基础医学与临床 ›› 2022, Vol. 42 ›› Issue (2): 274-281.doi: 10.16352/j.issn.1001-6325.2022.02.026

• 研究论文 • 上一篇    下一篇

二氢杨梅素对实验性结肠炎小鼠调节性B细胞的影响

李小丽1, 陈慧玲1, 王小春2, 韩悌云1,3*, 张德奎1*   

  1. 1.兰州大学第二医院 消化内科 甘肃 兰州 730000;
    2.甘肃省人民医院 消化内科, 甘肃 兰州 730000;
    3.兰州大学第二医院 消化疾病重点实验室, 甘肃 兰州 730000
  • 收稿日期:2020-12-29 修回日期:2021-06-13 出版日期:2022-02-05 发布日期:2022-01-24
  • 通讯作者: * hantiyun@163.com; sczdk1972@163.com
  • 基金资助:
    兰州大学第二医院“萃英科技创新计划”项目(CY2018-ZD01)

Effect of dihydromyricetin on regulatory B cells in experimental colitis mice

LI Xiao-li1, CHEN Hui-ling1, WANG Xiao-chun2, HAN Ti-yun1,3*, ZHANG De-kui1*   

  1. 1. Department of Gastroenterology, Lanzhou University Second Hospital, Lanzhou 730000;
    2. Department of Gastroenterology, Gansu Provincial Hospital, Lanzhou 730000;
    3. Key Laboratory of Digestive Diseases, Lanzhou University Second Hospital, Lanzhou 730000,China
  • Received:2020-12-29 Revised:2021-06-13 Online:2022-02-05 Published:2022-01-24
  • Contact: * hantiyun@163.com; sczdk1972@163.com

摘要: 目的 探讨二氢杨梅素(DHM)对实验性结肠炎小鼠的作用及其对调节性B细胞(Breg)的影响。方法 将小鼠随机分为正常组、DSS(葡聚糖硫酸钠)组、DSS+DHM组、DSS+mesalazine组(造模采用4% DSS连续饮用7 d)。7 d后,每天分别给予灌胃水(0.2 mL/d)、DHM(40 mg/kg·d)和美沙拉嗪(520 mg/kg·d),连续灌胃7 d,记录小鼠每天疾病活动性指数(DAI),第14天脱颈处死小鼠,记录结肠长度,结肠组织切片行HE染色;流式细胞测量术检测脾脏单细胞悬液中CD19+B淋巴细胞和调节性B细胞(CD19+CD5+CD1d+); RT-qPCR检测脾脏细胞中IL-10、TGF-β、IL-6和IFN-γ的mRNA水平;CCK8法检测脾脏细胞增殖。选择DHM(40 μmol/L)作用脾脏细胞悬液24 h,流式细胞测量术检测Breg(CD19+CD5+CD1d+)细胞。结果 与对照组比较,DSS组DAI评分、病理学评分(炎症严重程度、炎症程度、隐窝损伤等)显著升高,结肠长度明显缩短(P<0.05);脾脏中CD19+ 及Breg细胞比例明显降低(P<0.05);炎性因子显著升高(P<0.05);抗炎因子明显降低(P<0.05);与DSS组比较,DSS+DHM组、DSS+Mesalazine组能明显逆转上述情况(P<0.05);在体外实验中,与对照组相比,经不同浓度DHM干预后,脾脏细胞增殖明显,Breg细胞比例明显增加(P<0.05)。结论 DHM对小鼠结肠炎具有明显的治疗作用,可能与促进调节性B细胞增殖及降低抗炎因子IL-10和TGF-β有关。

关键词: 二氢杨梅素(DHM), 调节B细胞, 实验性结肠炎, 细胞因子

Abstract: Objective To explore effects of Dihydromyricetin (DHM) on experimental colitis and regulatory B cells(Breg) of mice. Methods The mice were randomly divided into control group, DSS group, DSS+DHM group and DSS+ mesalazine group (4% DSS for 7 days). After 7 days, the mice were respectively gavaged with water (0.2 mL/d),DHM (40 mg/kg·d) and mesalazine (520 mg/kg·d) for 7 consecutive days and the disease activity index (DAI) was continuously recorded. The animals were sacrificed on the 14th day and the colon length was measured. The colon tissue sections were microscopied with HE staining. CD19+B lymphocytes and Breg(CD19+CD5+CD1d+) were determined by flow cytometry in splenic single cell suspension; real-time fluorescent quantitative PCR (RT-qPCR) was used to detect the relative mRNA level of IL-10,TGF-β,IL-6 and IFN-γin spleen cells. The effect of DHM on the proliferation of spleen cells was measured by CCK8. DHM (40 μmol/L) was selected to act on the spleen cell suspension for 24 h. flow cytometry was used to detect the ratio of Breg (CD19+CD5+CD1d+). Results Compared with the normal group, DAI and pathological score of DSS group were significantly higher, and the colon length was significantly decreased(P<0.05); The proportion of CD19+ B cells vs. Breg (CD19+CD5+CD1d+) was significantly decreased, the inflammatory factors were significantly increased, and the anti-inflammatory factors were obviously decreased (P<0.05). Compared with DSS group, DSS+DHM group and DSS+Mesalazine group significantly reversed the situation (P<0.05); It was fond in vitro experiment that DHM significantly promoted the proliferation of spleen cells and Breg (P<0.05). Conclusions DDHM has obvious therapeutic effect in experimental colitis mice, which may be related to the promotion of Breg proliferation and of secretion of anti-inflammatory factors IL-10 and TGF-β.

Key words: dihydromyricetin(DHM), regulatory B cell(Breg), experimental colitis, cytokine

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