摘要
目的:基于Janus激酶2(JAK2)/信号转导及转录激活因子3(STAT3)信号通路探究益正方对肺脾气虚型反复呼吸道感染(RRTI)模型大鼠免疫功能的影响。方法:60只SD大鼠分为对照组,RRTI组,低、中、高剂量益正方组(10、20和30 g/kg益正方生药灌胃),以及JAK2抑制剂AG490组(5 mg/kg AG490腹腔注射),每组各10只。除对照组外,其余组大鼠采用疲劳结合饮食失节法及刨花加烟叶烟熏法构建肺脾气虚型RRTI模型。观察大鼠一般状态;检测大鼠肺功能[1秒用力呼气容积(FEV1)、用力肺活量(FVC)和呼气峰流速(PEF)];计算大鼠脾脏和胸腺指数;HE染色观察肺组织病理学改变;ELISA法检测血清中免疫球蛋白A(IgA)、IgM、IgG、肿瘤坏死因子α(TNF-α)、白细胞介素4(IL-4)和IL-17水平,以及支气管肺泡灌洗液(BALF)中分泌型IgA(sIgA)水平;流式细胞术检测外周血中CD3^(+)、CD4^(+)和CD8^(+)T淋巴细胞比例;Western blot法检测肺组织中JAK2/STAT3信号通路相关蛋白表达。结果:与对照组相比,RRTI组大鼠一般状态较差,且肺组织存在明显病理损伤,同时肺损伤评分,FEV1、FVC、FEV1/FVC和PEF水平,脾脏和胸腺指数,血清IgA、IgM、IgG和IL-4水平,BALF sIgA水平,以及外周血CD3^(+)、CD4^(+)、CD8^(+)和CD4^(+)/CD8^(+)T淋巴细胞水平均显著降低,而血清TNF-α和IL-17水平,以及肺组织中p-JAK2/JAK2和p-STAT3/STAT3比值均显著升高(P<0.05);与RRTI组相比,低、中、高剂量益正方组和AG490组大鼠一般状态有所改善,肺组织病理损伤有不同程度减轻,同时肺损伤评分,FEV1、FVC、FEV1/FVC和PEF水平,脾脏和胸腺指数,血清IgA、IgM、IgG和IL-4水平,BALF sIgA水平,以及外周血CD3^(+)、CD4^(+)、CD8^(+)和CD4^(+)/CD8^(+)T淋巴细胞水平均显著升高,而血清TNF-α和IL-17水平,以及肺组织中p-JAK2/JAK2和p-STAT3/STAT3比值均显著降低(P<0.05),且益正方各治疗组呈剂量依赖性。结论:益正方能够缓解肺脾气虚型RRTI大鼠肺损伤并改善免疫功能,其作用机制可能与抑制JAK2/STAT3信号通路激活、减轻全身炎症反应有关。
AIM:To explore the effect of Yizheng Prescription on immune function in rats with repeated respi⁃ratory tract infection(RRTI)of lung-spleen Qi deficiency type and its molecular mechanism.METHODS:Sixty SD rats were divided into control group,RRTI group,low-,medium-and high-dose Yizheng Prescription groups(intragastric ga⁃vage of 10,20 and 30 g/kg Yizheng Prescription crude drug),and Janus kinase 2(JAK2)inhibitor AG490 group(intra⁃peritoneal injection of 5 mg/kg AG490),with 10 rats in each group.Except for the control group,lung-spleen Qi deficien⁃cy type RRTI model was established in the rats of the other groups using fatigue,diet disorder and wood shaving^(+)tobacco smoke method.The general state of the rats was observed.The lung function[forced expiratory volume in one second(FEV1),forced vital capacity(FVC)and peak expiratory flow rate(PEF)]was measured.The spleen and thymus indexes were calculated.The pathological changes of lung tissues were observed by HE staining.The levels of immunoglobulin A(IgA),IgM,IgG,tumor necrosis factor-α(TNF-α),interleukin-4(IL-4)and IL-17 in serum,and secretory IgA(sIgA)in bronchoalveolar lavage fluid(BALF)were detected by ELISA.The percentages of CD3^(+),CD4^(+)and CD8^(+)T-lympho⁃cytes in peripheral blood were detected by flow cytometry.The expression levels of JAK2/signal transducer and activator of transcription 3(STAT3)signaling pathway-related proteins in lung tissues were detected by Western blot.RESULTS:Compared with control group,the rats in RRTI group had poorer general status and obvious pathological damage in the lung tissue.Meanwhile,the lung injury score,the FEV1,FVC,FEV1/FVC and PEF levels,spleen and thymus indexes,serum IgA,IgM,IgG and IL-4 levels,BALF sIgA levels,and peripheral blood CD3^(+),CD4^(+),CD8^(+)and CD4^(+)/CD8^(+)T-lym⁃phocytes in RRTI group were significantly decreased,while serum TNF-αand IL-17 levels,and ratios of p-JAK2/JAK2 and p-STAT3/STAT3 in lung tissue were significantly increased(P<0.05).Compared with RRTI group,the general state of the rats in low-,medium-and high-dose Yizheng Prescription groups and JAK2 inhibitor AG490 group was improved,and the pathological damage of lung tissue was attenuated to varying degrees.Meanwhile,the lung injury score,the FEV1,FVC,FEV1/FVC and PEF levels,spleen and thymus indexes,serum IgA,IgM,IgG and IL-4 levels,BALF sIgA levels,and peripheral blood CD3^(+),CD4^(+),CD8^(+)and CD4^(+)/CD8^(+)T-lymphocytes were significantly increased,while serum TNF-αand IL-17 levels and ratios of p-JAK2/JAK2 and p-STAT3/STAT3 in lung tissue were significantly decreased(P<0.05).The effects in Yizheng Prescription groups were dose-dependent.CONCLUSION:Yizheng Prescription alleviates lung injury and improves immune function in RRTI rats of lung-spleen Qi deficiency type.Its mechanism may be related to inhibiting the activation of JAK2/STAT3 signaling pathway,and reducing systemic inflammatory response.
作者
田新磊
朱珊
赵文锦
祝志朋
周怡锦
TIAN Xinlei;ZHU Shan;ZHAO Wenjin;ZHU Zhipeng;ZHOU Yijin(Department of Pediatrics,Henan Hospital of Traditional Chinese Medicine,Zhengzhou 450002,China;Department of Pediatrics,The Second Clinical Medical College of Henan University of Traditional Chinese Medicine,Zhengzhou 450046,China.)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2023年第5期855-862,共8页
Chinese Journal of Pathophysiology
基金
河南省卫生健康委国家中医临床研究基地科研专项(No.2021JDZX2048)
第七批全国老中医药专家学术经验继承工作项目
河南省中医药科学研究专项课题(No.2022ZY1072)。
