金银花水提物对致敏小鼠肠道黏膜炎症和sIgA抗体反应的免疫调控

Effects of Lonicera on the immunoregulation of intestinal mucosal inflammatory response and sIgA response in ovalbumin sensitized BALB/c mice

目的观察中药金银花对卵清蛋白致敏的BALB/c小鼠肠道黏膜炎症和sIgA反应系统的影响。方法以BALB/c雌鼠为实验对象,按照给药浓度分为100%(H)、50%(M)、25%(L)金银花浓度组,激发后不给药组(Ch),正常生理盐水组(NS)。应用ELISA法测定小肠黏膜总IgA、OVA特异性IgA含量(OVA-IgA);免疫组织化学法检测小肠固有层(LP)IgA+浆细胞,集合淋巴小结(PP)中膜表面IgA+(smIgA+)淋巴细胞数量;RT-PCR法检测IL-4mRNA表达;组织原位杂交法检测小肠PP中IL-4、肠黏膜TGF-β1、TNF-αmRNA表达。结果经H、M浓度金银花干预后,过敏小鼠肠道黏液中总IgA、OVA-IgA分泌抑制,肠道LP中IgA+浆细胞数、PP中smIgA+淋巴细胞数、小肠LP及PP中IL-4mRNA表达均降低、肠黏膜TGF-βmRNA升高(P均<0.01),H、M、L浓度金银花尚可以剂量依赖方式降低小肠TNF-αmRNA表达(P均<0.01)。结论过敏小鼠肠黏膜炎症反应与前炎症因子和抑制性炎症因子失平衡有关;金银花可用于治疗过敏小鼠肠道免疫炎症。

Objectives To evaluate the therapeutic effects of Lonicera extract from water solution （Lon ext） on the intestinal mucosal inflammatory response and slgA response in ovalbumin （OVA） sensitized BALB/c mice. Methods Female BALB/c mice aged at 6 weeks fed on OVA-free diet, were randomly divided into 5 groups by the concentration of drugs used in intragastric treatments. Eight mice were included in each group with 100% （H） , 50% （M）, 25% （L）, of the Lon ext and no drug （Ch） administered after being sensitized and challenged with OVA. The mice receiving normal saline （NS） without sensitization and challenge were set as the negative control. The levels of total IgA and OVA-specific IgA （OVA-IgA） in the intestinal mucus of the mice were determined by ELISA. The number of IgA^＋ plasmaeytes in Lamina propria （LP） and IgA^＋ lymphoeytes on the surface membrane （sm） of Peyer＇ s patches （PP） were determined by immunohistochemistry. The IL-4 mRNA expression was determined by RT-PCR. In situ hybridization was used to determine the mRNA expression of IL-4 in small intestine PP, and TGF-β1 and TNF-α in intestinal mucosa. Results When the allergic mice were treated with the H or M Lonicera extract, the total and OVAspecific IgA secreted in intestinal mucsa was suppressed. The amount of IgA^＋ plasmacytes in LP and the smIgA^＋ lymphocytes in PP and the mRNA expression of IL-4 in LP and in PP were reduced, while the mRNA expression of TGF- β was notably upregulated （P 〈 0.01） . In group H, M and L, Lon ext reduced TNF-α mRNA expression in a dosedependent manner （P〈 0.01） . Conclusions The intestinal mucosal inflammatory responses in the ovalbumin-induced allergic mice may be related to the imbalance of anti-inflammatory and pro-inflammatory cytokines. Lon ext can be used to treat intestinal mucosal inflammation in ovalbumin sensitized mice.