甘草内生菌代谢物的抗炎作用

Anti-inflammatory Effect in the Metabolites of Endophytes from Glycyrrhiza Uralensis

关键词:  甘草  内生菌代谢物  抗炎  RAW264.7  大鼠

Key words:  Glycyrrhiza uralensis  the metabolites of endophytes  anti-inflammatory  RAW264.7  rats

摘要:

目的 研究甘草内生菌代谢物的体内外抗炎作用。方法 96孔板中接种巨噬细胞RAW264.7培养，模型组加入1 mg·L^-1脂多糖（LPS），各受试药物组分别加入10，20，40 mg·L^-1甘草水提液浸膏或内生菌发酵物，2 h后再加入LPS1 mg·L^-1培养，检测NO、IL-1β、PGE₂的含量。Wistar大鼠90只，随机分为9组，空白组、模型组、阳性对照组给予生理盐水0.01 mL·g^-1，甘草水煎液组给予甘草水煎液0.95 g·kg^-1，各内生菌代谢物组给予甘草内生菌发酵物0.95 g·kg^-1，每日1次，连续灌胃给药5 d。末次给药后1 h，除空白组外，各组大鼠尾静脉注射1 mg·kg^-1体质量LPS，制备急性肺炎模型，空白组注射等量生理盐水，阳性对照组按5 mg·kg^-1体质量腹腔注射地塞米松给药。6 h后大鼠股动脉取血，检测外周血白细胞计数及血清中TNF-α、IL-6、IL-10的含量。结果 与LPS模型组比较，40 mg·L^-1时，甘草浸膏组、5组内生菌代谢物组（JTZB006、JTZB018、JTZB059、JTZB060、JTZB063）巨噬细胞RAW264.7分泌的NO、PGE₂、IL-1β的含量显著降低（P<0.05或P<0.01）；与同一质量浓度（40 mg·L^-1）的甘草浸膏组比较，JTZB006、JTZB060组巨噬细胞RAW264.7分泌的PGE₂的含量显著降低（P<0.05）。与模型组比较，阳性对照组、甘草水煎液组、3组内生菌代谢物组（JTZB006、JTZB060、JTZB063）大鼠外周血中白细胞数量及血清中TNF-α、IL-6的含量均显著降低，IL-10的含量显著升高（P<0.05或P<0.01）；与甘草水煎液组比较，JTZB006、JTZB060、JTZB063组IL-10的含量显著升高（P<0.05或P<0.01）。结论 3株甘草内生菌代谢物（JTZB006、JTZB060、JTZB063）对LPS诱导的炎症模型具有体外及体内抗炎作用。

Abstract:

OBJECTIVE To study anti-inflammatory effect of the metabolites of endophytes isolated from Glycyrrhiza uralensis in vivo and in vitro.METHODS RAW264.7 cells were inoculated onto a 96-well plate. The model group was added with 1 mg·L^-1 LPS for culture, and each test drug group was added with Glycyrrhiza water extracts or the fermentation products of endophytes (10, 20, 40 mg·L^-1) for 2 h of treatment, then LPS (1 mg·L^-1) was added for stimulation. The content of nitric oxide (NO), Interleukin-1β (IL-1β) and Prostaglandin E₂ (PGE₂) were measured. Ninety Wistar rats were randomly divided into 9 groups, and each group was given the corresponding drug by gavage administration once per day for 5 d. Except the blank group, the other groups were intravenously injected with LPS (1 mg·kg^-1) to establish an acute pneumonia model, while the blank group was injected with the same volume of normal saline. After 6 h, blood was collected from rat femoral artery to detect peripheral white blood cell counts and the contents of serum TNF-α, IL-6 and IL-10.RESULTS Compared with LPS model group, when the mass concentration was maintained at 40 mg·L^-1, the contents of NO, PGE₂ and IL-1β were significantly reduced in extractum Glycyrrhiza group and endophytes fermentation products groups(JTZB006, JTZB018, JTZB059, JTZB060, JTZB063)(P<0.05 or P<0.01). The contents of PGE₂ in JTZB006, JTZB060 were significantly lower than those of extractum Glycyrrhiza group at 40 mg·L^-1. Compared with model group, the white blood cell counts and the contents of TNF-α and IL-6 were significantly reduced in positive control group, Glycyrrhiza water decoction group, 3 groups of the fermentation products of endophytes. The contents of IL-10 in JTZB006, JTZB060 and JTZB063 significantly higher than those of Glycyrrhiza water decoction group in rat serum (P<0.05 or P<0.01).CONCLUSION JTZB006, JTZB060 and JTZB063 have anti-inflammatory effects in LPS induced inflammatory model.