Article(id=1322924175182143883, tenantId=1146029695717560320, journalId=1255847919539208197, issueId=1322923782029062261, articleNumber=null, orderNo=null, doi=10.13193/j.issn.1673-7717.2025.06.037, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=null, receivedDateStr=null, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1793339302105, onlineDateStr=2026-10-30, pubDate=1749484800000, pubDateStr=2025-06-10, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1793339302105, onlineIssueDateStr=2026-10-30, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1793339302105, creator=13701087609, updateTime=1793339302105, updator=13701087609, issue=Issue{id=1322923782029062261, tenantId=1146029695717560320, journalId=1255847919539208197, year='2025', volume='43', issue='6', pageStart='1', pageEnd='258', issueExtLink='null', onlineDate='null', pubDate='1749484800000', pubDateStr='2025-06-10', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1793339208371, creator='13701087609', updateTime=1793340556755, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1322929437859258468, tenantId=1146029695717560320, journalId=1255847919539208197, issueId=1322923782029062261, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1322929437859258469, tenantId=1146029695717560320, journalId=1255847919539208197, issueId=1322923782029062261, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=183, endPage=188, ext={EN=ArticleExt(id=1322924175366693260, articleId=1322924175182143883, tenantId=1146029695717560320, journalId=1255847919539208197, language=EN, title=Exploring Mechanism of Intestinal Inflammation in Rats with Spleen Deficiency Treated with Acupuncture Based on LncRNA(UCA1)Regulation of miRNA-331/BRD4 Axis, columnId=1256183365644968140, journalTitle=Chinese Archives of Traditional Chinese Medicine, columnName=Platform of Provincial Project, runingTitle=null, highlight=null, articleAbstract=
Objective

The aim of this study is to investigate the regulatory mechanism of LncRNA(UCA1)on the miRNA-331-3p/bromine domain protein 4(BRD4)axis through acupuncture,in order to ameliorate colonic inflammation in rats with spleen deficiency.Additionally,this study seeks to explore the contemporary mechanism of“strengthening spleen and benefiting Qi”acupuncture therapy in improving symptoms of spleen deficiency and loose stools.

Methods

Sixty male SD rats were randomly assigned to four groups:normal group,model group,non-meridian non-acupuncture group and acupuncture group,with 15 rats in each group.With the exception of the normal group,the remaining three groups were utilized to establish a rat model of spleen deficiency through the compound method of overwork and irregular diet.Following successful modeling,the acupuncture group re-ceived electro-acupuncture at“Zusanli(ST36)”and“Pishu(BL20)”,while the non-meridian non-acupuncture group was treated with electro-acupuncture at the tail tip as a control.The treatment was administered once daily for 20 minutes over a period of 14 days.After treatment,serum and colonic tissues were collected from the rats.The expressions of LncRNA(UCA1)and miRNA-331-3p/BRD4 axis-related genes in rat colon tissue were detected by qPCR.The contents of interleukin-1β(IL-1β)and interleukin-6(IL-6)in rat serum were detected by ELISA.The pathological morphological changes of rat colon tissue were observed by HE staining.The staining of BRD4 in colon was observed by immunohistochemistry method.The immunoactivities of IL-1β,IL-6,tumor necrosis factor-α(TNF-α)and NFκB p65 expression levels were detected by immunofluorescence histochemistry.The apoptosis in colon tissues of each group was detected by TUNEL staining.

Results

Compared with those of the normal group,the expression levels of miRNA-331-3p in colon tissues of rats in the model group and non-meridian non-acupuncture group were significantly decreased(P<0.05),and the mRNA expression levels of LncRNA(UCA1),BRD4,nuclear factor kappa-B(NF-κB)p65,IL-1β,IL-6,IL-8 and TNF-αwere increased(P<0.05).Compared with those of the model group,the expression levels of miRNA-331-3p in the colon tissue of rats were significantly increased(P<0.05)in the acupuncture group,and the mRNA expression levels of LncRNA(UCA1),BRD4,NF-κB p65,IL-1β,IL-6,IL-8 and TNF-αwere decreased(P<0.05).Compared with those of the normal group,the expression levels of IL-1βand IL-6 in the serum of rats in the model group were significantly increased(P<0.05),and the expression levels of IL-1βand IL-6 in the colon tissue of rats in the acupuncture group were decreased(P<0.05)compared with those in the model group.Immunohistochemical results showed that the expression of BRD4 in the colon tissue of rats in the model group was increased compared with that in the normal group(P<0.05).Compared with that in the model group,the expression of BRD4 in the acupuncture group was decreased(P<0.05).The immunofluorescence histochemistry results showed that the mean absorbance values of IL-1β,IL-6,TNF-αand NF-κB p65 immunofluorescence in colon tissues of the model group and non-meridian non-acupuncture group were increased compared with those of the normal group(P<0.05).Compared with those of the model group,the mean absorbance values of IL-1β,IL-6,TNF-αand NF-κB p65 immunofluorescence in colon tissues of rats in the acupuncture group were decreased(P<0.05).In addition,HE staining results showed that acupuncture could effectively reduce inflammatory cell infiltration and loss of cupped cells in colonic mucosal tissues.TUNEL staining results showed that acupuncture could improve colonic inflammation and inhibit apoptosis of colonic mucosal cells in spleen deficiency rats.

Conclusion

Electro-acupuncture at“Zusanli(ST36)”and“Pishu(BL20)”can interfere with the LncRNA(UCA1)regulating miRNA-331-3p/BRD4 axis,inhibit the expressions of intestinal inflammation and pro-inflammatory mediators,and then inhibit the apoptosis of colonic mucosal cells and improve the immunity of the body,thus improving the symptoms of spleen deficiency and loose stools.

, authors=Jidong LIU1, 2, Jiahao ZANG1, Jianbo WANG1, 2, Yi QU1, 2, Yanan XUE1, 2, authorsList=Jidong LIU, Jiahao ZANG, Jianbo WANG, Yi QU, Yanan XUE, authorCompany=null, correspAuthors=null, 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, fund=null), CN=ArticleExt(id=1322924178768273823, articleId=1322924175182143883, tenantId=1146029695717560320, journalId=1255847919539208197, language=CN, title=从LncRNA-UCA1探讨针刺调节miRNA-331-3p/BRD4轴改善脾虚大鼠肠道炎症作用机制, columnId=1256183369033965799, journalTitle=中华中医药学刊, columnName=省级项目平台, runingTitle=null, highlight=null, articleAbstract=
目的

从长链非编码RNA尿路上皮癌相关基因1(LncRNA-UCA1)探讨针刺调节miRNA-331-3p/溴结构域蛋白4(BRD4)轴改善脾虚大鼠肠道炎症作用机制,探讨“健益脾气”针刺疗法改善脾虚便溏症状现代作用机制。

方法

将60只SD雄性大鼠以随机数字表法分为正常组、模型组、非经非穴组、针刺组,每组15只。除正常组外,其余3组采用劳倦过度和不规律饮食复合方法建立脾虚证大鼠模型。造模成功后,针刺组电针“足三里”穴及“脾俞”穴干预治疗,非经非穴组给予电针尾尖作为对照干预,每日1次/20 min,共治疗14 d。治疗结束后,取大鼠血清及结肠组织。qPCR法检测大鼠结肠组织LncRNA-UCA1、miRNA-331-3p/BRD4轴相关基因的表达;ELISA法检测大鼠血清中白介素-1β(IL-1β)、白细胞介素-6(IL-6)的含量;HE染色法观察大鼠结肠组织病理形态变化;免疫组化法观察结肠中BRD4、白细胞介素-8(IL-8)的表达情况;免疫荧光法观察IL-1β、IL-6、肿瘤坏死因子-α(TNF-α)及核因子-κBp65(NFκBp65)的表达水平;TUNEL染色法观察各组结肠组织凋亡情况。

结果

与正常组比较,模型组、非经非穴组大鼠结肠组织miRNA-331-3p的表达水平明显下降(P<0.05),LncRNA-UCA1、BRD4、NF-κB p65、IL-1β、IL-6、IL-8、TNF-α的mRNA表达水平有所升高(P<0.05);与模型组比较,针刺组大鼠结肠组织miRNA-331-3p的表达水平明显升高(P<0.05),LncRNA-UCA1、BRD4、NF-κB p65、IL-1β、IL-6、IL-8、TNF-α的mRNA表达水平有所降低(P<0.05)。与正常组比较,模型组大鼠血清中促炎因子IL-1β、IL-6表达水平明显升高(P<0.05),与模型组比较,针刺组大鼠结肠组织中促炎因子IL-1β、IL-6的表达水平有所降低(P<0.05)。免疫组化法结果显示与正常组比较,模型组大鼠结肠组织BRD4、IL-8表达有所升高(P<0.05);与模型组比较,针刺组的BRD4表达有所降低(P<0.05)。免疫荧光结果显示与正常组比较,模型组、非经非穴组结肠组织IL-1β、IL-6、TNF-α、NF-κB p65免疫荧光平均吸光度值升高(P<0.05);与模型组比较,针刺组大鼠结肠组织中IL-1β、IL-6、TNF-α、NF-κB p65免疫荧光平均吸光度值降低(P<0.05)。另外HE染色结果显示,针刺治疗可有效减轻结肠黏膜组织中炎症细胞浸润和杯状细胞的损失;TUNEL染色结果显示,针刺治疗可以改善脾虚大鼠结肠炎症,抑制结肠黏膜细胞凋亡。

结论

电针“足三里”“脾俞”穴可以干预LncRNA-UCA1调节miRNA-331-3p/BRD4轴,抑制肠道炎症和促炎症介质的表达,进而抑制结肠黏膜细胞凋亡,提高机体免疫力,从而改善脾虚便溏症状。

, authors=刘继东1, 2, 张家豪1, 王建波1, 2, 曲怡1, 2, 薛亚楠1, 2, authorsList=刘继东, 张家豪, 王建波, 曲怡, 薛亚楠, authorCompany=null, correspAuthors=null, authorNote=

刘继东(1976-),男(锡伯族),辽宁沈阳人,副教授,博士,研究方向:针灸作用机制。

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刘继东(1976-),男(锡伯族),辽宁沈阳人,副教授,博士,研究方向:针灸作用机制。

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刘继东(1976-),男(锡伯族),辽宁沈阳人,副教授,博士,研究方向:针灸作用机制。

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注:A.IL-1β表达水平;B.IL-6表达水平。*与正常组比较,P<0.05;#与模型组比较,P<0.05。

, figureFileSmall=aFl25nWArN34dMlok2uuAQ==, figureFileBig=eGysvTSksHtzhRvl2dU/XQ==, tableContent=null), ArticleFig(id=1322924182991938002, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322924175182143883, language=EN, label=null, caption=null, figureFileSmall=02coWT6e0M/+KyY+VShhyQ==, figureFileBig=UDqVK7ITtvElcrp/waDDbw==, tableContent=null), ArticleFig(id=1322924183054852563, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322924175182143883, language=CN, label=图2, caption=各组大鼠肠黏膜组织病理观察(HE), figureFileSmall=02coWT6e0M/+KyY+VShhyQ==, figureFileBig=UDqVK7ITtvElcrp/waDDbw==, tableContent=null), ArticleFig(id=1322924183126155732, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322924175182143883, language=EN, label=null, caption=null, figureFileSmall=SV3FAGAe264jjKYiMvgkGw==, figureFileBig=EJete70ImCf7xqs1+Z1bcA==, tableContent=null), ArticleFig(id=1322924183197458901, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322924175182143883, language=CN, label=图3, caption=各组大鼠结肠组织BRD4、IL-8的表达(400×)

注:*与正常组比较,P<0.05;#与模型组比较,P<0.05。

, figureFileSmall=SV3FAGAe264jjKYiMvgkGw==, figureFileBig=EJete70ImCf7xqs1+Z1bcA==, tableContent=null), ArticleFig(id=1322924183277150678, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322924175182143883, language=EN, label=null, caption=null, figureFileSmall=0QUBiU/CXEbbTKzvNCOiiw==, figureFileBig=A6F5Ym+yRnBOCQ6jJJHrFw==, tableContent=null), ArticleFig(id=1322924183352648151, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322924175182143883, language=CN, label=图4, caption=各组大鼠结肠组织IL-6、IL-1β免疫荧光染色图, figureFileSmall=0QUBiU/CXEbbTKzvNCOiiw==, figureFileBig=A6F5Ym+yRnBOCQ6jJJHrFw==, tableContent=null), ArticleFig(id=1322924183419757016, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322924175182143883, language=EN, label=null, caption=null, figureFileSmall=gEQhxUUsq3wYSsC9BveXCw==, figureFileBig=FnKgDccp3sjuLXp5PBTlHQ==, tableContent=null), ArticleFig(id=1322924183474282969, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322924175182143883, language=CN, label=图5, caption=各组大鼠结肠组织TNF-α、NF-κB p65免疫荧光染色图, figureFileSmall=gEQhxUUsq3wYSsC9BveXCw==, figureFileBig=FnKgDccp3sjuLXp5PBTlHQ==, tableContent=null), ArticleFig(id=1322924183537197530, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322924175182143883, language=EN, label=null, caption=null, figureFileSmall=Ki/NPceewz6Z4qzE1AJKFw==, figureFileBig=dncYEmn7wk15F4+xVaWT8A==, tableContent=null), ArticleFig(id=1322924183604306395, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322924175182143883, language=CN, label=图6, caption=各组大鼠结肠上皮黏膜细胞凋亡情况(200×), figureFileSmall=Ki/NPceewz6Z4qzE1AJKFw==, figureFileBig=dncYEmn7wk15F4+xVaWT8A==, tableContent=null), ArticleFig(id=1322924183683998172, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322924175182143883, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
基因引物序列(5′→3′)产物长度/bp
miRNA-331-3p上游:UCAUUUUACCCUCCACCAGGGGG(加尾法)
LncRNA-UCA1上游:GAGCCCTGTCCTTTGATG279
下游:GATAGACTGTGACCCGATA
BRD4上游:GAGGAAAGAAACAGGGACA77
下游:TCTGCGGAGAAGTTGATG
NFκBp65上游:ACAACTATGAGATGAACTCCGGG114
下游:GATAGCAGTGGGCTGTCTCC
IL-6上游:ATCAAGAGTCAACTGGCTCAACTCA142
下游:TGTTGGGCGCACATAGCTTATC
IL-8上游:CCCCCATGGTTCAGAAGATTG125
下游:TTGTCAGAAGCCAGCGTTCAC
TNFα上游:CCACGCTCTTCTGTCTACTG145
下游:GCTACGGGCTTGTCACTC
IL-1β上游:TGTGATGTTCCCATTAGAC131
下游:AATACCACTTGTTGGCTTA
β-actin上游:CCTCTATGCCAACACAGT155
下游:AGCCACCAATCCACACAG
), ArticleFig(id=1322924183751107037, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322924175182143883, language=CN, label=表1, caption=

引物序列

, figureFileSmall=null, figureFileBig=null, tableContent=
基因引物序列(5′→3′)产物长度/bp
miRNA-331-3p上游:UCAUUUUACCCUCCACCAGGGGG(加尾法)
LncRNA-UCA1上游:GAGCCCTGTCCTTTGATG279
下游:GATAGACTGTGACCCGATA
BRD4上游:GAGGAAAGAAACAGGGACA77
下游:TCTGCGGAGAAGTTGATG
NFκBp65上游:ACAACTATGAGATGAACTCCGGG114
下游:GATAGCAGTGGGCTGTCTCC
IL-6上游:ATCAAGAGTCAACTGGCTCAACTCA142
下游:TGTTGGGCGCACATAGCTTATC
IL-8上游:CCCCCATGGTTCAGAAGATTG125
下游:TTGTCAGAAGCCAGCGTTCAC
TNFα上游:CCACGCTCTTCTGTCTACTG145
下游:GCTACGGGCTTGTCACTC
IL-1β上游:TGTGATGTTCCCATTAGAC131
下游:AATACCACTTGTTGGCTTA
β-actin上游:CCTCTATGCCAACACAGT155
下游:AGCCACCAATCCACACAG
), ArticleFig(id=1322924183830798814, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322924175182143883, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
组别n造模前体质量治疗前体质量治疗后体质量
正常组15215.40±6.69402.20±17.76483.40±29.62
模型组15212.40±6.77181.80±5.26*223.00±12.53*
非经非穴组15217.20±4.77185.20±13.77*236.60±9.99*
针刺组15211.60±7.77194.40±13.24*284.00±10.93*#
), ArticleFig(id=1322924183902101983, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322924175182143883, language=CN, label=表2, caption=

各组大鼠体质量比较

, figureFileSmall=null, figureFileBig=null, tableContent=
组别n造模前体质量治疗前体质量治疗后体质量
正常组15215.40±6.69402.20±17.76483.40±29.62
模型组15212.40±6.77181.80±5.26*223.00±12.53*
非经非穴组15217.20±4.77185.20±13.77*236.60±9.99*
针刺组15211.60±7.77194.40±13.24*284.00±10.93*#
), ArticleFig(id=1322924183985988064, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322924175182143883, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
组别n造模前抓力/(N)治疗前抓力/(N)治疗后抓力/(N)结肠长度/(cm)
正常组15887.40±27.371020.10±68.121278.00±22.318.88±0.34*
模型组15874.00±19.65798.20±23.91*807.00±54.14*7.30±0.42*
非经非穴组15891.80±25.58775.40±45.32*785.40±26.47*7.43±0.57*#
针刺组15893.40±16.89803.40±36.86*869.40±23.45*#8.08±0.15
), ArticleFig(id=1322924184061485537, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322924175182143883, language=CN, label=表3, caption=

各组大鼠抓力及结肠长度比较

, figureFileSmall=null, figureFileBig=null, tableContent=
组别n造模前抓力/(N)治疗前抓力/(N)治疗后抓力/(N)结肠长度/(cm)
正常组15887.40±27.371020.10±68.121278.00±22.318.88±0.34*
模型组15874.00±19.65798.20±23.91*807.00±54.14*7.30±0.42*
非经非穴组15891.80±25.58775.40±45.32*785.40±26.47*7.43±0.57*#
针刺组15893.40±16.89803.40±36.86*869.40±23.45*#8.08±0.15
), ArticleFig(id=1322924184136983010, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322924175182143883, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
组别nmiRNA-331-3pUCA1BRD4NFκB P65
正常组151.00±0.001.00±0.001.00±0.001.00±0.00
模型组150.68±0.32*1.80±0.12*2.02±0.38*1.97±0.25*
非经非穴组150.70±0.59*1.88±0.11*2.13±0.49*1.97±0.24*
针刺组150.87±0.95*#1.30±0.11*#1.41±0.80#1.37±0.19#
), ArticleFig(id=1322924184220869091, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322924175182143883, language=CN, label=表4, caption=

针刺对脾虚大鼠UCA1及miRNA-331-3p/BRD4轴mRNA表达影响

, figureFileSmall=null, figureFileBig=null, tableContent=
组别nmiRNA-331-3pUCA1BRD4NFκB P65
正常组151.00±0.001.00±0.001.00±0.001.00±0.00
模型组150.68±0.32*1.80±0.12*2.02±0.38*1.97±0.25*
非经非穴组150.70±0.59*1.88±0.11*2.13±0.49*1.97±0.24*
针刺组150.87±0.95*#1.30±0.11*#1.41±0.80#1.37±0.19#
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组别nIL-1β mRNAIL-6 mRNAIL-8 mRNATNF-αmRNA
正常组151.00±0.001.00±0.001.00±0.001.00±0.00
模型组152.22±0.54*1.39±0.22*2.10±0.55*1.88±0.23*
非经非穴组151.81±0.25*1.34±0.16*1.86±0.10*1.43±0.20*#
针刺组151.42±0.19#0.97±0.16#1.19±0.30#1.14±0.27#
), ArticleFig(id=1322924184376058341, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322924175182143883, language=CN, label=表5, caption=

针刺对脾虚大鼠IL-1β、IL-6、IL-8、TNF-α的mRNA表达影响(±s)

, figureFileSmall=null, figureFileBig=null, tableContent=
组别nIL-1β mRNAIL-6 mRNAIL-8 mRNATNF-αmRNA
正常组151.00±0.001.00±0.001.00±0.001.00±0.00
模型组152.22±0.54*1.39±0.22*2.10±0.55*1.88±0.23*
非经非穴组151.81±0.25*1.34±0.16*1.86±0.10*1.43±0.20*#
针刺组151.42±0.19#0.97±0.16#1.19±0.30#1.14±0.27#
), ArticleFig(id=1322924184443167206, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322924175182143883, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
组别nIL-1βIL-6TNF-αNFκB P65
正常组15102.90±2.5298.34±4.80105.44±2.1487.58±3.05
模型组15131.89±6.14*146.54±3.66*135.30±2.38*120.96±4.54*
非经非穴组15128.25±9.64*148.83±6.58*131.57±2.56*111.51±2.80*#
针刺组15113.54±3.76#124.41±3.78*#115.86±2.60*#98.42±4.28*#
), ArticleFig(id=1322924184514470375, tenantId=1146029695717560320, journalId=1255847919539208197, articleId=1322924175182143883, language=CN, label=表6, caption=

各组大鼠中IL-1β、IL-6、TNF-α及NF-κB p65平均荧光强度比较(±s)

, figureFileSmall=null, figureFileBig=null, tableContent=
组别nIL-1βIL-6TNF-αNFκB P65
正常组15102.90±2.5298.34±4.80105.44±2.1487.58±3.05
模型组15131.89±6.14*146.54±3.66*135.30±2.38*120.96±4.54*
非经非穴组15128.25±9.64*148.83±6.58*131.57±2.56*111.51±2.80*#
针刺组15113.54±3.76#124.41±3.78*#115.86±2.60*#98.42±4.28*#
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从LncRNA-UCA1探讨针刺调节miRNA-331-3p/BRD4轴改善脾虚大鼠肠道炎症作用机制
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刘继东 1, 2 , 张家豪 1 , 王建波 1, 2 , 曲怡 1, 2 , 薛亚楠 1, 2
中华中医药学刊 | 省级项目平台 2025,43(6): 183-188
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中华中医药学刊 |省级项目平台 2025 , 43 (6) : 183 -188
从LncRNA-UCA1探讨针刺调节miRNA-331-3p/BRD4轴改善脾虚大鼠肠道炎症作用机制
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2.辽宁中医药大学中医脏象理论及应用教育部重点实验室,辽宁 沈阳 110847, bio={"content":"

刘继东(1976-),男(锡伯族),辽宁沈阳人,副教授,博士,研究方向:针灸作用机制。

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刘继东(1976-),男(锡伯族),辽宁沈阳人,副教授,博士,研究方向:针灸作用机制。

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刘继东1, 2, 张家豪1, 王建波1, 2, 曲怡1, 2, 薛亚楠1, 2
作者信息
  • 1.辽宁中医药大学,辽宁 沈阳 110847
  • 2.辽宁中医药大学中医脏象理论及应用教育部重点实验室,辽宁 沈阳 110847
作者简介:

刘继东(1976-),男(锡伯族),辽宁沈阳人,副教授,博士,研究方向:针灸作用机制。

Exploring Mechanism of Intestinal Inflammation in Rats with Spleen Deficiency Treated with Acupuncture Based on LncRNA(UCA1)Regulation of miRNA-331/BRD4 Axis
Jidong LIU1, 2, Jiahao ZANG1, Jianbo WANG1, 2, Yi QU1, 2, Yanan XUE1, 2
Affiliations
  • 1.Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China
  • 2.Key Laboratory of The Theory and Application of Viscera in Chinese Medicine,Ministry of Education,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China
出版时间: 2025-06-10 doi: 10.13193/j.issn.1673-7717.2025.06.037
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目的

从长链非编码RNA尿路上皮癌相关基因1(LncRNA-UCA1)探讨针刺调节miRNA-331-3p/溴结构域蛋白4(BRD4)轴改善脾虚大鼠肠道炎症作用机制,探讨“健益脾气”针刺疗法改善脾虚便溏症状现代作用机制。

方法

将60只SD雄性大鼠以随机数字表法分为正常组、模型组、非经非穴组、针刺组,每组15只。除正常组外,其余3组采用劳倦过度和不规律饮食复合方法建立脾虚证大鼠模型。造模成功后,针刺组电针“足三里”穴及“脾俞”穴干预治疗,非经非穴组给予电针尾尖作为对照干预,每日1次/20 min,共治疗14 d。治疗结束后,取大鼠血清及结肠组织。qPCR法检测大鼠结肠组织LncRNA-UCA1、miRNA-331-3p/BRD4轴相关基因的表达;ELISA法检测大鼠血清中白介素-1β(IL-1β)、白细胞介素-6(IL-6)的含量;HE染色法观察大鼠结肠组织病理形态变化;免疫组化法观察结肠中BRD4、白细胞介素-8(IL-8)的表达情况;免疫荧光法观察IL-1β、IL-6、肿瘤坏死因子-α(TNF-α)及核因子-κBp65(NFκBp65)的表达水平;TUNEL染色法观察各组结肠组织凋亡情况。

结果

与正常组比较,模型组、非经非穴组大鼠结肠组织miRNA-331-3p的表达水平明显下降(P<0.05),LncRNA-UCA1、BRD4、NF-κB p65、IL-1β、IL-6、IL-8、TNF-α的mRNA表达水平有所升高(P<0.05);与模型组比较,针刺组大鼠结肠组织miRNA-331-3p的表达水平明显升高(P<0.05),LncRNA-UCA1、BRD4、NF-κB p65、IL-1β、IL-6、IL-8、TNF-α的mRNA表达水平有所降低(P<0.05)。与正常组比较,模型组大鼠血清中促炎因子IL-1β、IL-6表达水平明显升高(P<0.05),与模型组比较,针刺组大鼠结肠组织中促炎因子IL-1β、IL-6的表达水平有所降低(P<0.05)。免疫组化法结果显示与正常组比较,模型组大鼠结肠组织BRD4、IL-8表达有所升高(P<0.05);与模型组比较,针刺组的BRD4表达有所降低(P<0.05)。免疫荧光结果显示与正常组比较,模型组、非经非穴组结肠组织IL-1β、IL-6、TNF-α、NF-κB p65免疫荧光平均吸光度值升高(P<0.05);与模型组比较,针刺组大鼠结肠组织中IL-1β、IL-6、TNF-α、NF-κB p65免疫荧光平均吸光度值降低(P<0.05)。另外HE染色结果显示,针刺治疗可有效减轻结肠黏膜组织中炎症细胞浸润和杯状细胞的损失;TUNEL染色结果显示,针刺治疗可以改善脾虚大鼠结肠炎症,抑制结肠黏膜细胞凋亡。

结论

电针“足三里”“脾俞”穴可以干预LncRNA-UCA1调节miRNA-331-3p/BRD4轴,抑制肠道炎症和促炎症介质的表达,进而抑制结肠黏膜细胞凋亡,提高机体免疫力,从而改善脾虚便溏症状。

足三里  /  脾俞  /  脾虚  /  肠道炎症  /  LncRNA-UCA1  /  miRNA-331/BRD4
Objective

The aim of this study is to investigate the regulatory mechanism of LncRNA(UCA1)on the miRNA-331-3p/bromine domain protein 4(BRD4)axis through acupuncture,in order to ameliorate colonic inflammation in rats with spleen deficiency.Additionally,this study seeks to explore the contemporary mechanism of“strengthening spleen and benefiting Qi”acupuncture therapy in improving symptoms of spleen deficiency and loose stools.

Methods

Sixty male SD rats were randomly assigned to four groups:normal group,model group,non-meridian non-acupuncture group and acupuncture group,with 15 rats in each group.With the exception of the normal group,the remaining three groups were utilized to establish a rat model of spleen deficiency through the compound method of overwork and irregular diet.Following successful modeling,the acupuncture group re-ceived electro-acupuncture at“Zusanli(ST36)”and“Pishu(BL20)”,while the non-meridian non-acupuncture group was treated with electro-acupuncture at the tail tip as a control.The treatment was administered once daily for 20 minutes over a period of 14 days.After treatment,serum and colonic tissues were collected from the rats.The expressions of LncRNA(UCA1)and miRNA-331-3p/BRD4 axis-related genes in rat colon tissue were detected by qPCR.The contents of interleukin-1β(IL-1β)and interleukin-6(IL-6)in rat serum were detected by ELISA.The pathological morphological changes of rat colon tissue were observed by HE staining.The staining of BRD4 in colon was observed by immunohistochemistry method.The immunoactivities of IL-1β,IL-6,tumor necrosis factor-α(TNF-α)and NFκB p65 expression levels were detected by immunofluorescence histochemistry.The apoptosis in colon tissues of each group was detected by TUNEL staining.

Results

Compared with those of the normal group,the expression levels of miRNA-331-3p in colon tissues of rats in the model group and non-meridian non-acupuncture group were significantly decreased(P<0.05),and the mRNA expression levels of LncRNA(UCA1),BRD4,nuclear factor kappa-B(NF-κB)p65,IL-1β,IL-6,IL-8 and TNF-αwere increased(P<0.05).Compared with those of the model group,the expression levels of miRNA-331-3p in the colon tissue of rats were significantly increased(P<0.05)in the acupuncture group,and the mRNA expression levels of LncRNA(UCA1),BRD4,NF-κB p65,IL-1β,IL-6,IL-8 and TNF-αwere decreased(P<0.05).Compared with those of the normal group,the expression levels of IL-1βand IL-6 in the serum of rats in the model group were significantly increased(P<0.05),and the expression levels of IL-1βand IL-6 in the colon tissue of rats in the acupuncture group were decreased(P<0.05)compared with those in the model group.Immunohistochemical results showed that the expression of BRD4 in the colon tissue of rats in the model group was increased compared with that in the normal group(P<0.05).Compared with that in the model group,the expression of BRD4 in the acupuncture group was decreased(P<0.05).The immunofluorescence histochemistry results showed that the mean absorbance values of IL-1β,IL-6,TNF-αand NF-κB p65 immunofluorescence in colon tissues of the model group and non-meridian non-acupuncture group were increased compared with those of the normal group(P<0.05).Compared with those of the model group,the mean absorbance values of IL-1β,IL-6,TNF-αand NF-κB p65 immunofluorescence in colon tissues of rats in the acupuncture group were decreased(P<0.05).In addition,HE staining results showed that acupuncture could effectively reduce inflammatory cell infiltration and loss of cupped cells in colonic mucosal tissues.TUNEL staining results showed that acupuncture could improve colonic inflammation and inhibit apoptosis of colonic mucosal cells in spleen deficiency rats.

Conclusion

Electro-acupuncture at“Zusanli(ST36)”and“Pishu(BL20)”can interfere with the LncRNA(UCA1)regulating miRNA-331-3p/BRD4 axis,inhibit the expressions of intestinal inflammation and pro-inflammatory mediators,and then inhibit the apoptosis of colonic mucosal cells and improve the immunity of the body,thus improving the symptoms of spleen deficiency and loose stools.

Zusanli(ST36)  /  Pishu(BL20)  /  spleen deficiency  /  intestinal inflammation  /  LncRNA-UCA1  /  miRNA-331/BRD4
刘继东, 张家豪, 王建波, 曲怡, 薛亚楠. 从LncRNA-UCA1探讨针刺调节miRNA-331-3p/BRD4轴改善脾虚大鼠肠道炎症作用机制. 中华中医药学刊, 2025 , 43 (6) : 183 -188 . DOI: 10.13193/j.issn.1673-7717.2025.06.037
Jidong LIU, Jiahao ZANG, Jianbo WANG, Yi QU, Yanan XUE. Exploring Mechanism of Intestinal Inflammation in Rats with Spleen Deficiency Treated with Acupuncture Based on LncRNA(UCA1)Regulation of miRNA-331/BRD4 Axis[J]. Chinese Archives of Traditional Chinese Medicine, 2025 , 43 (6) : 183 -188 . DOI: 10.13193/j.issn.1673-7717.2025.06.037
《素问·灵兰秘典论篇》曰:“脾胃者,仓廪之官,五味出焉。”李中梓《医宗必读》也写道:“一有此身,必资谷气,谷入于胃,洒陈于六腑而气至,和调于五脏而血生,而人资之以为生者也,故曰后天之本在脾。”脾把饮食物化生为人体所需精微,并将这些精微吸收转输至全身,人体脏腑赖脾胃后天濡养,气血化生得源而获充足的营养滋润[1]。而脾胃虚弱,不能受纳水谷并运化精微,纳化停滞,清浊杂混而下,则成便溏。李东垣《脾胃论》曰:“历观诸篇而参考之,则元气之充足,皆由脾胃之气无所伤,而后能滋养元气。”《内经》也记载:“正气存内,邪不可干”,脾胃虚弱,气血元气生化乏源,机体抗邪功能降低,免疫系统失衡,则会诱发肠道炎症[2]。现代免疫学认为,免疫防御系统是机体抵抗各种致病微生物侵袭,维持机体协调平衡性的重要生理功能,脾虚引起便溏,免疫防御被破坏,导致肠道菌群失调,容易引起致病菌的增加,诱发肠炎变成本虚标实的症状。故此,脾虚是诱发肠道炎症的一个重要因素。现代研究表明,肠上皮细胞凋亡进程可由多种炎症因子[如白介素-1β(IL-1β)、白细胞介素-6(IL-6)]活性增加而驱动[3]。研究还发现在炎症部位可观察到结肠上皮黏膜细胞的凋亡增加,这会破坏结肠黏膜结构完整和屏障功能,进一步加强结肠炎症反应[4]。
长非编码RNA(LncRNA)多数位于细胞质中,是一种长度超过200个核苷酸,因缺少可译框架而没有蛋白质编码功能的RNA[5]。LncRNAs已被证明参与多种炎症过程,并通过与核酸和蛋白质相互作用、引诱miRNAs、激活或抑制单个分子而与免疫疾病的发病机制有关[6]。已发现UCA1与多种炎症及肿瘤密切相关并在其发生中发挥作用,可发挥促炎功能[7]。已知微小RNA(miRNAs)结合靶mRNA的3’非翻译区(3’-UTR)并调节其表达,多种mRNA可由单个miRNA同时靶向,从而调节各个基因的基因表达[8]。miRNAs已被证明可以调节多种生物活性,包括炎症、细胞死亡和癌症进展[9]。已发现miR-331-3p参与多种疾病的炎症反应过程且UCA1通过调节miR-331-3p可以改善肠道炎症[10]。BRD4是溴和末端外(BET)家族的重要成员,其溴结构域可以结合乙酰化组蛋白和转录因子以调节多种病理生理活性,包括癌症和炎症[11]。BRD4是参与炎症反应的重要溴和末端外蛋白且BRD4是调节炎症的NF-κB通路的关键调节因子[12]。现代研究发现BRD4是miR-331-3p的直接靶点,BRD4受miR-331-3p调节,BRD4的消耗逆转了miR-331-3p抑制对LPS处理的FHC中的细胞活力、细胞凋亡和炎症的影响[10]。
脾虚失健,运化失司,T淋巴细胞功能下降,从而促使机体免疫能力降低,进一步加重便溏。研究表明脾虚和肠道炎症有密切的联系,且于临床上广泛应用针灸改善脾虚症状,但其作用机制与是否调控相关肠道炎症尚缺乏充足的理论及系统的研究。本实验通过观察电针脾虚大鼠干预LncRNA-UCA1调节miRNA-331-3p/BRD4轴的治疗作用,探讨其健运脾气、抵抗炎症、抑制细胞凋亡的效果,为临床针灸治疗脾虚提供科学实验依据。
SPF级SD大鼠60只,8周龄,体质量180~200 g,购于辽宁长生生物技术股份有限公司,动物生产许可证号[SCXK(辽)2020-0001]。饲养环境:辽宁中医药大学实验动物中心,使用许可证号[SYXK(辽)2019-0004],湿度为(45±5)%,温度为(22±2)℃。依据随机数字表法将大鼠分为4组,为正常组、模型组、非经非穴组和针刺组,每组15只。实验过程中严格按照中华人民共和国科技部2006年颁布的《关于善待实验动物的指导性意见》对动物进行处置。本实验通过辽宁中医药大学动物伦理审查批准实施[2021-016]。
Stratagene Mx3000p荧光PCR仪(Agilent Technologies),M8数字切片扫描成像系统(Precipoint),针灸针(0.19 mm×10 mm,苏州医疗用品厂有限公司),电子针疗仪(苏州医疗用品厂有限公司),酶标仪(赛默飞世尔科技公司),TNF-α一抗(华安生物,ER65189),miRNA提取试剂盒(29122,康为世纪),miRNA反转试剂盒(25722,康为世纪),miRNA扩增试剂盒(02422,康为世纪),免疫组化试剂盒(KIT-9710,福州迈新生物技术开发有限公司),组织自发荧光淬灭剂(G1221,武汉赛维尔生物科技有限公司)。
采用劳倦过度和不规律饮食复合造模法建立SD大鼠模型。适应性饲养1周后开始造模。正常组每日自由进食及饮水,其余3组先饱食1 d,再禁食2 d,3 d为1个周期,并每日力竭游泳(尾部系5%体质量保险丝),每日1次,前后相差10 min,水温保持在25℃,水深保持在50 cm,持续21 d。以大鼠出现食欲不振、形体消瘦、倦怠神疲、毛发失荣、大便不调(便溏),扎堆嗜睡等症状,符合《中医实验动物模型方法学》中对于脾虚大鼠的评定标准。
造模成功后,针刺组选取“足三里”及“脾俞”进行电针治疗,定位参考中国针灸学会实验动物研究会制定的《动物针灸穴位图谱》,固定后,采用疏密波、频率2 Hz/100 Hz,输出电压2 V,20 min/d,持续14 d。非经非穴组确定大鼠尾尖为非经非穴点,给予同等条件下的电针处理。正常组和模型组大鼠在每天同一时间固定,不给予电针干预。
取腹主动脉血:以上4组均用10%水合氯醛进行腹腔麻醉注射(3 mL/kg体质量),酒精消毒,充分暴露腹腔,棉球剥离开脏器,分离出粉色腹主动脉,右手持取血针,针尖接近平行刺入腹主动脉,回血后,取血针另一端连接真空抗凝采血管,收集3/4管后拔出。
取结肠:暴露腹腔并剪取结肠,0.9%氯化钠溶液冲洗肠腔,取部分结肠置于4%多聚甲醛固定,另将剩余结肠置于-80℃保存。
实时定量荧光PCR观察大鼠结肠组织LncRNA-UCA1、miRNA-331-3p、BRD4、P65、IL-1β、IL-6、IL-8、TNF-α mRNA表达:于-80℃取出结肠组织,并使用Trizol法提取总RNA,后逆转录为cDNA;miRNA则按miRNA提取及转录试剂盒说明书提取并转录为cDNA,设定条件:95℃,10 min;95℃,15 s;60℃,1 min,40个循环,扩增所用引物见表1,荧光定量PCR仪进行检测,溶解反应按Mx3000p自动设定的条件进行。2-△△CT法对PCR结果进行定量分析。
ELISA检测大鼠血清中IL-1β、IL-6的含量:室温血液自然凝固15 min,离心20 min(3000 r/min)后,收集上清液加样酶标板,标准品梯度稀释加入标准品孔,空白孔加入40μL样品稀释液再加入10μL待测样品,充分混匀,封板后以37℃温育30 min,弃去液体并甩干,每孔加满20倍稀释洗涤液,静置30 s弃去(重复5次)后拍干。加入酶标试剂温育并洗涤。每孔先后加入显色剂A、B各50μL,37℃避光显色15 min每孔加终止液50μL。450 nm波长(酶标仪)下测定吸光度,计算IL-1β、IL-6的表达量。
HE染色法观察结肠组织形态学变化:将4%多聚甲醛固定好的结肠组织取出,乙醇梯度脱水,二甲苯透明,常规石蜡包埋、切片,经脱蜡、清洗后进行HE染色,显微镜下观察组织形态学变化。
免疫组化染色法观察结肠BRD4表达:取出4%多聚甲醛固定好的结肠组织,石蜡包埋,切片4μm厚,放入60℃恒温箱中加热3 h,取出冷却至室温。将切片脱蜡、水化,水流冲洗。置于柠檬酸缓冲液中95~100℃下孵育20 min,以用于抗原提取。静置冷却至室温后,分别用阻断内源性过氧化物酶、非特异染色阻断剂孵育10 min。一抗,4℃孵育过夜。分别用IgG聚合物、加链霉菌抗生物素蛋白-过氧化氢酶室温孵育10 min,DBA显色。采用生物显微镜阅片,Image J软件对图像进行光密度分析。
免疫荧光染色法观察结肠IL-1β、IL-6、TNF-α、NFκBp65的表达:取结肠石蜡切片,经过脱蜡后,将切片充分浸泡在柠檬酸钠抗原修复液内,95~100℃下孵育20 min,取出自然冷却至室温。切片甩干,滴加组织自发荧光淬灭剂A液,室温孵育30 min,BSA牛血清白蛋白封闭40 min。一抗,4℃孵育过夜。荧光二抗避光孵育50 min,洗涤后加入DAPI,避光孵育10 min,最后用荧光显微镜观察。采集图像后用Image J软件进行图像分析。
TUNEL染色法观察结肠组织细胞凋亡:取4%多聚甲醛固定好的样本,石蜡包埋后切片处理,按照TUNEL染色试剂盒操作步骤检测结肠组织细胞凋亡。荧光显微镜观察采集图像及分析。
采用SPSS 26.0软件进行统计分析,数据采用均数±标准差()表示,所有数据均行正态性及方差齐性检验,均满足正态分布。组间比较使用单因素方差分析。以P<0.05为差异有统计学意义。
见表2、表3。
与正常组比较,模型组、非经非穴组大鼠结肠组织miRNA-331-3p的表达水平明显下降(P<0.05),LncRNA-UCA1、BRD4、P65、IL-1β、IL-6、IL-8、TNF-α的mRNA表达水平有所升高(P<0.05);与模型组比较,针刺组大鼠结肠组织miRNA-331-3p的表达水平明显升高(P<0.05),LncRNA-UCA1、BRD4、P65、IL-1β、IL-6、IL-8、TNF-α的mRNA表达水平有所降低(P<0.05)。见表4、表5。
与正常组比较,模型组、非经非穴组大鼠结肠组织中促炎因子IL-1β、IL-6表达水平明显升高(P<0.05),与模型组比较,针刺组大鼠结肠组织中促炎因子IL-1β、IL-6的表达水平有所降低(P<0.05)。见图1。
正常组中结肠组织隐窝边缘规则,杯状细胞和隐窝结构正常;模型组、非经非穴组中炎性细胞增加,隐窝边缘不规则,且杯状细胞减少;针刺组可见炎性细胞有所减少,杯状细胞数量和隐窝结构得到一定程度的恢复。见插页ⅩⅩⅦ图2。
与正常组相比,模型组、非经非穴组大鼠结肠组织中BRD4、IL-8的表达量明显增加,差异有统计学意义(P<0.05);与模型组相比,针刺组大鼠结肠组织中BRD4、IL-8的表达量有所减少,差异有统计学意义(P<0.05)。见插页ⅩⅩⅦ图3。
由插页ⅩⅩⅧ图4、插页ⅩⅩⅨ图5及表6可见,阳性表达呈现绿色,细胞核表达呈现蓝色,Merge为两者共表达。与正常组比较,模型组及非经非穴组IL-1β、IL-6、TNF-α、NF-κB p65的免疫荧光表达有所升高(P<0.05)。与模型组比较,针刺组IL-1β、IL-6、TNF-α、NF-κB p65的免疫荧光表达有所降低(P<0.05)。
正常组中绿色荧光标记的凋亡细胞数量最少,凋亡细胞与总细胞重合最少,凋亡阳性细胞表达率最低;与正常组比较,模型组、非经非穴组结肠组织细胞凋亡情况最为严重,凋亡细胞与总细胞重合最多,凋亡阳性细胞表达率升高(P<0.05);与模型组比较,针刺组细胞凋亡率下降明显,凋亡细胞与总细胞重合均减少(P<0.05)。见插页ⅩⅩⅩ图6。
《医宗必读·肾为先天脾为后天论》曰:“脾为中宫之土,土为万物之母[13]。”脾中央土以灌四傍,主运化水谷,布散精微,为“后天之本”,脾将饮食水谷化为精微,并将其转输至全身。《素问·阴阳应象大论篇》云:“清气在下,则生飧泄;浊气在上,则生腹胀。”《儒门事亲》亦云:“故湿之气……其化在天为雨,在地为泥,在人为脾,甚则为泄。”脾胃虚弱,无法受纳水谷并运化精微,水谷停滞,清浊混杂而下,遂成便溏。《景岳全书·泄泻》载:“泄泻之本无不由于脾胃,若饮食失节……不能输化,致合污下降,而泻利作矣。”论述了脾胃与便溏的紧密的联系。脾胃亏虚,脏腑不得充养,机体的抵抗力降低[14]。且便溏日久,使肠道屏障功能逐渐减弱,无法发挥正常功能,机体免疫系统作用减弱,则易产生肠道炎症[15]。肠道炎症会破坏肠黏膜的完整性且降低肠黏膜屏障功能,进而促进肠黏膜细胞凋亡[16]。
因此本研究以“从脾论治”入手,通过电针足三里、脾俞干预LncRNA-UCA1调节miRNA-331-3p/BRD4轴治疗脾虚肠道炎症的机制。足三里穴出自《灵枢·本输》,为足阳明胃经合穴、胃之下合穴,属于“四总穴”“回阳九针”之一[17]。故此选择针刺足三里以健脾,符合从脾论治之法。脾俞穴出自《灵枢·背俞》,属足太阳膀胱经,脾之背俞穴。背俞穴广泛用于治疗相应的脏腑病证及有关的组织器官病证,故脾俞穴是治疗脾胃病的要穴,除可用于治疗背痛等局部病证外,还善于治疗脾胃疾患如腹胀、腹泻、痢疾、纳呆、水肿等[18]。
肠道轻度炎性反应目前被认为与脾虚有关,可以通过针刺调节脾虚,从而降低炎性,改善结肠黏膜功能[19]。本研究结果显示,大鼠体质量降低,抓力降低,病理改变呈现出轻度炎性反应,与有关研究结果一致。针刺干预后,大鼠体质量增加,抓力升高,肠道炎性反应减轻。相关实验表明针刺可调节脾虚,降低炎性反应,改善肠道功能及完整性,减轻便溏症状[20]。
LncRNAs已被证明与DNA或RNA特异性碱基配对,这使它们能够调节基因表达并参与几种生物学过程[21]。据报道LncRNA在各种场合参与炎症活动,且已经在肠道炎症中鉴定出LncRNA的失调[22]。诸多研究表明,miRNAs可能与肠道炎症的进展有关,其机制可能通过控制肠道炎症相关基因参与炎症的发病[23]。现代研究揭示了miR-331-3p被UCA1靶向并反向调节,本研究中也证明了这一点[10]。据报道,BRD4是炎性疾病中的重要调节因子且BRD4是miR-331-3p的直接靶点,miR-331-3p的缺失会导致BRD4的高表达来促使炎症发生。NF-κB是一种转录因子,控制着一系列参与炎症信号级联反应的促炎基因,在肠道炎症的发病机制中起着重要作用[24]。研究已表明,BRD4是调节炎症的NF-κB通路的关键调节因子,NF-κB信号可以被BRD4激活[25]。本实验结果表明BRD4可以通过激活肠道炎症中的NF-κB信号通路促进结肠上皮细胞损伤。
本研究探讨了电针治疗LncRNA-UCA1干预miR-331-3p/BRD4轴对细胞凋亡的影响。结果发现,BRD4的过度表达及miR-331-3p低表达会促进结肠上皮细胞凋亡,而BRD4的表达下调及miR-331-3p高表达则表现出相反的作用。此外,炎症结肠中促炎细胞因子(IL-1β、IL-6等)的水平显著升高,会破坏肠黏膜完整性并且降低肠黏膜屏障功能,进而促进肠黏膜细胞凋亡。
总之,本研究首先证明电针治疗LncRNA-UCA1干预miR-331-3p/BRD4轴在肠道炎症的调节中起重要作用。LncRNA-UCA1的过表达通过抑制结肠miR-331-3p并加强BRD4的表达从而促进炎症。本研究显示电针足三里、脾俞可以抑制LncRNA-UCA1的表达,进一步抑制BRD4并促进miR-331-3p的表达,从而抑制炎症因子的表达。LncRNAUCA1、miR-331-3p和BRD4在炎症性结肠病变中的差异表达可能为针刺治疗脾虚提供一个有希望的生物标志物。LncRNA-UCA1干预miR-331-3p/BRD4轴是脾虚治疗的潜在治疗靶点。
  • 辽宁省教育厅高等学校基本科研项目(LJKZ0886)
  • 辽宁省科学技术计划项目(2021-MS-248)
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2025年第43卷第6期
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doi: 10.13193/j.issn.1673-7717.2025.06.037
  • 首发时间:2026-10-30
  • 出版时间:2025-06-10
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辽宁省教育厅高等学校基本科研项目(LJKZ0886)
辽宁省科学技术计划项目(2021-MS-248)
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    1.辽宁中医药大学,辽宁 沈阳 110847
    2.辽宁中医药大学中医脏象理论及应用教育部重点实验室,辽宁 沈阳 110847
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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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