI: 10.16438/j.0513-4870.2021-0676
Juan LUO, Jie HU, Yu-han LIU, Xue BAI, Yu-min CHEN, Yu-ru LI, Ya-fang ZHAO, Hui KONG, Yan ZHAO, Hui-hua QU. The effect of Moutan cortex carbonisata nano-components on the blood-cooling and hemostatic[J]. Acta Pharmaceutica Sinica, 2021 , 56 (8) : 2093 -2101 . DOI: 10.16438/j.0513-4870.2021-0676
丹皮(Moutan cortex, MC) 是毛茛科植物牡丹Paeonia suffruticosa Andr.的皮, 具有清热凉血、活血化瘀功效, 主要用于治疗热入营血、吐血衄血等; 丹皮炭是丹皮的制炭炮制品, 始见于元代的《十药神书》载:“烧灰存性, 研极细末, 用纸包, 碗盖于地上一夕, 出火毒”。丹皮炭归心、肝、肾经, 具有凉血止血功效, 常用于血热出血。经过炮制后的炭药可增强或出现止血作用, 目前对其止血机制的研究主要集中在炭素、小分子、鞣质和钙离子等物质基础的变化; Li等[1]研究显示鞣质和乙酸乙酯部分是丹皮炭中止血作用的主要有效部分; Zhu等[2]研究显示丹皮酚的含量降低, 丹皮酚主要用于活血作用, 从而可能增加了丹皮炭的止血作用。目前国内外对于丹皮炭的研究主要集中在制炭前后化学成分的变化[3, 4], 缺乏与药效直接关联的物质基础研究。
中药炭药始载于《五十二病方》, 历经千年而不衰, 具有明确的疗效。如荆芥炭(茎类)、荷叶炭(叶类)、蒲黄炭(花粉类)、黄柏炭(皮类) 和地榆炭(根类) 等不同基源, 制炭之后虽然在止血作用强弱上存在差异, 作用于人体内的部位有偏向, 如吐血(胃、肺)、便血(直肠) 和衄血(鼻) 等, 但是几乎所有炭药都显现出趋同的止血作用, 即所谓的炒炭止血[5, 6]。这种神奇的“趋同作用”, 促使研究者将目光着眼于这些炭药的共性之处——“高温炭化”制备过程。在前期实验研究中发现, 经高温炭化后的炭药中存在一种粒径小于10 nm的纳米颗粒, 如枳实炭[7], 姜炭[8]、山楂炭[9]和黄柏炭[10]等; 这种纳米颗粒的粒径大小、光学特性和表面基团等性质是与纳米材料中“碳点”相类似的新物质。因此, 本团队将这一类物质命名为“纳米类成分(nano-components)”。
本实验通过从丹皮炭中提取纯化出了丹皮炭纳米类成分(Moutan cortex carbonisata nano-components, MCC-NCs), 利用透射电子显微镜(transmission electron microscopy, TEM)、紫外- 可见吸收光谱(UV-vis absorption, UV-vis)、荧光光谱(fluorescence spectroscopy, FL)、傅里叶变换红外光谱(Fourier transform infrared spectroscopy, FTIR) 和X射线光电子能谱(X-ray pho‐ toelectron spectroscopy, XPS) 对其粒径、结构、光学特征、元素组成和表面官能团等理化性质进行了全面的表征。此外, 利用动物血热出血模型评估了MCC-NCs的凉血止血作用。
药品与试剂药材丹皮  (北京仟草药业有限公司); 云南白药粉(云南白药集团股份有限公司); 干酵母粉(安琪酵母股份有限公司); 无水乙醇(北京化工公司)。实验所用均为去离子水。
仪器  马弗炉(北京中科博科技有限公司); 水浴锅(金坛市华元开城实验仪器厂); 过滤器、微孔滤膜(天津市津腾实验设备有限公司); 透析袋(北京瑞达恒辉科技发展有限公司); Tecnai G2 20透射电子显微镜(美国FEI公司); 红外光谱仪、X射线光电子能谱仪(Escalab 250Xi) (赛默飞科技有限公司); ZL6000全自动血流变分析仪(北京众驰伟业公司); CA-550全自动凝血分析仪、XI-800血液分析仪(日本希斯美康有限公司)。
MCC-NCs的制备  称量丹皮药材240.0 g放置在坩埚中, 用锡箔纸包裹后放入马弗炉中, 设置煅烧程序为: 第一阶段70 ℃保持25 min, 第二阶段350 ℃保持60 min。自然降温至室温, 取出, 粉碎成粉末。称取粉末60.0 g, 加水2 L, 煎煮3次, 滤过, 滤液合并, 浓缩, 微孔滤膜滤过, 滤液置透析袋内, 透析7天(每12 h换水1次), 收集透析后溶液, 质量浓度为1 g·mL-1 (以丹皮炭计), 保存在4 ℃待用, 制备过程中所用水均为去离子水, 制备过程如图 1所示。
MCC-NCs的表征  利用TEM分析MCC-NCs的形貌特征及粒径大小分布: 用去离子水稀释MCC-NCs, 以0.22 μm的微孔滤膜滤过, 并超声2 h后将样品点在铜网上, 室温阴干。利用UV-vis和FL分析MCC-NCs的光学特征, 主要包括样品的紫外吸收光谱、最大激发波长(λEX)、最大发射波长(λEM) 及荧光量子产率。采用XPS测试分析样本的元素组成和元素配位情况。将不同条件制备的纳米类成分的水溶液冷冻干燥获得粉末, 以固体粉末进行XPS分析, 表征纳米类成分的元素组成和元素配位情况。采用FTIR测试材料的红外吸收光谱, 分析样本的表面基团。将不同条件制备的纳米类成分溶液烘干, 研磨成细末, 于红外灯下将粉末按照1∶100的比例与存放于干燥箱中的溴化钾粉末混合均匀, 压片, 用傅立叶变换红外光谱仪对其进行扫描, 表征纳米类成分的表面基团。
相对荧光量子产率以硫酸奎宁作为参比, 其是一个已知量子产率的标准物质。将MCC-NCs与硫酸奎宁溶液(折射率为1.33) 稀释成适宜浓度, 于相同激发波长下扫描获取两种物质的发射荧光光谱, 分别计算其荧光光谱的积分面积。根据公式(1) 计算MCC-NCs的量子产率:
$\begin{array}{*{20}{l}}{{Q_{{\rm{NCs}}}} = {Q_{\rm{R}}}}\end{array} \times \frac{{{I_{{\rm{NCs}}}}}}{{{I_{\rm{R}}}}} \times \frac{{{A_{\rm{R}}}}}{{{A_{{\rm{NCs}}}}}} \times \frac{{\eta _{{\rm{NCs}}}^2}}{{\eta _{\rm{R}}^2}}$
其中, Q代表所测物质的量子产率; I为在相同激发波长下得到的发射光谱的积分面积; A代表在相同波长条件下被测物质的吸收度; η为溶剂的折射率; “NCs”与“R”分别代表MCC-NCs与硫酸奎宁。为了保证实验结果的准确性, 吸收度测定时保证两种溶液的吸光度都在0.05以下。
造模和给药  大鼠(SPF级SD雄性, 体重200 ± 5.0 g, 北京斯贝福生物技术有限公司), 所有动物实验均符合北京中医药大学伦理委员会的规定。大鼠随机分为6组, 每组10只, 分别为空白组、模型组、阳性药组(云南白药粉)、MCC-NCs高剂量、中剂量和低剂量组; 阳性药组灌胃云南白药(0.25 g·kg-1); MCC-NCs组灌胃MCC-NCs溶液高、中和低剂量(1.25、0.63、0.31 g·kg-1); 空白组和模型组每天给予相同体积的去离子水。所有动物每天给药1次, 连续7天。
血热出血模型造模除空白组外, 在第7天时, 其余组动物均皮下注射20% 的干酵母液(10 mL·kg-1), 5 h后灌胃无水乙醇1 mL, 1 h后再给相应的受试药物。
检测指标  在第8天给药1 h后, 将各组动物用4% 水合氯醛麻醉后, 腹主动脉取血, 用肝素钠抗凝管采取全血, 3 500 r·min-1离心10 min, 检测全血黏度和全血高、中、低切黏度; 用乙二胺四乙酸抗凝管采取全血, 检测红细胞比容(hematocrit, HCT)、红细胞分布宽度(red cell volume distribution width, RDW)、血红蛋白含量(hemoglobin, Hb) 和红细胞计数(red blood cell, RBC)。此外, 用3.8%枸橼酸钠(1∶9) 抗凝管采取全血, 3 000 r·min-1离心10 min, 取血浆检测凝血4项活化部分凝血活酶时间(activated partial thromboplastin time, APTT)、凝血酶时间(thrombin time, TT)、凝血酶原时间(prothrombin time, PT) 和纤维蛋白原(fibrinogen, FIB) 水平。
统计学分析  采用SPSS20.0软件对数据进行统计分析, 符合正态分布的计量资料以均数±标准差(x ± s) 表示, 采用方差分析比较多组间数据, P<0.05为差异有统计学意义。
在TEM图谱中可以清楚地看到MCC-NCs的形貌特征(图 2A), 分散均匀, 形状近似球形, 无明显的聚集; 使用Image J软件对100多个颗粒进行统计分析, 粒径分布在0.80~2.8 nm, 平均直径为1.73 ± 0.45 nm (图 2B); 高分辨TEM图谱显示MCC-NCs的晶格间距为0.26 nm (图 2C); 所制备的MCC-NCs在269 nm处有一个弱吸收带(图 2D), 该吸收带归属于π-π*电子跃迁[11]。此外, 在最大激发波长342 nm时, 制备的MCC-NCs的荧光最大发射光谱在442 nm处(图 2E); 用FTIR测定MCC-NCs的表面基团特征(图 2F), 在3 431 cm-1处较强的特征吸收表明-OH基团的存在, 在2 919和2 850 cm-1处的特征吸收分别表现为-CH3和-CH2。约1 635 cm-1处的强谱带归属于-C=O基团, 1 384 cm-1处的峰表示C-N和C=C键的伸缩振动[12]。在最大激发波长为342 nm时, 其量子产率为7.18%。
在MCC-NCs的XPS光谱(图 3A) 中, 284、531和400 eV处的峰值分别对应C1s、O1s和N1s光谱。光谱结果表明, MCC-NCs由C、O和N元素组成, 原子比例分别为68.08%、28.92% 和3.00%。C1s区域的高分辨率光谱显示C元素的官能团有C=C (284.1 eV)、C-O/C-N (285.4 eV) 和O-C=O (287.5 eV), 如图 3B所示。O1s区域的高分辨率扫描显示了532.2 eV时C=O和531.1 eV时C-O-C的不同化学环境(图 3C)。此外, N1s的高分辨率光谱对应于N-H在398.9 eV和C-N在400.0 eV的化学环境(图 3D)。
图 4所示, 与空白组比较, 血热出血模型组的全血高、中、低黏度和血浆黏度均升高, 且有统计学差异(P<0.05); 与模型组比较, 阳性药组和MCC-NCs组的高剂量的全血高、中、低切黏度均降低, 且有统计学差异(P<0.05); MCC-NCs组的中剂量和低剂量在1和5 mPa·s时黏度降低, 且有统计学差异(P<0.05); 与模型组的血浆黏度比较, 发现阳性药组、MCC-NCs组的高剂量和中剂量均降低, 且有统计学差异(P<0.05)。
与空白组比较, 血热出血模型组的HCT、RDW、Hb和RBC显著升高, 且有统计学差异(P<0.05); 与模型组比较, 阳性药组动物的HCT、RDW、Hb和RBC显著降低(P<0.05), MCC-NCs组高剂量、中剂量和低剂量的HCT、Hb和RBC显著降低(P<0.05), MCC-NCs组高剂量中的RDW显著降低(P<0.05), 说明高剂量的效果最佳。结果如图 5所示。
图 6所示, 与空白组比较, 血热出血模型组的动物APTT、TT和FIB均升高, 且有统计学差异(P<0.05); 与模型组比较, 阳性药组和MCC-NCs组高、中和低剂量的APTT、TT和FIB均降低, 且有统计学差异(P<0.05)。
图 7所示, 从肺、胃组织图中肉眼可见空白组的颜色、形态正常, 无病变现象; 模型组的肺出现明显淤血, 胃组织有大小不一的黏膜出血病变; 与模型组比较, 云南白药和MCC-NCs组大鼠肺、胃组织出血现象相对减少。从肺组织病理学检查结果发现, 空白组的大鼠肺组织中肺泡结构清晰正常, 无出血、炎症浸润现象; 模型组的肺泡结构变形, 伴有肺间质出血, 肺泡大量液体流失, 炎性细胞浸润; 云南白药和MCC-NCs组大鼠的肺组织的损伤明显减轻, 出血和炎性细胞浸润减小。从胃组织病理学检查结果发现, 空白组的大鼠胃组织结构正常, 模型组的胃黏膜层是黏膜出血性; 云南白药和MCC-NCs组大鼠的胃黏膜出血症状明显减轻。
中药炭药的应用时间轴线跨越了两千多年, 药用种类不断被丰富, 治疗范围也不断被拓展, 用以治疗各种出血疾病和其他各种疾病。现在对于炭药的研究主要集中在炮制前后小分子成分含量的变化, 但某些成分在炮制前后的升降趋势并不相同[13]; 同时, 对于炭药止血的作用有效成分研究并没有完全统一, 利用药理药效为“眼睛”, 以提取分离纯化技术分析炭药止血的有效成分, 有人认为是钙、铁、镁等离子发挥作用[14]、鞣质在炭药止血功效中发挥作用[15]或酮类成分的变化与其止血作用有关[16]。因此, 利用新的技术, 从一个全新的角度来阐释炭药止血的物质基础非常有必要。
本研究以天然产物丹皮为炭源, 提取分离了一种类似于“碳点”的物质, 本团队将其命名为MCC-NCs, 利用TEM、UV-vis、PL、FITR和XPS对MCC-NCs粒径、结构、光学特征、元素组成和表面官能团等表征鉴定, 发现MCC-NCs尺寸分布在0.80~2.8 nm, 晶格间距为0.26 nm, 主要由C、O和N元素组成, 在其表面还有丰富的官能团, 如-OH、-C=O、C=C等。纳米颗粒是现在研究的热点之一, 其生物活性近年来得到了广泛的研究。纳米颗粒具有低毒性、光化学稳定性、易于功能化、高生物相容性和亲水性等特点[17]; 同时, 有报道称纳米颗粒对体内血液生化和血液学分析无显著影响, 也未引起主要器官的组织病理学异常[18]; 尽管纳米颗粒给药方式不同, 但其都可以高效快速地从体内排出[19]。这些优势使得纳米颗粒在药物载体[20, 21]、生物标记[22]和生物成像[23]等领域广泛被应用。
丹皮炭具有清热、凉血、和血、消瘀等功效, 主治热入血分、发斑、惊痫等。干酵母皮下注射可以激活体内内源性的致热源细胞产生并释放致热源, 引起机体炎性发热、温度升高[24]。乙醇摄入会导致胃细胞坏死和血管损伤, 从而导致胃出血、坏死[25]。利用干酵母联合乙醇致血热出血模型, 可以较好地模拟中医中的外邪入侵、热入血分、津液消耗、血液黏稠的演变过程[26]。本研究实验结果发现, 与空白组比较, 模型组动物的全血高、中、低黏度和血浆黏度均升高; 血液参数HCT、RDW、Hb和RBC显著升高。通过肉眼观察肺、胃组织, 发现肺组织有明显的瘀血, 胃组织有不同大小的黏膜出血性病变; 组织病理血检查结果发现, 肺泡结构变形, 伴有肺间质出血, 肺泡大量液体流失, 炎性细胞浸润; 胃黏膜出血性病变, 这表明造模成功。然而, 预处理MCC-NCs可显著地降低全血高、中、低黏度和血浆黏度; 降低HCT和RDW, 减少Hb和RBC含量(P<0.05), 这表明MCC-NCs可以改善血热出血模型动物的血液流变性, 表现出一定的凉血作用; 同时, 有效地降低了HCT、RDW、Hb和RBC参数; 病理学检查结果发现, MCC-NCs可以明显地减轻肺组织的损伤, 减小出血和炎性细胞浸润。此外, MCC-NCs可以明显地减轻胃黏膜出血症状。以上结果表明, MCC-NCs具有一定的止血作用, 这与先前报道一致[27]
凝血过程主要包括内在凝血途径、外在凝血途径和凝血共同途径, PT用来关联外在凝血途径的情况, APTT与内在凝血途径相关。血浆中共同的凝血途径和促进FIB向纤维蛋白的转化, 与TT和FIB水平有关[28]。本研究发现, MCC-NCs组高、中和低剂量的APTT、TT和FIB均降低(P<0.05), 这表明MCC-NCs止血作用主要与内源性凝血途径或纤维蛋白原系统的激活有关。
综上所述, 本研究首次从丹皮炭中提取分离出一种新的物质MCC-NCs, 并通过MCC-NCs对血热出血模型的治疗作用, 表明MCC-NCs具有一定的凉血止血作用, 这为探索中药炭药止血的物质基础提供了新的策略。
作者贡献: 罗娟负责实验方案设计与实施、结果分析及文章撰写; 胡杰和刘育含参与实验施行与数据采集; 白雪、陈玉民、李郁茹和赵亚芳参与实验实施和数据采集; 孔慧和赵琰负责实验设计、论文修改和基金获得; 屈会化负责实验监督及论文审阅。
利益冲突: 所有作者均声明不存在利益冲突。
  • 中央高校基本科研业务费专项资金资助项目(2019-JYB-TD-001)
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2021年第56卷第8期
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doi: 10.16438/j.0513-4870.2021-0676
  • 接收时间:2021-05-08
  • 首发时间:2025-12-18
  • 出版时间:2021-08-12
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  • 收稿日期:2021-05-08
  • 修回日期:2021-05-27
基金
中央高校基本科研业务费专项资金资助项目(2019-JYB-TD-001)
作者信息
    1.北京中医药大学中医学院, 北京 100029
    2.北京中医药大学中药学院, 北京 100029
    3.北京中医药大学中医药研究院, 北京 100029

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*孔慧, Tel: 86-10-64286705, E-mail: ;
赵琰, E-mail:
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2种不同金属材料的力学参数

Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)

Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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