Article(id=1153429496603594993, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153429493357203682, articleNumber=null, orderNo=null, doi=10.19812/j.cnki.jfsq11-5956/ts.20241129003, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1732809600000, receivedDateStr=2024-11-29, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1752928621673, onlineDateStr=2025-07-19, pubDate=1741968000000, pubDateStr=2025-03-15, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1752928621673, onlineIssueDateStr=2025-07-19, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1752928621673, creator=13701087609, updateTime=1752928621673, updator=13701087609, issue=Issue{id=1153429493357203682, tenantId=1146029695717560320, journalId=1149652044408987649, year='2025', volume='16', issue='5', pageStart='1', pageEnd='326', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1752928620900, creator=13701087609, updateTime=1758690311058, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1177595773500932351, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153429493357203682, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1177595773500932352, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153429493357203682, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=119, endPage=126, ext={EN=ArticleExt(id=1153429497106911489, articleId=1153429496603594993, tenantId=1146029695717560320, journalId=1149652044408987649, language=EN, title=Mechanism of Matricaria chamomile L. extract on lipopolysaccharide induced acute lung injury, columnId=1151895322591638525, journalTitle=Journal of Food Safety & Quality, columnName=Special Topic: Functional Foods and Functional Components, runingTitle=null, highlight=null, articleAbstract=

Objective To investigate the protective effects of Matricarla chamomilla L. extract on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice and explore the mechanism by which Matricarla chamomilla L. extract alleviates ALI. Methods Forty-eight male SPF Kunming mice were randomly divided into control group, LPS group, dexamethasone (DEX) group, and low, middle and high doses of Matricarla chamomilla L. extract groups. The mice in each group were first continuously administered by gastric gavage for 7 days, and dexamethasone was administered intraperitoneally in the dexamethasone group 5 mg/kg. Matricarla chamomilla L. extract was administered 85, 170 and 340 mg/(kg·d) for gastric lavage in the low, medium and high dose groups, respectively. Mice of all groups, except those of control group, were induced into ALI mouse model by intratracheal instillation of LPS. Subsequently, the mice were subjected to the determination of their levels of the inflammatory factors tumor necrosis factor-α (TNF-α), interleukin 6 (IL-6) and interleukin 1β (IL-1β) in bronchoalveolar lavage fluid (BALF) and perform Giemsa staining and white blood cell count on the BALF sediment. The content and activities of malondialdehyde (MDA) and superoxide dismutase (SOD) in serum were detected. Lung tissue was taken and the pathological morphology of the inferior lobes of the right lung was observed. The expression levels of Toll-like receptor 4 (TLR4), recombinant myeloid differentiation factor 88 (MyD88), nuclear factor-kappa B (NF-κB) in lung tissue were detected. Results Compared with the blank group, the alveolar lavage fluid of LPS model group mice TNF-α, IL-6, IL-1β, White blood cell count and serum MDA content significantly increased (P<0.05), serum SOD significantly decreased (P<0.05), HE staining showed severe interstitial inflammation infiltration in the lungs, structural damage and abnormal morphology of lung tissue, protein expression levels of TLR4, MyD88, NF-κBp65 in lung tissue of mices increased (P<0.05). Compared with the LPS model, the levels in mice of each treatment group TNF-α, white blood cell count, and serum MDA contents were significantly reduced (P<0.05), while serum SOD content was significantly increased (P<0.05). HE staining showed a decrease in inflammatory cell infiltration in mouse lung tissue, clearer alveolar spaces, more complete lung tissue morphology, and protein expression levels of TLR4, MyD88 and NF-κBp65 in lung tissue of mice decreased (P<0.05). Conclusion Matricarla chamomilla L. extract can alleviate LPS induced pulmonary inflammation and overactivation of oxidative stress, thereby alleviating ALI in mice. Among them, a dose of 170 mg/(kg·d) has a better effect, and pathway such as TLR4/MyD88/NF-κB.

, correspAuthors=Jing-Jing LUO, 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=Jing WANG, Zi-Fan CAI, Yu LI, Xuan-Le ZHU, Jing-Jing LUO), CN=ArticleExt(id=1153429525468795778, articleId=1153429496603594993, tenantId=1146029695717560320, journalId=1149652044408987649, language=CN, title=洋甘菊提取物对脂多糖诱导的小鼠急性肺损伤的机制研究, columnId=1151895323909124661, journalTitle=食品安全质量检测学报, columnName=本期专题:功能性食品与功能性成分, runingTitle=null, highlight=null, articleAbstract=

目的 研究洋甘菊提取物对脂多糖(lipopolysaccharide, LPS)诱导小鼠急性肺损伤(acute lung injury, ALI)的保护作用, 并探索洋甘菊提取物减轻ALI的作用机制。方法 本研究将48只雄性SPF级昆明小鼠随机分为6组, 分别是空白组、模型组、阳性对照组、洋甘菊提取物低剂量、中剂量、高剂量组, 每组8只。各组小鼠连续灌胃给药7 d, 阳性对照组腹腔注射给药地塞米松(dexamethasone, DEX) 5 mg/kg; 洋甘菊提取物低、中、高剂量组分别灌胃85、170和340 mg/(kg·d)洋甘菊提取物。除空白组由气管向肺内滴注生理盐水外, 其余各组均由气管向肺内滴注LPS的方式建立ALI模型。检测小鼠肺泡灌洗液(bronchoalveolar lavage fluid, BALF)中细胞因子肿瘤坏死因子α (tumor necrosis factor-α, TNF-α)、白细胞介素-6 (interleukin 6, IL-6)和白细胞介素-1β (interleukin 1β, IL-1β)、并对BALF沉淀物进行吉姆萨染色及白细胞计数; 摘眼球取血, 检测血清中丙二醛(malondialdehyde, MDA)和超氧化物歧化酶(superoxide dismutase, SOD)的含量; 取右肺下叶组织, 观察病理形态; 检测小鼠肺组织中Toll样受体4 (Toll-like receptor 4, TLR4)、髓样分化因子(recombinant myeloid differentiation factor 88, MyD88)、核因子κB (nuclear factor-kappa B, NF-κB)等表达水平。结果 与空白组相比, LPS模型组小鼠BALF中TNF-α、IL-6、IL-1β、白细胞计数和血清中MDA含量显著升高(P<0.05), 血清中SOD显著降低(P<0.05)。HE染色可见, 肺间质炎症浸润严重, 肺组织结构破坏和形态不正常, 肺组织的TLR4、MyD88、NF-κBp65表达水平升高(P<0.05)。与LPS模型相比, 各给药组小鼠BALF中的TNF-α、白细胞计数和血清中MDA含量显著降低(P<0.05), 血清中SOD显著升高(P<0.05), HE染色可见, 小鼠肺组织炎性细胞浸润减少, 肺泡腔较清晰, 肺组织形态较完整, 肺组织的TLR4、MyD88、NF-κBp65表达水平降低(P<0.05)。结论 洋甘菊提取物可以缓解LPS所诱导的肺部炎症反应与氧化应激的过度激活, 从而减轻小鼠ALI, 其中170 mg/(kg·d)剂量作用较好, 洋甘菊提取物可能通过调节TLR4/MyD88/NF-κB信号通路发挥作用。

, correspAuthors=罗晶晶, authorNote=null, correspAuthorsNote=
* 罗晶晶(1984—), 女, 硕士, 高级实验师, 主要研究方向为心血管疾病基础研究。E-mail:
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王婧(2002—), 女, 主要研究方向为心血管疾病基础研究。E-mail:

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王婧(2002—), 女, 主要研究方向为心血管疾病基础研究。E-mail:

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王婧(2002—), 女, 主要研究方向为心血管疾病基础研究。E-mail:

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注: #. 模型组与空白组比较, 差异显著(P<0.05); *. 各用药组与模型组比较, 差异显著(P<0.05)。下同。

, figureFileSmall=BVO4JSYteu0WJaDQD9z+Dw==, figureFileBig=kjVqemSM4xlKoDsAgHh+7g==, tableContent=null), ArticleFig(id=1177619631264121807, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429496603594993, language=EN, label=Fig.2, caption=Staining in BALF of mice in each group (Giemsa staining, 400×), figureFileSmall=G16bR2GxOrntM+WlYgY/fQ==, figureFileBig=HwpY9xTvcjTeT+EsRVCpUg==, tableContent=null), ArticleFig(id=1177619631335424977, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429496603594993, language=CN, label=图2, caption=各组小鼠BALF染色(吉姆萨染色, 400×), figureFileSmall=G16bR2GxOrntM+WlYgY/fQ==, figureFileBig=HwpY9xTvcjTeT+EsRVCpUg==, tableContent=null), ArticleFig(id=1177619631452865491, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429496603594993, language=EN, label=Fig.3, caption=Number of white blood cells in BALF of mice in each group (n=6), figureFileSmall=dJt+JlY4X4Ipq3vMmptXOw==, figureFileBig=EQeX0N2Sph2Z3AWckDcXdA==, tableContent=null), ArticleFig(id=1177619631624831957, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429496603594993, language=CN, label=图3, caption=各组小鼠BALF中白细胞数(n=6), figureFileSmall=dJt+JlY4X4Ipq3vMmptXOw==, figureFileBig=EQeX0N2Sph2Z3AWckDcXdA==, tableContent=null), ArticleFig(id=1177619631733883863, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429496603594993, language=EN, label=Fig.4, caption=Pathological changes of lung tissue sections (HE staining) in mice in each group (200×), figureFileSmall=XizV4essDSz90OYyc5hk0g==, figureFileBig=+A888AqZlncrPLT0ijgPXg==, tableContent=null), ArticleFig(id=1177619631851324376, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429496603594993, language=CN, label=图4, caption=各组小鼠肺组织切片(HE染色)病理学变化(200×), figureFileSmall=XizV4essDSz90OYyc5hk0g==, figureFileBig=+A888AqZlncrPLT0ijgPXg==, tableContent=null), ArticleFig(id=1177619631951987674, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429496603594993, language=EN, label=Fig.5, caption=MDA, SOD content of serum in mice in each group (n=6), figureFileSmall=CToI34/N3yS7W9VApO555Q==, figureFileBig=1g60kGaAIwyZUYvI9ST+YQ==, tableContent=null), ArticleFig(id=1177619632052650972, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429496603594993, language=CN, label=图5, caption=各组小鼠血清中MDA、SOD含量(n=6), figureFileSmall=CToI34/N3yS7W9VApO555Q==, figureFileBig=1g60kGaAIwyZUYvI9ST+YQ==, tableContent=null), ArticleFig(id=1177619632144925662, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429496603594993, language=EN, label=Fig.6, caption=Protein expression levels of TLR4, MyD88, NF-κBp65 and p-NF-κB-p65 in lung tissues of mice in each group (n=3), figureFileSmall=+2OJ8zbDazLV3C3fV+n7UA==, figureFileBig=Hn5udJ/e6oalID64ftN41Q==, tableContent=null), ArticleFig(id=1177619632237200352, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429496603594993, language=CN, label=图6, caption=各组小鼠肺组织中TLR4、MyD88、NF-κBp65和p-NF-κB-p65蛋白表达量(n=3)

注: a. TLR4、MyD88、NF-κBp65、p-NF-κB-p65蛋白Western blotting条带; b、c、d、e分别表示TLR4、MyD88、NF-κBp65、p-NF-κB-p65蛋白灰度值分析。

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洋甘菊提取物对脂多糖诱导的小鼠急性肺损伤的机制研究
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王婧 1 , 蔡子凡 1 , 李彧 1 , 朱轩乐 2 , 罗晶晶 3, *
食品安全质量检测学报 | 本期专题:功能性食品与功能性成分 2025,16(5): 119-126
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食品安全质量检测学报 | 本期专题:功能性食品与功能性成分 2025, 16(5): 119-126
洋甘菊提取物对脂多糖诱导的小鼠急性肺损伤的机制研究
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王婧1 , 蔡子凡1, 李彧1, 朱轩乐2, 罗晶晶3, *
作者信息
  • 1.新疆医科大学第二临床医学院, 乌鲁木齐 830017
  • 2.新疆医科大学第一临床医学院, 乌鲁木齐 830017
  • 3.新疆医科大学基础医学院, 乌鲁木齐 830017
  • 王婧(2002—), 女, 主要研究方向为心血管疾病基础研究。E-mail:

通讯作者:

* 罗晶晶(1984—), 女, 硕士, 高级实验师, 主要研究方向为心血管疾病基础研究。E-mail:
Mechanism of Matricaria chamomile L. extract on lipopolysaccharide induced acute lung injury
Jing WANG1 , Zi-Fan CAI1, Yu LI1, Xuan-Le ZHU2, Jing-Jing LUO3, *
Affiliations
  • 1. Second Clinical College, Xinjiang Medical University, Urumqi 830017, China
  • 2. First Clinical College of Xinjiang Medical University, Urumqi 830017, China
  • 3. School of Basic Medical Sciences, Xinjiang Medical Universtity, Urumqi 830017, China
出版时间: 2025-03-15 doi: 10.19812/j.cnki.jfsq11-5956/ts.20241129003
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目的 研究洋甘菊提取物对脂多糖(lipopolysaccharide, LPS)诱导小鼠急性肺损伤(acute lung injury, ALI)的保护作用, 并探索洋甘菊提取物减轻ALI的作用机制。方法 本研究将48只雄性SPF级昆明小鼠随机分为6组, 分别是空白组、模型组、阳性对照组、洋甘菊提取物低剂量、中剂量、高剂量组, 每组8只。各组小鼠连续灌胃给药7 d, 阳性对照组腹腔注射给药地塞米松(dexamethasone, DEX) 5 mg/kg; 洋甘菊提取物低、中、高剂量组分别灌胃85、170和340 mg/(kg·d)洋甘菊提取物。除空白组由气管向肺内滴注生理盐水外, 其余各组均由气管向肺内滴注LPS的方式建立ALI模型。检测小鼠肺泡灌洗液(bronchoalveolar lavage fluid, BALF)中细胞因子肿瘤坏死因子α (tumor necrosis factor-α, TNF-α)、白细胞介素-6 (interleukin 6, IL-6)和白细胞介素-1β (interleukin 1β, IL-1β)、并对BALF沉淀物进行吉姆萨染色及白细胞计数; 摘眼球取血, 检测血清中丙二醛(malondialdehyde, MDA)和超氧化物歧化酶(superoxide dismutase, SOD)的含量; 取右肺下叶组织, 观察病理形态; 检测小鼠肺组织中Toll样受体4 (Toll-like receptor 4, TLR4)、髓样分化因子(recombinant myeloid differentiation factor 88, MyD88)、核因子κB (nuclear factor-kappa B, NF-κB)等表达水平。结果 与空白组相比, LPS模型组小鼠BALF中TNF-α、IL-6、IL-1β、白细胞计数和血清中MDA含量显著升高(P<0.05), 血清中SOD显著降低(P<0.05)。HE染色可见, 肺间质炎症浸润严重, 肺组织结构破坏和形态不正常, 肺组织的TLR4、MyD88、NF-κBp65表达水平升高(P<0.05)。与LPS模型相比, 各给药组小鼠BALF中的TNF-α、白细胞计数和血清中MDA含量显著降低(P<0.05), 血清中SOD显著升高(P<0.05), HE染色可见, 小鼠肺组织炎性细胞浸润减少, 肺泡腔较清晰, 肺组织形态较完整, 肺组织的TLR4、MyD88、NF-κBp65表达水平降低(P<0.05)。结论 洋甘菊提取物可以缓解LPS所诱导的肺部炎症反应与氧化应激的过度激活, 从而减轻小鼠ALI, 其中170 mg/(kg·d)剂量作用较好, 洋甘菊提取物可能通过调节TLR4/MyD88/NF-κB信号通路发挥作用。

洋甘菊提取物  /  脂多糖  /  急性肺损伤  /  保护作用

Objective To investigate the protective effects of Matricarla chamomilla L. extract on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice and explore the mechanism by which Matricarla chamomilla L. extract alleviates ALI. Methods Forty-eight male SPF Kunming mice were randomly divided into control group, LPS group, dexamethasone (DEX) group, and low, middle and high doses of Matricarla chamomilla L. extract groups. The mice in each group were first continuously administered by gastric gavage for 7 days, and dexamethasone was administered intraperitoneally in the dexamethasone group 5 mg/kg. Matricarla chamomilla L. extract was administered 85, 170 and 340 mg/(kg·d) for gastric lavage in the low, medium and high dose groups, respectively. Mice of all groups, except those of control group, were induced into ALI mouse model by intratracheal instillation of LPS. Subsequently, the mice were subjected to the determination of their levels of the inflammatory factors tumor necrosis factor-α (TNF-α), interleukin 6 (IL-6) and interleukin 1β (IL-1β) in bronchoalveolar lavage fluid (BALF) and perform Giemsa staining and white blood cell count on the BALF sediment. The content and activities of malondialdehyde (MDA) and superoxide dismutase (SOD) in serum were detected. Lung tissue was taken and the pathological morphology of the inferior lobes of the right lung was observed. The expression levels of Toll-like receptor 4 (TLR4), recombinant myeloid differentiation factor 88 (MyD88), nuclear factor-kappa B (NF-κB) in lung tissue were detected. Results Compared with the blank group, the alveolar lavage fluid of LPS model group mice TNF-α, IL-6, IL-1β, White blood cell count and serum MDA content significantly increased (P<0.05), serum SOD significantly decreased (P<0.05), HE staining showed severe interstitial inflammation infiltration in the lungs, structural damage and abnormal morphology of lung tissue, protein expression levels of TLR4, MyD88, NF-κBp65 in lung tissue of mices increased (P<0.05). Compared with the LPS model, the levels in mice of each treatment group TNF-α, white blood cell count, and serum MDA contents were significantly reduced (P<0.05), while serum SOD content was significantly increased (P<0.05). HE staining showed a decrease in inflammatory cell infiltration in mouse lung tissue, clearer alveolar spaces, more complete lung tissue morphology, and protein expression levels of TLR4, MyD88 and NF-κBp65 in lung tissue of mice decreased (P<0.05). Conclusion Matricarla chamomilla L. extract can alleviate LPS induced pulmonary inflammation and overactivation of oxidative stress, thereby alleviating ALI in mice. Among them, a dose of 170 mg/(kg·d) has a better effect, and pathway such as TLR4/MyD88/NF-κB.

Matricarla chamomilla L. extract  /  lipopolysaccharide  /  acute lung injury  /  protective effect
王婧, 蔡子凡, 李彧, 朱轩乐, 罗晶晶. 洋甘菊提取物对脂多糖诱导的小鼠急性肺损伤的机制研究. 食品安全质量检测学报, 2025 , 16 (5) : 119 -126 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20241129003
Jing WANG, Zi-Fan CAI, Yu LI, Xuan-Le ZHU, Jing-Jing LUO. Mechanism of Matricaria chamomile L. extract on lipopolysaccharide induced acute lung injury[J]. Journal of Food Safety & Quality, 2025 , 16 (5) : 119 -126 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20241129003
急性肺损伤(acute lung injury, ALI)是呼吸系统类疾病中常见的一种, 其是由多种致病因素引起肺部出现以炎症反应、肺水肿和肺泡毛细血管屏障损伤为病理特征, 以呼吸衰竭综合症和进行性低氧血症为临床特征的综合病征[1], 最终发展为急性呼吸窘迫综合征(acute respiratory distress syndrome, ARDS)[2]。尽管机械通气的结果有所改善, 但ARDS的死亡率仍然高达35%~55%[3-4]。因其死亡率较高, 且发病机制尚不明确, 故难以进行精准治疗。目前, 其临床治疗主要包括保护性机械通气, 且尚无有效的药物治疗[5-6]。因此, 建立一个合适的中草药预防和保护动物ALI的模型对于疾病诊断及新药研发具有重要意义。
洋甘菊(Matricarla chamomilla L.)属于菊科一年生草本植物[7], 又称母菊, 是新疆传统明方祖卡木颗粒中主要的药材之一, 其治疗呼吸系统疾病的历史悠久[8], 在古埃及、希腊、罗马和我国医学中都有记载[9-10]。洋甘菊的花或全草都可以用药, 性凉、味辛、微苦, 具有散气消炎、止咳平喘、祛风湿和清热解毒等功效, 可用于肺热咳喘、湿痹肿痛、感冒发热、咽喉肿痛等症状[11], 其提取物的植物化学成分, 目前已鉴定出301多种成分, 其中主要化学成分为多糖、挥发油、黄酮、香豆素等[12-15]。研究表明, 洋甘菊提取物具有多种生物活性, 如抗癌、抗感染、抑菌、解痉、抗炎、抗氧化、降脂、降糖、抗过敏、抗抑郁和神经保护作用等药理作用[16-19]。但洋甘菊是否能够减轻ALI的炎症反应, 减缓损伤后的组织纤维化发生, 目前鲜见报道。
本研究主要从ALI的发病机制入手, 包括炎症/抗炎反应失衡、氧化应激。研究发现, 炎症在ALI的发病机制中发挥着关键作用, 重要的是, 抑制炎症可降低ALI的损伤严重程度[20-21]。也有研究表明, Toll样受体4 (Toll-like receptor 4, TLR4)/髓样分化因子(recombinant myeloid differentiation factor 88, MyD88)/核因子κB (nuclear factor-kappa B, NF-κB)信号通路在参与ALI局部损伤、炎症反应等方面均起着重要的作用, TLR4触发下游MyD88/NF-κB信号, 通过产生各种促炎细胞因子, 加重肺损伤[22-23], 其被认为是与ALI密切相关的信号通路之一[24]。因此, 本研究拟基于TLR4/MyD88/NF-κB信号通路研究洋甘菊提取物对脂多糖(lipopolysaccharide, LPS)诱导小鼠ALI的影响及其作用机制, 以期为治疗ALI药物的进一步研发和转化提供科学佐证。
7~8周龄, 体重20~23 g雄性SPF级昆明小鼠48只, 购自新疆医科大学动物实验中心[生产许可证SCXK(新): 2018-0002], 自由进食常规饲料和饮水。
洋甘菊醇提取物(批号: 20230405, 西安塞奥生物科技有限公司)。
LPS(批号: 324T031)、羧甲基纤维素钠(批号: 104X021)(索莱宝生物科技有限公司); 白细胞介素-1β (interleukin 1β, IL-1β)试剂盒(批号: E20230620-20174B)、肿瘤坏死因子-α (tumor necrosis factor-α, TNF-α)试剂盒(批号: E20230620-20852B)、白细胞介素-6 (interleukin 6, IL-6)试剂盒(批号: E20230620-20188B)、丙二醛(malondialdehyde, MDA)试剂盒(批号: S6915N469944W)(上海分计生物科技有限公司); 总超氧化物歧化酶(total superoxide dismutase, T-SOD)检测盒(批号: GY012PH45711, 武汉伊莱瑞特生物科技有限公司); BCA蛋白浓度测定试剂盒(批号: G2026)、TLR4抗体、MyD88抗体、NF-κB p65抗体、p-NF-κB p65抗体、Actin抗体(武汉塞维尔生物科技有限公司)。
Olympus显微镜(日本Olympus光学显微镜公司); Thermo Fisher rmultiskan FC酶标仪(美国Thermo Fisher Scientific公司); SVE-2全套电泳仪(武汉塞维尔生物科技有限公司); D3024R台式高速冷冻离心机[大龙兴创实验仪器(北京)股份公司]。
选用雄性SPF级昆明小鼠48只, 随机分为空白组、模型组、地塞米松(dexamethasone, DEX)阳性对照组、洋甘菊提取物低(Matricaria chamomile L.-Low, MC-L)、中(Matricaria chamomile L.-Medium, MC-M)、高(Matricaria chamomile L.-High, MC-H)剂量组, 共6组, 每组8只。模型建立前按MC-L组灌胃给药85 mg/(kg·d)、MC-M组灌胃给药170 mg/(kg·d)、MC-H组灌胃给药340 mg/(kg·d), 连续灌胃给药7 d; 阳性对照组腹腔注射DEX 5 mg/kg, 每日一次, 连续3 d; 空白组和模型组灌胃等量生理盐水。各组小鼠连续灌胃给药7 d后, 用10%乌拉坦加5%水合氯醛腹腔注射麻醉小鼠, 仰卧固定于手术台。参照文献[25]方法造模, 剃去小鼠颈部正中毛发, 酒精消毒, 颈部正中开口2 cm, 暴露及分离气管, 采用往气管内注射5 mg/kg LPS制备ALI模型, 空白组气道内给予等量生理盐水, 酒精消毒伤口并缝合皮肤。术后观察各组小鼠的呼吸和身体恢复状况。在造模24 h后, 取血、处死并采集肺泡灌洗液(bronchoalveolar lavage fluid, BALF)和肺脏组织。
将各组小鼠的颈部皮肤切开, 分离气管并做插管。将胸部打开, 寻找右支气管并用丝线结扎, 用PBS缓冲液将左肺进行肺泡灌洗3次, 0.5 mL/次, BALF收集并离心(4000 r/min, 10 min)后, 取上清液在-80 ℃条件下保存, 用于测量细胞因子。按照酶联免疫吸附测定(enzyme-linked immuno sorbent assay, ELISA)试剂盒说明书, 测定BALF中TNF-α、IL-6和IL-1β
BALF沉淀物用50 μmol/L的PBS缓冲液重悬, 吹打混匀后, 进行吉姆萨染色并对各组进行白细胞计数。
取右肺下叶固定于10%多聚甲醛中24 h, 将固定好的组织脱水处理, 将组织放入模块石蜡中包埋, 待蜡块冷却后, 将蜡块切成4 μm厚的蜡片, 载玻片捞片, 将载玻片置于60 ℃烤片机上烤片过夜, 通过二甲苯和梯度乙醇脱水处理, 用苏木精-伊红(hematoxylin and eosin staining, HE)染色, 透明、封片后用显微镜观察并拍片。
各组小鼠通过摘眼球取血, 室温静置2 h, 在离心机中以4000 r/min离心10 min, 分离血清, -80 ℃保存, 按MDA和SOD试剂盒测定血清中MDA、SOD的含量。
取适量小鼠肺组织, 用预冷的PBS洗涤2~3次, 加入10倍组织体积裂解液(含蛋白酶抑制剂和磷酸酶抑制剂)匀浆, 4 ℃、12000 r/min离心10 min, 收集上清蛋白裂解液, BCA法定量, 蛋白样本经SDS-PAGE凝胶电泳, 然后转膜, 封闭。洗涤后加入混合ECL发光试剂显影, 采用Image J 1.8.0软件分析蛋白条带灰度值, 计算目的条带灰度值与内参条带灰度值的比值。
数据通过GraphPad Prism 9.5软件统计分析, 各组小鼠各观测指标采用单因素方差分析, P<0.05为差异显著。
与空白组比较, LPS处理后的模型组小鼠BALF中TNF-α、IL-6、IL-1β含量均升高(P<0.05); 与模型组比较, 洋甘菊低、中、高剂量组和阳性对照组小鼠BALF中TNF-α含量均降低(P<0.05), 洋甘菊低、中剂量组和阳性对照组小鼠BALF中IL-6含量均降低(P<0.05), 洋甘菊高剂量组小鼠BALF中IL-6含量有降低趋势, 无统计学意义(P>0.05), 洋甘菊低、中、高剂量组和阳性对照组小鼠BALF中IL-1β含量有降低趋势, 无统计学意义(P>0.05)。表明洋甘菊醇提取物能够改善ALI的“炎症风暴”, 见图1
与空白组比较, LPS处理后的模型组小鼠BALF中白细胞计数显著升高(P<0.05); 与模型组比较, 洋甘菊低、中、高剂量组和阳性对照组小鼠BALF白细胞计数均降低(P<0.05), 见图23
图4所示, 空白组小鼠肺间质无炎性细胞浸润, 也未见纤维组织增生, 肺泡间隔未见增厚, 肺泡腔清晰, 肺组织完整; 与空白组相比, 模型组小鼠肺间质炎症浸润严重, 肺组织结构破坏, 部分肺泡融合、肺泡腔内可见炎性细胞聚集和充血, 肺组织形态不完整, 失去肺的正常形态; 与模型组相比, 各给药组小鼠肺组织损伤均有缓解, 有效减少肺组织炎性细胞浸润, 肺泡腔较清晰, 肺组织形态较完整。
与空白组比较, 模型组小鼠血清中MDA的含量升高(P<0.05), SOD含量降低(P<0.05); 与模型组比较, 洋甘菊低、中、高剂量组和阳性对照组小鼠血清MDA的含量降低(P<0.05), SOD含量升高(P<0.05), 见图5
图6显示, 与空白组比较, 模型组肺组织的TLR4、MyD88、NF-κBp65表达水平升高(P<0.05); 与模型组比较, 洋甘菊低、中剂量组小鼠肺组织的TLR4、MyD88、NF-κBp65表达水平降低(P<0.05), 其余各组蛋白表达水平有降低趋势, 无统计学意义(P>0.05)。
洋甘菊是一种重要的药用植物和香料原料, 多以其花序或全草入药, 含有丰富的生物活性物质。洋甘菊醇提取液富含的酚酸、香豆素类和黄酮类化合物, 具有抗氧化、抗炎、保肝护肝、抗血管增生、抗过敏活性、抗变应性和抗病毒等功效[26-27]。基于洋甘菊证明的广泛药理活性, 已经研究了其在多个领域的可能用途, 其中最重要的应用是在药用领域。事实上, 对动物模型和患者的几项研究表明, 这种植物对多种疾病都有治疗作用, 包括胃肠道疾病[28]、普通感冒[29]、肝脏疾病[30]、神经精神和呼吸系统问题等[31]
呼吸系统类的疾病是临床常见病之一, 其中常见的呼吸系统类疾病为ALI, 其是因多种致病因素, 如创伤、脓毒血症、细菌、呼吸道病毒等引起肺部出现炎症反应与氧化应激过度激活的过程[32-34], 炎症反应与氧化应激密切相关, 炎症失衡诱发氧化相关基因的过度表达及活性氧过量生成, 继发氧化应激损伤, 加重肺结构细胞凋亡[35]
目前, 普遍认为在ALI的发病中炎症细胞所介导的炎症反应起重要作用[36]。TNF-α能够诱导肺内皮细胞活化、白细胞迁移、粒细胞脱颗粒和毛细血管渗漏, 积聚的水肿液进一步阻碍了肺泡细胞的灌流和氧气交换, 引起ARDS[37]。HSIAO等[38]研究显示, IL-1β参与早期ALI中多形核中性粒细胞(plymorphonuclear neutrophil, PMN)的募集, 与早期炎症反应有关。IL-6与ALI关系密切, IL-6低表达可对ALI产生保护作用[39-40]。在本研究中, LPS诱导的ALI小鼠BALF中TNF-α、IL-1β、IL-6的浓度均增加, 洋甘菊提取物干预组小鼠的BALF中TNF-α、IL-1β、IL-6浓度降低, 表明洋甘菊提取物有可以减轻LPS诱导的ALI所引起炎症反应的作用。
ALI发病机制还与氧化应激有关, 同时许多研究表明洋甘菊正丁醇提取物对肺损伤疾病中肺组织MDA水平有一定降低作用, 对肺组织SOD水平有一定升高作用[41]。本研究通过检测MDA和SOD含量再次印证洋甘菊提取物通过抗氧化对ALI小鼠肺组织的保护机制, 结果显示, LPS诱导的ALI小鼠血清中MDA的浓度增加, SOD的浓度降低, 洋甘菊提取物使LPS诱导的ALI小鼠的血清中MDA浓度降低, SOD浓度增加, 表明洋甘菊提取物可以减轻ALI所引起的氧化应激。
有研究报道, TLR4/MyD88/NF-κB信号通路参与机体免疫应答、炎症反应、调节细胞凋亡、应激反应[42]。同时在LPS诱导的ALI模型中TLR4、MyD88、NF-κBp65、p-NF-κB-p65表达增加与肺损伤等脏器功能障碍相关[43]。当有病原相关分子如LPS刺激时, NF-κB上游最关键的激酶IKB激酶β (IKKβ)发生磷酸化, 导致p65/p50二聚体转移到细胞核, 并促进促炎细胞因子(如TNF-α、IL-1β等)表达[44]。TLR4被激活时, 导致TLR4同型二聚体通过激活磷脂酰肌醇-3-激酶(PI3K)和蛋白激酶B (Akt或PKB)触发促炎信号[45]。Akt激酶作用于NF-κB信号通路, 诱导IL-6、TNF-α、IL-1β等细胞因子的大量产生, 促进ALI的发展[46]。本研究中, 模型组小鼠肺组织中TLR4、MyD88、NF-κBp65、p-NF-κB-p65的表达增加, 与既往研究[47]中TLR4/MyD88/NF-κB通路参与LPS诱导的ALI的结果吻合。洋甘菊提取物使ALI小鼠肺组织中TLR4、MyD88、NF-κBp65表达降低, 表明洋甘菊提取物使TLR4/MyD88/NF-κB通路受抑制, 减少NF-κB入核, 进而抑制NF-κB对多种炎症因子表达的促进作用, 最终实现减轻炎症反应及氧化应激反应的效应。说明洋甘菊提取物抑制炎症细胞浸润, 调控炎症反应及氧化应激的机制可能与阻断TLR4/MyD88/NF-κB通路有关。
综上所述, 洋甘菊提取物可以减轻LPS诱导的小鼠ALI, 其分子机制可能是通过抑制TLR4/MyD88/NF-κB通路介导的炎症反应和氧化应激反应起作用。但本研究仅限于动物实验模型, 其结果用于临床可信性需进一步验证, 且洋甘菊提取物通过TLR4/MyD88/NF-κB通路参与ALI的调控机制仍有待今后进一步的探索。
洋甘菊提取物可以减轻LPS诱导的小鼠ALI所导致的炎症反应和氧化应激, 其中170 mg/(kg·d)剂量的洋甘菊提取物作用较好。洋甘菊提取物[170 mg/(kg·d)], 能有效减轻炎症反应, 抑制BALF中的炎症细胞募集, 降低TNF-α、IL-6、IL-1β、MDA浓度, 增加SOD浓度, 并通过抑制TLR4/MyD88/NF-κB通路对ALI起保护作用可用于预防呼吸系统类疾病, 减轻LPS导致的ALI, 是抗生素替代药物的优选。研究表明[48], 洋甘菊成分是安全、有效的, 因此可以用于其他动物模型及临床中使用, 但洋甘菊精油易刺激黏膜, 其醇和水提取物的组织通透性和溶解性差, 生物利用度低[49]。为了提高洋甘菊的安全性和有效性, 必须利用其他新型药物剂型, 如洋甘菊成分可以封装在脂质载体中, 如纳米乳剂、纳米胶囊、脂质体等, 这将提高其临床可接受性和良好的医学应用。
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doi: 10.19812/j.cnki.jfsq11-5956/ts.20241129003
  • 接收时间:2024-11-29
  • 首发时间:2025-07-19
  • 出版时间:2025-03-15
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  • 收稿日期:2024-11-29
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    1.新疆医科大学第二临床医学院, 乌鲁木齐 830017
    2.新疆医科大学第一临床医学院, 乌鲁木齐 830017
    3.新疆医科大学基础医学院, 乌鲁木齐 830017

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* 罗晶晶(1984—), 女, 硕士, 高级实验师, 主要研究方向为心血管疾病基础研究。E-mail:
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2种不同金属材料的力学参数

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

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