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Article(id=1212695796219298084, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1212693246539649865, articleNumber=1001-2494(2024)19-1813-12, orderNo=null, doi=10.11669/cpj.2024.19.005, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1715184000000, receivedDateStr=2024-05-09, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1767058808616, onlineDateStr=2025-12-30, pubDate=1728316800000, pubDateStr=2024-10-08, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1767058808616, onlineIssueDateStr=2025-12-30, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1767058808616, creator=13701087609, updateTime=1767058808616, updator=13701087609, issue=Issue{id=1212693246539649865, tenantId=1146029695717560320, journalId=1190317699101192196, year='2024', volume='59', issue='19', pageStart='1781', pageEnd='1880', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1767058200723, creator=13701087609, updateTime=1767059042003, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1212696775207600634, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1212693246539649865, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1212696775207600635, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1212693246539649865, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=1813, endPage=1824, ext={EN=ArticleExt(id=1212695796441596198, articleId=1212695796219298084, tenantId=1146029695717560320, journalId=1190317699101192196, language=EN, title=System Biology Analysis Reveals the Associated Pathway of Shengqing Huazhuo Prescription in Regulating Metabolic Abnormalities, columnId=null, journalTitle=Chinese Pharmaceutical Journal, columnName=null, runingTitle=null, highlight=null, articleAbstract=

OBJECTIVE To explore the mechanism of Shengqing Huazhuo (SQHZ) prescription in regulating simple obesity rats with spleen deficiency and dampness obstruction. METHODS The rat model of simple obesity with spleen deficiency and dampness obstruction was established, and the effects of SQHZ prescription on body weight, liver histomorphology and expression level of blood lipid metabolism in obesity rats were comprehensively evaluated. Ultra-high performance liquid chromatography-Q/Exactive mass spectrometry (UHPLC-QE-MS) was used to identify the blood components of SQHZ prescription, and the potential action pathways and targets were predicted by the network pharmacological platform. Non-targeted metabonomics was used to analyze the metabolomics of rat serum. The abundance of intestinal flora in rat faeces was detected by 16S rDNA amplicon sequencing technology. Finally, a multi-scale and multi-dimensional network was constructed by summarizing the KEGG pathways of the three, which can be used for overall visualization and deep analysis. RESULTS SQHZ prescription significantly reduced the body weight of obesity rats, and improved blood lipid levels and liver fat accumulation. Twenty-seven blood components of SQHZ prescription were identified by UHPLC-QE-MS technology. Network pharmacological analysis revealed the pathways related to metabolism of SQHZ prescription, mainly involving steroid hormone biosynthesis and ovarian steroidogenesis. Serum metabolomics analysis obtained 10 key differential metabolites, which involved metabolic pathways such as biosynthesis of unsaturated fatty acids and tryptophan metabolism. Intestinal microbiota sequencing results showed that SQHZ prescription could regulate the composition and improve the structure of intestinal microbiota in obesity rats, and the metabolic pathways involved are the biosynthesis of steroid hormones and unsaturated fatty acids. CONCLUSION SQHZ prescription not only shows significant therapeutic effects in treating simple obesity rats with spleen deficiency and dampness obstruction, but also has the ability to regulate serum metabolism and intestinal microbial community structure.

, correspAuthors=Hao CHEN, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, 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=Li ZHOU, Fei YI, Lifang WANG, Rui LI, Qin YANG, Yonggui SONG, Hao CHEN), CN=ArticleExt(id=1212695799352443251, articleId=1212695796219298084, tenantId=1146029695717560320, journalId=1190317699101192196, language=CN, title=系统生物学分析揭示升清化浊方调控代谢异常的关联路径, columnId=1190352405612040510, journalTitle=中国药学杂志, columnName=论著, runingTitle=null, highlight=null, articleAbstract=

目的 探讨升清化浊方调控大鼠脾虚湿阻型单纯性肥胖的作用机制。方法 建立脾虚湿阻型单纯性肥胖大鼠模型,综合评价升清化浊方对肥胖大鼠体质量、肝脏组织形态学以及血脂代谢表达水平的影响。利用超高效液相色谱-四级杆静电场轨道阱质谱法(ultra high performance liquid chromatography-Q/Exactive mass spectrometry, UHPLC-QE-MS)联用技术,鉴定升清化浊方的入血成分,并通过网络药理学平台预测其潜在作用靶点和通路。同时,采用非靶向代谢组学对大鼠血清进行代谢组学分析。通过16S rDNA扩增子测序技术,对大鼠粪便中的肠道菌群丰度进行检测。最后,对三者的KEGG通路汇总,构建了一个多尺度、多维度网络,用于整体可视化和深度分析。结果 升清化浊方给药后显著降低了肥胖大鼠的体质量,同时改善了血脂水平和肝脏脂肪蓄积情况。通过UHPLC-QE-MS技术鉴定了升清化浊方中的27个入血成分。网络药理学分析揭示了升清化浊方与代谢相关的通路,主要涉及到类固醇激素生物合成和卵巢类固醇生成等。血清代谢组学分析得到了10个关键差异代谢物,其涉及到的代谢途径是不饱和脂肪酸的生物合成、色氨酸代谢等。肠道菌群测序结果显示,升清化浊方能够调节肥胖大鼠的肠道菌群组成并改善结构,其涉及到的代谢途径是类固醇激素、不饱和脂肪酸的生物合成。结论 升清化浊方不仅在治疗大鼠脾虚湿阻型单纯性肥胖方面表现出显著疗效,还具备调节血清代谢和肠道微生物群落结构的能力。

, correspAuthors=陈浩, authorNote=null, correspAuthorsNote=
* 陈浩,男,博士,副教授,副主任药师 研究方向:中医药方证结合及中药炮制学研究 Tel:(0791)87119019
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周丽,女,硕士,讲师 研究方向:中医药数据挖掘和中医药防治单纯性肥胖疾病研究

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周丽,女,硕士,讲师 研究方向:中医药数据挖掘和中医药防治单纯性肥胖疾病研究

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周丽,女,硕士,讲师 研究方向:中医药数据挖掘和中医药防治单纯性肥胖疾病研究

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Gut, 2020, 69(10): 1796-1806., articleTitle=Adipose tissue derived bacteria are associated with inflammation in obesity and type 2 diabetes, refAbstract=null)], funds=[Fund(id=1212795881917694223, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212695796219298084, awardId=82360966, language=CN, fundingSource=国家自然科学基金项目资助(82360966), fundOrder=null, country=null), Fund(id=1212795882005774609, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212695796219298084, awardId=GJJ2200959, language=CN, fundingSource=江西省教育厅科学技术研究项目资助(GJJ2200959), fundOrder=null, country=null), Fund(id=1212795882102243604, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212695796219298084, awardId=202211433, language=CN, fundingSource=江西省卫生健康委员会科技计划项目资助(202211433), fundOrder=null, country=null), Fund(id=1212795882223878424, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212695796219298084, awardId=202310412024, 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ext=[AuthorCompanyExt(id=1212795876934860855, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212695796219298084, companyId=1212795876930666550, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=4 The Affiliated Hospital of Jiangxi University of Chinese Medicine, Nanchang 330006, China), AuthorCompanyExt(id=1212795876943249464, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212695796219298084, companyId=1212795876930666550, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=4 江西中医药大学附属医院, 南昌 330006)])], figs=[ArticleFig(id=1212795880365801701, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212695796219298084, language=EN, label=Fig.1, caption=Technique flow chart of the associated pathway of SQHZ Prescription in regulating metabolic abnormalities, figureFileSmall=7uMh6LNw5c7Hx9AbCvDh5w==, figureFileBig=oZPnZq/WydKtvlrjbIWyzg==, tableContent=null), ArticleFig(id=1212795880453882089, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212695796219298084, language=CN, label=图1, caption=升清化浊方调控代谢异常关联路径技术流程图, figureFileSmall=7uMh6LNw5c7Hx9AbCvDh5w==, figureFileBig=oZPnZq/WydKtvlrjbIWyzg==, tableContent=null), ArticleFig(id=1212795880579711213, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212695796219298084, language=EN, label=Fig.2, caption=Effects of Shengqing Huazhuo(SQHZ) prescription on simple obesity rats with spleen deficiency and dampness obstruction

A-body weight changes of rats in normal group and model group for 16 weeks of feeding; B-body weight changes in each group during 6 weeks of administration; C-results of TC, TG, LDL-C and HDL-C in each group; D-liver oil red O staining sections of rats in each group; 1)P<0.01, 2)P<0.001, vs normal group;3)P<0.05, 4)P<0.01, 5)P<0.001, vs model group; PC-positive control; SQHZ-Shengqing Huazhuo.n=8,$\stackrel{-}{x}$±s

, figureFileSmall=03SWvVhTwJEQ5CFc/S7vdQ==, figureFileBig=vgrJ1t7l3z/2iLYS5XLuKA==, tableContent=null), ArticleFig(id=1212795880676180206, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212695796219298084, language=CN, label=图2, caption=升清化浊方对脾虚湿阻型单纯性肥胖大鼠的影响结果图

A-正常组(N)与模型组(M)大鼠饲养16周的体质量变化图;B-给药6周期间各组体质量变化图;C-各组血清总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)结果;D-各组大鼠肝油红O染色切片图;与正常组比较,1)P<0.01,2)P<0.001;与模型组比较,3)P<0.05,4)P<0.01,5)P<0.001;PC-阳性对照;SQHZ-升清化浊方。n=8,$\stackrel{-}{x}$±s

, figureFileSmall=03SWvVhTwJEQ5CFc/S7vdQ==, figureFileBig=vgrJ1t7l3z/2iLYS5XLuKA==, tableContent=null), ArticleFig(id=1212795880747483376, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212695796219298084, language=EN, label=Fig.3, caption=Chromatogram of UHPLC-QE-MS analysis of blood components of SQHZ prescription and network pharmacology map

A-negative ion diagram of drug-containing serum detection; B-positive ion diagram of drug-containing serum detection; C-venn diagram; D-KEGG pathway diagram.

, figureFileSmall=/943lgAAoWXUxqyiAvWy/g==, figureFileBig=5cdiNiustkizDDuF/qnQVQ==, tableContent=null), ArticleFig(id=1212795880843952371, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212695796219298084, language=CN, label=图3, caption=升清化浊方入血成分UHPLC-QE-MS法分析色谱图和网络药理学分析图

A-含药血清检测负离子图;B-含药血清检测正离子图;C-Venn图;D-KEGG通路图。

, figureFileSmall=/943lgAAoWXUxqyiAvWy/g==, figureFileBig=5cdiNiustkizDDuF/qnQVQ==, tableContent=null), ArticleFig(id=1212795880932032758, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212695796219298084, language=EN, label=Fig.4, caption=Metabolomics analysis results of SQHZ prescription

A-PCA score plot of each group; B-normal group and model group OPLS-DA score plot; C-normal group and model group OPLS-DA model replacement test diagram; D-volcano map of differential metabolites screening between normal group and model group; E-volcano map of differential metabolites screening between model group and SQHZ group; F-KEGG pathway diagram.

, figureFileSmall=1Pmho0JxbxBiY2WIcsINZQ==, figureFileBig=/39G3PhUrv6VrM7aqUrn3A==, tableContent=null), ArticleFig(id=1212795881036890359, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212695796219298084, language=CN, label=图4, caption=升清化浊方代谢组学分析结果

A-各组PCA得分图;B-正常组与模型组OPLS-DA得分图;C-正常组与模型组 OPLS-DA 模型置换检验图;D-正常组与模型组差异代谢物筛选的火山图;E-模型组与升清化浊方组差异代谢物筛选的火山图;F-KEGG 通路图。

, figureFileSmall=1Pmho0JxbxBiY2WIcsINZQ==, figureFileBig=/39G3PhUrv6VrM7aqUrn3A==, tableContent=null), ArticleFig(id=1212795881141747960, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212695796219298084, language=EN, label=Fig.5, caption=The results of intestinal flora analysis of SQHZ prescription

A-OPLS-DA score plot of each group; B-microbiota grade abundance curve of each group; C-microbiota α diversity index of each group. n=6,$\stackrel{-}{x}$±s; D-heat map of species correlation at the phylum level of each group; E-heat map of species correlation at the genus level of each group; F-pathway analysis of intestinal flora in model group and SQHZ group.

, figureFileSmall=qk8pQPmvQjNvBBEfVCN5eQ==, figureFileBig=ePG7us8Km4dthhrdY5zJag==, tableContent=null), ArticleFig(id=1212795881213051130, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212695796219298084, language=CN, label=图5, caption=升清化浊方肠道菌群分析结果

A-各组OPLS-DA得分图;B-各组微生物群等级丰度曲线;C-各组微生物群α多样性指数。n=6,$\stackrel{-}{x}$±s;D-各组门水平物种相关性热图;E-各组属水平物种相关性热图;F-模型组与升清化浊方组肠道菌群通路分析。

, figureFileSmall=qk8pQPmvQjNvBBEfVCN5eQ==, figureFileBig=ePG7us8Km4dthhrdY5zJag==, tableContent=null), ArticleFig(id=1212795881280159998, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212695796219298084, language=EN, label=Fig.6, caption=Correlation network diagram and KEGG pathway analysis between Serum pharmacochemistry and serum metabolomics and intestinal flora of SQHZ prescription

A-correlation network diagram of intestinal flora and differential metabolites; B-the correlation network diagram among serum pharmacochemistry, serum metabolomics and intestinal flora; C-KEGG pathway diagram.

, figureFileSmall=FYqUk0WflKLaDSm2V1cxOw==, figureFileBig=NnZgA+iKf+Ww/2jVQL8rPg==, tableContent=null), ArticleFig(id=1212795881364046079, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212695796219298084, language=CN, label=图6, caption=升清化浊方血清药物化学与血清代谢组学、肠道菌群之间的相关性网络图及KEGG通路分析图

A-肠道菌群与差异代谢物的相关性网络图;B-血清药物化学与血清代谢组学、肠道菌群三者之间的相关性网络图;C-KEGG 通路图。

, figureFileSmall=FYqUk0WflKLaDSm2V1cxOw==, figureFileBig=NnZgA+iKf+Ww/2jVQL8rPg==, tableContent=null), ArticleFig(id=1212795881443737858, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212695796219298084, language=EN, label=Tab.1, caption=

The basic information of 27 components of SQHZ prescription

, figureFileSmall=null, figureFileBig=null, tableContent=
No. CAS No. Compound Composite score Molecular formula m/z tR/s
1 572-30-5 Quercetin-3-arabinoside 1.00 C20H18O11 435.09 116.97
2 77-95-2 D-(-)-quinic acid 1.00 C7H12O6 191.06 25.45
3 1447-88-7 Hispidulin 1.00 C16H12O6 299.06 336.76
4 548-77-6 Tectorigenin 1.00 C16H12O6 301.07 309.15
5 121-34-6 Vanillic acid 1.00 C8H8O4 169.05 195.82
6 40957-83-3 Glycitein 0.99 C16H12O5 283.06 411.05
7 482-36-0 Quercetin-3-O-galactoside 0.99 C21H20O12 463.09 94.65
8 552-58-9 Eriodictyol 0.98 C15H12O6 289.07 154.94
9 476-66-4 Ellagic acid 0.97 C14H6O8 301.00 81.83
10 - (2S,3R,4S,5S,6R)-2-[(2R,3R,4S,5S,6R)-4,5-dihydroxy-2-[[(3S,5R,8R,10R,12R,13R,14R,17S)-12-hydroxy-17-(2-hydroxy-6-methylhept-5-en-2-yl)-4,4,8,10,14-pentamethyl-2,3,5,6,7,9,11,12,13,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-3-yl]oxy]-6-(hydroxymethyl)oxan-3-yl]oxy-6-(hydroxymethyl)oxane-3,4,5-triol 0.97 C42H72O13 783.49 346.06
11 491-67-8 Baicalein 0.97 C15H10O5 271.06 190.72
12 22688-79-5 Quercetin-3-glucuronide 0.97 C21H18O13 477.07 95.90
13 485-72-3 Formononetin 0.95 C16H12O4 269.08 313.42
14 - Coumaroyl quinic acid 0.94 C16H18O8 337.09 56.55
15 61153-77-3 Licoflavone A 0.93 C20H18O4 323.13 246.95
16 3804-70-4 Columbianetin 0.90 C14H14O4 245.08 160.73
17 371113-07-4 Sayaendoside 0.90 C19H28O10 455.13 91.14
18 20243-59-8 Hydroxygenkwanin 0.89 C16H12O6 299.06 310.00
19 - 11,22-dimethyl (6E,17E)-3,14-dioxo-8,19-bis({[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy})-4,9,15,20-tetraoxatricyclo[16.4.0.07,12]docosa-6,10,17,21-tetraene-11,22-dicarboxylate 0.87 C13H18O6 293.10 259.72
20 26575-95-1 Alisol B 23-acetate 0.87 C32H50O5 515.37 581.15
21 181020-33-7 Estragonoside 0.85 C22H22O12 479.12 188.15
22 20633-93-6 Fortunellin 0.85 C28H32O14 593.19 182.12
23 479-90-3 Artemetin 0.84 C20H20O8 389.12 382.83
24 775-01-9 3,4-Dihydroxymandelic acid 0.83 C8H8O5 183.03 104.16
25 39763-33-2 Dehydrovomifoliol 0.83 C13H18O3 223.13 334.56
26 - Kaempferol-3-O-glucoside-7-O-rhamnoside 0.81 C27H30O15 593.15 215.05
27 668420-44-8 Pagoside 0.80 C24H28O11 515.15 284.06
), ArticleFig(id=1212795881548595460, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212695796219298084, language=CN, label=表1, caption=

升清化浊方27个入血成分的基本信息

, figureFileSmall=null, figureFileBig=null, tableContent=
No. CAS No. Compound Composite score Molecular formula m/z tR/s
1 572-30-5 Quercetin-3-arabinoside 1.00 C20H18O11 435.09 116.97
2 77-95-2 D-(-)-quinic acid 1.00 C7H12O6 191.06 25.45
3 1447-88-7 Hispidulin 1.00 C16H12O6 299.06 336.76
4 548-77-6 Tectorigenin 1.00 C16H12O6 301.07 309.15
5 121-34-6 Vanillic acid 1.00 C8H8O4 169.05 195.82
6 40957-83-3 Glycitein 0.99 C16H12O5 283.06 411.05
7 482-36-0 Quercetin-3-O-galactoside 0.99 C21H20O12 463.09 94.65
8 552-58-9 Eriodictyol 0.98 C15H12O6 289.07 154.94
9 476-66-4 Ellagic acid 0.97 C14H6O8 301.00 81.83
10 - (2S,3R,4S,5S,6R)-2-[(2R,3R,4S,5S,6R)-4,5-dihydroxy-2-[[(3S,5R,8R,10R,12R,13R,14R,17S)-12-hydroxy-17-(2-hydroxy-6-methylhept-5-en-2-yl)-4,4,8,10,14-pentamethyl-2,3,5,6,7,9,11,12,13,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-3-yl]oxy]-6-(hydroxymethyl)oxan-3-yl]oxy-6-(hydroxymethyl)oxane-3,4,5-triol 0.97 C42H72O13 783.49 346.06
11 491-67-8 Baicalein 0.97 C15H10O5 271.06 190.72
12 22688-79-5 Quercetin-3-glucuronide 0.97 C21H18O13 477.07 95.90
13 485-72-3 Formononetin 0.95 C16H12O4 269.08 313.42
14 - Coumaroyl quinic acid 0.94 C16H18O8 337.09 56.55
15 61153-77-3 Licoflavone A 0.93 C20H18O4 323.13 246.95
16 3804-70-4 Columbianetin 0.90 C14H14O4 245.08 160.73
17 371113-07-4 Sayaendoside 0.90 C19H28O10 455.13 91.14
18 20243-59-8 Hydroxygenkwanin 0.89 C16H12O6 299.06 310.00
19 - 11,22-dimethyl (6E,17E)-3,14-dioxo-8,19-bis({[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy})-4,9,15,20-tetraoxatricyclo[16.4.0.07,12]docosa-6,10,17,21-tetraene-11,22-dicarboxylate 0.87 C13H18O6 293.10 259.72
20 26575-95-1 Alisol B 23-acetate 0.87 C32H50O5 515.37 581.15
21 181020-33-7 Estragonoside 0.85 C22H22O12 479.12 188.15
22 20633-93-6 Fortunellin 0.85 C28H32O14 593.19 182.12
23 479-90-3 Artemetin 0.84 C20H20O8 389.12 382.83
24 775-01-9 3,4-Dihydroxymandelic acid 0.83 C8H8O5 183.03 104.16
25 39763-33-2 Dehydrovomifoliol 0.83 C13H18O3 223.13 334.56
26 - Kaempferol-3-O-glucoside-7-O-rhamnoside 0.81 C27H30O15 593.15 215.05
27 668420-44-8 Pagoside 0.80 C24H28O11 515.15 284.06
), ArticleFig(id=1212795881640870151, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212695796219298084, language=EN, label=Tab.2, caption=

Basic information of 10 key metabolites in simple obesity rats with spleen deficiency and dampness obstruction treated with SQHZ prescription

, figureFileSmall=null, figureFileBig=null, tableContent=
No. Metabolite tR/s m/z CAS No. Type SQHZ-M M-N Chemical structure
formula
VIP Trend VIP Trend
1 Uridine 114.99 279.04 58-96-8 NEG 2.02 ↑ 1.47 ↓
2 L-Kynurenine 194.57 209.09 2922-83-0 POS 2.01 ↓ 1.45 ↑
3 Biliverdin 442.00 581.24 114-25-0 NEG 2.20 ↓ 1.76 ↑
4 Pseudouridine 115.70 243.06 1445-07-4 NEG 1.79 ↑ 1.50 ↓
5 Prostaglandin B2 479.35 333.21 13367-85-6 NEG 1.84 ↑ 2.00 ↓
6 γ-Linolenic acid 622.12 277.22 506-26-3 NEG 1.65 ↑ 1.66 ↓
7 Docosahexaenoic acid 631.79 327.23 6217-54-5 NEG 1.75 ↑ 1.33 ↓
8 12-KETE 526.77 317.21 108437-64-5 NEG 2.24 ↑ 1.98 ↓
9 Docosapentaenoic acid 655.11 329.25 24880-45-3 NEG 2.02 ↓ 1.95 ↑
10 Adrenic acid 671.82 331.27 28874-58-0 NEG 2.32 ↓ 1.87 ↑
), ArticleFig(id=1212795881733144842, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1212695796219298084, language=CN, label=表2, caption=

升清化浊方治疗脾虚湿阻型单纯性肥胖大鼠体内10种关键代谢物的基本信息

, figureFileSmall=null, figureFileBig=null, tableContent=
No. Metabolite tR/s m/z CAS No. Type SQHZ-M M-N Chemical structure
formula
VIP Trend VIP Trend
1 Uridine 114.99 279.04 58-96-8 NEG 2.02 ↑ 1.47 ↓
2 L-Kynurenine 194.57 209.09 2922-83-0 POS 2.01 ↓ 1.45 ↑
3 Biliverdin 442.00 581.24 114-25-0 NEG 2.20 ↓ 1.76 ↑
4 Pseudouridine 115.70 243.06 1445-07-4 NEG 1.79 ↑ 1.50 ↓
5 Prostaglandin B2 479.35 333.21 13367-85-6 NEG 1.84 ↑ 2.00 ↓
6 γ-Linolenic acid 622.12 277.22 506-26-3 NEG 1.65 ↑ 1.66 ↓
7 Docosahexaenoic acid 631.79 327.23 6217-54-5 NEG 1.75 ↑ 1.33 ↓
8 12-KETE 526.77 317.21 108437-64-5 NEG 2.24 ↑ 1.98 ↓
9 Docosapentaenoic acid 655.11 329.25 24880-45-3 NEG 2.02 ↓ 1.95 ↑
10 Adrenic acid 671.82 331.27 28874-58-0 NEG 2.32 ↓ 1.87 ↑
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系统生物学分析揭示升清化浊方调控代谢异常的关联路径
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周丽 1 , 易菲 2 , 王丽芳 2 , 李蕊 2 , 杨琴 1 , 宋永贵 3 , 陈浩 2, 4, *
中国药学杂志 | 论著 2024,59(19): 1813-1824
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中国药学杂志 | 论著 2024, 59(19): 1813-1824
系统生物学分析揭示升清化浊方调控代谢异常的关联路径
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周丽1, 易菲2, 王丽芳2, 李蕊2, 杨琴1, 宋永贵3, 陈浩2, 4, *
作者信息
  • 1 江西中医药大学计算机学院, 南昌 330004
  • 2 江西中医药大学药学院, 南昌 330004
  • 3 江西中医药大学实验动物科技中心, 南昌 330004
  • 4 江西中医药大学附属医院, 南昌 330006

    • 周丽,女,硕士,讲师 研究方向:中医药数据挖掘和中医药防治单纯性肥胖疾病研究

通讯作者:

* 陈浩,男,博士,副教授,副主任药师 研究方向:中医药方证结合及中药炮制学研究 Tel:(0791)87119019
System Biology Analysis Reveals the Associated Pathway of Shengqing Huazhuo Prescription in Regulating Metabolic Abnormalities
Li ZHOU1, Fei YI2, Lifang WANG2, Rui LI2, Qin YANG1, Yonggui SONG3, Hao CHEN2, 4, *
Affiliations
  • 1 School of Computer, Jiangxi University of Chinese Medicine, Nanchang 330004, China
  • 2 School of Pharmacy, Jiangxi University of Chinese Medicine, Nanchang 330004, China
  • 3 Experimental Animal Science and Technology Center, Jiangxi University of Chinese Medicine, Nanchang 330004, China
  • 4 The Affiliated Hospital of Jiangxi University of Chinese Medicine, Nanchang 330006, China
出版时间: 2024-10-08 doi: 10.11669/cpj.2024.19.005
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摘要
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目的 探讨升清化浊方调控大鼠脾虚湿阻型单纯性肥胖的作用机制。方法 建立脾虚湿阻型单纯性肥胖大鼠模型,综合评价升清化浊方对肥胖大鼠体质量、肝脏组织形态学以及血脂代谢表达水平的影响。利用超高效液相色谱-四级杆静电场轨道阱质谱法(ultra high performance liquid chromatography-Q/Exactive mass spectrometry, UHPLC-QE-MS)联用技术,鉴定升清化浊方的入血成分,并通过网络药理学平台预测其潜在作用靶点和通路。同时,采用非靶向代谢组学对大鼠血清进行代谢组学分析。通过16S rDNA扩增子测序技术,对大鼠粪便中的肠道菌群丰度进行检测。最后,对三者的KEGG通路汇总,构建了一个多尺度、多维度网络,用于整体可视化和深度分析。结果 升清化浊方给药后显著降低了肥胖大鼠的体质量,同时改善了血脂水平和肝脏脂肪蓄积情况。通过UHPLC-QE-MS技术鉴定了升清化浊方中的27个入血成分。网络药理学分析揭示了升清化浊方与代谢相关的通路,主要涉及到类固醇激素生物合成和卵巢类固醇生成等。血清代谢组学分析得到了10个关键差异代谢物,其涉及到的代谢途径是不饱和脂肪酸的生物合成、色氨酸代谢等。肠道菌群测序结果显示,升清化浊方能够调节肥胖大鼠的肠道菌群组成并改善结构,其涉及到的代谢途径是类固醇激素、不饱和脂肪酸的生物合成。结论 升清化浊方不仅在治疗大鼠脾虚湿阻型单纯性肥胖方面表现出显著疗效,还具备调节血清代谢和肠道微生物群落结构的能力。

血清药物化学  /  超高效液相色谱-四级杆静电场轨道阱质谱法  /  升清化浊方  /  血清代谢组学  /  肠道菌群

OBJECTIVE To explore the mechanism of Shengqing Huazhuo (SQHZ) prescription in regulating simple obesity rats with spleen deficiency and dampness obstruction. METHODS The rat model of simple obesity with spleen deficiency and dampness obstruction was established, and the effects of SQHZ prescription on body weight, liver histomorphology and expression level of blood lipid metabolism in obesity rats were comprehensively evaluated. Ultra-high performance liquid chromatography-Q/Exactive mass spectrometry (UHPLC-QE-MS) was used to identify the blood components of SQHZ prescription, and the potential action pathways and targets were predicted by the network pharmacological platform. Non-targeted metabonomics was used to analyze the metabolomics of rat serum. The abundance of intestinal flora in rat faeces was detected by 16S rDNA amplicon sequencing technology. Finally, a multi-scale and multi-dimensional network was constructed by summarizing the KEGG pathways of the three, which can be used for overall visualization and deep analysis. RESULTS SQHZ prescription significantly reduced the body weight of obesity rats, and improved blood lipid levels and liver fat accumulation. Twenty-seven blood components of SQHZ prescription were identified by UHPLC-QE-MS technology. Network pharmacological analysis revealed the pathways related to metabolism of SQHZ prescription, mainly involving steroid hormone biosynthesis and ovarian steroidogenesis. Serum metabolomics analysis obtained 10 key differential metabolites, which involved metabolic pathways such as biosynthesis of unsaturated fatty acids and tryptophan metabolism. Intestinal microbiota sequencing results showed that SQHZ prescription could regulate the composition and improve the structure of intestinal microbiota in obesity rats, and the metabolic pathways involved are the biosynthesis of steroid hormones and unsaturated fatty acids. CONCLUSION SQHZ prescription not only shows significant therapeutic effects in treating simple obesity rats with spleen deficiency and dampness obstruction, but also has the ability to regulate serum metabolism and intestinal microbial community structure.

serum pharmacochemistry  /  UHPLC-QE-MS  /  Shengqing Huazhuo prescription  /  serum metabolomics  /  intestinal flora
周丽, 易菲, 王丽芳, 李蕊, 杨琴, 宋永贵, 陈浩. 系统生物学分析揭示升清化浊方调控代谢异常的关联路径. 中国药学杂志, 2024 , 59 (19) : 1813 -1824 . DOI: 10.11669/cpj.2024.19.005
Li ZHOU, Fei YI, Lifang WANG, Rui LI, Qin YANG, Yonggui SONG, Hao CHEN. System Biology Analysis Reveals the Associated Pathway of Shengqing Huazhuo Prescription in Regulating Metabolic Abnormalities[J]. Chinese Pharmaceutical Journal, 2024 , 59 (19) : 1813 -1824 . DOI: 10.11669/cpj.2024.19.005
正文
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肥胖是以全身脂肪组织过度增生为主要表征的代谢性疾病。长期的肥胖不仅影响体形美观,还会增加心脑血管、高血压、抑郁以及癌症等多种疾病的患病风险。据相关研究,全球近40%的成年人存在超重或肥胖问题[1]。针对全球肥胖发病率居高不下的难题,课题组在前期临床经验的基础上,结合数据挖掘的结果,研发出升清化浊方(已授权国家发明专利,所含中药均在国家药食同源目录中),为肥胖症患者提供一种防治肥胖有效的药食同源配方[2]。该方精选人参、荷叶、茯苓为君药;杜仲叶、炒决明子、火麻仁、山楂为臣药;桑葚、淡竹叶、铁皮石斛、玫瑰花为佐药。方剂的设计重在调理脾胃之气,在升清化浊、利水渗湿的前提下,配以益气补脾和活血的中药,共同发挥化湿化痰化浊、活血、益气补脾和润肠通便的作用,最终达到降脂减肥的目的。
在当前研究中,中药复方面临的主要难题是药效物质不清晰和关键作用靶点不明确,针对这些问题,本研究首先构建脾虚湿阻型单纯性肥胖大鼠模型,对该方进行药效学研究。其次,采用血清药物化学和网络药理学方法,鉴定该方中的潜在活性成分并预测其作用靶点。又进行了16S rRNA测序和非靶向血清代谢组学检测。运用系统生物学思维[3],构建血清药物化学、血清代谢组学、肠道菌群三者的关联性网络,并对其代谢通路做进一步分析,深入阐释升清化浊方治疗脾虚湿阻型单纯性肥胖的关键药效物质基础及代谢通路,从而揭示升清化浊方调控代谢异常的相关联路径,研究思路见图1。
1 材料与仪器
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1.1 实验动物
SPF级SD雄性大鼠41只,1.5月龄,体质量(190±20)g,购自江西中医药大学,实验动物生产许可证号:SCXK(赣)2018-0003,动物使用许可证:SYXK(赣)2022-0002。大鼠喂养地点为江西中医药大学实验动物科技中心,其相对湿度为45%~55%、其环境温度为20~24 ℃。江西中医药大学实验动物伦理委员会审批动物福利伦理号:JZLLSC20230174。
1.2 药物与试剂
升清化浊方的处方为:人参、荷叶、茯苓、杜仲叶、炒决明子、火麻仁、山楂、桑葚、淡竹叶、铁皮石斛、玫瑰花。本实验所用升清化浊汤委托江西中医药大学附属医院药学部制备(升清化浊方的质量浓度:5.937 5 g·mL-1)。奥利司他(批号:2112106A,杭州中美华东制药有限公司);胆固醇测定试剂盒(TC,批号:20220524)、甘油三酯测定试剂盒(TG,批号:20220106)、高密度脂蛋白胆固醇测定试剂盒(HDL-C,批号:20220916,宁波普瑞柏生物技术股份有限公司)。
1.3 主要仪器
YM-080S型超声仪(深圳市方奥微电子有限公司);Orbitrap Exploris 120型高分辨质谱、Vanquish型超高效液相(美国赛默飞世尔科技公司);ACQUITY UPLC BEH C18色谱柱(2.1 mm×100 mm,1.7 μm,美国沃特世公司);Agilent 2100型生物分析仪(美国安捷伦科技有限公司);罗氏modular p800全自动生化仪(瑞士罗氏诊断产品有限公司)。
2 方法
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2.1 体内外成分分析及网络药理学分析
2.1.1 动物实验分组、给药及血清样本的制备
取6只大鼠随机分为空白血清组和含药血清组,每组各3只大鼠。含药血清组的大鼠灌胃给予高剂量升清化浊汤7 d,与此同时空白血清组灌胃等量的生理盐水。在最后一次给药1.5 h后,用质量分数2%固体质量分数腹腔注射,给药剂量40 mg·kg-1,深度麻醉大鼠取血。全血3 500 r·min-1离心15 min,取上层血清,-20 ℃储存备用。
取400 μL含药血清组的血清样本,加入盐酸40 μL,混匀,静置;加入乙腈1.6 mL,离心,取上清液,氮气吹干;加入体积分数80%甲醇150 μL复溶,离心,取上清液倒进样瓶中上机检测。空白血清样本重复上述操作。
2.1.2 供试品溶液的制备
委托江西中医药大学附属医院药学部制备升清化浊方水煎剂,作为实验提取物。将该方水煎剂离心后,取上清液300 μL,加入适量水或甲醇稀释,再超声,离心,经0.22 μm滤膜过滤。每个样本各取200 μL,形成供试品提取物样本。
2.1.3 UHPLC-QE-MS分析
色谱条件:UHPLC BEH C18色谱柱(2.1 mm×100 mm, 1.7 μm)。流动相:体积分数0.1%甲酸水(A)-体积分数0.1%甲酸乙腈(B)。多步线性洗脱梯度程序如下:0~11 min,15%~75%B;11~12 min,75%~98%B;12~14 min,98%B;14~14.1 min,98%~15%B;14.1~16 min,15%B。质谱条件:Orbitrap Exploris 120型质谱仪通过Xcalibur软件,进行一级、二级质谱数据采集。
2.1.4 网络药理学分析
使用Similarity ensemble approach数据库(https://sea.bkslab.org/)获取升清化浊方的潜在作用靶点,并从疾病相关数据库中收集与肥胖相关的靶点。通过韦恩图(Venn)分析,找到两者共有的核心靶点。这些核心靶点被用于String数据库中来构建蛋白质相互作用网络。最后,通过DAVID数据库(https://david.ncifcrf.gov/)进行KEGG通路富集分析,以P<0.05表示差异有统计学意义。
2.2 动物分组与给药
从35只大鼠中随机选取8只,设为正常(normal,N)组,其余27只大鼠采用课题组前期复制造模法[4]造模10周,造模结束后,从这27只中选取24只已造模成功的大鼠,随机分为模型(model,M)组、阳性对照(positive control, PC)组、升清化浊方(Shengqing Huazhuo, SQHZ)组,每组8只。分组结束后第2天,升清化浊方组灌胃给药23.75 g·kg-1的生药量[按照60 kg成人临床日服用量为114 g,据体质量换算成大鼠等效剂量为23.75 g·kg-1(114 g/60 kg×6.25×2=23.75 g·kg-1);阳性对照组灌胃0.037 6 g·kg-1的奥利司他胶囊水溶剂;模型组和正常组分别灌胃等量生理盐水。每组大鼠每日灌胃2 mL,连续6周。
2.3 大鼠体质量测定
每周在相同的时间点测定大鼠体质量。
2.4 全自动生化分析仪检测血脂四项
将每组鼠血清放置于全自动生化仪中进行血脂四项检测。
2.5 肝脏油红O染色
将肝脏组织进行冷冻切片,在体积分数10%甲醛中固定10~15 min,用自来水洗净,滴加油红O染液染色2~5 min,自来水冲洗1~3 min,用苏木素再进行复染,然后用自来水再冲洗1~3 min,最后使用甘油明胶进行封片。
2.6 血清代谢组学分析
2.6.1 UHPLC-QE-MS分析
色谱条件Vanquish超高效液相色谱仪,通过Waters ACQUITY UPLC HSS T3 (2.1 mm×100 mm,1.8 μm)液相色谱柱对目标化合物进行色谱分离。液相色谱A相为水相,含5 mmol·L-1乙酸铵和5 mmol·L-1乙酸,B相为乙腈。样品盘温度4 ℃,进样体积2 μL。质谱条件:Orbitrap Exploris 120质谱仪通过Xcalibu软件,进行一级、二级质谱数据采集。
2.6.2 代谢物提取
取正常组、模型组、升清化浊方组每组6只大鼠的血清进行检测,首先在各血清样品中加入300 μL提取液,然后涡旋混匀30 s,再冰水浴超声10 min,-40 ℃静置1 h。将样品放在4 ℃冰箱中解冻,离心15 min,取上清液进样瓶中上机检测。
2.6.3 数据处理与分析
将得到的数据导入SIMCA软件(V16.0.2),进行主成分分析(principal component analysis, PCA)、正交偏最小二乘判别分析(partial least squares discriminant analysis, OPLS-DA),依据t检验(P<0.05)且变量投影重要性(variable importance in the projection, VIP)值>1为标准筛选差异代谢物[5]。将差异代谢物用R软件在KEGG通路数据库下载相关通路。图形采用R包绘制。
2.7 肠道菌群分析
取正常组、模型组、升清化浊组每组6只大鼠的粪便进行检测,随后使用十六烷基三甲基溴化铵(CTAB)提取了不同粪便样本中的DNA,再对细菌DNA进行聚合酶链反应(PCR)扩增,扩增区域为V3-V4[6]上游引物序列:5'-CCTACGGGNGGCWGCAG-3'、下游引物序列:5'-GACTACHVGGGTATCTAATCC-3';Archae[7]上游引物序列:5'-GYGCASCAGKCGMGAAW-3'、下游引物序列:5'-GGACTACHVGGGTWTCTAAT-3';V4上游引物序列:5'-GTGYCAGCMGCCGCGGTAA-3'、下游引物序列:(5'-GGACTACHVGGGTWTCTAAT-3');V4-V5上游引物序列:5'-GTGCCAGCMGCCGCGG-3'、下游引物序列:5'-CCGTCAATTCMTTTRAGTTT-3'。随后样品在Illumina NovaSeq平台上进行测序。
2.8 相关性分析
将入血成分、重点差异代谢物以及肠道菌属进行两两之间的Pearson相关性分析,以|r|>0.7,P<0.05为筛选条件[8],得到肠道菌属、重点差异代谢物两者之间的关联性网络图,以及入血成分、重点差异代谢物、肠道菌属三者之间的关联性网络图。
2.9 统计学方法
采用SPSS 21.0统计软件对实验数据进行分析。若计量资料满足正态分布,用平均值±标准差表示,多组之间比较使用单因素方差分析(One-way ANOVA),方差齐性时两两比较使用最小显著性差异(LSD)法检验,方差不齐时使用Games-Howell法;若计量资料不满足正态分布,采用非参数检验。以P<0.05表示差异有统计学意义。
3 结果
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3.1 升清化浊方对脾虚湿阻型单纯性肥胖大鼠体质量、血脂四项和肝脏脂滴的影响
与正常组相比,模型组大鼠的体质量与血清总胆固醇(total cholesterol,TC)、甘油三酯(triglycerides,TG)及低密度脂蛋白胆固醇(low-density lipoprotein cholesterol,LDL-C)等血脂指标显著增加,高密度脂蛋白胆固醇(high-density lipoprotein cholesterol,HDL-C)显著降低。经过给药治疗后,相较于模型组,升清化浊方组和阳性对照组大鼠的体质量与血清TC、TG及LDL-C等血脂指标都显著降低,HDL-C都有所升高,见图2A~C。这些结果表明升清化浊方治疗肥胖有很好的疗效。与正常组大鼠相比,模型组脂滴浸润现象明显。经升清化浊方给药干预后,相较于模型组,升清化浊方组和阳性对照组大鼠脂滴浸润现象明显降低(图2D)。表明升清化浊方能改善肝脏脂肪蓄积情况。
3.2 入血成分及潜在靶点分析
通过UHPLC-QE-MS法对升清化浊方提取物、空白血清、含药血清进行比较分析。本课题组以蛋白质-蛋白质相互作用(PPI)SEA数据库和相似性得分大于0.8为筛选条件[9],鉴定出升清化浊方27个入血成分。升清化浊方的含药血清正负离子流图(图3A~B),27个入血成分基本信息,见表1。这些成分可能是对机体起主要作用的潜在活性成分。
通过SEA数据库得到升清化浊方27个入血成分作用靶点共503个。从疾病相关数据库收集靶点,去重后得到与肥胖相关的靶点有2 964个。利用Venn作图工具,得到了201个治疗肥胖疾病的潜在靶点(图3C)。将String平台中获取的潜在靶点蛋白互作关系数据导入Cytoscape 3.9.0软件绘制PPI网络图,共有201个节点、1 753条边。依据度值前五为筛选条件,得到甘油醛-3-磷酸脱氢酶(GAPDH)、丝氨酸/苏氨酸蛋白激酶1(Akt1)、白细胞介素6(IL-6)、非受体酪氨酸激酶(SRC)、表皮生长因子受体(EGFR)为关键靶点。
利用DAVID平台对升清化浊方治疗肥胖的潜在靶点进行KEGG富集分析,主要涉及疾病通路、炎症通路和代谢通路等。其中,疾病相关通路有癌症通路、酒精性肝病、糖尿病并发症等通路;炎症相关通路包括PI3K-Akt信号通路、HIF-1信号通路等;代谢相关通路包括类固醇激素生物合成、卵巢类固醇生成。KEGG通路按P值前二十绘制气泡图(图3D)。
3.3 升清化浊方对脾虚湿阻型单纯性肥胖大鼠代谢组学的影响
分别将各组大鼠血清样本进行PCA,得到各组正负离子模式下的代谢轮廓图(图4A),结果发现,正常组和模型组的代谢物完全分开,血清代谢轮廓差异明显,证明脾虚湿阻型单纯性肥胖大鼠的身体机能及代谢水平也发生了较大的变化,而升清化浊方组的代谢水平靠近正常组,证明升清化浊方可以调节脾虚湿阻型单纯性肥胖大鼠的代谢水平。进一步采用OPLS-DA(图4B~C),结果发现,正常组与模型组的散点相距较远,验证了PCA的分析结果,进一步证明了脾虚湿阻型单纯性肥胖大鼠模型造模成功。执行VIP>1,独立样本t检验P<0.05的筛选条件,筛选出差异代谢物(图4D~E)。
将升清化浊方组与模型组之间的差异代谢物,通过R软件在KEGG网站进行通路分析来寻找相关的代谢途径,见图4F。结果发现,升清化浊方治疗脾虚湿阻型单纯性肥胖的相关代谢通路中,有7条代谢通路分别为不饱和脂肪酸的生物合成、花生四烯酸代谢、嘧啶代谢、核苷酸代谢、亚油酸代谢、色氨酸代谢、卟啉代谢。其中涉及到10个重点差异代谢物,即关键差异代谢物为:尿苷(uridine)、左旋犬尿氨酸(L-kynurenine)、胆绿素(biliverdin)、假尿苷(pseudouridine)、前列腺素B2(prostaglandin B2)、γ-亚麻酸(γ-linolenic acid)、二十二碳六烯酸(docosahexaenoic acid,DHA)、二十二碳五烯酸(docosapentaenoic acid)、二十二碳四烯酸(adrenic acid)、12-oxo-5Z,8Z,10E,14Z-二十碳四烯酸(12-KETE)。10个关键代谢物的基本信息,见表2。
3.4 升清化浊方对脾虚湿阻型单纯性肥胖大鼠肠道菌群群落结构的影响
3.4.1 OPLS-DA分析
OPLS-DA通过可视化的低维空间(通常是二维)重新排列样品,最大化展示组与组之间的关系。结果模型组离正常组之间的距离远,表明两者之间的微生物组成结构差异性大,再次证明脾虚湿阻型单纯性肥胖大鼠模型造模成功(图5A);升清化浊组离正常组距离近,甚至有一部分重合,表明这两者之间的微生物组成结构相似,差异性小,证明升清化浊方可以调节脾虚湿阻型单纯性肥胖大鼠的肠道菌群组成并改善肠道菌群结构。同时Chao1、Shannon和Simpson指数显著升高,表明升清化浊方可有效增加肠道微生物群的丰富度及多样性(图5B~C)。
3.4.2 物种组成差异分析
分析门水平和属水平的差异菌群(图5D~E)。正常组中的厚壁菌门(Firmicutes)、拟杆菌门(Bacteroidota)、疣微菌门(Verrucomicrobiota)、变形杆菌门(Proteobacteria)丰度所占比例较高。与正常组相比,模型组的厚壁菌门、拟杆菌门相对丰度显著降低,疣微菌门、变形杆菌门相对丰度显著升高。给药干预后,与模型组相比,升清化浊方组的厚壁菌门、拟杆菌门相对丰度显著升高,疣微菌门、变形杆菌门相对丰度显著降低。在属水平,正常组的阿克曼菌属(Akkermansia)、杜波西菌属(Dubosiella)、唾液联合乳杆菌属(Ligilactobacillus)、乳酸杆菌属(Lactobacillus)丰度所占的比例较高。与正常组相比,模型组的杜波西菌属、唾液联合乳杆菌属、乳酸杆菌属相对丰度显著降低,阿克曼菌属相对丰度显著升高。给药干预后,与模型组相比,升清化浊方组的杜波西菌属、唾液联合乳杆菌属、乳酸杆菌属相对丰度显著升高,阿克曼氏菌属相对丰度显著降低。
3.4.3 KEGG通路分析
利用PICRUSt2功能对模型组与升清化浊方组的菌群进行KEGG通路预测,见图5F。结果发现升清化浊方治疗脾虚湿阻型单纯性肥胖涉及80条通路,其中有36条通路具有显著性(P<0.05)。包括:HIF-1信号通路、类固醇激素的生物合成、FoxO信号通路、PI3K-Akt信号通路、糖尿病并发症中的AGE-RAGE信号通路并发症、卵巢类固醇生成、MAPK信号通路、不饱和脂肪酸的生物合成等通路。
3.5 升清化浊方的关联性分析
肠道菌群和代谢组学之间的关联性网络分析表明,有5种重点差异代谢物与9种肠道菌属有着显著相关性(图6A)。血清药物化学与血清代谢组学、肠道菌群三者之间的关联性网络分析表明,有27种入血成分,6种重点差异代谢物,21种肠道菌属被整合到网络中(图6B)。在网络的综合分析中,每条边都表示两个指标之间的显著相关性(|r|>0.7,P<0.05)。
随后将入血成分、差异代谢物以及肠道菌群所涉及的KEGG通路根据P值进行排序,并选取前30条通路进行绘图结果,见图6C。结果显示,30条通路中有20条是入血成分富集出的代谢通路、3条是差异代谢物所富集出的代谢通路、7条是肠道菌群所富集出的代谢通路。并且其中有2条通路与代谢密切相关:类固醇激素生物合成、不饱和脂肪酸的生物合成。
4 讨论
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研究发现,当人体脾、胃、肾和肺等脏腑的运化疏泄失调时,容易导致痰湿在体内聚积,从而引发肥胖症。在中医的痰湿观念下,通过利水渗湿、升清化浊和益胃生津等中药治疗,可以有效解决痰湿引起的肥胖问题。前期临床发现该方通过其中医调节机制,对脾虚湿阻型单纯性肥胖患者有良好的减肥降脂作用。试验结果显示,该方能有效缓解脾虚湿阻型单纯性肥胖大鼠的体质量、血脂(血清TC、TG及LDL-C水平)过高以及肝脏脂肪蓄积等肥胖问题。本研究对该方的作用机制做了进一步探讨。
本研究采用系统生物学的方法进行分析,并通过关联性网络将血清药物化学与血清代谢组学、肠道菌群进行关联。由肠道菌群与差异代谢物的相关性网络图可知DHA与苏黎世杆菌属(Turicibacter)呈显著正相关。研究表明多不饱和脂肪酸(polyunsaturated fatty acids,PUFA)可恢复肠道微生态平衡并缓解炎症[10]。二十碳五烯酸(eicosapentaenoic, EPA)和DHA作为n-3 PUFAs的代表成分,能够改善啮齿动物的胰岛素抵抗。苏黎世杆菌属的细菌是哺乳动物肠道微生物群的重要成员,与膳食脂肪和体质量的改变有关。研究表明苏黎世杆菌属菌株能够改变宿主胆汁酸和脂质代谢的基因,并将苏黎世杆菌属细菌定位为宿主脂肪生物学的调节[11]。Zhuang等[12]研究发现,经过DHA和EPA喂养的雌性小鼠会提高巴恩斯菌属(Barnesiella)、乳酸杆菌属、梭菌属(Clostridium Xl)、苏黎世杆菌属等菌属的相对丰度。在本研究中,升清化浊方作用于脾虚湿阻型单纯性肥胖大鼠可以提高其DHA的水平,并影响苏黎世杆菌属等丰度。入血成分-差异代谢物-肠道菌属三者之间的关联性分析可知,升清化浊方的27种入血成分可以通过调节肠道菌属和代谢物来达到防治肥胖的效果。本研究在多组学关联发现的科学基础上,同时参考了相关文献资料。例如,其中金柑苷和圣草酚都已被证实具有抗炎作用[13-14];3,4-二羟基扁桃酸是一种去甲肾上腺素代谢物,具有强大的抗氧化潜力[15]。重要的是,23-乙酰泽泻醇B不仅能有效降低小鼠体质量,还能调节肠道菌群的生物多样性,包括减少致病菌,增加有益菌,还能降低花生四烯酸代谢的炎症含量[16]。鞣花酸的摄入可以有效地减轻肥胖并改善肥胖介导的代谢并发症等[17]。作为呋喃香豆素的一员,已有大量研究表明二氢欧山芹素具有抗氧化、抗增殖、抗炎和抗一氧化氮生成等多种生物活性[18]。艾黄素是一种有价值的5-羟基-3,6,7,3',4'-五甲氧基黄酮,存在于多种不同的药用植物中,具有良好的口服生物利用度和药理价值,具有抗炎、抗癌等多种生物活性[19]。这些成分大多具有抗肥胖、抗炎、抗氧化等作用,与肠道菌群、代谢通路的结果密切相关。经过对相关文献[13-19]的深入查阅和分析,本研究进一步验证了药物有效成分调控肠道菌群及其代谢产物的能力。
随后对入血成分、差异代谢物以及肠道菌群所涉及的KEGG通路进行了排序,结果显示代谢方面的通路主要集中在类固醇激素生物合成和不饱和脂肪酸的生物合成。不饱和脂肪酸可分为单不饱和脂肪酸、多不饱和脂肪酸和反式脂肪酸,在代谢组学的研究中,其差异代谢产物主要集中在多不饱和脂肪酸。研究表明,n-3 PUFA [包括 EPA、DHA和α-亚麻酸(α-linolenic acid, ALA)] 是动物必需脂肪酸。n-3 PUFA不能由人体合成,必须通过饮食直接获取或由摄入的ALA转化而来[20]。因此,膳食补充剂或药物制剂对于提供足够的多不饱和脂肪酸至关重要[21]。此外,富含n-3 PUFA的饮食与减少或预防脂肪堆积、胰岛素抵抗、炎症、高血压、动脉粥样硬化、肥胖、心血管疾病和2型糖尿病密切相关[22-24]。不仅如此,还有研究表明n-6/n-3 PUFA比例过高时,会影响拟杆菌门与厚壁菌门的比例[25]。而厚壁菌和拟杆菌会促进短链脂肪酸的产生,是机体肠道中绝对的优势菌门[26]。此外,变形杆菌门外膜主要由脂多糖(lipopolysaccharides, LPS)组成,通常在肥胖等代谢性疾病中水平较高。研究表明,肥胖等代谢疾病会引起LPS在体内循环,聚集形成内毒素血症,引起肠道损伤[27]。本研究发现,升清化浊方显著降低了变形杆菌门的相对丰度,增加了厚壁菌门和拟杆菌门的相对丰度,表明升清化浊方可能是通过影响不饱和脂肪酸的生物合成来调节脾虚湿阻型单纯性肥胖大鼠菌群失衡的现象,减少肠道中的致病菌,增加有益菌。
综上所述,升清化浊方对大鼠脾虚湿阻型单纯性肥胖具有显著的治疗效果,主要得益于其调节肠道微生物群落结构和血清代谢的能力。研究结果揭示,升清化浊方的27种入血成分与大鼠的差异代谢物和菌群存在相关性,并且不饱和脂肪酸的生物合成是主要的代谢通路。这些成分可能通过调节PUFA代谢途径发挥其治疗作用。然而PUFA影响单纯性肥胖大鼠的具体作用机制还有待进一步研究。未来可以进一步探讨肠道菌群与PUFA之间的相互作用,为升清化浊方治疗大鼠单纯性肥胖的作用机制研究提供新的思路与方法。
基金
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  • 国家自然科学基金项目资助(82360966)
  • 江西省教育厅科学技术研究项目资助(GJJ2200959)
  • 江西省卫生健康委员会科技计划项目资助(202211433)
  • 大学生创新创业训练计划项目资助(202310412024)
  • 江西中医药大学大学生创新创业训练计划项目资助(X202310412274)
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参考文献 引证文献
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2024年第59卷第19期
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doi: 10.11669/cpj.2024.19.005
  • 接收时间：2024-05-09
  • 首发时间：2025-12-30
  • 出版时间：2024-10-08
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  • 收稿日期：2024-05-09
基金
国家自然科学基金项目资助(82360966)
江西省教育厅科学技术研究项目资助(GJJ2200959)
江西省卫生健康委员会科技计划项目资助(202211433)
大学生创新创业训练计划项目资助(202310412024)
江西中医药大学大学生创新创业训练计划项目资助(X202310412274)
作者信息
    1 江西中医药大学计算机学院, 南昌 330004
    2 江西中医药大学药学院, 南昌 330004
    3 江西中医药大学实验动物科技中心, 南昌 330004
    4 江西中医药大学附属医院, 南昌 330006

通讯作者:

* 陈浩,男,博士,副教授,副主任药师 研究方向:中医药方证结合及中药炮制学研究 Tel:(0791)87119019
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https://castjournals.cast.org.cn/joweb/zgyxzz/CN/10.11669/cpj.2024.19.005
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2种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
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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