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Article(id=1320435150077412025, tenantId=1146029695717560320, journalId=1319373162502258839, issueId=1320435149154669015, articleNumber=null, orderNo=null, doi=10.16016/j.2097-0927.202511042, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1762444800000, receivedDateStr=2025-11-07, revisedDate=1767456000000, revisedDateStr=2026-01-04, acceptedDate=null, acceptedDateStr=null, onlineDate=1792745872277, onlineDateStr=2026-10-23, pubDate=1769702400000, pubDateStr=2026-01-30, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1792745872277, onlineIssueDateStr=2026-10-23, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1792745872277, creator=13701087609, updateTime=1792745872277, updator=13701087609, issue=Issue{id=1320435149154669015, tenantId=1146029695717560320, journalId=1319373162502258839, year='2026', volume='48', issue='2', pageStart='107', pageEnd='250', issueExtLink='null', onlineDate='null', pubDate='1769702400000', pubDateStr='2026-01-30', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1792745872057, creator='13701087609', updateTime=1792749692151, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1320451171886682715, tenantId=1146029695717560320, journalId=1319373162502258839, issueId=1320435149154669015, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1320451171886682716, tenantId=1146029695717560320, journalId=1319373162502258839, issueId=1320435149154669015, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=116, endPage=127, ext={EN=ArticleExt(id=1320435152086483643, articleId=1320435150077412025, tenantId=1146029695717560320, journalId=1319373162502258839, language=EN, title=Curcumin alleviates indomethacin-ciprofloxacin combination-induced small intestinal injury in mice by modulating Treg/Th17 balance and synergistically treats severe traumatic infections, columnId=1320435151990014650, journalTitle=Journal of Army Medical University, columnName=Military Medicine, runingTitle=null, highlight=null, articleAbstract=
Objective

To investigate the effect and underlying mechanisms of curcumin (CUR) on small intestinal injuries associated with severe traumatic infections after combined treatment of ciprofloxacin (CIP) and indomethacin (IND).

Methods

Adult male C57BL/6 mice (8 weeks old) were randomly divided into (10 animals for survival rate observation, 10 for other indicators) control group, model group [hemorrhage+ fracture+Pseudomonas aeruginosa (PA) infection], IND+CIP treatment group, and IND+ CIP+CUR treatment group. The survival rates within 72 h post-modelling were observed across all groups. At 24 h post-modelling, peripheral blood samples, small intestinal tissues and mesenteric lymph nodes (MLN) were harvested. Small intestinal length, hemorrhagic status, and adhesions were also observed grossly in each group. Bacterial load was determined via dilute plating. HE staining was used to observe histopathological alterations in intestinal tissues across groups. ELISA was employed to measure the contents of inflammatory cytokines, IL-1β, IL-6, IL-10 and TNF-α in the serum and small intestinal homogenate of each group. Liver, renal, and cardiac functions were evaluated by measuring key biochemical indicators. Flow cytometry was applied to measure the percentages of IFN-γ, IL-4, TGF-β, and IL-17 positive cells within CD4+ T cells in the MLN. Real-time quantitative polymerase chain reaction (qPCR) was performed to detect the mRNA expression of forkhead box protein P3(FOXP3) and retinoic acid receptor-related orphan receptor gamma (RORγ)-t in each group.

Results

Both the IND+CIP treatment group and the IND+CIP+CUR treatment group significantly improved survival rates in mice with severe traumatic infections (P<0.05). CUR addition reduced bacterial load in the mice (P<0.01). The IND+CIP treatment group exhibited small intestinal injury, which worsened over time, manifested by elevated contents of TNF-α (P<0.01), IL-6 (P<0.05), and IL-1β (P<0.05) and decreased IL-10 (P<0.01) in the small intestine tissues, accompanied with pathological damage in the small intestine. The IND+CIP+CUR treatment group had significantly reversed small intestinal injury, reduced inflammatory factor levels (P<0.01), and restored normal small intestinal structure, showing no significant differences in relevant blood biochemical indicators when compared to the IND+CIP treatment group. The expression of FOXP3 was increased and that of RORγ-t was decreasedin CD4+ T cells from the IND+CIP+CUR treatment group than the IND+CIP treatment group (both P<0.01).

Conclusion

The combination of CUR with IND and CIP can effectively alleviate severe traumatic infections, while antagonize small intestinal damage observed in IND and CIP therapy, with no significant adverse effects on vital organ functions. This mechanism may be related to the balance between regulatory T (Treg) cells and T helper 17 (Th17) cells.

, authors=Daiqi CHEN1, 2, Jing YU2, Yu XIA2, Guochang LIU2, Haiyan WANG2, Suiyan LI1, Jun YAN2, authorsList=Daiqi CHEN, Jing YU, Yu XIA, Guochang LIU, Haiyan WANG, Suiyan LI, Jun YAN, authorCompany=null, correspAuthors=Suiyan LI, Jun YAN, authorNote=null, correspAuthorsNote=null, copyrightStatement=© 2026 Journal of Army Medical University, copyrightOwner=Editorial Office of Journal of Army Medical University, 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=1320435154028446411, articleId=1320435150077412025, tenantId=1146029695717560320, journalId=1319373162502258839, language=CN, title=姜黄素通过调节Treg/Th17平衡减轻吲哚美辛-环丙沙星联用所致小鼠小肠损伤并协同治疗严重创伤感染, columnId=1320435152170369724, journalTitle=陆军军医大学学报, columnName=论著·军事医学, runingTitle=null, highlight=null, articleAbstract=
目的

探讨姜黄素（curcumin，CUR）对吲哚美辛（indomethacin， IND）联用环丙沙星（ciprofloxacin， CIP）治疗严重创伤感染中小肠损伤的作用及其机制。

方法

将8周龄雄性C57BL/6小鼠完全随机分为4组（生存率观察，n=10；其余指标，n=8）：对照组、创伤感染组［失血+骨折+铜绿假单胞菌（Pseudomonas aeruginosa，PA）感染］、IND+CIP治疗组和IND+CIP+CUR治疗组。观察各组小鼠建模后72 h内生存率；收集建模24 h后外周血、小肠组织及肠系膜淋巴结（mesenteric lymph node， MLN），观察各组小肠长度变化、出血状况和粘连情况；稀释涂布平板法检测各组细菌载量；HE染色观察各组肠道组织病理学改变；ELISA分别检测各组血清和小肠匀浆炎症因子水平（IL-1β、IL-6、IL-10和TNF-α）；血液生化指标检测各组肝功、肾功和心功能重要指标。流式细胞术检测各组MLN中CD4+T细胞中IFN-γ、IL-4、TGF-β和IL-17阳性百分比；qPCR检测各组叉头状蛋白P3（forkhead box protein P3， FOXP3）和维甲酸受体相关孤儿受体γ（retinoic acid receptor-related orphan receptor γ， RORγ）-t mRNA表达。

结果

IND+CIP治疗组和IND+CIP+CUR治疗组均能显著提高严重创伤感染小鼠生存率（P<0.05），CUR联用能够降低小鼠细菌载量（P<0.01）。IND+CIP治疗组会引发小肠损伤，并随时间推移呈现加重趋势，表现为小肠TNF-α（P<0.01）、IL-6（P<0.01）和IL-1β水平升高（P<0.05），IL-10水平降低（P<0.05），并伴有小肠病理损伤；IND+CIP+CUR治疗组能显著逆转小肠损伤，降低炎性因子水平（P<0.01），使小肠结构恢复正常，相关血液生化指标与IND+CIP治疗组无明显差异。与IND+CIP治疗组相比，IND+CIP+CUR治疗组CD4+T细胞中FOXP3表达增高（P<0.01），RORγ-t表达降低（P<0.01）。

结论

CUR联用IND和CIP可有效治疗严重创伤感染，同时拮抗IND和CIP疗法中出现的小肠损伤，并对重要脏器功能无明显副作用，其机制可能与调节性T细胞（regulatory T cell， Treg）-辅助性T细胞17（helper T cell 17， Th17）平衡有关。

, authors=陈代琦1, 2, 余静2, 夏雨2, 刘国昌2, 王海燕2, 李遂焰1, 严军2, authorsList=陈代琦, 余静, 夏雨, 刘国昌, 王海燕, 李遂焰, 严军, authorCompany=null, correspAuthors=李遂焰, 严军, authorNote=null, correspAuthorsNote=
严 军，E-mail：
李遂焰，E-mail：
YAN Jun, E-mail：
LI Suiyan, E-mail：
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2Department of Special War Wound， State Key Laboratory of Trauma and Chemical Poisoning， Daping Hospital， Army Medical University （Third Military Medical University）， Chongqing， China, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null), CN=AuthorExt(id=1320435157392278234, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1320435150077412025, authorId=1320435156633109206, language=CN, stringName=陈代琦, firstName=null, middleName=null, lastName=null, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=1, 2, address=1西南交通大学生命科学与工程学院，四川 成都
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Phytother Res， 2022， 36（4）： 1708-1723., articleTitle=Curcumin regulates the homeostasis of Th17/Treg and improves the composition of gut microbiota in type 2 diabetic mice with colitis, refAbstract=null)], funds=null, companyList=[AuthorCompany(id=1320435154619843276, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1320435150077412025, xref=1, ext=[AuthorCompanyExt(id=1320435154854724301, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1320435150077412025, companyId=1320435154619843276, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1School of Life Science and Engineering， Southwest Jiaotong University， Chengdu， Sichuan), AuthorCompanyExt(id=1320435154867307214, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1320435150077412025, companyId=1320435154619843276, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1西南交通大学生命科学与工程学院，四川 成都)]), AuthorCompany(id=1320435156524057298, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1320435150077412025, xref=2, ext=[AuthorCompanyExt(id=1320435156532445907, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1320435150077412025, companyId=1320435156524057298, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2Department of Special War Wound， State Key Laboratory of Trauma and Chemical Poisoning， Daping Hospital， Army Medical University （Third Military Medical University）， Chongqing， China), AuthorCompanyExt(id=1320435156545028820, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1320435150077412025, companyId=1320435156524057298, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2陆军军医大学大坪医院特殊环境战伤防治研究室， 创伤与化学中毒全国重点实验室，重庆)])], figs=[ArticleFig(id=1320435163276886788, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1320435150077412025, language=EN, label=Figure 1, caption=Survival rates in severe traumatic infected mice after drug combination treatment, figureFileSmall=FilXJFGgwnnzjktzSCIJuA==, figureFileBig=ll5YOCIgSefm6CP2Slipvw==, tableContent=null), ArticleFig(id=1320435163352384261, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1320435150077412025, language=CN, label=图1, caption=药物联用后严重创伤感染小鼠生存率

A： PA concentration exploration （n=8）； B： Survival rates of mice in each combination therapy group （n=8）. TI： Trauma-infection group； PA： Pseudomonas aeruginosa； IND： Indomethacin； CIP： Ciprofloxacin； CUR： Curcumin

, figureFileSmall=FilXJFGgwnnzjktzSCIJuA==, figureFileBig=ll5YOCIgSefm6CP2Slipvw==, tableContent=null), ArticleFig(id=1320435163776008966, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1320435150077412025, language=EN, label=Figure 2, caption=Antibacterial efficacy after drug combination treatment, figureFileSmall=ESE5OBJ2OSgOGAYzI12OmA==, figureFileBig=j5vjSSMVb9PEvMdFcDja9A==, tableContent=null), ArticleFig(id=1320435163864089351, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1320435150077412025, language=CN, label=图2, caption=药物联用后抑菌效果

A： Blood agar plate （n=8）； B： 10-1 Blood agar plate colony count （n=8）； C： 10-2 Blood agar plate colony count （n=8）. TI： Trauma-infection group； IND： Indomethacin； CIP： Ciprofloxacin； CUR： Curcumin

, figureFileSmall=ESE5OBJ2OSgOGAYzI12OmA==, figureFileBig=j5vjSSMVb9PEvMdFcDja9A==, tableContent=null), ArticleFig(id=1320435163939586824, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1320435150077412025, language=EN, label=Figure 3, caption=Pathological changes in the small intestine in severe traumatic infected mice after drug combination treatment, figureFileSmall=2vev/ItR55mEglcH9cKERQ==, figureFileBig=ni4a41uFbxAoDsakIp2Z1Q==, tableContent=null), ArticleFig(id=1320435165571171081, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1320435150077412025, language=CN, label=图3, caption=药物联用后严重创伤感染小鼠小肠病理改变

A： Length of the small intestine （n=8）； B： Representative images of the small intestine by HE staining （n=8）； C： Histogram of Chiu’s score （n=8）. TI： Trauma-infection group； IND： Indomethacin； CIP： Ciprofloxacin； CUR： Curcumin

, figureFileSmall=2vev/ItR55mEglcH9cKERQ==, figureFileBig=ni4a41uFbxAoDsakIp2Z1Q==, tableContent=null), ArticleFig(id=1320435165676028682, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1320435150077412025, language=EN, label=Figure 4, caption=Serum secretion levels of inflammatory cytokines in severe traumatic infected mice after drug combination （n=6）, figureFileSmall=oxX68KuHCX5AqQVt6nVLLQ==, figureFileBig=fZb/ggvIUWf0NzBJhIKDZw==, tableContent=null), ArticleFig(id=1320435165755720459, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1320435150077412025, language=CN, label=图4, caption=药物联用后严重创伤感染小鼠血清炎性因子含量水平 （n=6）

A~C： Histogram of serum levels of TNF-α （A）， IL-6 （B）， IL-10 （C） and IL-1β （D）. TI： Trauma-infection group； IND： Indomethacin； CIP： Ciprofloxacin； CUR： Curcumin

, figureFileSmall=oxX68KuHCX5AqQVt6nVLLQ==, figureFileBig=fZb/ggvIUWf0NzBJhIKDZw==, tableContent=null), ArticleFig(id=1320435165839606540, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1320435150077412025, language=EN, label=Figure 5, caption=Small intestine secretion levels of inflammatory cytokines in severe traumatic infected mice after drug combination （n=6）, figureFileSmall=h3xE59bLe95dVR7mr+y/ZA==, figureFileBig=rWyIab9gD7+NiZhTeywwyg==, tableContent=null), ArticleFig(id=1320435165898326797, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1320435150077412025, language=CN, label=图5, caption=药物联用后严重创伤感染小鼠小肠炎性因子含量水平 （n=6）

A~C： Histogram of TNF-α （A）， IL-6 （B）， IL-10 （C） and IL-1β （D） contents in small intestine tissues. TI： Trauma-infection group； IND： Indomethacin； CIP： Ciprofloxacin； CUR： Curcumin

, figureFileSmall=h3xE59bLe95dVR7mr+y/ZA==, figureFileBig=rWyIab9gD7+NiZhTeywwyg==, tableContent=null), ArticleFig(id=1320435165965435662, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1320435150077412025, language=EN, label=Figure 6, caption=Subsets changes of CD4+ T cells in severe traumatic infected mice after drug combination treatment （n=3）, figureFileSmall=32lz7k/Ttnv3gcgdOCSymQ==, figureFileBig=vm+pfZj7ykVbvBgBzmcNfA==, tableContent=null), ArticleFig(id=1320435166032544527, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1320435150077412025, language=CN, label=图6, caption=药物联用后各组小鼠CD4+T细胞亚群变化（n=3）

A~D： Flow cytometry results and histograms of IFN-γ （A）， IL-4 （B）， TGF-β （C） and IL-17 （D） expression in CD4+ cells.TI： Trauma-infection group； IND： Indomethacin； CIP： Ciprofloxacin； CUR： Curcumin

, figureFileSmall=32lz7k/Ttnv3gcgdOCSymQ==, figureFileBig=vm+pfZj7ykVbvBgBzmcNfA==, tableContent=null), ArticleFig(id=1320435166103847696, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1320435150077412025, language=EN, label=Figure 7, caption=Changes in Treg-Th17 balance in severe traumatic infected mice after drug combination treatment （n=8）, figureFileSmall=Bsk1o1059/NG9h1VP/h+xw==, figureFileBig=4F+mFZ/dUgsnLlhb9oYZmw==, tableContent=null), ArticleFig(id=1320435166183539473, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1320435150077412025, language=CN, label=图7, caption=药物联用后严重创伤感染小鼠Treg-Th17平衡变化 （n=8）

A， B： Relative mRNA expression of RORγ-t （A） and FOXP3 （B） in mesenteric lymph nodes. TI： Trauma-infection group； IND： Indomethacin； CIP： Ciprofloxacin； CUR： Curcumin； RORγ-t： Retinoic acid receptor-related orphan receptor γ-t； FOXP3： Forkhead box protein P3

, figureFileSmall=Bsk1o1059/NG9h1VP/h+xw==, figureFileBig=4F+mFZ/dUgsnLlhb9oYZmw==, tableContent=null), ArticleFig(id=1320435166246454034, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1320435150077412025, language=EN, label=Table 1, caption=

Blood biochemical indicators in each group of mice (x¯±s)

, figureFileSmall=null, figureFileBig=null, tableContent=
IndicatorCONTIIND+CIPIND+CIP+CUR
TP/（g/L）54.70±4.2448.6±1.8447.25±1.6346.60±2.55
Alb/（g/L）29.35±1.0623.35±0.9222.95±1.0622.15±1.63
Glb/（g/L）25.35±3.1825.25±0.9224.30±0.5724.45±0.92
A/G10.17±0.110.93±0.010.95±0.020.91±0.04
TBil/（μmol/L）20.40±0.141.35±0.641.45±0.491.70±0.71
DBil/（μmol/L）0.40±0.140.20±0.140.30±0.280.30±0.00
IBil/（μmol/L）2.00±0.001.15±0.781.15±0.211.40±0.71
AST/（U/L）138.90±8.77699.75±244.31549.80±206.05540.85±255.62
ALT/（U/L）55.30±0.42109.05±3.04115.95±44.4887.25±3.89
AST/ALT2.51±0.146.36±2.024.77±0.026.27±3.21
ALP/（U/L）234.20±9.33133.40±54.59164.25±78.14123.60±32.67
GGT/（U/L）1.55±1.060.60±0.140.75±0.070.75±0.35
LDH710.30±31.402 823.20±1 646.712 073.45±1 099.901 686.30±747.69
PA9.90±5.099.30±7.6410.05±5.599.80±7.21
Urea/（mmol/L）11.72±0.3211.23±0.6216.81±1.7013.76±9.65
Crea/（μmol/L）14.20±6.0813.90±8.2013.90±3.8212.00±10.89
U/C197.52±141.96279.50±70.05234.11±155.78256.20±200.64
CK/（U/L）1 699.05±119.57546 7.10±3 136.10b2 856.65±618.79a3 264.70±2 419.86a
CK-MB/（U/L）356.55±21.85543.70±125.44436.90±50.20424.55±98.36
α-HBDH/（U/L）232.00±15.561112.50±707.81814.50±451.84625.50±294.86
), ArticleFig(id=1320435166334534419, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1320435150077412025, language=CN, label=表1, caption=

各组小鼠血液生化指标 (x¯±s)

, figureFileSmall=null, figureFileBig=null, tableContent=
IndicatorCONTIIND+CIPIND+CIP+CUR
TP/（g/L）54.70±4.2448.6±1.8447.25±1.6346.60±2.55
Alb/（g/L）29.35±1.0623.35±0.9222.95±1.0622.15±1.63
Glb/（g/L）25.35±3.1825.25±0.9224.30±0.5724.45±0.92
A/G10.17±0.110.93±0.010.95±0.020.91±0.04
TBil/（μmol/L）20.40±0.141.35±0.641.45±0.491.70±0.71
DBil/（μmol/L）0.40±0.140.20±0.140.30±0.280.30±0.00
IBil/（μmol/L）2.00±0.001.15±0.781.15±0.211.40±0.71
AST/（U/L）138.90±8.77699.75±244.31549.80±206.05540.85±255.62
ALT/（U/L）55.30±0.42109.05±3.04115.95±44.4887.25±3.89
AST/ALT2.51±0.146.36±2.024.77±0.026.27±3.21
ALP/（U/L）234.20±9.33133.40±54.59164.25±78.14123.60±32.67
GGT/（U/L）1.55±1.060.60±0.140.75±0.070.75±0.35
LDH710.30±31.402 823.20±1 646.712 073.45±1 099.901 686.30±747.69
PA9.90±5.099.30±7.6410.05±5.599.80±7.21
Urea/（mmol/L）11.72±0.3211.23±0.6216.81±1.7013.76±9.65
Crea/（μmol/L）14.20±6.0813.90±8.2013.90±3.8212.00±10.89
U/C197.52±141.96279.50±70.05234.11±155.78256.20±200.64
CK/（U/L）1 699.05±119.57546 7.10±3 136.10b2 856.65±618.79a3 264.70±2 419.86a
CK-MB/（U/L）356.55±21.85543.70±125.44436.90±50.20424.55±98.36
α-HBDH/（U/L）232.00±15.561112.50±707.81814.50±451.84625.50±294.86
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姜黄素通过调节Treg/Th17平衡减轻吲哚美辛-环丙沙星联用所致小鼠小肠损伤并协同治疗严重创伤感染
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陈代琦 1, 2 , 余静 2 , 夏雨 2 , 刘国昌 2 , 王海燕 2 , 李遂焰 1 , 严军 2
陆军军医大学学报 | 论著·军事医学 2026,48(2): 116-127
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陆军军医大学学报 |论著·军事医学 2026 , 48 (2) : 116 -127
姜黄素通过调节Treg/Th17平衡减轻吲哚美辛-环丙沙星联用所致小鼠小肠损伤并协同治疗严重创伤感染
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陈代琦1, 2, 余静2, 夏雨2, 刘国昌2, 王海燕2, 李遂焰1 , 严军2
作者信息
  • 1西南交通大学生命科学与工程学院，四川 成都
  • 2陆军军医大学大坪医院特殊环境战伤防治研究室， 创伤与化学中毒全国重点实验室，重庆
通讯作者:
严 军，E-mail：
李遂焰，E-mail：
YAN Jun, E-mail：
LI Suiyan, E-mail：
Curcumin alleviates indomethacin-ciprofloxacin combination-induced small intestinal injury in mice by modulating Treg/Th17 balance and synergistically treats severe traumatic infections
Daiqi CHEN1, 2, Jing YU2, Yu XIA2, Guochang LIU2, Haiyan WANG2, Suiyan LI1 , Jun YAN2
Affiliations
  • 1School of Life Science and Engineering， Southwest Jiaotong University， Chengdu， Sichuan
  • 2Department of Special War Wound， State Key Laboratory of Trauma and Chemical Poisoning， Daping Hospital， Army Medical University （Third Military Medical University）， Chongqing， China
出版时间: 2026-01-30 doi: 10.16016/j.2097-0927.202511042
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摘要
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目的

探讨姜黄素（curcumin，CUR）对吲哚美辛（indomethacin， IND）联用环丙沙星（ciprofloxacin， CIP）治疗严重创伤感染中小肠损伤的作用及其机制。

方法

将8周龄雄性C57BL/6小鼠完全随机分为4组（生存率观察，n=10；其余指标，n=8）：对照组、创伤感染组［失血+骨折+铜绿假单胞菌（Pseudomonas aeruginosa，PA）感染］、IND+CIP治疗组和IND+CIP+CUR治疗组。观察各组小鼠建模后72 h内生存率；收集建模24 h后外周血、小肠组织及肠系膜淋巴结（mesenteric lymph node， MLN），观察各组小肠长度变化、出血状况和粘连情况；稀释涂布平板法检测各组细菌载量；HE染色观察各组肠道组织病理学改变；ELISA分别检测各组血清和小肠匀浆炎症因子水平（IL-1β、IL-6、IL-10和TNF-α）；血液生化指标检测各组肝功、肾功和心功能重要指标。流式细胞术检测各组MLN中CD4+T细胞中IFN-γ、IL-4、TGF-β和IL-17阳性百分比；qPCR检测各组叉头状蛋白P3（forkhead box protein P3， FOXP3）和维甲酸受体相关孤儿受体γ（retinoic acid receptor-related orphan receptor γ， RORγ）-t mRNA表达。

结果

IND+CIP治疗组和IND+CIP+CUR治疗组均能显著提高严重创伤感染小鼠生存率（P<0.05），CUR联用能够降低小鼠细菌载量（P<0.01）。IND+CIP治疗组会引发小肠损伤，并随时间推移呈现加重趋势，表现为小肠TNF-α（P<0.01）、IL-6（P<0.01）和IL-1β水平升高（P<0.05），IL-10水平降低（P<0.05），并伴有小肠病理损伤；IND+CIP+CUR治疗组能显著逆转小肠损伤，降低炎性因子水平（P<0.01），使小肠结构恢复正常，相关血液生化指标与IND+CIP治疗组无明显差异。与IND+CIP治疗组相比，IND+CIP+CUR治疗组CD4+T细胞中FOXP3表达增高（P<0.01），RORγ-t表达降低（P<0.01）。

结论

CUR联用IND和CIP可有效治疗严重创伤感染，同时拮抗IND和CIP疗法中出现的小肠损伤，并对重要脏器功能无明显副作用，其机制可能与调节性T细胞（regulatory T cell， Treg）-辅助性T细胞17（helper T cell 17， Th17）平衡有关。

姜黄素  /  吲哚美辛  /  环丙沙星  /  小肠损伤  /  调节性T细胞-辅助性T细胞17平衡
Objective

To investigate the effect and underlying mechanisms of curcumin (CUR) on small intestinal injuries associated with severe traumatic infections after combined treatment of ciprofloxacin (CIP) and indomethacin (IND).

Methods

Adult male C57BL/6 mice (8 weeks old) were randomly divided into (10 animals for survival rate observation, 10 for other indicators) control group, model group [hemorrhage+ fracture+Pseudomonas aeruginosa (PA) infection], IND+CIP treatment group, and IND+ CIP+CUR treatment group. The survival rates within 72 h post-modelling were observed across all groups. At 24 h post-modelling, peripheral blood samples, small intestinal tissues and mesenteric lymph nodes (MLN) were harvested. Small intestinal length, hemorrhagic status, and adhesions were also observed grossly in each group. Bacterial load was determined via dilute plating. HE staining was used to observe histopathological alterations in intestinal tissues across groups. ELISA was employed to measure the contents of inflammatory cytokines, IL-1β, IL-6, IL-10 and TNF-α in the serum and small intestinal homogenate of each group. Liver, renal, and cardiac functions were evaluated by measuring key biochemical indicators. Flow cytometry was applied to measure the percentages of IFN-γ, IL-4, TGF-β, and IL-17 positive cells within CD4+ T cells in the MLN. Real-time quantitative polymerase chain reaction (qPCR) was performed to detect the mRNA expression of forkhead box protein P3(FOXP3) and retinoic acid receptor-related orphan receptor gamma (RORγ)-t in each group.

Results

Both the IND+CIP treatment group and the IND+CIP+CUR treatment group significantly improved survival rates in mice with severe traumatic infections (P<0.05). CUR addition reduced bacterial load in the mice (P<0.01). The IND+CIP treatment group exhibited small intestinal injury, which worsened over time, manifested by elevated contents of TNF-α (P<0.01), IL-6 (P<0.05), and IL-1β (P<0.05) and decreased IL-10 (P<0.01) in the small intestine tissues, accompanied with pathological damage in the small intestine. The IND+CIP+CUR treatment group had significantly reversed small intestinal injury, reduced inflammatory factor levels (P<0.01), and restored normal small intestinal structure, showing no significant differences in relevant blood biochemical indicators when compared to the IND+CIP treatment group. The expression of FOXP3 was increased and that of RORγ-t was decreasedin CD4+ T cells from the IND+CIP+CUR treatment group than the IND+CIP treatment group (both P<0.01).

Conclusion

The combination of CUR with IND and CIP can effectively alleviate severe traumatic infections, while antagonize small intestinal damage observed in IND and CIP therapy, with no significant adverse effects on vital organ functions. This mechanism may be related to the balance between regulatory T (Treg) cells and T helper 17 (Th17) cells.

curcumin  /  indomethacin  /  ciprofloxacin  /  intestinal injury  /  regulatory T cell -T helper 17 cell balance
陈代琦, 余静, 夏雨, 刘国昌, 王海燕, 李遂焰, 严军. 姜黄素通过调节Treg/Th17平衡减轻吲哚美辛-环丙沙星联用所致小鼠小肠损伤并协同治疗严重创伤感染. 陆军军医大学学报, 2026 , 48 (2) : 116 -127 . DOI: 10.16016/j.2097-0927.202511042
Daiqi CHEN, Jing YU, Yu XIA, Guochang LIU, Haiyan WANG, Suiyan LI, Jun YAN. Curcumin alleviates indomethacin-ciprofloxacin combination-induced small intestinal injury in mice by modulating Treg/Th17 balance and synergistically treats severe traumatic infections[J]. Journal of Army Medical University, 2026 , 48 (2) : 116 -127 . DOI: 10.16016/j.2097-0927.202511042
正文
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严重创伤可导致机体炎症反应和免疫失衡，甚至引起严重感染和脓毒症［1］。造成的损害可导致机体防御屏障破坏和病原体入侵，从而引发感染和一系列复杂的创伤后事件，其中引起的肠道损伤可能在多器官衰竭的发展中发挥关键作用［2］，是使病情恶化的驱动力。作为最主要的营养吸收器官，以小肠损伤为主的肠道损伤不仅会影响患者的预后康复，且肠道内大量的细菌可能穿过破损的屏障进入血液循环，引发第2次全身性炎症反应［3］。
非甾体抗炎药（nonsteroidal anti-inflammatory drugs，NSAIDs）联用抗生素是“菌毒炎并治”防治严重创伤感染的有效策略［4-6］。研究发现，吲哚美辛（indomethacin， IND）联用环丙沙星（ciprofloxacin， CIP）可提高创伤感染小鼠存活率，降低炎症反应程度，增强抑菌作用，且这种药物联用策略对重要器官（肝、肾、心脏）的功能没有造成重大损害［4］。进一步研究还发现，IND联用CIP可能通过磷酸肌醇3-激酶（phosphoinositide 3-kinase，PI3K）/蛋白激酶B（protein kinase B，AKT）信号通路调节巨噬细胞功能改善严重创伤感染小鼠预后［5］。NSAIDs和抗生素联用策略在临床回顾性研究中也得到了验证，NSAIDs和抗生素联用患者的脓毒症发生率低于单用抗生素患者［6］。然而，NSAIDs存在严重的消化道不良反应，并且会诱导小肠损伤［7-9］，而在严重创伤、烧伤、重症等病理条件下，甚至会出现肠黏膜屏障功能受损情况［10-11］。因此，NSAIDs引发的小肠副作用在一定程度上限制了NSAIDs联用抗生素策略的使用。
姜黄素（curcumin， CUR）源自膳食香料姜黄，是一种具有多种生物学和药理活性的多酚化合物，具有抗炎、抗氧化、免疫调节、抗肿瘤和神经保护等作用［12-13］。CUR可在常见的肠道疾病中发挥有益作用，如姜黄素可通过调节Toll样受体4（toll-like receptor 4， TLR4）/核因子κB（nuclear factor kappa-B， NF-κB）/激活蛋白1（activator protein 1， AP-1）信号通路抑制IL-1、IL-6、IL-8和TNF-α等促炎因子产生，改善炎症性肠病（inflammatory bowel disease， IBD）和溃疡性结肠炎（ulcerative colitis， UC）患者症状［14-15］。另外，CUR对免疫细胞也具有调节作用，CUR可以通过调节树突状细胞（dendritic cell， DC）和调节性T细胞（regulatory T cell， Treg）稳态，改变肠道菌群组成，改善因葡聚糖硫酸钠（dextran sulfate sodium salt， DSS）诱发的结肠炎病情［16］。然而，CUR在“菌毒炎并治”策略中对NSAIDs引发小肠损伤的作用和机制仍不清楚。
本研究旨在探讨CUR联用IND和CIP治疗严重创伤感染的有效性和安全性，尤其是确认CUR是否在其中可以拮抗IND引发的小肠损伤，并探讨CUR的作用机制，为严重创伤感染防治过程中“菌毒炎并治”策略的完善和优化提供新思路。
1　材料与方法
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1.1　实验动物与菌株
C57BL/6野生型（wild type，WT）雄性小鼠，8周龄，体质量20~30 g，购自斯贝福生物技术有限公司（中国北京）。实验前置于陆军军医大学大坪医院实验动物中心SPF条件下分笼饲养，确保温度25 ℃、湿度50%、正常昼夜明暗交替环境（光照时间8:00-20:00），自由进食饮水。将小鼠完全随机分成4组：对照组（control group， CON）、创伤组（trauma-infection group， TI）［失血+骨折+铜绿假单胞菌（pseudomonas aeruginosa， PA）感染］、IND+CIP治疗组和IND+CIP+CUR治疗组。其中，生存率观察实验每组10只，在确定最佳模型菌液浓度后，采用该浓度重新构建模型进行其余指标检测实验，每组8只。PA由陆军军医大学基础医学院微生物学教研室馈赠。本实验获陆军军医大学实验动物福利伦理审查委员会批准（AMUWEC20237369）。
1.2　实验试剂
IND和CUR购自美国Sigma公司；羧甲基纤维素（carboxymethylcellulose， CMC）购自麦克林（中国）；小鼠ELISA试剂盒购自中国BOSTER公司；PBS购自中国BOSTER公司；TRIzol Reagent购自日本TaKaRa公司；Prime Script RT reagent Kit with gDNA Eraser和SYBR Premix Ex Taq Ⅱ购自日本TaKaRa公司；引物由上海捷瑞生物工程有限公司合成；红细胞裂解液购自北京天根生物公司；多聚甲醛购自北京索莱宝公司；LB固体、液体培养基购自上海碧云天生物技术公司；0.9%生理盐水购自四川科伦药业公司；Biotin-Antibody Cocktail和Anti-Biotin Microbeads购自德国Miltenyi公司；RPMI1640培养基购自武汉博士德生物工程有限公司；胎牛血清购自以色列BI公司；三抗（青霉素、链霉素和两性霉素B）购自上海达特希尔生物科技公司；70 µm细胞筛网购自无锡NEST公司；细胞因子刺激试剂盒（500×）购自美国Invitrogen公司；PE-IFN-γ、PE/Cy7-IL-4、APC-TGF-β、PerCP/Cy5.5-IL-17、FITC-CD 4及相应染料的同型对照均购买自美国Biolegend公司；Fixation/Permeabilization solution和Perm/Wash buffer购自美国eBioscience公司。
1.3　严重创伤感染模型的建立
将所获PA菌液用无菌一次性接种环蘸取少许菌液在LB固体培养基上划线接种，然后将平板倒置放入37 ℃培养箱中培养24 h。待培养基长出肉眼可见的单菌落后，在生物安全柜内，用接种环将单个菌落挑入含15 mL LB液体培养基的50 mL离心管中，菌和培养基的比例为1∶1 000。随后将装有培养基的离心管放入恒温摇床中，摇床调为37 ℃，加速离心力30×g，摇菌过夜（16 h，以菌体浑浊为佳）。菌液摇好后放入高速离心机中，在加速离心力300 ×g下离心10 min，接着在生物安全柜中弃去菌液上清，加入生理盐水重悬（吹打混匀）。用移液枪吸取200 μL菌液置于96孔板中，在酶标仪中测量光密度值（计数），根据测量结果加入适量生理盐水稀释至1.25×108 CFU/mL浓度待用。
采用“失血/骨折复合腹腔注射PA”方式构建小鼠严重创伤感染模型。实验前首先称量每只小鼠体质量，并记录。实验时抓取小鼠的颈部头皮，并固定尾巴，轻轻按压需要摘取的眼部皮肤，使眼球充血突出；常规眼眶取血，失血滴数按体质量×0.6来计算；达到相应放血量后使用医用脱脂棉按压放血眼眶进行止血；随后立即通过腹腔注射向小鼠体内补液生理盐水300 μL（防止失血性休克）；再用止血钳夹致小鼠双后肢股骨骨折，然后将小鼠放回鼠笼，继续用垫料（木屑）按压止血；2 h后向小鼠体内腹腔注射适量铜绿假单胞菌溶液。观察小鼠生存率，调整菌液用量（50~12.5 μL），使伤后72 h内小鼠死亡率稳定在70%左右，即成功建立小鼠严重创伤感染模型。
1.4　生存率观察
按1.3方法建模。IND+CIP治疗组和IND+CIP+CUR治疗组在模型建立后立即进行首次给药：IND剂量为10 mg/kg，溶剂为CMC，给药方式为灌胃；CUR剂量为200 mg/kg，溶剂为CMC，给药方式为腹腔注射；CIP剂量为10 mg/kg，溶剂为生理盐水，给药方式为腹腔注射。每24 h补药1次，每隔1 h观察各组小鼠生存状况，共观察72 h。
1.5　细菌载量计数
按1.3方法建模。24 h后取材，收集小鼠眼眶外周血。采用稀释涂布平板法，在生物安全柜内将小鼠全血倍比稀释10-1、10-2；充分混匀后，吸取各浓度100 μL到血平板上（每组各浓度3个平板），用一次性塑料涂布棒快速将其向平板各个方向均匀涂抹开，使长出单菌落；接着把血平板放入37 ℃培养箱，培养18~24 h；待平板长出肉眼可见的单菌落后，对菌落进行计数。
1.6　小肠组织病理分析
按1.3方法建模。24 h后取材，取小肠组织记录小肠长度，用生理盐水冲洗除去血液和粪渣，一部分用4%多聚甲醛固定（自盲肠端15 cm处截取2~3 cm），室温固定24 h。将各组固定后的小肠组织流水过夜，然后按50%、70%、80%、95%、95%、100%、100%酒精梯度进行脱水，氯仿透明2 h，浸石蜡1 h（共2次），然后进行包埋、切片、HE染色、封片，显微镜观察组织病理结果并拍照，同时按照Chiu’s肠黏膜损伤评分法进行分级评分。根据光镜下肠黏膜损伤情况，评分一共分为5级：绒毛正常评分为0级；绒毛顶端下间歇增宽评分为1级；绒毛顶端上皮脱落、破溃评分为2级；绒毛顶端破坏扩展到基底部评分为3级；上皮完全脱落评分为4级；固有层崩溃，出现溃疡及出血点评分为5级。
1.7　ELISA检测
按1.3方法建模。24 h后取各组小鼠眼眶血，室温静置30 min。加速离心力300 ×g离心10 min，吸取上清黄色澄清液体，即血清；取各组部分小鼠小肠组织，去除脂肪系膜组织，加生理盐水用匀浆器研磨制成10%匀浆，在低温加速离心力300 ×g下离心10 min，取上清。采用ELISA法检测各组血清和小肠匀浆中的IL-1β、IL-6、IL-10和TNF-α水平。
1.8　血液生化指标检测
按1.3方法建模。24 h后取各组小鼠眼眶血制备血清；每组血清至少500 μL。进行血液生化指标检测，检测指标肝功：总蛋白（total protein， TP）、白蛋白（albumin， Alb）、球蛋白（globulin， Glb）、白/球比值（albumin/globulin， A/G）、总胆红素（total bilirubin， TBil）、直接胆红素（direct bilirubin， DBil）、间接胆红素（indirect bilirubin， IBil）、天门冬氨酸氨基转移酶（aspartate aminotransferase， AST）、丙氨酸氨基转移酶（alanine aminotransferase， ALT）、AST/ALT（AS：AL）、碱性磷酸酶（alkaline phosphatase， ALP）、L-γ-谷氨酰基转移酶（gamma-glutamyltransferase， GGT）；肾功：尿素、肌酐（creatinine， Crea）、尿素/肌酐（urea/creatinine ratioU/C）；心功：肌酸激酶（creatine kinase， CK）、肌酸激酶同工酶-MB（creatine kinase isoenzyme-MB， CK-MB）、α-羟丁酸脱氢酶（hydroxybutyrate dehydrogenase， α-HBDH）。
1.9　CD4+ T细胞亚群百分比检测
按1.3方法建模，24 h后取各组小鼠肠系膜淋巴结（mesenteric lymph node， MLN），研磨后离心裂红，计数重悬，每组分装至1.5 mL EP管内，每管1 mL培养基，按试剂盒说明书每管内加入2 μL细胞因子刺激试剂（500×）；涡旋混合，在37 ℃，5%CO2下培养4~6 h。细胞孵育后，加速离心力300 ×g离心10 min去上清；每孔内加入200 μL大鼠血清，混匀后室温封闭20 min；加入FITC-CD4抗体冰浴20 min；加入适量PBS洗涤后离心；各管加入100 μL Fixation/Permeabilization试剂，混匀过夜；第2天用1 mL 1×BD Perm/Wash buffer洗2次（300 ×g下离心10 min，弃上清）；每管加入200 μL大鼠血清，混匀后室温封闭20 min；按组别每管加入1 μL抗体（PE-IFN-γ；PE/Cy7-IL-4；APC-TGF-β；PerCP/Cy5.5-IL-17）；冰浴20 min，加入1 mL 1×BD Perm/Wash buffer洗1次，混匀，300 ×g下离心10 min，弃上清，每管各加入500 μL 1×BD Perm/Wash buffer/PBS，混匀，采用流式细胞术检测各组CD4+细胞中IFN-γ、IL-4、TGF-β和IL-17阳性细胞百分比。
1.10　Treg和辅助性T细胞17（helper T cell 17， Th17）标志物基因表达
按1.3方法建模，24 h后取各组小鼠MLN，常规提取总RNA，并逆转录成cDNA。采用qPCR检测各组小鼠MLN中叉头状蛋白P3（forkhead box protein P3， FOXP3）和维甲酸受体相关孤儿受体γ（retinoic acid receptor-related orphan receptor γ， RORγ）-t mRNA表达。
1.11　统计学分析
采用GraphPad Prism 10.1.2软件进行统计和作图，计量资料用x¯±s表示。组间差异采用单因素方差分析，P<0.05表示差异有统计学意义。
2　结果
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2.1　药物联用均可提高严重创伤感染小鼠生存率
生存率观察实验结果显示，向失血复合骨折小鼠腹腔注射不同剂量体积的1.25×108 CFU/mL PA菌液，发现注射剂量12.5 µL菌液（即最佳建模剂量）后小鼠死亡率为70%左右（P<0.01），即成功构建小鼠严重创伤感染模型（图1A），后续模型构建皆采用此浓度。与模型组相比，药物联用组（IND+CIP治疗组和IND+CIP+CUR治疗组）生存率明显提高（P<0.05，图1B），且两种药物联用组在提升模型小鼠生存率方面效果相当。
2.2　CUR协同IND和CIP联用可增强抑菌效果
细菌载量实验结果显示，当全血稀释10倍时，模型组平均载菌量为132.3个，IND+CIP治疗组平均载菌量为35个，IND+CIP+CUR治疗组平均载菌量为11个菌落；当全血稀释100倍时，TI组平均载菌量为15.6个，IND+CIP治疗组平均载菌量为3.3个；IND+CIP+CUR治疗组平均载菌量为1.3个菌落（图2A）。此结果提示，CUR协同IND和CIP联用可增强对PA的抑制作用，其效果优于IND+CIP治疗组（P<0.01，图2B、C）。
2.3　CUR减轻严重创伤感染小鼠小肠损伤
HE染色及Chiu’s肠黏膜损伤评分结果显示，与CON组相比，TI组小鼠出现便血、腹水、肠道组织粘连等严重病情，并伴有小肠长度缩短（图3A）。模型组小肠上皮固有层分离（图3B），小肠损伤评分较CON组明显升高（P<0.01），主要表现为：小肠溃疡和炎症细胞浸润、小肠结构不完整；IND+CIP治疗组与TI组情况相似，但IND+CIP治疗组小肠病理评分高于创伤感染组（P<0.01）。相反，IND+CIP+CUR治疗组小肠损伤评分降低（P<0.01，图3C），小肠长度恢复正常且小肠结构较为完整。表明，CUR协同IND联用CIP可有效拮抗小肠损伤。
2.4　两种药物配组抑制血清炎性因子能力无明显差异
ELISA检测结果显示，与TI组相比，IND+CIP治疗组与IND+CIP+CUR治疗组均可降低促炎因子IL-1β、IL-6和TNF-α水平（P<0.05），提高抗炎因子IL-10水平（P<0.05，图4）；两种药物配组之间无统计学差异。提示CUR协同IND和CIP联用有抑制全身性炎症能力，但对IND联用CIP的抗炎效果没有明显影响。
2.5　CUR降低小肠匀浆炎性因子水平
ELISA检测结果显示，与CON组相比，TI组与IND+CIP治疗组小肠匀浆中促炎细胞因子水平IL-1β、IL-6和TNF-α明显升高（P<0.01，图5A、B、D）。IND+CIP治疗组虽然能够降低血清炎症因子水平，但对于小肠局部炎症没有明显抑制作用。相反，IND+CIP+CUR治疗组小肠匀浆中IL-1β、IL-6和TNF-α水平显著降低（P<0.01）。另外，TI组出现代偿性IL-10水平升高，虽然两组药物配组与Con组比小肠匀浆IL-10水平均升高，但IND+CIP+CUR治疗组高于IND+CIP治疗组（P<0.05，图5C），结合IND+CIP治疗组的促炎因子水平明显高于IND+CIP+CUR治疗组（P<0.01，图5A、B、D），提示CUR可能通过抑制小肠促炎细胞因子、促进抗炎因子水平来减轻肠道损伤。
2.6　两种药物配组不影响重要脏器功能
本研究选择心脏、肝脏和肾脏为重要脏器代表，探讨各药物配组是否会对小鼠的心功、肝功和肾功能造成损害，结果见表1。与CON组相比，IND+CIP治疗组和IND+CIP+CUR治疗组肝功、肾功指标无统计学差异，在心功指标CK中，与CON组相比，TI组CK指标升高（P<0.01），而两种药物治疗组降低且逐渐趋于CON组水平。提示两种药物配组不影响小鼠重要脏器功能。
2.7　CUR调控Treg-Th17平衡
流式细胞检测结果显示，与TI组相比，IND+CIP治疗组和IND+CIP+CUR治疗组MLN CD4+T细胞中INF-γ、IL-4和IL-17阳性细胞百分比降低（P<0.01），TGF-β阳性细胞百分比升高（P<0.01）；与IND+CIP治疗组相比，IND+CIP+CUR治疗组MLN CD4+T细胞中INF-γ、IL-4和IL-17阳性细胞百分比更低（P<0.01），而TGF-β阳性细胞百分比显著升高（P<0.01）。提示CUR协同IND和CIP联用可抑制Th1和Th2百分比，特别是促进Treg分化，进而影响Treg-Th17平衡，拮抗小肠损伤，见图6。
qPCR检测结果显示，与IND+CIP治疗组相比，IND+CIP+CUR治疗组MLN中FOXP3 mRNA表达升高（P<0.01），RORγ-t mRNA表达降低。再次验证了CUR协同IND和CIP联用可能通过调控Treg-Th17平衡以延缓药物联用过程中出现的小肠损伤（P<0.01，图7）。
3　讨论
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严重创伤感染伴随着全身炎症反应与免疫失衡，肠道作为重要的免疫与代谢器官，其损伤不仅影响营养吸收，还可能加剧全身性炎症反应与感染扩散。本研究在前期证实IND与CIP联用可提高创伤感染小鼠生存率的基础上，研究聚焦NSAIDs引起小肠损伤的重要问题，进一步探讨了CUR作为辅助用药在联合方案中的作用，并从免疫学角度阐释其作用机制，为“菌毒炎并治”策略提供更加安全有效的实验依据。
3.1　规避NSAIDs诱发肠道损伤副作用是NSAIDs联用抗生素防治严重创伤感染策略的新问题
严重创伤并发症的常规防治策略往往重视抗生素的使用，而对伤后炎症发展和控制以及对伤员预后的影响认识不足。抗生素的合理应用虽然可以有效杀灭细菌，但与此同时，死亡细菌释放的内、外毒素、细菌DNA以及胞壁成分可引发机体过度炎症反应，进而导致多器官功能障碍与死亡［17-18］。因此，以抗感染和抗炎并举为特征的细菌、毒素和炎症防治即“菌毒炎并治”策略逐渐成为严重创伤并发症防治新途径。
本课题组前期研究工作发现，IND和CIP联用大幅度逆转严重创伤感染小鼠死亡率，降低模型小鼠血清炎性因子，显著增强模型小鼠体内的抑菌作用，并得到临床回顾性研究证实［4-6］。但是，该方案无法预防肠道局部损伤，甚至表现出加重趋势。如何规避NSAIDs（包括IND）诱发肠道损伤的副作用仍是NSAIDs联用抗生素防治严重创伤感染策略必须正视的问题。基于上述考虑，本团队利用反向网络药理学筛选了NSAIDs引发小肠损伤的拮抗药物，发现CUR存在NSAIDs作用靶点，并通过实验室工作验证。可见，CUR可能作为辅助药物参与以NSAIDs联用抗生素为基础的“菌毒炎并治”策略。
3.2　CUR协同策略可确保新药物配组的有效性和安全性
本研究在IND联用CIP方案中引入CUR，构建新型三联疗法（IND+CIP+CUR），旨在评估CUR协同IND和CIP联用的疗效。新型三药配组（IND+CIP+CUR）疗效必须不低于传统两药配组（IND+CIP），才具备进一步探讨CUR新价值的研究意义。本研究发现，加入CUR的药物配组可增强原有IND和CIP联用对PA的抑制作用，并显著抑制小肠炎症，体现了超越原有两药联用的效果。而且，与IND和CIP联用相比，CUR协同IND和CIP联用同样具有降低严重创伤感染小鼠死亡率的功能，而且对模型小鼠重要脏器功能无不良影响。上述发现表明，CUR可确保协同IND和CIP联用治疗严重创伤感染的有效性和安全性，符合“菌毒炎并治”策略中候选辅助药物的基本条件。
3.3　CUR协同策略可有效拮抗肠道损伤
严重创伤感染后缺血/缺氧因子、促炎因子、应激细胞因子和内毒素的释放通过多种信号通路诱发肠道屏障功能障碍，导致创伤后休克，全身炎症反应综合征、细菌易位、脓毒症、高分解代谢、多器官功能障碍综合征或其他并发症［19-20］。因此，预防严重创伤感染后肠道损伤是一项重大挑战。CUR是一种源自姜黄根茎的天然多酚化合物，因其抗炎、抗氧化及免疫调节活性且在多种疾病模型中展现出治疗潜力而被人们认识［21］。尤其在肠道相关疾病中，CUR被证实可通过调控多条信号通路，抑制促炎因子释放，维持肠道菌群稳态，从而发挥保护作用［22-23］。本研究发现，与传统两药配组（IND+CIP）相比，新型三药配组（IND+CIP+CUR）能够显著恢复小肠长度和小肠绒毛的完整形态，降低小肠组织损伤评分，并抑制小肠组织促炎细胞因子IL-1β、IL-6和TNF-α水平。该发现表明，CUR协同策略能够在确保传统两药配组（IND+CIP）疗效的基础上有效拮抗肠道损伤，进一步优化治疗结局。
3.4　CUR可促进肠道Treg细胞分化进而调节Treg/Th17平衡
MLN作为肠道关键淋巴组织，是肠道免疫应答核心枢纽和CD4⁺T细胞驻留场所之一。它负责接收来自肠道组织的抗原信号，并在此启动和调节适应性免疫反应。肠道中Treg-Th17失衡是结肠炎的典型特征［24］。Treg通过分泌抑制性细胞因子如TGF-β等，负向调控免疫反应，维持肠道免疫耐受与稳态修复；Th17细胞则主要分泌IL-17等促炎因子，驱动中性粒细胞浸润与组织炎症。因此，上调肠道Treg以恢复Treg-Th17平衡已被确定为治疗结肠炎的可行策略［25］。研究发现CUR能调节重症肌无力小鼠模型脾脏和MLN细胞Th17/Treg失衡表现为CD4+T细胞中FOXP3比例升高，IL-17A比例降低并提高了小鼠血液中TGF-β1的表达［26］，且CUR对溃疡性结肠炎小鼠MLN中CD4+FOXP3比例以及FOXP3 mRNA的表达有促进作用［27］。
本研究发现，与模型组和传统两药配组相比，新型三药配组（IND+CIP+CUR）MLN CD4⁺T细胞中Treg百分比和Treg标志物FOXP3 mRNA表达均明显升高，Th17百分比和标志物RORγ-t mRNA表达则明显降低。表明CUR协同策略能够促进肠道Treg分化，改变严重创伤感染引发的Treg-Th17失衡，可能是最终缓解小肠炎症反应和组织损伤的重要机制。
3.5　不足与展望
本研究揭示了CUR协同IND和CIP联用策略可通过调节Treg/Th17平衡以减轻肠道损伤，为严重创伤感染防治提供了新视野，但研究主要聚焦于T细胞免疫应答，尚未能全面阐明CUR对肠道固有层中其他关键免疫细胞（如巨噬细胞、DC）的影响，以及这些细胞与T细胞之间的相互作用网络。其次，CUR对肠道物理屏障功能（如紧密连接蛋白表达）的直接保护作用未被评估，其调节免疫与修复屏障的因果关系有待进一步明确。
综上，本研究揭示的新型三药配组（IND+CIP+CUR）方案是一种极具潜力的治疗策略。它不仅成功维持了传统两药配组（IND+CIP）方案在提升严重创伤感染小鼠生存率、炎症控制和细菌抑制方面的优势，更通过协同增强抗菌效果，并通过Treg-Th17平衡的调节，有效缓解肠道损伤，具有重要的临床转化潜力。未来研究可进一步探讨CUR对其他免疫细胞亚群及肠道屏障功能的影响，全面揭示其在严重创伤感染防治中的肠道保护机制。

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doi: 10.16016/j.2097-0927.202511042
  • 接收时间：2025-11-07
  • 首发时间：2026-10-23
  • 出版时间：2026-01-30
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  • 收稿日期：2025-11-07
  • 修回日期：2026-01-04
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    1西南交通大学生命科学与工程学院，四川 成都
    2陆军军医大学大坪医院特殊环境战伤防治研究室， 创伤与化学中毒全国重点实验室，重庆

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鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
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