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Article(id=1156264149886821299, tenantId=1146029695717560320, journalId=1146123166801305609, issueId=1156264148657886112, articleNumber=null, orderNo=null, doi=10.12404/j.issn.1671-1815.2403806, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1716307200000, receivedDateStr=2024-05-22, revisedDate=1734278400000, revisedDateStr=2024-12-16, acceptedDate=null, acceptedDateStr=null, onlineDate=1753604455682, onlineDateStr=2025-07-27, pubDate=1740672000000, pubDateStr=2025-02-28, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1753604455682, onlineIssueDateStr=2025-07-27, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1753604455682, creator=13701087609, updateTime=1753604455682, updator=13701087609, issue=Issue{id=1156264148657886112, tenantId=1146029695717560320, journalId=1146123166801305609, year='2025', volume='25', issue='6', pageStart='2193', pageEnd='2636', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1753604455388, creator=13701087609, updateTime=1753771257443, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1156963767234945803, tenantId=1146029695717560320, journalId=1146123166801305609, issueId=1156264148657886112, language=EN, specialIssueTitle=, coverIllustrator=, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1156963767234945804, tenantId=1146029695717560320, journalId=1146123166801305609, issueId=1156264148657886112, language=CN, specialIssueTitle=, coverIllustrator=, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=2265, endPage=2273, ext={EN=ArticleExt(id=1156264150369166263, articleId=1156264149886821299, tenantId=1146029695717560320, journalId=1146123166801305609, language=EN, title=Mechanistic on the Inhibition of LPS-induced Nuclear Translocation of HMGB1 in Macrophages by Modulation of HDAC3 by Shenfu Injection, columnId=1156262732384031076, journalTitle=Science Technology and Engineering, columnName=Papers·Medicine, runingTitle=null, highlight=null, articleAbstract=

To observe the effects of LPS(lipopolysaccharide)-HDAC3(induced histone deacetylase 3) on the expression of HMGB1(high mobility histone B1) and nuclear translocation in RAW264.7 cells, and the intervention effect of SFI(Shenfu injection). RAW264.7 cells were induced by LPS to establish a cellular inflammatory injury model, and the cells were intervened with SFI at doses of 3, 6, and 12 μL/mL for 24 h. RT-qPCR(real-time fluorescence PCR) was used to detect the transcriptional levels of HDAC3, HMGB1, IL-1β, and TNF-α in the cells, and Western-blot was used to detect the protein expression of HMGB1 and HDAC3, and immune immunoassay to detect the protein expression of HMGB1 and HDAC3.HDAC3 protein expression, immunofluorescence to observe the effect of SFI on the subcellular localization of HMGB1. ELISA to detect the secretion levels of HMGB1, IL-1β, and TNF-α in the cell supernatant. And small interfering RNA(siRNA) after targeting to silence the HDAC3 in RAW264.7 cells. to observe the effect of SFI on HMGB1 subcellular localization. Compared with the control group, the transcription and expression of HDAC3 in RAW264.7 cells in the model group were significantly reduced (P<0.01), and the expression of HMGB1 was significantly elevated (P<0.01) and simultaneously migrated from the nucleus to the cytoplasm. The inflammatory factors in the supernatant of the cells, such as HMGB1, IL-1β and TNF-α, were significantly elevated (P<0.01). And compared with the model group, SFI (6, 12 μL/mL dose group) up-regulated the transcription and expression levels of HDAC3, down-regulated the transcription, expression, and nuclear translocation of the inflammatory factor HMGB1, and inhibited the secretion of HMGB1, IL-1β, and TNF-α in RAW264.7 cells. After targeted silencing of HDAC3, a large amount of HMGB1 was localized in the cytoplasm, and there was no significant change in protein localization after LPS stimulation, and SFI could not reverse the abnormal localization of HMGB1. SFI may inhibit LPS-induced extra-nuclear migration of HMGB1 in RAW264.7 cells by up-regulating HDAC3 expression, which in turn inhibited its downstream inflammatory response.

, correspAuthors=Xia LIU, 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=Xiao-long YANG, Wei GOU, Fei AI, Xia LIU, Chun-wei CHU, Xiang-yun CHEN, Jun-feng GUO), CN=ArticleExt(id=1156264163132433361, articleId=1156264149886821299, tenantId=1146029695717560320, journalId=1146123166801305609, language=CN, title=参附注射液调控HDAC3抑制LPS诱导的巨噬细胞HMGB1核转位的机制, columnId=1156262732526637414, journalTitle=科学技术与工程, columnName=论文·医药、卫生, runingTitle=null, highlight=null, articleAbstract=

为观察脂多糖(lipopolysaccharides, LPS)诱导的RAW264.7细胞中组蛋白去乙酰化酶3(histone deacetylase 3,HDAC3)对高迁移率组蛋白B1(high mobility group box-1 protein,HMGB1)表达和核移位的影响及参附注射液(Shenfu injection,SFI)的干预作用。通过LPS诱导RAW264.7细胞建立细胞炎症损伤模型,分别用3、6、12 μL/mL剂量SFI干预细胞24 h。实时荧光PCR法(RT-qPCR)检测细胞中HDAC3、HMGB1、白介素-1β(interleukin-1β,IL-1β)、肿瘤坏死因子α(tumor necrosis factor α,TNF-α)转录水平;Western-blot法检测HMGB1和HDAC3蛋白表达;免疫荧光法观察SFI对HMGB1亚细胞定位的影响;ELISA法检测细胞上清HMGB1、IL-1β和TNF-α分泌水平;小干扰RNA(small interfering RNA,siRNA)靶向沉默RAW264.7细胞中HDAC3后,免疫荧光法观察SFI对HMGB1亚细胞定位的影响。结果表明模型组与对照组比较,模型组RAW264.7细胞中HDAC3的转录和表达均显著降低(P<0.01),HMGB1表达显著升高(P<0.01)且同时从核内向胞浆迁移;细胞上清中炎症因子HMGB1、IL-1β和TNF-α明显升高(P<0.01);与模型组比较,SFI(6、12 μL/mL剂量组)上调RAW264.7细胞中HDAC3的转录和表达水平,下调炎性因子HMGB1的转录、表达、核移位,抑制HMGB1、IL-1β和TNF-α的分泌;靶向沉默HDAC3后,大量HMGB1定位于胞浆,经LPS刺激后蛋白定位无明显变化,且SFI不能逆转HMGB1的异常定位。可见SFI可能通过上调HDAC3表达从而抑制LPS诱导的RAW264.7细胞中HMGB1核外迁移,进而抑制了其下游的炎症反应。

, correspAuthors=刘霞, authorNote=null, correspAuthorsNote=
* 刘霞(1980—),女,汉族,贵州盘县人,博士,教授,博士研究生导师。研究方向:中医药治疗炎性相关疾病的物质基础及分子机制。E-mail:。
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杨晓龙(1994—),男,苗族,湖北恩施人,硕士研究生。研究方向:中医药治疗炎性相关疾病的物质基础及分子机制。 E-mail:。

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杨晓龙(1994—),男,苗族,湖北恩施人,硕士研究生。研究方向:中医药治疗炎性相关疾病的物质基础及分子机制。 E-mail:。

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杨晓龙(1994—),男,苗族,湖北恩施人,硕士研究生。研究方向:中医药治疗炎性相关疾病的物质基础及分子机制。 E-mail:。

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A quality evaluation strategy for Chinese medicinal preparations based on the characteristics of “component structure”[J]. Chinese Herbal Medicine, 2019, 50(17): 4003-4007., articleTitle=A quality evaluation strategy for Chinese medicinal preparations based on the characteristics of “component structure”, refAbstract=null)], funds=[Fund(id=1233422560378024512, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1156264149886821299, awardId=82260874, language=CN, fundingSource=国家自然科学基金地区基金(82260874), fundOrder=null, country=null), Fund(id=1233422560495465027, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1156264149886821299, awardId=2018, language=CN, fundingSource=贵州中医药大学博士启动基金[(2018), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1233422548004827983, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1156264149886821299, xref=1, ext=[AuthorCompanyExt(id=1233422548013216591, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1156264149886821299, companyId=1233422548004827983, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1 School of Basic Medical Sciences, Guizhou University of Traditional Chinese Medicine, Guiyang 550025, China), AuthorCompanyExt(id=1233422548021605200, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1156264149886821299, companyId=1233422548004827983, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1 贵州中医药大学基础医学院, 贵阳 550025)]), AuthorCompany(id=1233422548218737502, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1156264149886821299, xref=2, ext=[AuthorCompanyExt(id=1233422548222931807, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1156264149886821299, companyId=1233422548218737502, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2 The Second Clinical Medical College, Guizhou University of Traditional Chinese Medicine, Guiyang 550025, China), AuthorCompanyExt(id=1233422548235514721, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1156264149886821299, companyId=1233422548218737502, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2 贵州中医药大学第二临床医学院, 贵阳 550025)]), AuthorCompany(id=1233422548378121068, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1156264149886821299, xref=3, ext=[AuthorCompanyExt(id=1233422548386509678, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1156264149886821299, companyId=1233422548378121068, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=3 The Yangsheng College of Guizhou University of Traditional Chinese Medicine, Guiyang 550025, China), AuthorCompanyExt(id=1233422548394898287, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1156264149886821299, companyId=1233422548378121068, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=3 贵州中医药大学中医养生学院, 贵阳 550025)])], figs=[ArticleFig(id=1233422556280189292, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1156264149886821299, language=EN, label=Fig.1, caption=Effects of SFI on protein expression of HDAC3 and HMGB1 in LPS-inducted RAW264.7 cells, figureFileSmall=PNAKBvdGWBTrpSy7q4chQw==, figureFileBig=jGoLIYbSzDSP9J/CF5HeuA==, tableContent=null), ArticleFig(id=1233422556401824118, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1156264149886821299, language=CN, label=图1, caption=SFI对LPS诱导的RAW 264.7细胞中HMGB1和HDAC3的蛋白表达的影响, figureFileSmall=PNAKBvdGWBTrpSy7q4chQw==, figureFileBig=jGoLIYbSzDSP9J/CF5HeuA==, tableContent=null), ArticleFig(id=1233422556557013378, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1156264149886821299, language=EN, label=Fig.2, caption=SFI inhibited the translocation HMGB1 from nucleus to cytoplasm inLPS-inducted RAW264.7 cells, figureFileSmall=gC/Z49id3tfQIDoB7eY9GQ==, figureFileBig=8TNmmuTEo4aH+haZbI/YIQ==, tableContent=null), ArticleFig(id=1233422556661870986, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1156264149886821299, language=CN, label=图2, caption=SFI抑制LPS诱导的RAW264.7细胞中HMGB1从细胞核向细胞质转运

Control为对照组; LPS为脂多糖;Merge为合并图层激光共聚焦×630倍

, figureFileSmall=gC/Z49id3tfQIDoB7eY9GQ==, figureFileBig=8TNmmuTEo4aH+haZbI/YIQ==, tableContent=null), ArticleFig(id=1233422556829643157, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1156264149886821299, language=EN, label=Fig.3, caption=Western blot detection of HDAC3 expression in RAW264.7 cells, figureFileSmall=oX6/mXkSR+O48TrKETz2Wg==, figureFileBig=a3VyyJxBBM21dYfNSV/Y/g==, tableContent=null), ArticleFig(id=1233422556942889376, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1156264149886821299, language=CN, label=图3, caption=Western blot检测RAW264.7细胞中HDAC3的表达, figureFileSmall=oX6/mXkSR+O48TrKETz2Wg==, figureFileBig=a3VyyJxBBM21dYfNSV/Y/g==, tableContent=null), ArticleFig(id=1233422557060329898, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1156264149886821299, language=EN, label=Fig.4, caption=Effects of SFI on the subcellular localization of HMGB1 in LPS-induced RAW264.7 cells following silencing by HDAC3-siRNA was evaluated by immunofluorescence, figureFileSmall=9A7qIeEmtmHvxSAHPcwAAQ==, figureFileBig=zfQ+hd4Fut+52q2BTnkoiA==, tableContent=null), ArticleFig(id=1233422557173576118, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1156264149886821299, language=CN, label=图4, caption=HDAC3沉默后,免疫荧光法观察SFI对LPS诱导的RAW264.7细胞中HMGB1的亚细胞定位的影响

Control为对照组; LPS为脂多糖;Merge为合并图层;荧光显微镜×400倍

, figureFileSmall=9A7qIeEmtmHvxSAHPcwAAQ==, figureFileBig=zfQ+hd4Fut+52q2BTnkoiA==, tableContent=null), ArticleFig(id=1233422557274239421, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1156264149886821299, language=EN, label=Table 1, caption=

PCR primer sequence and length

, figureFileSmall=null, figureFileBig=null, tableContent=
基因 引物序列(5'-3') 产物长度/bp
β-Actin 上游:GAGGTATCCTGACCCTGAAGTA 104
下游:CACACGCAGCTCATTGTAGA
TNF-α 上游:CTACCTTGTTGCCTCCTCTTT 116
下游:GAGCAGAGGTTCAGTGATGTAG
HMGB1 上游:CTGATTGGCTGGAGGAATGT 99
下游:ATACCACCAAGTTTCCCATCTC
IL-β 上游:GCAGCATCTCGACAAGAGCTT 91
下游:GCTCCACGGGCAAGACATAG
HDAC3 上游:ATGCCTTCAACGTGGGTGAC 99
下游:TTAGCTGTGTTGCCCCTTGC
), ArticleFig(id=1233422557404262856, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1156264149886821299, language=CN, label=表1, caption=

PCR引物序列及长度

, figureFileSmall=null, figureFileBig=null, tableContent=
基因 引物序列(5'-3') 产物长度/bp
β-Actin 上游:GAGGTATCCTGACCCTGAAGTA 104
下游:CACACGCAGCTCATTGTAGA
TNF-α 上游:CTACCTTGTTGCCTCCTCTTT 116
下游:GAGCAGAGGTTCAGTGATGTAG
HMGB1 上游:CTGATTGGCTGGAGGAATGT 99
下游:ATACCACCAAGTTTCCCATCTC
IL-β 上游:GCAGCATCTCGACAAGAGCTT 91
下游:GCTCCACGGGCAAGACATAG
HDAC3 上游:ATGCCTTCAACGTGGGTGAC 99
下游:TTAGCTGTGTTGCCCCTTGC
), ArticleFig(id=1233422557538480599, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1156264149886821299, language=EN, label=Table 2, caption=

The sequences of siRNA-1、siRNA-2 and siRNA-3

, figureFileSmall=null, figureFileBig=null, tableContent=
组别 引物序列(5'-3')
siRNA-1 上游:AUUGGUAUCCUGGAGCUGCTT
下游:GCAGCUCCAGGAUACCAAUTT
siRNA-2 上游:AAUGCCUUCAACGUGGGUGTT
下游:CACCCACGUUGAAGGCAUUTT
siRNA-3 上游:UGUGCCCUUACGAGAUGGCTT
下游:GCCAUCUCGUAAGGGCACATT
siRNA NC 上游:UUCUCCGAACGUGUCACGUTT
下游:ACGUGACACGUUCGGAGAATT
), ArticleFig(id=1233422557664309725, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1156264149886821299, language=CN, label=表2, caption=

siRNA-1、siRNA-2和siRNA-3的引物序列

, figureFileSmall=null, figureFileBig=null, tableContent=
组别 引物序列(5'-3')
siRNA-1 上游:AUUGGUAUCCUGGAGCUGCTT
下游:GCAGCUCCAGGAUACCAAUTT
siRNA-2 上游:AAUGCCUUCAACGUGGGUGTT
下游:CACCCACGUUGAAGGCAUUTT
siRNA-3 上游:UGUGCCCUUACGAGAUGGCTT
下游:GCCAUCUCGUAAGGGCACATT
siRNA NC 上游:UUCUCCGAACGUGUCACGUTT
下游:ACGUGACACGUUCGGAGAATT
), ArticleFig(id=1233422557823693291, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1156264149886821299, language=EN, label=Table 3, caption=

Effects of SFI on mRNA expression of HDAC3、HDAC5、HMGB1、P300、IL-1β、TNF-α in LPS-inducted RAW264.7 cells($\stackrel{-}{\mathrm{x}}$±s,n=3)

, figureFileSmall=null, figureFileBig=null, tableContent=
组别 HDAC3 HMGB1 IL-1β TNF-α
空白组 1 1 1 1
模型组 0.397± 0.050## 2.347±0.231## 2.827±0.375## 2.037±0.320##
SFL组 0.420±0.053 1.573±0.297* 2.130±0.281* 1.657±0.273
SFM组 0.732±0.038 ** 1.010±0.079** 1.223±0.199** 1.263±0.110**
SFH组 0.703±0.140** 1.073±0.136** 1.200±0.108** 1.243±0.134**
), ArticleFig(id=1233422559266533875, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1156264149886821299, language=CN, label=表3, caption=

SFI对LPS诱导的RAW264.7细胞中HDAC3、HMGB1、IL-1β、TNF-α mRNA表达的影响($\stackrel{-}{\mathrm{x}}$±s,n=3)

, figureFileSmall=null, figureFileBig=null, tableContent=
组别 HDAC3 HMGB1 IL-1β TNF-α
空白组 1 1 1 1
模型组 0.397± 0.050## 2.347±0.231## 2.827±0.375## 2.037±0.320##
SFL组 0.420±0.053 1.573±0.297* 2.130±0.281* 1.657±0.273
SFM组 0.732±0.038 ** 1.010±0.079** 1.223±0.199** 1.263±0.110**
SFH组 0.703±0.140** 1.073±0.136** 1.200±0.108** 1.243±0.134**
), ArticleFig(id=1233422559409140221, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1156264149886821299, language=EN, label=Table 4, caption=

Effects of SFI on protein expression of HDAC3、HMGB1 in LPS-inducted RAW264.7 cells($\stackrel{-}{\mathrm{x}}$±s,n=3)

, figureFileSmall=null, figureFileBig=null, tableContent=
组别 HDAC3 HMGB1
空白组 0.909± 0.036 1.048± 0.192
模型组 0.311± 0.036## 2.721±0.206##
SFL组 0.391±0.021 2.411±0.076
SFM组 0.582±0.028** 1.810±0.256**
SFH组 0.742±0.037** 1.013±0.181**
), ArticleFig(id=1233422559543357963, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1156264149886821299, language=CN, label=表4, caption=

SFI对LPS诱导的RAW264.7细胞中HDAC3、HMGB1蛋白表达的影响($\stackrel{-}{\mathrm{x}}$±s,n=3)

, figureFileSmall=null, figureFileBig=null, tableContent=
组别 HDAC3 HMGB1
空白组 0.909± 0.036 1.048± 0.192
模型组 0.311± 0.036## 2.721±0.206##
SFL组 0.391±0.021 2.411±0.076
SFM组 0.582±0.028** 1.810±0.256**
SFH组 0.742±0.037** 1.013±0.181**
), ArticleFig(id=1233422559711130133, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1156264149886821299, language=EN, label=Table 5, caption=

Effect of SFI on inflammatory factors in the supernatant of LPS-induced RAW264.7 cells

, figureFileSmall=null, figureFileBig=null, tableContent=
组别 HMGB1/(μg·mL-1) IL-1β/(μg·mL-1) TNF-α/(μg·mL-1)
空白组 0.071±0.007 0.157± 0.004 0.080± 0.005
模型组 0.140±0.021## 0.226± 0.013## 0.162± 0.011##
SFL组 0.090±0.004** 0.185± 0.013* 0.122±0.013**
SFM组 0.070±0.019** 0.131± 0.046** 0.118±0.014**
SFH组 0.065±0.010** 0.118± 0.018** 0.109±0.009**
), ArticleFig(id=1233422559849542175, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1156264149886821299, language=CN, label=表5, caption=

SFI对LPS诱导的RAW 264.7细胞上清液中HMGB1、TNF-α和IL-1β炎症因子影响

, figureFileSmall=null, figureFileBig=null, tableContent=
组别 HMGB1/(μg·mL-1) IL-1β/(μg·mL-1) TNF-α/(μg·mL-1)
空白组 0.071±0.007 0.157± 0.004 0.080± 0.005
模型组 0.140±0.021## 0.226± 0.013## 0.162± 0.011##
SFL组 0.090±0.004** 0.185± 0.013* 0.122±0.013**
SFM组 0.070±0.019** 0.131± 0.046** 0.118±0.014**
SFH组 0.065±0.010** 0.118± 0.018** 0.109±0.009**
), ArticleFig(id=1233422560008925740, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1156264149886821299, language=EN, label=Table 6, caption=

Western blot detection of HDAC3 expression in RAW264.7 cells

, figureFileSmall=null, figureFileBig=null, tableContent=
组别 HDAC3
空白组 0.772± 0.032
NC组 0.699± 0.033##
HDAC3siRNA-1组 0.408±0.030**
HDAC3 siRNA-2组 0.386±0.023**
HDAC3 siRNA-3组 0.314±0.02${{3}^{\mathrm{*}\mathrm{*}}}^{\#}$
), ArticleFig(id=1233422560185086518, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1156264149886821299, language=CN, label=表6, caption=

Western blot检测RAW264.7细胞中HDAC3蛋白的表达

, figureFileSmall=null, figureFileBig=null, tableContent=
组别 HDAC3
空白组 0.772± 0.032
NC组 0.699± 0.033##
HDAC3siRNA-1组 0.408±0.030**
HDAC3 siRNA-2组 0.386±0.023**
HDAC3 siRNA-3组 0.314±0.02${{3}^{\mathrm{*}\mathrm{*}}}^{\#}$
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参附注射液调控HDAC3抑制LPS诱导的巨噬细胞HMGB1核转位的机制
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杨晓龙 1 , 苟玮 1 , 艾飞 2 , 刘霞 3, * , 褚春薇 1 , 陈向云 1 , 郭俊峰 1
科学技术与工程 | 论文·医药、卫生 2025,25(6): 2265-2273
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科学技术与工程 | 论文·医药、卫生 2025, 25(6): 2265-2273
参附注射液调控HDAC3抑制LPS诱导的巨噬细胞HMGB1核转位的机制
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杨晓龙1 , 苟玮1, 艾飞2, 刘霞3, * , 褚春薇1, 陈向云1, 郭俊峰1
作者信息
  • 1 贵州中医药大学基础医学院, 贵阳 550025
  • 2 贵州中医药大学第二临床医学院, 贵阳 550025
  • 3 贵州中医药大学中医养生学院, 贵阳 550025

    • 杨晓龙(1994—),男,苗族,湖北恩施人,硕士研究生。研究方向:中医药治疗炎性相关疾病的物质基础及分子机制。 E-mail:。

通讯作者:

* 刘霞(1980—),女,汉族,贵州盘县人,博士,教授,博士研究生导师。研究方向:中医药治疗炎性相关疾病的物质基础及分子机制。E-mail:。
Mechanistic on the Inhibition of LPS-induced Nuclear Translocation of HMGB1 in Macrophages by Modulation of HDAC3 by Shenfu Injection
Xiao-long YANG1 , Wei GOU1, Fei AI2, Xia LIU3, * , Chun-wei CHU1, Xiang-yun CHEN1, Jun-feng GUO1
Affiliations
  • 1 School of Basic Medical Sciences, Guizhou University of Traditional Chinese Medicine, Guiyang 550025, China
  • 2 The Second Clinical Medical College, Guizhou University of Traditional Chinese Medicine, Guiyang 550025, China
  • 3 The Yangsheng College of Guizhou University of Traditional Chinese Medicine, Guiyang 550025, China
出版时间: 2025-02-28 doi: 10.12404/j.issn.1671-1815.2403806
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摘要
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为观察脂多糖(lipopolysaccharides, LPS)诱导的RAW264.7细胞中组蛋白去乙酰化酶3(histone deacetylase 3,HDAC3)对高迁移率组蛋白B1(high mobility group box-1 protein,HMGB1)表达和核移位的影响及参附注射液(Shenfu injection,SFI)的干预作用。通过LPS诱导RAW264.7细胞建立细胞炎症损伤模型,分别用3、6、12 μL/mL剂量SFI干预细胞24 h。实时荧光PCR法(RT-qPCR)检测细胞中HDAC3、HMGB1、白介素-1β(interleukin-1β,IL-1β)、肿瘤坏死因子α(tumor necrosis factor α,TNF-α)转录水平;Western-blot法检测HMGB1和HDAC3蛋白表达;免疫荧光法观察SFI对HMGB1亚细胞定位的影响;ELISA法检测细胞上清HMGB1、IL-1β和TNF-α分泌水平;小干扰RNA(small interfering RNA,siRNA)靶向沉默RAW264.7细胞中HDAC3后,免疫荧光法观察SFI对HMGB1亚细胞定位的影响。结果表明模型组与对照组比较,模型组RAW264.7细胞中HDAC3的转录和表达均显著降低(P<0.01),HMGB1表达显著升高(P<0.01)且同时从核内向胞浆迁移;细胞上清中炎症因子HMGB1、IL-1β和TNF-α明显升高(P<0.01);与模型组比较,SFI(6、12 μL/mL剂量组)上调RAW264.7细胞中HDAC3的转录和表达水平,下调炎性因子HMGB1的转录、表达、核移位,抑制HMGB1、IL-1β和TNF-α的分泌;靶向沉默HDAC3后,大量HMGB1定位于胞浆,经LPS刺激后蛋白定位无明显变化,且SFI不能逆转HMGB1的异常定位。可见SFI可能通过上调HDAC3表达从而抑制LPS诱导的RAW264.7细胞中HMGB1核外迁移,进而抑制了其下游的炎症反应。

参附注射液  /  HMGB1  /  HDAC3  /  巨噬细胞  /  炎症  /  内毒素休克

To observe the effects of LPS(lipopolysaccharide)-HDAC3(induced histone deacetylase 3) on the expression of HMGB1(high mobility histone B1) and nuclear translocation in RAW264.7 cells, and the intervention effect of SFI(Shenfu injection). RAW264.7 cells were induced by LPS to establish a cellular inflammatory injury model, and the cells were intervened with SFI at doses of 3, 6, and 12 μL/mL for 24 h. RT-qPCR(real-time fluorescence PCR) was used to detect the transcriptional levels of HDAC3, HMGB1, IL-1β, and TNF-α in the cells, and Western-blot was used to detect the protein expression of HMGB1 and HDAC3, and immune immunoassay to detect the protein expression of HMGB1 and HDAC3.HDAC3 protein expression, immunofluorescence to observe the effect of SFI on the subcellular localization of HMGB1. ELISA to detect the secretion levels of HMGB1, IL-1β, and TNF-α in the cell supernatant. And small interfering RNA(siRNA) after targeting to silence the HDAC3 in RAW264.7 cells. to observe the effect of SFI on HMGB1 subcellular localization. Compared with the control group, the transcription and expression of HDAC3 in RAW264.7 cells in the model group were significantly reduced (P<0.01), and the expression of HMGB1 was significantly elevated (P<0.01) and simultaneously migrated from the nucleus to the cytoplasm. The inflammatory factors in the supernatant of the cells, such as HMGB1, IL-1β and TNF-α, were significantly elevated (P<0.01). And compared with the model group, SFI (6, 12 μL/mL dose group) up-regulated the transcription and expression levels of HDAC3, down-regulated the transcription, expression, and nuclear translocation of the inflammatory factor HMGB1, and inhibited the secretion of HMGB1, IL-1β, and TNF-α in RAW264.7 cells. After targeted silencing of HDAC3, a large amount of HMGB1 was localized in the cytoplasm, and there was no significant change in protein localization after LPS stimulation, and SFI could not reverse the abnormal localization of HMGB1. SFI may inhibit LPS-induced extra-nuclear migration of HMGB1 in RAW264.7 cells by up-regulating HDAC3 expression, which in turn inhibited its downstream inflammatory response.

Shenfu injection  /  HMGB1  /  HDAC3  /  macrophage  /  inflammation  /  endotoxic shock
杨晓龙, 苟玮, 艾飞, 刘霞, 褚春薇, 陈向云, 郭俊峰. 参附注射液调控HDAC3抑制LPS诱导的巨噬细胞HMGB1核转位的机制. 科学技术与工程, 2025 , 25 (6) : 2265 -2273 . DOI: 10.12404/j.issn.1671-1815.2403806
Xiao-long YANG, Wei GOU, Fei AI, Xia LIU, Chun-wei CHU, Xiang-yun CHEN, Jun-feng GUO. Mechanistic on the Inhibition of LPS-induced Nuclear Translocation of HMGB1 in Macrophages by Modulation of HDAC3 by Shenfu Injection[J]. Science Technology and Engineering, 2025 , 25 (6) : 2265 -2273 . DOI: 10.12404/j.issn.1671-1815.2403806
正文
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内毒素休克是一种高致死性的全身炎症反应综合征,其致死率居高不下的主要原因与发病初期促炎因子迅速合成释放(免疫系统过度激活)有关[1]。临床数据发现血液中高迁移率族蛋白B1(high mobility group box-1 protein, HMGB1)水平显著升高与内毒素休克严重程度和死亡率成正相关[2],HMGB1可由活化的巨噬细胞释放,分泌出胞的HMGB1触发炎症级联放大效应,可能是导致脓毒症休克晚期致死性的关键因子[3-4]。既往研究表明,HMGB1的超乙酰化是导致其向核外迁移和分泌出胞重要条件,在组蛋白乙酰基转移酶(histone acetyltransferases,HATs)作用下高乙酰化出核,组蛋白去乙酰化酶(histone deacetylase,HDACs),如组蛋白去乙酰化酶3(histone deacetylase 3,HDAC3)的作用下去乙酰化而入核[5]。因此,通过抑制HMGB1的乙酰化水平,阻断HMGB1的分泌和释放,可作为治疗内毒素休克的治疗手段。
内毒素休克属于中医外感病“厥脱”范畴。参附注射液(Shenfu injection,SFI)以红参和附子相伍,共奏回阳救逆、益气固脱之功,临床上用于阳气脱失的厥脱证疗效显著[6]。课题组前期研究结果表明,SFI可有效抑制LPS诱导的巨噬细胞HMGB1核外迁移及分泌[7-8],但参附注射液是否通过调节HMGB1乙酰化水平(或去乙酰化水平)从而影响其分泌还有待进一步证实。基于此,本文研究选用LPS诱导小鼠RAW264.7巨噬细胞的方法建立炎症细胞模型。观察SFI对HDAC3和HMGB1的影响,旨在探索SFI是否通过调节HDAC3的表达从而抑制LPS介导的HMGB1核转位,以期补充和完善SFI治疗内毒素休克的机制。
1 实验材料
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1.1 细胞
RAW264.7小鼠巨噬细胞株,购于中国科学院上海生命科学院细胞资源中心。
1.2 药物
参附注射液(批号:Z51020664,华润三九药业有限公司,规格:10 mL/支,即用即取);LPS(美国Sigma公司,批号:039M4004V,来自大肠杆菌055:B5,用PBS配制成10 mg/mL的药液)。
1.3 主要试剂
胎牛血清(批号:2156874)、DMEM培养基(批号:8120111)、0.25%Trypsin-EDTA(批号:2085174);青-链霉素混合液(批号:15140-122)均购自Gibco公司;Cell Counting Kit-8检测试剂盒(东仁化学科技有限公司,批号:NN710);抗体HMGB1、HDAC3、GAPDH(均购自Abcam生物科技有限公司,批号分别为:2600-1、S1306、ab9485);HRP标记的二抗羊抗兔(武汉博士德生物工程有限公司,批号BA1054);Trizol(天根,批号:DP424);荧光定量PCR检测试剂盒(公司:Genecopies,批号:AOPR-1200);逆转录试剂盒公司:批号:DBI-2220);小鼠HMGB1、TNF-α、IL-1β酶联免疫吸附测定检测试剂盒(购自欣博盛生物科技有限公司;批号分别为:M173318-215a、M170318102a、M190325-318a);RIPA裂解液(批号:P0013C)、PMSF(批号:C1002)、BCA蛋白定量试剂盒(批号:P0010)、Triton X-100(ST795)、DAPI(C1002)均购自碧云天生物公司;PCR引物(由生工生物工程(上海)股份有限公司设计、合成,详见表1);HDAC3的化学合成双链小分子RNA(由锐博生物合成),如表2所示。
1.4 主要仪器
ABI荧光定量PCR仪(型号7500);Merinton微量核酸定量仪(型号:SMA4000);CO2培养箱(CLASS100型)、MK3型酶标仪(美国 Thermo 公司);低温离心机(德国Sigma公司,型号:3-18K);高压灭菌锅(德国Systec公司);德国Leica倒置显微镜;奥林巴斯BX53型生物显微镜;Bio-Rad蛋白凝胶电泳仪、Bio-Rad凝胶成像系统(美国BioRad);HI650型离心机(湖南湘仪实验室仪器开发有限公司);LSM800型激光共聚焦扫描显微镜(德国Leica公司,SM800型);全自动化学发光图像分析仪(上海天能科技有限公司,型号Tanon5200)。
2 实验方法
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2.1 细胞培养
RAW264.7细胞培养瓶中加入完全培养基(含1%双抗、100 μg/mL链霉素、100 U/mL青霉素和10%胎牛血清的杜氏改良Eagle(dulbecco’s modified eagle medium,DMEM)高糖培养基,以下简称为“基础培养基”),置于参数为37 ℃、5%CO2的恒温加湿培养箱中孵育。镜下观察细胞状态和生长情况,当细胞生长至融合度为70%~80%时,取对数生长期细胞用于后续实验。
2.2 免疫荧光观察SFI对RAW264.7细胞HMGB1核转位情况
2.2.1 细胞爬片制作及分组给药
爬片制作方法和给药浓度参照前期研究[8],将细胞以2×104/孔的密度滴加于置有盖玻片的24孔板中,随机分正常对照组、模型组和参附注射液(Shenfu injection, SFI)3、6、12 μL/mL组,每组设3个复孔,每孔培养基为0.5 mL。当细胞融合度为30%时,弃上清,加入无血清培养基,除正常组外,其余组均以终浓度为0.2 μg/mL的脂多糖(lipopolysaccharides, LPS)刺激30 min,随后分别以SFI 3、6、12 μL/mL干预细胞24 h。
2.2.2 免疫荧光标记法
爬片用磷酸盐缓冲液(phosphate buffered saline,PBS)洗3次;加4%的多聚甲醛固定15 min;PBS洗净后加0.5%Triton X-100,室温下通透20 min;PBS洗净,充分吸干液体后,滴山羊血清,室温下封闭30 min;吸去液体加一抗(HMGB1稀释比为1∶200),放入湿盒,于4 ℃冰箱中过夜;磷酸盐吐温缓冲液(phosphate buffered saline with tween 20,PBST)洗净液体,避光环境下(后续步骤均避光操作)加入Cy3标记的羊抗兔IgG(稀释比为1∶100),湿盒中室温孵育1 h;PBST洗净,DAPI染核,孵育室温5 min;PBST洗净,吸干多余液体,用含抗荧光淬灭剂的封片液封片,然后在荧光显微镜下观察HMGB1的亚细胞定位并采集图像。
2.3 SFI对RAW264.7细胞中HDAC3、HMGB1等的影响
2.3.1 RT-qPCR法检测细胞中HDAC3、HMGB1、IL-1β、TNF-α的mRNA水平
取对数期生长期细胞按1×105/mL的密度接种于6孔板,实验分组及给药同2.2.1节。经Trizol提取细胞总RNA,然后根据逆转录试剂盒上的说明,将其逆转录成cDNA。以该cDNA为模板进行定量聚合酶链反应。引物序列如表1所示。反应程序为:95 ℃预变性10 min;95 ℃变性10 s,55 ℃退火20 s,72 ℃延伸35 s,共计40个循环。循环结束后从55 ℃升高到95 ℃获取熔解曲线。熔解曲线分析和数据采用2-ΔΔCT方法进行分析[9]。
2.3.2 采用Western-blot法检测HMGB1和HDAC3蛋白表达量
细胞处理方法同2.3.1节,预冷PBS清洗细胞后,每孔加入120 μL/孔细胞裂解液[(含100∶1∶1的放射免疫沉淀法裂解缓冲液(radio immunoprecipitation assay,RIPA)、苯基甲磺酰氟(Phenylmethanesulfonyl fluoride, PMSF)(100 mmol/L)和磷酸酶抑制剂],摇匀置于冰上裂解30 min后,收集裂解液于离心管中,4 ℃离心机12 000 r/min离心5 min。取上清即为总蛋白,样品存放于-20 ℃备用。经双吡啶甲酸(bicinchoninic acid,BCA)法计算出总蛋白浓度后,将蛋白与5×上样缓冲液以体积比4∶1混合,置于沸水煮10 min变性,随后进行SDS-PAGE凝胶电泳、转膜[0.45 μm聚偏二氟乙烯膜(polyvinylidene fluoride,PVDF)]、封闭、TBST洗净分别加HMGB1、HDAC3和GAPDH一抗(稀释浓度为1∶1 000)4 ℃过夜、TBST洗净加相应辣根过氧化物酶(horseradish peroxidase, HRP)标记的二抗(稀释浓度为1∶5 000)37 ℃摇床2 h。TBST洗净后,利用ECL显色剂和化学发光分析仪对条带进行显影成像。用Image J软件计算条带灰度值,以目的蛋白(HMGB1、HDAC3)与内参GAPDH的比值作为标准化的目的蛋白表达量。
2.3.3 ELISA法检测细胞上清液中HMGB1、IL-1β、TNF-α分泌水平
收集各组细胞上清于提前标记的离心管,按照ELISA试剂盒说明书步骤,进行实验,用酶标仪检测HMGB1、TNF-α、IL-1β的水平。
2.4 SFI对LPS诱导的RAW264.7 HDAC3-/-细胞中HMGB1定位的影响
2.4.1 HDAC3-SiRNA转染片段筛选
HDAC3-SiRNA由锐博生物合成,共3个片段供筛选。细胞铺板后分空白对照组(CON组)、阴性对照siRNA组(NC组)、HDAC3-siRNA-1组、HDAC3-siRNA-2组、HDAC3-siRNA-3组,每组设3个复孔。当细胞达30%融合度时,将每孔液体换成基础培养基,用LipofectamineTM 2000作为转染试剂分别以上片段转入细胞中,6 h后换成完全培养基,继续培养24 h收集细胞,通过Western-blot的方法检测HDAC3蛋白表达量,选取沉默效果最优的片段进行后续转染实验。
2.4.2 观察SFI对敲减HDAC3后各组RAW264.7细胞中HDAC3的表达情况
将细胞接种于六孔板,随机分CON组、NC组、HDAC3-siRNA组、HDAC3-siRNA+LPS组和HDAC3-siRNA+LPS+SFI 6 μL/mL组。当细胞融合度为30%左右时,按上述分组进行细胞转染(方法同2.4.1节),培养6 h后,按分组要求给予LPS刺激和SFI干预(处理方法同2.2.1节)24 h后收集细胞。蛋白免疫印迹(western blot,WB)检测各组细胞HDAC3的表达情况。
2.4.3 观察SFI对敲减HDAC3后对各组RAW264.7细胞中HMGB1核转位的影响
制作细胞爬片并分组处理(方法同2.4.2节)。对细胞爬片进行免疫荧光染色(方法同2.2.2节),在荧光显微镜下观察并采集图像。
2.5 数据分析
所有统计均使用SPSS 22.0进行分析。通过GraphPad Prism 7软件绘制图形。各组之间的比较采用单因素方差分析进行评估,P<0.05视为具有统计学意义。
3 实验结果
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3.1 SFI上调LPS诱导的RAW 264.7细胞中的HDAC3 mRNA,同时抑制HMGB1、TNFα和mRNA
RT-qPCR结果提示如表3所示,与正常对照组比较,LPS可以显著下调HDAC3 mRNA水平,同时上调HMGB1、IL-1β和TNF-α mRNA水平(P<0.01)。与模型组比较,SFI 6 μL/mL和12 μL/mL可显著上调LPS诱导的RAW 264.7细胞中HDAC3的转录,同时抑制HMGB1、IL-1β和TNF-α的转录。
3.2 SFI上调LPS诱导的RAW 264.7细胞中的HDAC3表达,同时抑制HMGB1的表达
WB检测结果如图1和表4所示,LPS可以显著下调HDAC3的表达,同时上调HMGB1的表达,参附注射液6 μL/mL和12 μL/mL处理后,均可显著上调RAW 264.7细胞中HDAC3的表达,从而更接近于正常水平,同时抑制HMGB1的表达。
3.3 SFI可抑制LPS诱导的RAW264.7细胞中HMGB1从细胞核向细胞质易位
从图2可知,正常组中,Cy3标记的HMGB1为红色,并与DAPI着色的细胞核(蓝色)相重叠,即HMGB1主要定位于细胞核中。在LPS刺激下,HMGB1在细胞核和细胞质中均有发现,提示LPS可诱导HMGB1从细胞核向胞质转运。此外,可观察到HMGB1接近细胞膜的边缘,表明HMGB1可能从细胞质分泌到细胞的外部。在SFI干预组中,HMGB1主要位于细胞核中,与对照组相似。结果提示,SFI可以抑制HMGB1的核移位。
3.4 SFI可抑制LPS诱导的RAW264.7细胞上清中HMGB1、IL-1β、TNF-α的分泌
通过ELISA的方法检测细胞上清液中炎症因子的分泌情况,结果如表5所示。与正常组比较,模型组细胞上清中HMGB1、TNF-α和IL-1β分泌显著升高(P<0.01),与模型组比较,SFI可剂量依赖性的抑制炎症因子HMGB1、IL-1β、TNF-α的分泌。
3.5 SFI对LPS诱导的RAW264.7 HDAC3-/-细胞中HMGB1定位的影响
3.5.1 转染效率检测
分别用siRNA-1、siRNA-2和siRNA-3分别转染RAW264.7细胞后,经Western blot的方法检测HDAC3的蛋白表达量。结果如图3所示,与空白对照组和阴性对照组相比,HDAC3 siRNA-1组、HDAC3 siRNA-2组和HDAC3 siRNA-3组差异均有统计学意义(P<0.01),其中以HDAC3 siRNA-3组沉默效果最佳,因此,选取siRNA-3用于后续实验。
3.5.2 HDAC3沉默后参附注射液对HMGB1在细胞质中的转运无影响
为了确定HDAC3在HMGB1的核移位过程中是否起作用,课题组对HDAC3沉默后的RAW264.7细胞中HMGB1的定位进行了观察,结果如图4所示。HDAC3 siRNA-3组中,部分HMGB1从核转移到细胞质,经LPS进行刺激后,HMGB1的位置并未发生明显改变,SFI干预对HMGB1的转运也没有影响。
4 讨论与结论
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HMGB1是具有稳定核小体结构、修复及重组 DNA、调控基因转录功能的一类核内非组蛋白结构蛋白。核外迁移与其高乙酰化水平有关,抑制HMGB1乙酰化水平可能是抑制HMGB1向核外迁移的有效途径。HMGB1具有两个核定位簇,即第27~29和第181~183位赖氨酸位点,研究证实,在LPS的诱导下,单核/巨噬细胞核内的HMGB1的核定位簇发生高度乙酰化,使其亚细胞定位发生改变,由核内向胞浆迁移,进一步分泌出胞成为致炎因子[10]。乙酰化是一种动态的、可逆的翻译后修饰,组蛋白乙酰化水平均由HATs和HDACs维持,这两种可逆修饰在细胞的基因转录、凋亡、分化以及基因的激活或沉默过程中起着至关重要的作用[11-12]。研究证实,HATs/HDACs可对非组蛋白的赖氨酸位点进行修饰,增加HATs(如P300/CBP)或减少HDACs(如HDAC3)可促进HMGB1的高乙酰化和易位,表明HAT/HDAC在调节HMGB1的分泌中起关键作用[13]。
在感染或组织损伤时,先天免疫系统或适应性免疫系统会发生炎症反应,尽管炎症反应可清除大多数感染,但同样也可能会导致免疫过激,是导致内毒素休克的主要原因。免疫过激的与炎症因子风暴有关。巨噬细胞受到LPS刺激后,分泌的IL-1β、TNF-α等细胞因子可与巨噬细胞相互作用形成细胞因子网络,激活急性期反应,此时HMGB1可作为内源性危险信号以高乙酰化状态由细胞核中大量释放出胞[14],胞外HMGB1与其特异性受体结合,可激活NF-κB这一核心炎症通路,再次导致TNF-α、IL-1β和趋化因子(chemokines,CK)产生[6],这些促炎因子又可刺激单核巨噬细胞分泌HMGB1,几种势力形成强大的正反馈环路,引发炎症因子风暴。胞外HMGB1是炎症晚期级联放大反应中的关键一环。单核/巨噬细胞经炎性刺激后6~8 h开始主动分泌HMGB1,16~24 h达到高峰,且持续3 d处于较高水平[14]。与早期促炎因子相比,HMGB1出现较晚且持续时间更长,参与了内毒素休克病理生理大部分过程。HMGB1释放的延迟动力学与疾病的严重程度和随后的致命结果有相关性[3],HMGB1可诱发或促进内毒素休克[15-16],以HMGB1为靶标治疗炎性疾病有较长的治疗时间窗[17]。因此,阻断HMGB1的分泌和释放,或者阻断其与相应受体结合,已成为针对HMGB1为靶标的主要治疗手段。用HMGB1的特异性抗体以及其受体抑制物等治疗炎症也逐步进入研究领域,现已研发多种HMGB1抑制剂,如重组box A、特异性HMGB1抗体、丙酮酸乙酯等,有些已经进入临床试验阶段[18]。单味中药如当归、大黄,中药提取物如槲皮素、甘草甜素、丹参酮 IIA 磺酸钠及复方如血必净注射液等调控HMGB1治疗炎症也受到中外研究者的关注[19-20]。
SFI是临床治疗内毒素休克常用药[21],为宋《严氏济生方》参附汤的改制方。《本经》所载的365味药中仅有9对药既互为相使,又自成一方,参附汤中的人参附子就占1对,开创了回阳救逆方之先河。SFI的主要成分为红参和附片,对内毒素休克即阳气暴脱的厥脱证疗效卓越。现代药理表明,参附汤中的人参皂苷和乌头类生物碱可有效改善内毒素休克微循环、抑制炎症反应[23]。
本文研究发现了SFI对LPS诱导的巨噬细胞中HDAC3的表达有影响,并且可能通过增加HDAC3而阻断HMGB1核浆迁徙。本文研究为SFI在表观遗传学方面治疗内毒素休克提供了新的研究方向和研究靶点,丰富了中医药抗炎机制。目前对中药复方抗炎机制研究仍有很长的路要走,如中药复方制剂难以制定国际评价体系,是阻碍其国际化发展进程的瓶颈问题[24]。因此在今后的研究中,要进一步提取SFI在HDCAs抑制内毒素休克关键通道上的有效成分,开展动物和细胞实验和临床研究。以期提升药物国际认可度和临床优势,使回阳救逆类复方药有更为广阔的发展空间。
基金
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  • 国家自然科学基金地区基金(82260874)
  • 贵州中医药大学博士启动基金[(2018)
文献
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Yang Bing, Feng Liang, Jia Xiaobin. A quality evaluation strategy for Chinese medicinal preparations based on the characteristics of “component structure”[J]. Chinese Herbal Medicine, 2019, 50(17): 4003-4007.
2025年第25卷第6期
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doi: 10.12404/j.issn.1671-1815.2403806
  • 接收时间：2024-05-22
  • 首发时间：2025-07-27
  • 出版时间：2025-02-28
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  • 收稿日期：2024-05-22
  • 修回日期：2024-12-16
基金
国家自然科学基金地区基金(82260874)
贵州中医药大学博士启动基金[(2018)
作者信息
    1 贵州中医药大学基础医学院, 贵阳 550025
    2 贵州中医药大学第二临床医学院, 贵阳 550025
    3 贵州中医药大学中医养生学院, 贵阳 550025

通讯作者:

* 刘霞(1980—),女,汉族,贵州盘县人,博士,教授,博士研究生导师。研究方向:中医药治疗炎性相关疾病的物质基础及分子机制。E-mail:。
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https://castjournals.cast.org.cn/joweb/kxjsygc/CN/10.12404/j.issn.1671-1815.2403806
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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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