二氢杨梅素对实验性结肠炎小鼠调节性B细胞的影响  

Effect of dihydromyricetin on regulatory B cells in experimental colitis mice

作　　者：李小丽[1] 陈慧玲[1] 王小春[2] 韩悌云 张德奎[1] LI Xiao-li;CHEN Hui-ling;WANG Xiao-chun;HAN Ti-yun;ZHANG De-kui(Department of Gastroenterology, Lanzhou University Second Hospital, Lanzhou 730000;Department of Gastroenterology, Gansu Provincial Hospital, Lanzhou 730000;Key Laboratory of Digestive Diseases,Lanzhou University Second Hospital, Lanzhou 730000,China)

机构地区：[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

出　　处：《基础医学与临床》2022年第2期274-281,共8页Basic and Clinical Medicine

基　　金：兰州大学第二医院“萃英科技创新计划”项目(CY2018-ZD01)。

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(40μmol/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。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)wasused 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(40μmol/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.

关 键 词：二氢杨梅素(DHM) 调节B细胞 实验性结肠炎 细胞因子 

分 类 号：R574[医药卫生—消化系统]