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[屋尘螨第2组变应原T细胞融合表位对哮喘小鼠信号转导及转录激活因子6信号通路的影响]

[The effect of Dermatophagoides pteronyssinus group 2 allergen T cell fusion epitope on STAT6 signaling in mice with asthma].

作者信息

Zhan Xiao-dong, Duan Bin-bin, Tao Ning, Li Chao-pin

出版信息

Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2017 Feb;35(1):19-23.

Abstract

OBJECTIVE

To investigate the alterations of signal transducer and activator of transcription factor 6(STAT6) signaling in a mouse model of asthma receiving treatment with Dermatophagoides pteronyssinus group 2 allergen(Der p 2) T cell fusion epitope and the mechanisms of the specific immunotherapy.

METHODS

Thirty mice were randomly divided into three groups by the random number table method: the asthma group, the treatment group receiving immunotherapy with Der p 2 T cell fusion epitope, and the negative control group (PBS group)(n = 10 in each group). Mice in the asthma and the treatment groups received intraperitoneal (i. p.) injection of 100 μl Der p 2 solution [PBS containing 100 μg/ml Der p 2 and 2% Al(OH)3] on days 0,7 and 14, respetively, while mice in the PBS group received same volume of PBS containing 2% Al(OH)3. From day 21, 30-min steam inhalation of 0.5 μg/ml Der p 2 was applied to the asthma and treatment groups (once daily for 7 successive days), and the PBS group inhaled same volume of PBS. From day 25 to day 27, the mice in the treatment group received i. p. injection of 200 μl of Der p 2 T cell fusion epitope (100 μg/ml) while the PBS and the asthma groups received the same volume of PBS. Mice were sacrificed at 24 h after the last inhalation, the bronchoalveolar lavage fluid (BALF) collected, and the total protein was extracted from the lung tissue. The levels of IFN-γ, IL-4 and IL-13 in BALF were determined by ELISA. The expression of STAT6 and phosphorylated STAT6 (p-STAT6) in the lung tissue was detected by Western blotting. Data were analyzed with the one-way variance analysis (ANOVA) method.

RESULTS

The level of IFN-γ in the treatment group[(234.40 ± 24.46) pg/ml] was significantly higher than that in the asthma group[(155.80 ± 20.53) pg/ml](P < 0.01). The levels of IL-4 and IL-13 in the treatment group [(30.00 ± 5.50) pg/ml and (174.50 ± 25.99) pg/ml, respectively] were both significantly lower than those in the asthma group[(53.28 ± 8.26) pg/ml and (308.10 ± 28.32) pg/ml, respectively](P < 0.01). Similarly, the levels of STAT6 and p-STAT6 in the treatment group(0.803 ± 0.221 and 0.966 ± 0.323, respectively) were both significantly lower than those in the asthma group (1.669 ± 0.296 and 1.735 ± 0.298, respectively)(P < 0.01).

CONCLUSION

The Der p 2 T cell fusion epitope may function through inhibiting STAT6 to treat asthma in mice.

摘要

目的

研究在接受屋尘螨2类变应原(Der p 2)T细胞融合表位治疗的哮喘小鼠模型中信号转导及转录激活因子6(STAT6)信号通路的变化以及特异性免疫治疗的机制。

方法

采用随机数字表法将30只小鼠随机分为3组:哮喘组、接受Der p 2 T细胞融合表位免疫治疗的治疗组和阴性对照组(PBS组)(每组10只)。哮喘组和治疗组小鼠分别于第0、7和14天腹腔注射100 μl Der p 2溶液[含100 μg/ml Der p 2和2%氢氧化铝的PBS],而PBS组小鼠注射相同体积含2%氢氧化铝的PBS。从第21天起,哮喘组和治疗组小鼠进行30分钟0.5 μg/ml Der p 2的蒸汽吸入(连续7天,每天1次),PBS组吸入相同体积的PBS。从第25天至第27天,治疗组小鼠腹腔注射200 μl Der p 2 T细胞融合表位(100 μg/ml),而PBS组和哮喘组注射相同体积的PBS。在末次吸入后24小时处死小鼠,收集支气管肺泡灌洗液(BALF),并从肺组织中提取总蛋白。采用酶联免疫吸附测定法(ELISA)测定BALF中干扰素-γ(IFN-γ)、白细胞介素-4(IL-4)和白细胞介素-13(IL-13)的水平。采用蛋白质免疫印迹法检测肺组织中STAT6和磷酸化STAT6(p-STAT6)的表达。数据采用单因素方差分析(ANOVA)方法进行分析。

结果

治疗组IFN-γ水平[(234.40±24.46)pg/ml]显著高于哮喘组[(155.80±20.53)pg/ml](P<0.01)。治疗组IL-4和IL-13水平[分别为(30.00±5.50)pg/ml和(174.50±25.99)pg/ml]均显著低于哮喘组[分别为(53.28±8.26)pg/ml和(308.10±28.32)pg/ml](P<0.01)。同样,治疗组STAT6和p-STAT6水平(分别为0.803±0.221和0.966±0.323)均显著低于哮喘组(分别为1.669±0.296和1.735±0.298)(P<0.01)。

结论

Der p 2 T细胞融合表位可能通过抑制STAT6发挥作用来治疗小鼠哮喘。

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