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Article(id=1218291752058929453, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1218291750003724554, articleNumber=1001-2494(2024)13-1211-08, orderNo=null, doi=10.11669/cpj.2024.13.005, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1677513600000, receivedDateStr=2023-02-28, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1768392988457, onlineDateStr=2026-01-14, pubDate=1720368000000, pubDateStr=2024-07-08, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1768392988457, onlineIssueDateStr=2026-01-14, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1768392988457, creator=13701087609, updateTime=1768392988457, updator=13701087609, issue=Issue{id=1218291750003724554, tenantId=1146029695717560320, journalId=1190317699101192196, year='2024', volume='59', issue='13', pageStart='1173', pageEnd='1272', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1768392987967, creator=13701087609, updateTime=1768394537396, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1218298248834503031, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1218291750003724554, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1218298248838697336, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1218291750003724554, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=1211, endPage=1218, ext={EN=ArticleExt(id=1218291752281227568, articleId=1218291752058929453, tenantId=1146029695717560320, journalId=1190317699101192196, language=EN, title=Effect of Curcumin on Gastric Tissue Morphology and Inflammatory Microenvironment in Mice with Gastric Epithelial Dysplasia, columnId=null, journalTitle=Chinese Pharmaceutical Journal, columnName=null, runingTitle=null, highlight=null, articleAbstract=

OBJECTIVE To observe the effects of curcumin on the gastric tissue morphology and inflammatory microenvironment in mice with gastric epithelia dysplasia (GED), and to evaluate the preventive and therapeutic effects of curcumin on GED. METHODS Normal control group, model group, low, medium, and high dose groups of curcumin, and positive control group were set up. Except for the normal control group, all other groups were induced to establish GED animal models using a compound factor modeling method. After 8 weeks of modeling, the low, medium, and high dose groups of curcumin and the positive control group were respectively given curcumin at doses of 38, 75, 150 mg·kg-1 and vitacoenzyme 350 mg/kg for intervention, continued for 8 weeks. At the end of the experiment, the gastric levels of pepsinogen PGⅠ, IFN-γ, IL-1β, IL-6, and the relative expression levels of p-JAK2, p-STAT3, Cyclin D1 in each experimental group were detected. HE staining and PCNA antibody immunohistochemistry were performed to observe the gastric mucosal tissue morphology and cell proliferation in each group. The pathological scores of GED and inflammatory cell infiltration were evaluated, and the proliferation index (PI) was calculated. RESULTS Compared with the normal control group, the serum concentration of PGⅠ in mice in the model group decreased, while the levels of IL-1β, IFN-γ, and IL-6 in gastric tissue increased (P<0.05). The scores of GED and gastric mucosal inflammatory cell infiltration, as well as the proliferation index (PI), all increased (P<0.01). Histopathological observations revealed changes such as epithelial dysplasia and inflammatory cell infiltration in the gastric mucosa, enhanced cell proliferation activity, and upregulation of p-JAK2, p-STAT3, and Cyclin D1 expression (P<0.05). Compared with the model group, the medium and high dose groups of curcumin showed an increase in serum PG1 concentration, a decrease in the inflammatory factors IFN-γ and IL-6 levels in gastric tissue (P<0.05), a decrease in GED and gastric mucosal inflammatory cell infiltration scores, as well as PI (P<0.05 or 0.01). The curcumin treatment alleviated gastric mucosal dysplasia and inflammatory cell infiltration, inhibited cell proliferation activity, and downregulated the expression of p-JAK2, p-STAT3, and Cyclin D1 (P<0.05). In the low dose group of curcumin, the expression of p-JAK2 protein was downregulated, and in the high dose group, IL-1β decreased (P<0.05). CONCLUSION Curcumin has a preventive and alleviating effect on the epithelial dysplasia of the gastric mucosa in mice, and can improve the tissue morphology of the gastric mucosa in GED mice. Its mechanism of action may be related to the downregulation of the JAK2/STAT3/Cyclin D1 pathway signaling, inhibiting excessive cell proliferation.

, correspAuthors=Yanhua Song, 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=Yong XIA, Bin CAO, Caiju XU, Yanhua Song), CN=ArticleExt(id=1218291755431149944, articleId=1218291752058929453, tenantId=1146029695717560320, journalId=1190317699101192196, language=CN, title=姜黄素对胃黏膜上皮异型增生小鼠的胃组织形态和炎症微环境的影响, columnId=1190352405612040510, journalTitle=中国药学杂志, columnName=论著, runingTitle=null, highlight=null, articleAbstract=

目的 观察姜黄素对胃黏膜上皮异型增生(gastric epithelia dysplasia,GED)小鼠的胃组织形态及炎症微环境的影响,评估姜黄素对GED的预防和缓解作用。方法 设正常对照组、模型组、姜黄素低、中、高剂量组和阳性对照组,除正常对照组外,其余各组均采用复合因素造模法造模,诱导GED动物模型。造模8周后,姜黄素低、中、高剂量组及阳性对照组分别给予姜黄素38、75、150 mg·kg-1和维霉素350 mg·kg-1进行干预,持续8周。实验期末,检测各实验组小鼠胃蛋白酶原(PGⅠ)、干扰素-γ(IFN-γ)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)含量及磷酸化酪氨酸激酶2(p-JAK2)、信号转导与转录激活因子3(p-STAT3)、细胞周期蛋白D1(cyclin D1)相对表达量。苏木素-伊红(hematoxylin-eosin,HE)染色和增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)抗体免疫组化染色,观察各组小鼠胃黏膜组织形态及细胞增殖情况,对GED和炎症细胞浸润进行组织病理学评分,计算细胞增殖指数(proliferation index,PI)。结果 与正常对照组比较,模型组小鼠血清PGⅠ浓度下降,胃组织中IL-1β、IFN-γ、IL-6含量上升(P<0.05),GED和胃黏膜炎症细胞浸润评分及PI均上升(P<0.01),组织病理学观察发现胃黏膜出现异型增生、炎症细胞浸润等改变,细胞增殖活性增强,p-JAK2、p-STAT3、Cyclin D1表达上调(P<0.05)。与模型组比较,姜黄素中、高剂量组血清PG1浓度上升,胃组织中炎症因子IFN-γ、IL-6含量降低(P<0.05),GED和胃黏膜炎症细胞浸润评分及PI均下降(P<0.05或0.01),胃黏膜异型增生、炎症细胞浸润缓解,细胞增殖活性受到抑制,p-JAK2、p-STAT3、Cyclin D1表达下调(P<0.05)。姜黄素低剂量组p-JAK2蛋白表达量下调,高剂量组IL-1β下降(P<0.05)。结论 姜黄素对小鼠GED有预防和缓解作用,可以改善GED小鼠胃黏膜的组织形态,其作用机制可能与下调JAK2/STAT3/Cyclin D1通路的信号转导,抑制细胞过度增殖有关。

, correspAuthors=宋燕华, authorNote=null, correspAuthorsNote=
* 宋燕华,女,硕士,副主任医师 研究方向:毒理学Tel:(0571)87115254
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夏勇,男,学士,主任技师 研究方向:毒理学与实验病理学

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夏勇,男,学士,主任技师 研究方向:毒理学与实验病理学

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夏勇,男,学士,主任技师 研究方向:毒理学与实验病理学

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DOI:10.1155/2015/176726., articleTitle=Innate immunity components and cytokines in gastric mucosa in children with Helicobacter pylori infection, refAbstract=null), Reference(id=1218312854457995834, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218291752058929453, doi=null, pmid=null, pmcid=null, year=2021, volume=13, issue=12, pageStart=16667, pageEnd=16683, url=null, language=null, rfNumber=[35], rfOrder=34, authorNames=ZHUANG M, DING X, SONG W, journalName=Aging, refType=null, unstructuredReference=ZHUANG M, DING X, SONG W, et al. Correlation of IL-6 and JAK2/STAT3 signaling pathway with prognosis of nasopharyngeal carcinoma patients[J]. 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Cell Cycle, 2018, 17(2):225-239., articleTitle=VEGFC/VEGFR3 signaling regulates mouse spermatogonial cell proliferation via the activation of AKT/MAPK and cyclin D1 pathway and mediates the apoptosis by affecting caspase 3/9 and Bcl-2, refAbstract=null)], funds=null, companyList=[AuthorCompany(id=1218312848036516250, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218291752058929453, xref=null, ext=[AuthorCompanyExt(id=1218312848040710555, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218291752058929453, companyId=1218312848036516250, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou 310051,China), AuthorCompanyExt(id=1218312848049099164, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218291752058929453, companyId=1218312848036516250, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=浙江省疾病预防控制中心, 杭州 310051)])], figs=[ArticleFig(id=1218312849563242970, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218291752058929453, language=EN, label=Fig.1, caption=The ELISA measurement results of serum PGⅠ content in each experimental group of mice. n=10,$\bar{x}±s$

1)P<0.05, compared with control group;2)P<0.05, compared with model group.

, figureFileSmall=Yq/PFnbj91Dx4iSLopSJLA==, figureFileBig=JuSQj9BpG0dvGrK3Sj/6Qw==, tableContent=null), ArticleFig(id=1218312849655517660, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218291752058929453, language=CN, label=图1, caption=姜黄素各实验组小鼠血清胃蛋白酶原(PGⅠ)含量酶联免疫吸附测定法(ELISA)测定结果. n=10,$\bar{x}±s$

与正常对照组比较,1)P<0.05; 与模型组比较,2)P<0.05。

, figureFileSmall=Yq/PFnbj91Dx4iSLopSJLA==, figureFileBig=JuSQj9BpG0dvGrK3Sj/6Qw==, tableContent=null), ArticleFig(id=1218312849756180958, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218291752058929453, language=EN, label=Fig.2, caption=The observation resullts of hematoxylin-eosin(HE) staining of gastric tissue in each experimental group of mice(×200)

A-normal control group; B-model group; C-curcumin low dose group (38 mg·kg-1); D-curcumin medium dose group (76 mg·kg-1); E-curcumin high dose group(150 mg·kg-1) ; F-positive control group.

, figureFileSmall=6kVc0ShN2eVKe3XS+skqLg==, figureFileBig=5obtyZ5eaOmEJyauSbSSZA==, tableContent=null), ArticleFig(id=1218312849823289824, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218291752058929453, language=CN, label=图2, caption=姜黄素各实验组小鼠胃组织苏木素-伊红(HE)染色结果(×200)

A-正常对照组;B-模型组;C-姜黄素低剂量组(38 mg·kg-1);D-姜黄素中剂量组(76 mg·kg-1);E-姜黄素高剂量组(150 mg·kg-1);F-阳性对照组。

, figureFileSmall=6kVc0ShN2eVKe3XS+skqLg==, figureFileBig=5obtyZ5eaOmEJyauSbSSZA==, tableContent=null), ArticleFig(id=1218312849902981602, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218291752058929453, language=EN, label=Fig.3, caption=Histopathological score results of epithelial dysplasia and inflammatory cell infiltration in the gastric mucosa of mice in each experimental group. n=10

A-GED score scatter plot; B-inflammatory cell infiltration scorescatter plot; 1)P<0.01,compared with control group;2)P<0.05,3)P<0.01, compared with model group.

, figureFileSmall=eq2YBdgRuCsK66V7jwYprQ==, figureFileBig=zFyjYkQDr2KmcOJsX/JeyA==, tableContent=null), ArticleFig(id=1218312849986867684, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218291752058929453, language=CN, label=图3, caption=姜黄素各实验组小鼠胃黏膜上皮异性增生和炎症细胞增生组织病理学评分结果。n=10

A-GED评分散点图; B-炎症细胞浸润评分散点;与正常对照组比较,1)P<0.01;与模型组比较,2)P<0.05,3)P<0.01。

, figureFileSmall=eq2YBdgRuCsK66V7jwYprQ==, figureFileBig=zFyjYkQDr2KmcOJsX/JeyA==, tableContent=null), ArticleFig(id=1218312851266130405, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218291752058929453, language=EN, label=Fig.4, caption=The observation results of immunohistochemical staining of PCNA in gastric mucosa of mice in each experimental group(×200)

A-normal control group; B-model group; C-curcumin low dose group(38 mg·kg-1); D-curcumin medium dose group (76 mg·kg-1); E-curcumi n high dose group (150 mg·kg-1); F-positive control group.

, figureFileSmall=F9LPm8STeQAtKt4lcDr7Eg==, figureFileBig=DRC1pCWQxRKr/zmoXaT1fQ==, tableContent=null), ArticleFig(id=1218312851333239271, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218291752058929453, language=CN, label=图4, caption=姜黄素各实验组小鼠胃黏膜增殖细胞核抗原(PCNA)免疫组化染色(×200)

A-正常对照组; B-模型组; C-姜黄素低剂量组(38 mg·kg-1); D-姜黄素中剂量组(76 mg·kg-1); E-姜黄素高剂量组(150 mg·kg-1); F-阳性对照组。

, figureFileSmall=F9LPm8STeQAtKt4lcDr7Eg==, figureFileBig=DRC1pCWQxRKr/zmoXaT1fQ==, tableContent=null), ArticleFig(id=1218312851396153833, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218291752058929453, language=EN, label=Fig.5, caption=Comparison of gastric mucosal cell proliferation index among different experimental groups of mice. n=10,$\bar{x}±s$

1)P< 0.01, compared with control group;2)P<0.05,3)P<0.01, compared with model group.

, figureFileSmall=FNfX7j5PZOGCa4MrhcBXeQ==, figureFileBig=8kD7bUNfkjfLudW9K6vo7w==, tableContent=null), ArticleFig(id=1218312851454874090, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218291752058929453, language=CN, label=图5, caption=姜黄素各实验组小鼠胃黏膜细胞增殖指数比较。n=10,$\bar{x}±s$

与正常对照组比较,1)P<0.01;与模型组比较,2)P<0.5,3)P<0.01。

, figureFileSmall=FNfX7j5PZOGCa4MrhcBXeQ==, figureFileBig=8kD7bUNfkjfLudW9K6vo7w==, tableContent=null), ArticleFig(id=1218312851513594348, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218291752058929453, language=EN, label=Fig.6, caption=Bands of signal transduction proteins p-JAK2, p-STAT3, and cyclin D1 expression in gastric tissues of mice in each experimental group, figureFileSmall=gKSXytM2zWdS1NJG0YQL+A==, figureFileBig=PFsQ35yRZDDwMNBzoMTDnA==, tableContent=null), ArticleFig(id=1218312851589091822, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218291752058929453, language=CN, label=图6, caption=姜黄素各实验组小鼠胃组织中信号转导蛋白p-JAK2、p-STAT3、cyclin D1表达条带, figureFileSmall=gKSXytM2zWdS1NJG0YQL+A==, figureFileBig=PFsQ35yRZDDwMNBzoMTDnA==, tableContent=null), ArticleFig(id=1218312851668783600, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218291752058929453, language=EN, label=Fig.7, caption=Relative expression levels of signal transduction proteins in gastric tissues of mice in each experimental group. n=10,$\bar{x}±s$

A-comparison of relative expression levels of p-JAK2; B-comparison of relative expression levels of p-STAT3; C-comparison of relative expression levels of Cyclin D1; 1)P<0.05, compared with control group;2)P<0.05, compared with model group.

, figureFileSmall=t4g7wyjubqrL8dy6GycwJg==, figureFileBig=qX+EtU2YLxnZY1lWzS5niA==, tableContent=null), ArticleFig(id=1218312851735892466, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218291752058929453, language=CN, label=图7, caption=姜黄素各实验组小鼠胃组织中信号转导蛋相对表达量。 n=10,$\bar{x}±s$

A-p-JAK2相对表达量比较;B-p-STAT3相对表达量比较; C-Cyclin D1相对表达量比较;与正常对照组比较,1)P<0.05;与模型组比较,2)P<0.05。

, figureFileSmall=t4g7wyjubqrL8dy6GycwJg==, figureFileBig=qX+EtU2YLxnZY1lWzS5niA==, tableContent=null), ArticleFig(id=1218312851790418420, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218291752058929453, language=EN, label=Tab.1, caption=

Comparison of IFN-γ,IL-1β and IL-6 levels in gastric tissues of mice in each experimental group. n=10,$\bar{x}±s$

, figureFileSmall=null, figureFileBig=null, tableContent=
Group c(IFN-γ)
/×10-6 μg·g-1		 c(IL-1β)
/×10-6 μg·g-1		 c(IL-6)
/×10-6 μg·g-1
Control 325 ±27 471 ±83 738 ±167
Model 477 ±501) 1 132 ±1221) 1 639 ±2731)
38 mg·kg-1 CUR 475 ±49 1 142 ±103 1 498 ±266
75 mg·kg-1 CUR 407 ±432) 1 046 ±112 1 193 ±3252)
150 mg·kg-1 CUR 383 ±372) 778 ±1082) 987 ±2522)
Positive control 450 ±52 929 ±1202) 1 431 ±239
), ArticleFig(id=1218312851865915894, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218291752058929453, language=CN, label=表1, caption=

姜黄素各实验组小鼠胃组织中干扰素-γ(IFN-γ)、白介素-1β(IL-1β)、白介素-6(IL-6)含量的比较。 n=10,$\bar{x}±s$

, figureFileSmall=null, figureFileBig=null, tableContent=
Group c(IFN-γ)
/×10-6 μg·g-1		 c(IL-1β)
/×10-6 μg·g-1		 c(IL-6)
/×10-6 μg·g-1
Control 325 ±27 471 ±83 738 ±167
Model 477 ±501) 1 132 ±1221) 1 639 ±2731)
38 mg·kg-1 CUR 475 ±49 1 142 ±103 1 498 ±266
75 mg·kg-1 CUR 407 ±432) 1 046 ±112 1 193 ±3252)
150 mg·kg-1 CUR 383 ±372) 778 ±1082) 987 ±2522)
Positive control 450 ±52 929 ±1202) 1 431 ±239
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姜黄素对胃黏膜上皮异型增生小鼠的胃组织形态和炎症微环境的影响
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夏勇 , 曹彬 , 徐彩菊 , 宋燕华 *
中国药学杂志 | 论著 2024,59(13): 1211-1218
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中国药学杂志 | 论著 2024, 59(13): 1211-1218
姜黄素对胃黏膜上皮异型增生小鼠的胃组织形态和炎症微环境的影响
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夏勇, 曹彬, 徐彩菊, 宋燕华*
作者信息
  • 浙江省疾病预防控制中心, 杭州 310051

    • 夏勇,男,学士,主任技师 研究方向:毒理学与实验病理学

通讯作者:

* 宋燕华,女,硕士,副主任医师 研究方向:毒理学Tel:(0571)87115254
Effect of Curcumin on Gastric Tissue Morphology and Inflammatory Microenvironment in Mice with Gastric Epithelial Dysplasia
Yong XIA, Bin CAO, Caiju XU, Yanhua Song*
Affiliations
  • Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou 310051,China
出版时间: 2024-07-08 doi: 10.11669/cpj.2024.13.005
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目的 观察姜黄素对胃黏膜上皮异型增生(gastric epithelia dysplasia,GED)小鼠的胃组织形态及炎症微环境的影响,评估姜黄素对GED的预防和缓解作用。方法 设正常对照组、模型组、姜黄素低、中、高剂量组和阳性对照组,除正常对照组外,其余各组均采用复合因素造模法造模,诱导GED动物模型。造模8周后,姜黄素低、中、高剂量组及阳性对照组分别给予姜黄素38、75、150 mg·kg-1和维霉素350 mg·kg-1进行干预,持续8周。实验期末,检测各实验组小鼠胃蛋白酶原(PGⅠ)、干扰素-γ(IFN-γ)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)含量及磷酸化酪氨酸激酶2(p-JAK2)、信号转导与转录激活因子3(p-STAT3)、细胞周期蛋白D1(cyclin D1)相对表达量。苏木素-伊红(hematoxylin-eosin,HE)染色和增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)抗体免疫组化染色,观察各组小鼠胃黏膜组织形态及细胞增殖情况,对GED和炎症细胞浸润进行组织病理学评分,计算细胞增殖指数(proliferation index,PI)。结果 与正常对照组比较,模型组小鼠血清PGⅠ浓度下降,胃组织中IL-1β、IFN-γ、IL-6含量上升(P<0.05),GED和胃黏膜炎症细胞浸润评分及PI均上升(P<0.01),组织病理学观察发现胃黏膜出现异型增生、炎症细胞浸润等改变,细胞增殖活性增强,p-JAK2、p-STAT3、Cyclin D1表达上调(P<0.05)。与模型组比较,姜黄素中、高剂量组血清PG1浓度上升,胃组织中炎症因子IFN-γ、IL-6含量降低(P<0.05),GED和胃黏膜炎症细胞浸润评分及PI均下降(P<0.05或0.01),胃黏膜异型增生、炎症细胞浸润缓解,细胞增殖活性受到抑制,p-JAK2、p-STAT3、Cyclin D1表达下调(P<0.05)。姜黄素低剂量组p-JAK2蛋白表达量下调,高剂量组IL-1β下降(P<0.05)。结论 姜黄素对小鼠GED有预防和缓解作用,可以改善GED小鼠胃黏膜的组织形态,其作用机制可能与下调JAK2/STAT3/Cyclin D1通路的信号转导,抑制细胞过度增殖有关。

姜黄素  /  胃黏膜上皮异型增生  /  炎症因子  /  磷酸化酪氨酸激酶2/信号转导与转录激活因子3/细胞周期蛋白D1

OBJECTIVE To observe the effects of curcumin on the gastric tissue morphology and inflammatory microenvironment in mice with gastric epithelia dysplasia (GED), and to evaluate the preventive and therapeutic effects of curcumin on GED. METHODS Normal control group, model group, low, medium, and high dose groups of curcumin, and positive control group were set up. Except for the normal control group, all other groups were induced to establish GED animal models using a compound factor modeling method. After 8 weeks of modeling, the low, medium, and high dose groups of curcumin and the positive control group were respectively given curcumin at doses of 38, 75, 150 mg·kg-1 and vitacoenzyme 350 mg/kg for intervention, continued for 8 weeks. At the end of the experiment, the gastric levels of pepsinogen PGⅠ, IFN-γ, IL-1β, IL-6, and the relative expression levels of p-JAK2, p-STAT3, Cyclin D1 in each experimental group were detected. HE staining and PCNA antibody immunohistochemistry were performed to observe the gastric mucosal tissue morphology and cell proliferation in each group. The pathological scores of GED and inflammatory cell infiltration were evaluated, and the proliferation index (PI) was calculated. RESULTS Compared with the normal control group, the serum concentration of PGⅠ in mice in the model group decreased, while the levels of IL-1β, IFN-γ, and IL-6 in gastric tissue increased (P<0.05). The scores of GED and gastric mucosal inflammatory cell infiltration, as well as the proliferation index (PI), all increased (P<0.01). Histopathological observations revealed changes such as epithelial dysplasia and inflammatory cell infiltration in the gastric mucosa, enhanced cell proliferation activity, and upregulation of p-JAK2, p-STAT3, and Cyclin D1 expression (P<0.05). Compared with the model group, the medium and high dose groups of curcumin showed an increase in serum PG1 concentration, a decrease in the inflammatory factors IFN-γ and IL-6 levels in gastric tissue (P<0.05), a decrease in GED and gastric mucosal inflammatory cell infiltration scores, as well as PI (P<0.05 or 0.01). The curcumin treatment alleviated gastric mucosal dysplasia and inflammatory cell infiltration, inhibited cell proliferation activity, and downregulated the expression of p-JAK2, p-STAT3, and Cyclin D1 (P<0.05). In the low dose group of curcumin, the expression of p-JAK2 protein was downregulated, and in the high dose group, IL-1β decreased (P<0.05). CONCLUSION Curcumin has a preventive and alleviating effect on the epithelial dysplasia of the gastric mucosa in mice, and can improve the tissue morphology of the gastric mucosa in GED mice. Its mechanism of action may be related to the downregulation of the JAK2/STAT3/Cyclin D1 pathway signaling, inhibiting excessive cell proliferation.

curcumin  /  gastric epithelial dysplasia  /  inflammatory factor  /  JAK2/STAT3/Cyclin D1
夏勇, 曹彬, 徐彩菊, 宋燕华. 姜黄素对胃黏膜上皮异型增生小鼠的胃组织形态和炎症微环境的影响. 中国药学杂志, 2024 , 59 (13) : 1211 -1218 . DOI: 10.11669/cpj.2024.13.005
Yong XIA, Bin CAO, Caiju XU, Yanhua Song. Effect of Curcumin on Gastric Tissue Morphology and Inflammatory Microenvironment in Mice with Gastric Epithelial Dysplasia[J]. Chinese Pharmaceutical Journal, 2024 , 59 (13) : 1211 -1218 . DOI: 10.11669/cpj.2024.13.005
正文
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胃癌(GC)是一种常见的恶性肿瘤,患者的死亡率较高,5年生存率低于30%,晚期预后较差 [1-2]。GC的发生是一个慢性的病理过程,包括慢性胃炎、异型增生和GC等多个阶段[3]。胃黏膜上皮异型增生(gastric epithelia dysplasia,GED)属于GC前病变,是胃黏膜癌变前的最后一个病理阶段。GED患者的恶变风险很高,GC发生率接近80%[2]。 在GED阶段进行干预、阻止其向恶性转化对于早期预防GC具有重要意义[4]。然而,阻止从GED到GC的转化至今仍然是一个挑战。炎症微环境和过度增殖是慢性胃炎向GED转化的2个重要特征[5-6],炎症微环境与GC的发生、发展高度相关[7-8]。中药用于治疗胃病已有悠久的历史。许多从植物中提取的活性成分对胃肠道疾病显示出良好的治疗效果。姜黄素(curcumin,Cur)是从草本植物中提取的天然多酚类化合物,具有抗炎、抗肿瘤等多种生物活性。本研究采用复合因素造模法,建立小鼠GED疾病模型,用姜黄素进行干预,观察其对GED小鼠的组织形态、细胞过度增殖及炎症微环境的影响,探讨姜黄素对GED的预防作用及潜在机制,以期为临床治疗提供新思路和理论支持。
1 材料与方法
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1.1 实验动物
SPF级ICR雄性小鼠60只,体质量( 21.3±1.3) g,购于浙江维通利华实验动物有限公司。实验动物饲养于屏障环境,温度范围22~25 ℃, 相对湿度范围40%~70%。饲养许可证号SYXK(浙)2023-00420。本研究遵循3R原则并经浙江省疾病预防控制中心伦理委员会批准(2023-001),动物实验遵照ZJPF08-40《实验动物管理程序》进行。
1.2 试剂与仪器
维霉素片(vitacoenzyme,VCE)(规格:每片0.2 g,批号:20230715,洛阳伊龙药业有限公司);姜黄素(规格:95%,批号:JH230925, 渭南东江天成实业有限公司);增殖细胞核抗原(PCNA)抗体[货号:GM087929,基因科技(上海)股份有限公司];二抗及DAB显色试剂盒套装(批号:QHL 220322,艾普迪实验器材制造(上海)有限公司); 小鼠胃蛋白酶原Ⅰ(pepsinogen Ⅰ,PGI)测定试剂盒(批号: JL37198,上海将来实业股份有限公司);辣根过氧化酶标记IgG抗体(货号:#7074)、抗Cyclin D1抗体(货号:#2978)(美国CST公司);抗p-JAK2 抗体(货号:AF3024)、抗p-STAT3抗体(货号:#AF3293)(美国 Affinity 公司)。免疫组化染色机[Autostainer 720型,艾普迪实验器材制造(上海)有限公司]; 病理数字切片扫描仪(Pannoramic SCAN型,匈牙利3DHISTECH公司),组织匀浆机(Precellys 24型,法国BERTIN公司),化学发光仪(FluoChem E型,美国Santa Clara公司)。
1.3 分组及给药
设正常对照组、模型组、姜黄素低、中、高剂量组和阳性对照组,采用本实验室开发的完全随机化分组程序进行随机化分组[9],每组10只动物。采用自由饮用和灌胃法给予相应药液或溶媒,灌胃容量5 mL·kg-1。有研究表明,姜黄素在50 mg·kg-1剂量下,对大鼠胃黏膜损伤有保护作用[10-11],折算成小鼠的等效剂量约为71 mg·kg-1,故设中剂量组给药剂量为75 mg·kg-1,并以2倍的剂量间距,上、下各设1个剂量组[12-13]。从实验开始首日起,参照文献[14-15],采用复合因素造模法诱导GED,对各组小鼠(除正常对照组外)进行造模,用200 mg·L-1N-甲基-N'-硝基-N-亚硝基胍(MNNG)溶液[16]代替饮水供小鼠自由饮用,每次饱食1 d后,禁食1 d,禁食日早晨撤除饲料,下午灌胃给予20 mmol·L-1去氧胆酸钠溶液(用体积分数30%乙醇溶液配制)灌胃,灌胃后继续禁食,至次日晨给予饲料。正常对照组给予等量的溶媒(蒸馏水),不作给药和禁食处理。自实验开始后第9周起,除继续进行造模处理外,姜黄素低、中、高剂量组每日上午分别灌胃给予38、75和150 mg·kg-1姜黄素,阳性对照组每日上午给予350 mg·kg-1维霉素进行干预 [17],模型组给予等量蒸馏水,正常对照组处理同模型组,持续8周。实验期末,各组小鼠禁食过夜,次日眼眶采血法采集血液样本,二氧化碳麻醉箱处死小鼠,采集胃组织样本进行指标检测及组织病理学检查。
1.4 小鼠血清胃蛋白酶原(PGⅠ)含量测定
血液样本在室温放置2 h后,1 000 r·min-1离心20 min,取上清,用酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA)测定小鼠血清中PGⅠ含量,具体操作按照试剂盒说明书的方法进行。
1.5 小鼠胃组织HE染色及组织病理学形态观察
将小鼠胃沿胃大弯对称剖剪开,在解剖板上摊平,生理盐水冲净,滤纸吸干,切取腺胃部胃组织2条(包含病变部位),转移至脱水盒中摊平,固定48 h,包埋切片,苏木素-伊红(HE)染色,参照文献分别对胃组织标本的GED和炎症细胞浸润进行评分[18-20],GED按严重程度分无、轻微、轻、中、重5级,分别计0、1、2、3、4分,炎症细胞浸润按10个高倍镜视野中炎症细胞数量平均值分5级:≤10、≤25、≤50、≤100、>100分别计0、1、2、3、4分;胃黏膜出现腺体异型增生、慢性炎细胞浸润等病理改变,组织病理学评分高于正常对照组且有统计学意义为模型建立[15,17,20]。
1.6 小鼠胃黏膜组织增殖细胞核抗原表达情况观察
胃组织石蜡切片,经脱蜡、水化,水化后的组织切片经3%H2O2孵育、抗原修复、血清封闭、PCNA抗体孵育(1:150稀释)、二抗孵育、3,3-二氨基联苯胺四盐酸盐显色、苏木素复染后,在高倍镜下观察并评分。PCNA抗原表达情况通过计算每个腺体的阳性细胞数来评估,PCNA阳性细胞胞核呈棕褐色, 每张切片上随机取20个腺体, 在高倍镜视野下,计数每个腺体中的阳性细胞数并计算其平均值,即为细胞增殖指数(proliferation index,PI)[18]。
1.7 小鼠胃组织中IFN-γ、IL-1β、IL-6含量测定
将胃组织放至碎冰中冷却,剪碎后按质量与体积1:10比例加入预冷的pH 7.2 磷酸缓冲液,匀浆后取上清液测定干扰素-γ(interferon-γ,IFN-γ), 白介素-1β(interleukin-1β,IL-1β)及白介素-6(interleukin-6,IL-6)含量,结果以每克胃组织中的炎症因子含量表示(μg·g-1)。
1.8 Western blot 法测定胃组织中信号转导蛋白表达水平
按100 mg·mL-1的比例,称取胃组织加入预冷的(radio-immunoprecipitation assay buffer,RIPA)裂解液,置碎冰中裂解后匀浆,取上清测蛋白浓度,加入上样缓冲液,置于金属浴变性,经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、湿法转膜后,封闭2 h,一抗β-actin抗体、p-JAK2抗体、p-STAT3抗体、Cyclin D1抗体均于4 ℃下孵育过夜,次日用二抗辣根过氧化酶标记IgG抗体孵育2 h,经化学发光反应显影后拍摄条带,Alpha View SA 3.30 软件分析目标条带的灰度值,结果以目的蛋白相对表达量表示。
1.9 统计分析
使用SPSS 23.0软件进行统计。计量资料用平均值±标准差描述($\bar{x}±s$),采用单因素方差分析统计,实验组间的两两比较采用Q检验;等级资料用中位数和四分位数间距表示(M[IQR]),采用秩和检验统计。检验水准:α=0.05。
2 结果
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2.1 各实验组小鼠血清PGⅠ含量比较
各实验组小鼠血清PGⅠ含量差异有统计学意义(F检验,P<0.01)。进一步分析发现,与正常对照组比较,模型组小鼠血清PGⅠ含量明显降低,差异有统计学意义(Q检验,P<0.05),提示胃黏膜受到损伤;与模型组比较,姜黄素中、高剂量组小鼠血清PGⅠ含量有恢复上升趋势(Q检验,P<0.05),阳性对照组小鼠血清PGⅠ含量亦有上升(Q检验,P<0.05)。见图1。
2.2 各实验组小鼠胃组织病理学观察及评分结果
正常对照组小鼠胃黏膜结构完整,胃腺呈高柱状,排列整齐,形态规则。胃黏膜下层可见少量淋巴细胞散在,未见异型增生(图2A);模型组与低剂量组小鼠胃腺异型增生,累及黏膜深层,腺体排列紊乱、腺腔扩张、扭曲或内折叠,细胞假复层多见,黏膜固有层及黏膜下层可见大量炎症细胞浸润,胃黏膜增生、化生,糜烂伴局部充血(图2B、C);中剂量组小鼠胃黏膜异型增生腺体数量较少,主要累及黏膜浅层和中层,黏膜固有层及黏膜下层炎症细胞数量减少, 局部胃黏膜增生、糜烂、充血,程度较轻(图2D); 高剂量及阳性对照组小鼠胃黏膜腺体异型增生少见或未见,主要累及黏膜浅层。黏膜层及黏膜下层, 炎症细胞明显减少,胃黏膜基本完整(图2E、F)。
各实验组小鼠GED及炎症细胞浸润评分结果见图3。图中给出了各组小鼠病理评分分值的散点分布及每组的评分中位数和四分位间距。模型组与阴性对照组比较,小鼠GED及炎症细胞浸润评分升高(H检验,P<0.01);姜黄素中、 高剂量组与模型组小鼠比较,GED及炎症细胞浸润评分均降低, 差异有统计学意义(H检验,P<0.01或0.05);阳性对照组与模型组小鼠比较, GED评分下降,差异有统计学意义(H检验,P<0.05)。
2.3 各实验组小鼠胃黏膜PCNA抗原表达情况比较
从图4A中可观察到胃黏膜中细胞核被标记成棕褐色的PCNA阳性细胞,以正常对照组最为少见,模型组和低剂量组数量最多(图4B、C),中剂量组次之(图4D),高剂量组明显减少(图4E),阳性对照组PCNA阳性细胞数量与中剂量组接近(图4F)。统计结果表明,与正常对照组比较,模型组小鼠PI明显升高,差异有统计学意义(H检验,P<0.01),与模型组比较,姜黄素中、高剂量组小鼠PI降低,差异有统计学意义(H检验,P<0.05或0.01),阳性对照组小鼠PI降低,差异有统计学意义(H检验,P<0.05)。提示姜黄素对胃黏膜腺上皮细胞的过度增殖有一定的抑制作用,见图5。
2.4 各实验组小鼠胃组织中IFN-γ、IL-1β及IL-6含量的比较
各实验组小鼠胃组织中IFN-γ、IL-1β、IL-6含量差异有统计学意义(F检验,P<0.01)。与正常对照组比较,模型组小鼠IFN-γ、IL-1β、IL-6含量均升高(Q检验,P<0.05);与模型组小鼠比较, 姜黄素高剂量组小鼠IFN-γ、IL-1β、IL-6含量均降低,姜黄素中剂量组小鼠IFN-γ、L-6含量降低,差异均有统计学意义(Q检验,P<0.05);阳性对照组与模型组小鼠比较, IL-1β含量降低(Q检验,P<0.05), 见表1。
2.5 各实验组小鼠p-JAK2、p-STAT3、Cyclin D1蛋白相对表达量比较
各实验组小鼠p-JAK2、p-STAT3、Cyclin D1蛋白相对表达量差异有统计学意义(F检验,P<0.01), 与正常对照组比较,模型组小鼠p-JAK2、p-STAT3、Cyclin D1蛋白相对表达量均上调(Q检验,P<0.05),与模型组小鼠比较, 姜黄素中、高剂量组小鼠磷酸化酪氨酸激酶2(p-JAK2)、信号转导与转录激活因子3(p-STAT3)、细胞周期蛋白D1(Cyclin D1)相对表达水平均下调(Q检验,P<0.05),姜黄素低剂量组小鼠p-JAK2蛋白相对表达水平均下调(Q检验,P<0.05), 见图6~7。
3 讨论
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GC的发生机制复杂,实施一级预防十分困难,GED是GC发展的前期病变,如能早期预防或干预治疗,可以有效降低恶变风险。探索干预GED的有效方法可能是预防GC的关键[6]。构建动物模型是开展GED研究的基础。为模拟人类的发病机制,本研究采用了MNNG+脱氧胆酸钠+乙醇+饥饱失常复合因素造模法。MNNG是一种化学诱变剂,其作用是模拟通过食物过量摄入亚硝酸盐引起的致癌作用,MNNG能够直接作用于DNA,形成烷基化DNA加合物,诱发癌症[21-22]。胆汁反流也是引起GED的重要原因之一,给予小鼠脱氧胆酸钠是为了模拟胆汁反流造成胃黏膜损伤。脱氧胆酸钠是一种阴离子去垢剂,能破坏细胞膜结构,诱发炎症反应,引起胃黏膜慢性炎症[23]。 乙醇的作用是模拟过量饮酒对胃黏膜的损害,乙醇可破坏黏液屏障和黏膜细胞,引起胃黏膜充血、水肿、糜烂,诱导炎性细胞的聚集和活化,引发炎性反应,乙醇是GC的独立危险因素,能促进其他致癌物更有效地渗透组织。乙醇可代谢转化成乙醛,具有致癌性[24]。饥饱失常可导致胃肠功能紊乱,加重胃黏膜损伤。
GED与细胞的过度增殖高度相关, PCNA含量变化有明显的周期性, 在静止期很少, S期达高峰, 其表达水平越高,表明增殖活性越强,增殖越快,恶变风险越高[25]。抑制细胞过度增殖有望阻断GED的进展[20]。PG Ⅰ主要由胃黏膜腺体的主细胞和黏液细胞分泌,胃黏膜受损时,PG Ⅰ 的分泌量发生变化,血液中的PGⅠ浓度随之改变,与胃黏膜的损伤程度基本一致,在GC前病变研究中,血清PG Ⅰ常被用作血清学观察指标[26]。
在持续造模16周后,血清学检测发现模型组小鼠PGⅠ水平降低,提示小鼠胃黏膜受到损伤,免疫组化染色观察发现模型组小鼠胃黏膜PCNA阳性细胞明显增加,处于S期的细胞数远高于正常对照组,呈现过度增殖状态。HE染色观察发现模型小鼠胃黏膜出现异型增生、化生、糜烂、充血等改变,并伴有大量炎症细胞浸润, 模型组GED及炎细胞浸润评分高于正常对照组,且有统计学意义,说明GED模型建立[3,27]。
GED是GC前病变的主要病理阶段, 也是进展为GC之前的最后一个阶段。一旦达到这个阶段,GC的概率至少增加10倍[28-29]。预防或干预GED对于防止胃黏膜病变的恶性转化有重要意义。大量流行病学证据表明,胃黏膜上皮的慢性炎症在GED 及GC中普遍存在。炎症微环境的持续存在对于启动、维持和促进GED转化起着重要作用[30]。炎症因子和炎症细胞是炎症微环境的主要成分。炎症因子高表达可诱导炎症细胞的活化,介导炎症细胞向胃组织损伤部位迁移,启动和放大胃黏膜炎症反应,加重胃黏膜损伤,刺激细胞增殖、增加变异细胞过度增殖的风险[31] 。多项研究表明:IFN-γ高表达可导致小鼠胃黏膜自发性炎症、壁细胞和主细胞丢失及化生和异型增生[7];IL-1β可与其他炎症因子协同,促进炎症反应,引起胃黏膜炎症、糜烂、溃疡,高表达的IL-1β可引起腺体萎缩[32];IL-6能够促进炎症细胞的活化和炎症因子的释放,加剧胃黏膜的炎症反应,高表达的IL-6可能导致细胞增殖异常,IL-6异常升高对于评估胃黏膜损伤具有重要的参考价值[33]。炎症因子INF-γ、IL-1β、IL-6的表达水平,与胃黏膜的病变相关,常用于评估胃黏膜损伤的严重程度[34]。
在本研究中,模型组小鼠胃组织中可见大量炎症细胞浸润,炎症因子水平明显增高,提示存在炎症反应和胃黏膜损伤,相较于模型组,姜黄素干预组小鼠胃组织中INF-γ、IL-1β、IL-6均降低,血清PG1水平有所恢复,胃黏膜的GED、糜烂、充血等病理改变减轻,炎症细胞浸润评分降低,细胞过度增殖受到抑制。提示姜黄素能改善小鼠胃黏膜炎症微环境,对胃黏膜的炎症反应和黏膜损伤有缓解作用。
胃黏膜上皮细胞的过度增殖是导致GED的重要机制,胃黏膜上皮细胞的增殖受到多种因素的调控,包括细胞周期调控蛋白、信号通路、炎症因子等。当这些调控因素异常时,可能导致胃黏膜上皮细胞的增殖异常。 JAK2/STAT3/Cyclin D1信号通路可调控细胞周期调节蛋白,IL-6、IFN-γ是该信号通路的激活因子,在GED的炎症微环境中常呈高表达状态,因此,JAK2/STAT3/Cyclin D1可能是调控GED的重要通路之一[35]。
炎症因子IL-6、IFN-γ可通过活化JAK2,促进STAT3磷酸化并进入胞核,调控基因转录,上调Cyclin D1的表达,Cyclin D1促进细胞从G1期进入S期,加速细胞增殖[36]。Cyclin D1还可通过调控PCNA的磷酸化状态,加快细胞增殖过程。PCNA量的变化与DNA合成一致, 可客观反映细胞的增殖活性及增殖速度。
本研究结果显示,GED小鼠胃组织中JAK2、STAT3及Cyclin D1 蛋白相对表达量均明显升高,提示JAK2/STAT3/Cyclin D1信号通路被激活,上调了细胞周期调节蛋白Cyclin D1的表达水平,胃黏膜中PCNA阳性细胞数量增加,PI指数上升,说明Cyclin D1表达上调,促进细胞周期进程,加快G1/S期转换,提高了细胞的增殖速度。而在姜黄素干预组小鼠胃组织中JAK2、STAT3、Cyclin D1 蛋白相对表达量均有所下调,提示姜黄素对胃黏膜上皮细胞的过度增殖有抑制作用。
综上所述,姜黄素对小鼠胃黏膜上皮异型增生有预防和缓解作用,可以改善GED小鼠胃黏膜的组织形态及炎症微环境,降低炎症因子水平。其作用机制可能与其降低胃组织中的炎症因子INF-γ、IL-6水平,通过JAK2/STAT3/Cyclin D1通路的信号转导,下调周期调节蛋白的表达,抑制细胞过度增殖有关。
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doi: 10.11669/cpj.2024.13.005
  • 接收时间：2023-02-28
  • 首发时间：2026-01-14
  • 出版时间：2024-07-08
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  • 收稿日期：2023-02-28
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    浙江省疾病预防控制中心, 杭州 310051

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* 宋燕华,女,硕士,副主任医师 研究方向:毒理学Tel:(0571)87115254
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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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