摘要
目的探究蛇床子素对毛细支气管炎小鼠模型中Th1/Th2细胞比例失衡的影响及可能的作用机制。方法用呼吸道合胞病毒(RSV)滴鼻法构建毛细支气管炎小鼠模型。将60只SPF级雄性BALB/c小鼠(6~8周龄)随机为5组:对照组,模型组,Cuc组,蛇床子素组和蛇床子素+Cuc组,每组12只。Cuc组腹腔注射1 mg·kg^(-1)Cucurbitacin I,蛇床子素组给予80 mg·kg^(-1)蛇床子素灌胃,蛇床子素+Cuc组给予80 mg·kg^(-1)蛇床子素灌胃的同时腹腔注射1 mg·kg^(-1)Cucurbitacin I,正常组和模型组均给予0.5%CMC-Na灌胃和等量生理盐水腹腔注射,每天1次,连续7 d。用流式细胞仪检测外周血Th1/Th2细胞比例;用酶联免疫吸附测定法检测肺泡灌洗液(BALF)中白细胞介素(IL)-2、干扰素γ(IFN-γ)、IL-4、IL-13水平;计算各组小鼠的胸腺、脾指数;蛋白质印迹法检测肺组织Janus激酶2(JAK2)/信号转导与转录激活因子3(STAT3)通路相关蛋白的表达水平。结果正常组、模型组、Cuc组、蛇床子素组和蛇床子素+Cuc组外周血中Th1/Th2细胞比例分别为2.37±0.25,0.35±0.06,0.66±0.11,0.73±0.10和1.13±0.14;这5组BALF中IFN-γ含量分别为(6.43±1.01),(3.14±0.52),(4.35±0.69),(4.69±0.75)和(5.54±1.06)pg·mL^(-1);这5组BALF中IL-4含量分别为(11.07±1.36),(53.85±6.29),(34.36±4.28),(31.26±3.64)和(20.34±2.59)pg·mL^(-1);这5组肺组织中p-JAK2/JAK2比值分别为0.26±0.05,0.95±0.12,0.71±0.09,0.60±0.06和0.45±0.07;这5组肺组织中p-STAT3/STAT3比值分别为0.40±0.08,0.81±0.14,0.62±0.11,0.66±0.09和0.53±0.08。上述5个指标,模型组与正常组比较,差异均有统计学意义(均P<0.01);Cuc组和蛇床子素组与模型组比较,差异均有统计学意义(均P<0.05);且蛇床子素与Cucurbitacin I联合应用对Janus激酶2/信号转导与转录激活因子3(JAK2/STAT3)通路的抑制作用和重新平衡Th1/Th2细胞比例的促进作用优于两者单独应用。结论蛇床子素可重新平衡毛细支气管炎模型小鼠Th1/Th2细
Objective To explore the effect of osthole on the imbalance of Th1/Th2 cell ratio in a mouse model of bronchiolitis and its possible mechanism.Methods The respiratory syncytial virus(RSV)nasal drip method was used to construct a mouse model of bronchiolitis.Sixty SPF male BALB/c mice(6-8 weeks old)were randomly divided into five groups:control group,model group,cuc group,osthole group and osthole+cuc group,with 12 mice in each group.The cuc group was given 1 mg·kg^(-1) of cucurbitacin I by intraperitoneal injection;the osthole group was given 80 mg·kg^(-1) of osthole by gavage;the osthole+cuc group was given osthole by gavage and cucurbitacin I by intraperitoneal injection;the control group and model group were given 0.5%of CMC-Na by gavage and the same amount of normal saline by intraperitoneal injection,once daily for 7 consecutive days.Flow cytometry was used to detect the ratio of Th1/Th2 cells in the peripheral blood;Enzyme linked immunosorbent assay was used to detect the levels of interleukin(IL)-2,interferon-γ(IFN-γ),IL-4 and IL-13 in alveolar lavage fluid(BALF);the thymus and spleen indexes of mice in each group were calculated;Western blot was used to detect the expression of janus kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3)pathway related proteins in lung tissue.Results The ratio of Th1/Th2 cells in peripheral blood of control group,model group,cuc group,osthole group and osthole+cuc group was 2.37±0.25,0.35±0.06,0.66±0.11,0.73±0.10 and 1.13±0.14,respectively;the content of IFN-γin BALF of these five groups was(6.43±1.01),(3.14±0.52),(4.35±0.69),(4.69±0.75)and(5.54±1.06)pg·mL^(-1),respectively;the level of IL-4 in BALF of these five groups was(11.07±1.36),(53.85±6.29),(34.36±4.28),(31.26±3.64)and(20.34±2.59)pg·mL^(-1),respectively;the ratio of p-JAK2/JAK2 in lung tissues of these five groups was 0.26±0.05,0.95±0.12,0.71±0.09,0.60±0.06 and 0.45±0.07,respectively;the ratio of p-STAT3/STAT3 in lung tissues of these five groups was 0.40±0.08,0.81±0.
作者
胡亚静
鹿璨
张艳娣
闫红丽
田晶晶
HU Ya-jing;LU Can;ZHANG Yan-di;YAN Hong-li;TIAN Jing-jing(Teaching and Research Office of Basic Medicine,Hebei College of Science and Technology,Baoding 071000,Hebei Province,China;Scientific Research Office,Affiliated Hospital of Hebei University,Baoding 071000,Hebei Province,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2022年第8期801-806,共6页
The Chinese Journal of Clinical Pharmacology
基金
2020年度医学科学研究重点课题基金资助项目(20200269)。
