徐露;王娴;王宣璎;姚庆华;陈寅波;
XU Lu;WANG Xian;WANG Xuan-ying;YAO Qing-hua;CHEN Yin-bo;Cancer Hospital of University of Chinese Academy of Sciences(Zhejiang Cancer Hospital);the Second Clinical Medical College of Zhejiang Chinese Medical University;Sir Run Run Shaw Hospital Affiliated to Medical College of Zhejiang University;

• 1:中国科学院大学附属肿瘤医院(浙江省肿瘤医院)
• 2:浙江中医药大学第二临床医学院
• 3:浙江大学医学院附属邵逸夫医院

摘要(Abstract)：

探讨姜黄素介导白细胞介素6(interleukin-6,IL-6)/信号转导和转录激活因子3(signal transducer and activator of transcription 3,STAT3)信号通路修复结肠癌5-氟尿嘧啶(5-fluorouracil,5-FU)化疗引起的肠黏膜损伤的潜在作用机制。SD大鼠腹腔注射60 mg·kg~(-1)·d~(-1) 5-FU,连续注射4 d,制备5-FU化疗肠黏膜损伤模型,随机分为模型组(等体积生理盐水)和姜黄素低、中、高剂量组(50,100,200 mg·kg~(-1)),以正常SD大鼠为对照组(等体积生理盐水),各组连续4 d灌胃给药,并记录每日体质量及粪便变化情况。给药结束后,心脏取血,收集空肠组织。酶联免疫吸附(ELISA)法检测各组大鼠血清白细胞介素1β(interleukin-1β,IL-1β)、肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)表达水平,同时测定空肠组织伊文斯蓝(Evans blue,EB)浓度。苏木精-伊红(HE)染色法观察大鼠空肠组织病理状态并测量空肠绒毛长度及隐窝深度。免疫组化法测定紧密连接蛋白(claudin,occludin,ZO-1)阳性表达。Western blot法检测空肠组织IL-6,p-STAT3,E-cadherin,vimentin,N-cadherin蛋白表达水平。结果显示,姜黄素可升高大鼠体质量、粪便质量(P<0.05或P<0.01),降低粪便评分、EB浓度、IL-1β和TNF-α水平(P<0.05或P<0.01);可较大程度上维持空肠组织黏膜表面与绒毛结构的完整性,减轻病理改变,且呈剂量依赖性。同时姜黄素可升高occludin,claudin,ZO-1阳性表达(P<0.05或P<0.01),修复肠道屏障功能,下调空肠组织中IL-6,p-STAT3,vimentin,N-cadherin蛋白表达水平(P<0.05或P<0.01),上调E-cadherin蛋白表达水平(P<0.05)。由此可知,姜黄素可修复结肠癌5-FU化疗诱导的肠黏膜损伤,其作用机制可能与抑制IL-6/STAT3信号通路,进而抑制上皮间充质转化(epithelial-mesenchymal transition,EMT)进程有关。
This study aims to investigate the potential mechanism of curcumin in mediating interleukin-6(IL-6)/signal transducer and activator of transcription 3(STAT3) signaling pathway to repair intestinal mucosal injury induced by 5-fluorouracil(5-FU) chemotherapy for colon cancer. SD rats were intraperitoneally injected with 60 mg·kg~(-1)·d~(-1) 5-FU for 4 days to establish a model of intestinal mucosal injury. Then the rats were randomly divided into model group(equal volume of normal saline), curcumin low, medium and high dose groups(50, 100, 200 mg·kg~(-1)), and normal SD rats were used as control group(equal volume of normal saline). Each group received gavage administration for 4 consecutive days, and the changes of body weight and feces were recorded every day. After administration, blood was collected from the heart, and jejunum tissues were collected. The levels of serum interleukin-1β(IL-1β) and tumor necrosis factor-α(TNF-α) were detected by ELISA, and at the same time, the concentration of Evans blue(EB) in jejunum was measured. Hematoxylin-eosin(HE) staining was used to observe the pathological state of jejunum, and the length of jejunum villi and the depth of crypt were measured. The positive expression levels of claudin, occludin and ZO-1 were detected by immunohistochemistry. Western blot was used to detect the protein expression of IL-6, p-STAT3, E-cadherin, vimentin and N-cadherin in jejunum tissues. The results showed that, curcumin significantly increased body weight and fecal weight(P<0.05 or P<0.01), decreased fecal score, EB concentration, IL-1β and TNF-α levels(P<0.05 or P<0.01) in rats. In addition, curcumin maintained the integrity of mucosal surface and villi structure of jejunum to a large extent, and reduced pathological changes in a dose-dependent manner. Meanwhile, curcumin could increase the positive expression of occludin, claudin and ZO-1(P<0.05 or P<0.01), repair intestinal barrier function, downregulate the protein expression of IL-6, p-STAT3, vimentin and N-cadherin in jejunum tissues(P<0.05 or P<0.01), and upregulate the protein expression of E-cadherin(P<0.05). Therefore, curcumin could repair the intestinal mucosal injury induced by 5-FU chemotherapy for colon cancer, and the mechanism may be related to the inhibition of IL-6/STAT3 signal and the inhibition of epithelial-mesenchymal transition(EMT) process.

关键词(KeyWords)： 结肠癌;姜黄素;IL-6/STAT3信号通路;5-氟尿嘧啶(5-FU);肠黏膜损伤
colon cancer;curcumin;IL-6/STAT3 signaling pathway;5-fluorouracil(5-FU);intestinal mucosal injury

Abstract:

Keywords:

基金项目(Foundation): 浙江省中医药管理局项目(2019ZA018,2014ZB069)

作者(Author): 徐露;王娴;王宣璎;姚庆华;陈寅波;
XU Lu;WANG Xian;WANG Xuan-ying;YAO Qing-hua;CHEN Yin-bo;Cancer Hospital of University of Chinese Academy of Sciences(Zhejiang Cancer Hospital);the Second Clinical Medical College of Zhejiang Chinese Medical University;Sir Run Run Shaw Hospital Affiliated to Medical College of Zhejiang University;

Email:

DOI: 10.19540/j.cnki.cjcmm.20201106.401

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