芹菜发酵液对小鼠溃疡性结肠炎的保护作用研究

doi:10.16431/j.cnki.1671-7236.2021.11.007

摘要/Abstract

摘要： 本试验旨在研究发酵前后芹菜汁的主要功能成分变化和芹菜发酵液对葡聚糖硫酸钠（DSS）诱导的小鼠溃疡性结肠炎（UC）的防治及免疫调节作用。选取50只4周龄雄性C57BL/6小鼠随机分为空白组、模型组及低、中、高浓度芹菜发酵液组，每组10只。按10 μL/g BW的剂量标准给空白组和模型组小鼠灌胃无菌生理盐水，其他组小鼠则灌胃不同浓度的芹菜发酵液，持续7 d。从第8天开始，除空白组自由饮用无菌水外，其余组连续7 d自由饮用3% DSS溶液；在此过程，每日称量体重，进行疾病活动指数（DAI）评分。在第14天，通过摘眼球取血法获得全血，随后脱颈处死小鼠，解剖取出结肠组织测量长度并通过苏木素-伊红（HE）染色法制作病理切片，进行病理学分析。以流式细胞仪分选技术检测外周血中CD4⁺/CD8⁺T淋巴细胞比值，以酶联免疫吸附法（ELISA）检测血清中肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）、γ-干扰素（IFN-γ）和IL-10的含量。结果表明：①芹菜汁经发酵后，总酸、总糖、总多酚、类黄酮、维生素C和超氧化物歧化酶（SOD）等多种活性成分的含量均显著增加（P<0.05）。②与模型组相比，芹菜发酵液高浓度组能减少DSS引起的小鼠体重损失（P<0.01）、结肠缩短（P<0.01）和DAI降低（P<0.05）。③组织病理学分析结果表明，各发酵液组的UC症状均得到不同程度改善，肠腺结构相对完整，杯状细胞轻微减少，仅有少量淋巴细胞和中性粒细胞浸入。④流式细胞仪分析结果显示，相较于模型组，发酵液组全血CD4⁺/CD8⁺T淋巴细胞比值极显著升高（P<0.01）。⑤ELISA检测结果显示，与模型组相比，芹菜发酵液组的IL-6、TNF-α和IFN-γ含量极显著下调（P<0.01），IL-10的含量极显著上调（P<0.01）。综上，芹菜汁经发酵后主要功能活性成分均得到显著提高，并且发酵芹菜液对DSS诱导的小鼠UC具有一定的防治和免疫调节作用，其作用机制可能与维持外周血中CD4⁺与CD8⁺T淋巴细胞平衡，以及抑制促炎症因子（TNF-α、IL-6和IFN-γ）表达，促进抗炎症因子（IL-10）表达有关。

关键词: 芹菜发酵液; 葡聚糖硫酸钠; 溃疡性结肠炎; 疾病活动指数

Abstract: The purpose of this experiment was to study the changes in the main functional components of celery juice after fermentation and the effect of celery fermentation broth on the prevention and immunoregulation of dextran sodium sulfate (DSS) induced ulcerative colitis (UC) in mice. Fifty male 4-week-old C57BL/6 mice were randomly divided into blank group, model group, and low, medium, and high concentration of cetery fermentation broth groups, with 10 mice in each group. The mice in blank and model groups were given sterile normal saline, the others were given different concentrations of celery fermentation broth by gavage for 7 days, according to the dose standard of 10 μL/g BW. Beginning on the 8th day, except for the blank group to drink sterile water freely, the mice in other groups drunk 3% DSS solution for 7 days. During the induction process, the mice were weighed daily and scored by the disease activity index (DAI). On the 14th day, blood was obtained through the eyeball method, and then the mice were killed by dislocation of spine, and the colon tissue was taken to measure the length and made into pathological sections by the hematoxylin eosin (HE) staining method for histopathological analysis. CD4⁺ and CD8⁺ T lymphocyte ratios in peripheral blood of mice was detected by flow cytometry sorting technology. The serum levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interferon-γ (IFN-γ) and interleukin-10 (IL-10) were detected by enzyme-linked immunosorbent assay (ELISA). The results showed that:①After fermentation, the contents of various active ingredients such as total acid, total sugar, total polyphenols, flavonoids, vitamin C and superoxide dismutase (SOD) in celery juice increased significantly (P<0.05). ②Compared with the model group, high concentration group could reduce the weight loss (P<0.01), colon shortening (P<0.01) and DAI reduction (P<0.05) of mice induced by DSS. ③Histopathological analysis showed that the symptoms of UC in the fermentation broth groups were improved to varying degrees, the intestinal gland structure was relatively complete, the goblet cells were slightly reduced, and only a few lymphocytes and neutrophils were infiltrated. ④The results of flow cytometry analysis showed that CD4⁺/CD8⁺ T lymphocytes in the whole blood of the fermentation broth groups were increased (P<0.01) compared with the model group. ⑤The ELISA test results showed that the content of IL-6, TNF-α and IFN-γ in the fermentation broth groups were lower, and the content of IL-10 was higher than that of the model group (P<0.01). In summary, the main active components of celery juice after fermentation were generally higher than those of unfermented ones. Celery fermentation broth had preventive and immunoregulatory effects on DSS-induced UC in mice. The mechanism might be related to maintaining the balance of CD4⁺ and CD8⁺ T lymphocytes in the peripheral blood, inhibiting the expression of pro-inflammatory factors (IL-6, TNF-α and IFN-γ), and increasing the expression of anti-inflammatory factors (IL-10).

Key words: celery fermentation broth; dextran sodium sulfate; ulcerative colitis; disease activity index

中图分类号:

S716.6

赵冬, 曹金虎, 金慧琴, 单衍可, 刘晓明, 刘斐. 芹菜发酵液对小鼠溃疡性结肠炎的保护作用研究[J]. 中国畜牧兽医, 2021, 48(11): 3975-3984.

ZHAO Dong, CAO Jinhu, JIN Huiqin, SHAN Yanke, LIU Xiaoming, LIU Fei. Study on Protective Effect of Celery Fermentation Broth on Ulcerative Colitis in Mice[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(11): 3975-3984.

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