西替利嗪抑制哮喘小鼠肺组织JAK2-STAT3通路激活及肥大细胞活化

Cetirizine inhibits activation of JAK2-STAT3 pathway and mast cell activation in lung tissue of asthmatic mice

目的通过构建哮喘小鼠模型,探究西替利嗪对哮喘小鼠肺组织Janus激酶2/信号转导子和转录激活子3(JAK2/STAT3)通路及肥大细胞活化的影响。方法将60只无特定病原体(SPF)级BALB/c小鼠分为对照组、模型组、西替利嗪低剂量组(2 mg/kg)、西替利嗪高剂量组(5 mg/kg)、西替利嗪联合AG490组(5 mg/kg西替利嗪联合0.4 mg/kg STAT3抑制剂AG490),每组10只。通过OVA致敏并激发构建哮喘模型,各处理组每天连续灌胃相应量西替利嗪,西替利嗪联合AG490组还需每周1次腹腔注射0.4 mg/kg AG490;对照组灌胃等剂量生理盐水直至造模结束。ELISA检测小鼠血清中组胺水平;计数肺泡灌洗液(BALF)中总细胞数;HE染色观察肺组织病变情况、甲苯胺蓝染色观察小鼠肺组织肥大细胞脱颗粒情况,免疫组织化学染色法检测肺组织类胰蛋白酶表达,Western blot法检测JAK2、磷酸化的JAK2(p-JAK2)、STAT3、磷酸化的STAT3(p-STAT3)蛋白表达。结果与对照组相比,模型组小鼠血清中组胺水平、BALF细胞总数、嗜酸性粒细胞计数、肺组织病理损伤程度、类胰蛋白酶水平和p-JAK2/JAK2、p-STAT3/STAT3表达显著增加;相较于模型组,西替利嗪低剂量组、高剂量组、西替利嗪联合AG490组小鼠血清中组胺水平、BALF细胞总数、嗜酸性粒细胞计数、肺组织病理损伤程度、类胰蛋白酶、p-JAK2/JAK2、p-STAT3/STAT3表达显著降低;相较于高剂量组,西替利嗪联合AG490组血清中组胺水平BALF细胞总数、嗜酸性粒细胞计数、肺组织病理损伤程度、类胰蛋白酶、p-STAT3/STAT3表达显著降低,而p-JAK2/JAK2表达无明显变化。结论西替利嗪能够抑制哮喘小鼠肺组织JAK2/STAT3通路激活及肥大细胞活化。

Objective To investigate the effects of cetirizine on Janus kinase 2/signal transducer and activator of transcription 3(JAK2/STAT3) pathway and mast cell activation in asthmatic mice by establishing a mouse model of asthma. Methods Sixty SPF BALB/c mice(10 in each group) were divided into control group, model group, low-dose cetirizine group(2 mg/kg), high-dose cetirizine group(5 mg/kg), cetirizine combined with AG490 group(5 mg/kg cetirizine combined with 0.4 mg/kg AG490). The asthma model was established by ovalbumin(OVA) sensitization and eliciting. Each treatment group was given certain amount of cetirizine by gavage every day, and the cetirizine combined with AG490 group was given intraperitoneal injection of 0.4 mg/kg AG490 once a week;the control group was given the same dose of normal saline until the end of modeling. The level of histamine in serum was detected by ELISA;the total number of cells in bronchoalveolar lavage fluid(BALF) was counted;the pathological changes of lung tissue were detected by HE staining;the expression of tryptase in lung tissue was detected by immunohistochemical staining;the expressions of pathway related proteins were detected by Western blot analysis. Results Compared with those in the control group, the level of histamine in serum, the total number of BALF cell, the eosinophilia count, the degree of lung pathological injury, and the expressions of tryptase, p-JAK2/JAK2, and p-STAT3/STAT3 in the model group were significantly higher;compared with those in the model group, the level of histamine in serum, the totalnumber of BALF cell, the eosinophilia count, the degree of lung pathological injury, and the expressions of tryptase, p-JAK2/JAK2, and p-STAT3/STAT3 in low-dose group, high-dose group, and cetirizine combined with AG490 group were significantly lower;compared with those in the high-dose group, the level of histamine in serum, the total number of BALF cell, the eosinophilia count, the degree of lung pathological injury, and the expressions of tryptase and p-STAT3/STAT3 in cetirizine combined with AG490 group were significantly lower, however, the expression of p-JAK2/JAK2 had no significant change. Conclusion Cetirizine inhibits JAK2-STAT3 pathway activation and mast cell activation in lung tissue of asthmatic mice.