文献类型：期刊文献

中文题名：甘草内生菌的鉴定及抗炎作用研究

英文题名：Study on the identification and anti-inflammatory effect of endophyte from Glycyrrhiza uralensis in vitro and in vivo

作者：杨志军[1];邓毅[1];曼琼[1];杨秀娟[1];杨延泽[1];刘靓[1]

第一作者：杨志军

机构：[1]甘肃中医药大学药学院,兰州730000

第一机构：甘肃中医药大学药学院（西北中藏药协同创新中心办公室）

年份：2018

卷号：34

期号：7

起止页码：852

中文期刊名：中国临床药理学杂志

外文期刊名：The Chinese Journal of Clinical Pharmacology

收录：CSTPCD;;北大核心:【北大核心2017】；CSCD:【CSCD2017_2018】；

基金：国家自然科学基金资助项目(81360633)

语种：中文

中文关键词：甘草;内生菌;鉴定;抗炎

外文关键词：Glycyrrhiza uralensis ; endophyte ; identification ; anti - inflammation

摘要：目的对分离的甘草内生菌进行鉴定,并研究其抗炎作用。方法取对数生长期贴壁巨噬细胞RAW264.7,制成1×106cell·m L^(-1)的单细胞悬液,培养至细胞贴壁。空白组,加1640培养基培养;模型组,加1 mg·L^(-1)脂多糖(LPS)培养;对照组,分别加入10,20,40 mg·L^(-1)的甘草水提液浸膏;实验组,分别加入10,20,40 mg·L^(-1)的内生菌发酵物(JTYB006、JTYB012、JTYB029、JTYF023),药物组均预处理2 h,再加入1 mg·mL^(-1)LPS刺激培养;以Griess法检测一氧化氮(NO)含量,以酶联免疫吸附法检测白细胞介素-1β(IL-1β)、前列腺素-E2(PGE2)含量。按照体重将昆明种小鼠随机分为7组:模型组、对照Ⅰ组、对照Ⅱ组、实验Ⅰ~Ⅳ组。通过二甲苯致小鼠耳肿胀、冰醋酸致小鼠腹腔毛细血管通透性增高,评价甘草内生菌发酵物的体内抗炎作用。结果甘草内生细菌JTYB006、JTYB012、JTYB029分别属于芽孢杆菌(Bacillus.)3个不同种属,内生真菌JTYF023属于曲霉属赭曲霉。模型组的NO、PGE2、IL-1β的含量分别为(89.53±8.02)μmol·L^(-1),(420.85±57.85)pg·mL^(-1),(35.24±5.77)pg·m L^(-1),40 mg·L^(-1)实验Ⅰ~Ⅳ组(JTYB006、JTYB012、JTYB029、JTYF023)的NO含量分别为(65.60±4.31),(63.48±6.89),(62.30±5.92),(56.47±7.26)μmol·L^(-1),PGE2含量为(254.07±43.62),(295.40±49.13),(280.65±52.91),(296.42±54.90)pg·mL^(-1),IL^(-1)β的含量为(21.34±5.36),(20.65±4.78),(16.67±3.98),(22.54±5.30)pg·mL^(-1),实验组与模型组比较,差异均有统计学意义(P<0.05或P<0.01)。模型组小鼠耳肿胀度、腹腔冲洗液光密度值分别为17.99±1.60,0.28±0.02;实验Ⅰ、Ⅲ和Ⅳ组的小鼠耳肿胀度分别为15.05±2.38,13.68±2.69,15.36±2.03,腹腔冲洗液光密度值分别为0.24±0.04,0.18±0.05,0.21±0.05,实验组与模型组比较,差异均有统计学意义(P<0.05或P<0.01)。结论 3株(JTYB006、JTYB029、JTYF023)甘草内生菌发酵物具有体外及体内抗炎作用。
Objective To identify endophytes from Glycyrrhiza uralensis and study their anti -inflammatory effects in vivo and in vitro. Methods RAW264. 7 cells in the logarithm growth period were made 1 x 10^6 cell mg·L-1 to be single cell suspension. The blank group was added with 1640 culture medium, the model group was added with 1 mg mL-1 lipopolysaccharide （LPS）for culture, the control group was added with Glycyrrhiza water extracts（ 10,20,40 mg · L-1 ）, and experimental group was added with 10, 20,40mg·L-1 endophyte fermentation products （JTYB006, JTYB012, JTYB029, JTYF023 ） for 2 h of treatment, then1 mg·L-1LPS was added for stimulation. The content of nitric oxide （NO） were measured by Griess, and interleukin - 1β （ IL - 1β） and prostaglandin E2 （ PGE2 ） were measured by ELISA. Kunming mice were randomly divided into 7 groups ： model, control I , control Ⅱ , experimental I , experimental Ⅱ , experimental m, experimental IV. Then xylene - induced mouse ear edema test and Glacial acetic acid -induced mouse abdominal capillary permeability increase test were con- ducted. Results Three strains of endophytic bacterium are Bacillus, belonging to 3 species, and 1 strain of endophytic fungi is Aspergillus ochraeeus. The contents of NO, PGE2 and IL - 1β were （89.53 ± 8.02）μmol·L-1, （420. 85 ±57.85）μmol·L-1, （35.24 ±5.77）pg·m L-1in model group. When the mass concentration was maintained at 40 mg · L- 1, the contents of NO were （65.60 ± 4. 31 ）, （ 63.48 ± 6. 89 ）, （ 62.30 ± 5.92 ）, （ 56.47 ±7.26 ） μmol·L-1 ,the contents of PGE2 were（254.07 ±43.62）, （295.40 ±49. 13）, （280.65 ±52.91 ）, （296. 42 ±54. 90） pg·m L-1 ,the contents of IL - 113 were （21.34 ± 5.36）, （20. 65 ±4. 78）, （ 16.67 ±3.98）, （22.54 ± 5.30）pg·m L-1 in 4 experimental groups. Comparison between model group and experimental groups showed that the di- fferences were significant （P 〈 0.05 or P 〈 0. 01 ）. The ear swelling degree and the absorbance of abdominal Flushing fluid in model group were 17.99 ±1.60,0. 28 ±0. 02. The ear swelling degree were 15.05 ±2.38,13.68 ±2.69, 15.36 ±2. 03 ,the absorbance of abdominal Flushing fluid were 0. 24 ±0. 04,0. 18 ±0. 05,0. 21 ±0. 05 in experimental I , Ill, IV groups. By comparison between model group and experimental groups, the differences were significant （P 〈 0.05 or P 〈 0. 01 ）. Conclusion The fermentations of endophytes in 3 strains （JTYB006, JTYB029 and JTYF023 ） have anti - inflammatory effects both in vitro and in vivo.