目的 探讨矢车菊素-3,5-O-双葡萄糖苷（cyanidin-3,5-O-diglucoside,C35G）对改善五氟尿嘧啶（5-FU, 30 mg/kg）诱导化疗性小鼠肠黏膜炎作用机制。方法 将BALB/c雄性小鼠随机分成正常组、模型组、C35G组。以连续4 d腹腔注射5-FU诱导肠黏膜损伤小鼠模型,C35G低高剂量组连续7日分别灌胃C35G（50、150 mg/kg）。末次给药后,CO2处死小鼠,取血和结肠组织。采用酶联免疫吸附试法（ELISA）检测各组小鼠血清中肿瘤坏死因子（TNF-α）、白介素-1β（IL-1β）、白介素-10（IL-10）水平。苏木精-伊红（H＆E）、过碘酸-雪夫（PAS）、阿利新蓝（Alcian Blue）染色观察各组小鼠结肠组织病理变化、杯细胞分布及黏蛋白2（mucin2,MUC2）的分泌情况。Western blot 法检测结肠中Wnt、c-Myc蛋白表达水平。荧光实时定量PCR检测各组小鼠结肠组织中紧密连接蛋白mRNA表达水平。结果 与5-FU组相比,C35G处理能显著降低5-FU造成的小鼠结肠肠黏膜损伤,并显著降低小鼠血清中TNF-α与IL-1β水平（P<0.05）,且升高IL-10水平（P<0.01）。C35G能显著促进结肠组织中Wnt和c-Myc的蛋白表达,并显著提高紧密连接蛋白（Zo1、claudin1、claudin7、occludin）mRNA表达水平。结论 C35G对5-FU诱导的小鼠化疗性肠黏膜炎具有明显改善作用。
Abstract
Objective To explore the effect of cyanidin-3,5-O-diglucoside (C35G) on 5-fluorouracil (5-FU)-induced intestinal mucositis in mice. Methods BALB/c male mice were randomly divided into normal group, model group, low-dose and high-dose C35G groups. Except for the normal group, mice in other groups were treated with 5-FU to induce a mouse model of intestinal mucositis for 4 days. Mice in the C35G low-dose and high-dose groups were intragastrically administrated with C35G (50 and 150 mg/kg) for 7 consecutive days. Finally, all mice were euthanized, and blood and colon tissues were collected. The levels of TNF-α, IL-1β and IL-10 in the serum were detected by enzyme-linked immunosorbent assay (ELISA). HE, PAS, and alcian blue staining kits were used to stain the colon tissue sections, and the pathological changes of colon tissues, the content of goblet cells and the secretion of mucin 2 were observed. Western blot was used to detect the expression levels of Wnt and c-Myc in colon tissus. Real-time quantitative PCR was used to detect the expression levels of tight junction proteingenes. Results Compared with the 5-FU group, the preventive intervention of C35G could significantly reduce the colonic mucosal damages in mice exposed to 5-FU. In addition, C35G intervention could also significantly reduce the levels of TNF-α and IL-1β in the serum (P<0.05), increase the level of IL-10 (P<0.01), and significantly increase the expression levels of Wnt and c-Myc in colon tissues. At the same time, the mRNA expression levels of tight junction proteins (Zo1, claudin1, claudin7, occludin) were significantly increased. Conclusion C35G is effective in ameliorating 5-FU-induced intestinal mucositis in mice.
关键词
矢车菊素-3 /
5-O-双葡萄糖苷 /
五氟尿嘧啶 /
肠黏膜损伤 /
炎症
Key words
cyanidin-3 /
5-o-diglucoside /
5-fluorouracil /
intestinal mucosal damage /
inflammation
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基金
国家自然科学基金（No.81560530,81760589,81960590,82273630）,广西自然科学基金（No.2016GXNSFBA380172,2020GXNSFAA159160）,中组部“西部之光”访问学者计划（2023）,广西八桂青年拔尖人才支持计划（2023）,人力资源和社会保障部高层次留学回国人员资助计划（No.RSTH[2019]160）
