【作者】 李莉；

【导师】 汪雪峰；

【作者基本信息】 江苏大学 ， 临床检验诊断学（专业学位）， 2019， 硕士

【摘要】 第一部分日本血吸虫多肽SJMHE1对小鼠哮喘模型的作用研究[目的]观察日本血吸虫多肽SJMHE1对哮喘小鼠气道炎症的作用。[方法]择6～8周健康雄性BALB/C小鼠,随机分为4组:PBS组、卵清蛋白(OVA)组、OVA/PBS组,OVA/SJMHEI组。OVA致敏构建支气管哮喘模型,HE染色观察小鼠肺组织病理形态学变化,BALF计数细胞总数及嗜酸性粒细胞数,ELISA测定小鼠血清中OVA特异性IgE水平。[结果]与PBS组比较,OVA、OVA/PBS组肺组织炎性细胞的浸润增加,BALF中细胞总数及嗜酸性粒细胞数增加,血清OVA特异性IgE增多;与OVA、OVA/PBS组比较,OVA/SJMHE1组肺组织炎症细胞的浸润减少,BALF中细胞总数及嗜酸性粒细胞数减少,血清OVA特异性IgE无明显变化。[结论]日本血吸虫多肽SJMHE1可抑制哮喘小鼠气道炎症。第二部分日本血吸虫多肽SJMHE1对哮喘小鼠模型Th1/Th2/Th17/Treg调节的影响[目的]观察日本血吸虫多肽SJMHE1对哮喘小鼠模型Th1/Th2/Th17/Treg调节的影响[方法]择6～8周健康雄性BALB/C小鼠,随机分为4组:PBS组、卵清蛋白(OVA)组、OVA/PBS组,OVA/SJMHE1组。OVA致敏构建支气管哮喘模型,用qPCR(荧光定量PCR)的方法检测脾细胞和肺组织中促炎细胞因子和抗炎细胞因子变化,流式细胞术检测脾细胞中Th1、Th2、Th17、Treg细胞比例,免疫组织化学法检测肺组织中Foxp3蛋白表达水平,免疫印迹(Western b1ot)的方法测定肺组织中GATA3和RORγt蛋白表达水平。[结果]日本血吸虫多肽SJMHE1可调节哮喘小鼠脾细胞和肺组织中促炎和抗炎细胞因子的产生,降低哮喘小鼠脾细胞中Th2的百分率,增加小鼠脾细胞中Th1和Treg细胞的比例,同时增加小鼠脾细胞和肺组织中Foxp3表达量,降低小鼠肺组织中GATA3和RORγt的含量。[结论]日本血吸虫多肽SJMHE1抑制哮喘小鼠气道炎症的免疫机制可能是通过下调Th2和Th17反应并上调Th1和Treg反应。第三部分日本血吸虫多肽SJMHE1降低哮喘小鼠髓源性抑制细胞的数量[目的]观察日本血吸虫多肽SJMHE1对哮喘小鼠髓源性抑制细胞(MDSC)的影响。[方法]择～8周健康雄性BALB/C小鼠,随机分为4组:PBS组、卵清蛋白(OVA)组、OVA/PBS组,OVA/SJMHE1组。OVA致敏构建支气管哮喘模型,流式细胞术测定脾脏MDSC中多形核细胞型MDSC(PMN-MDSC)、单核细胞型MDSC(M-MDSC)所占比例,免疫组织化学染色法检测肺组织MDSC的表达。[结果]与PBS组比较,OVA、OVA/PBS组脾细胞中PMN-MDSC比例及肺组织MDSC数量增加,脾细胞M-MDSC 比例无明显变化;与OVA、OVA/PBS组比较,OVA/SJMHE1组脾细胞PMN-MDSC比例及肺组织MDSC数量减少,脾细胞M-MDSC比例无明显变化。[结论]SJMHE1可能通过减少MDSC数量抑制哮喘小鼠气道炎症。更多还原

【Abstract】 PART I Effect of schistosoma japonicum peptide SJMHE1 in asthmatic mice[Objective]To investigate the effect of schistosoma japonicum peptide SJMHE1 on airway inflammation in asthmatic mice[Method]Male Balb/c mice(6 to 8 weeks old)were randomly divided into four groups:PBS group,ovalbumin(OVA)group,OVA/PBS group and OVA/SJMHE1 group.Asthmatic mice were established using sensitization and challenging in OVA,OVA/PBS and OVA/SJMHE1 group.Hematoxylin and eosin(HE)staining was used for testing the lung pathological changes in mice.Serum anti-OVA-specific immunoglobulin E(IgE)level was detected by ELISA Hemocytometer was used to count the number of BALF cells[Result]Compared to the PBS group,OVA and OVA/PBS groups showed the increase of the inflammatory cell infiltration,the number of BALF cells and eosinophils,and the IgE product.Compared to OVA or OVA/PBS group,OVA/SJMHE1 group induced the decrease of the inflammatory cell infiltration,the number of BALF cells and eosinophils,whereas OVA/SJMHE1 group provided almost no reduction in serum IgE level in the spleen[Conclusion]Taken together,our results indicated that Schistosoma japonicum peptide SJMHE1 can reduce airway inflammation in asthmatic micePART II Schistosoma japonicum peptide SJMHE1 corrects the balance of Thl/Th2/Th17/Treg in asthmatic mice[Objective]The present study is designed to determine whether schistosoma japonicum peptide SJMHE1 has immunoregulatory effects on the(helper T(Th))Thl/Th2/Thl7/regulatory T cell(Treg)balance in asthmatic mice.[Method]Male Balb/c mice(6 to 8 weeks old)were randomly divided into four groups:PBS group,ovalbumin(OVA)group,OVA/PBS group and OVA/SJMHE1 group.Asthmatic mice were established using sensitization and challenging in OVA,OVA/PBS and OVA/SJMHE1 group.The production of pro-inflammatory and anti-inflammatory cytokins in splenocytes and lungs were measured by real time quantitative PCR(qPCR).The proportion of Thl/Th2/Thl7/Treg in the spleenocytes was detected by flow cytometry.Furthermore,The expression of GATA3 and RORyt in the lung tissue were measured by western blotting and the expression of Foxp3 in the lung tissue was detected by immunohistochemistry[Result]We showed that SJMHE1 treatment modulated the production of pro-inflammatory and anti-inflammatory cytokins in splenocytes and lungs of allergic mice,reduced the percent of Th2 cells,and increased the proportion of Thl and Treg cells,accompanying by the increase of Foxp3 expression,and the decrease of GATA3 and RORyt in lungs of allergic mice[Conclusion]The findings provide evidence that SJMHE1 can intervene the development of asthma for diminishing its airway inflammation in mice.The immunological mechanism may involve the down-regulation of Th2 and Th17 response and up-regulation of Thl and Treg responsePART III Schistosoma japonicum peptide SJMHE1 reduces the number of myeloid-derived suppressor cell in asthmatic mice[Objective]To investigate the effect of schistosoma japonicum peptide SJMHE1 on MDSC in asthmatic mice[Method]Male Balb/c mice(6 to 8 weeks old)were randomly divided into four groups:PBS group,ovalbumin(OVA)group,OVA/PBS group and OVA/SJMHE1 group.Asthmatic mice were established using sensitization and challenging in OVA,OVA/PBS and OVA/SJMHE1 group.The proportion of PMN-MDSCs and M-MDSC in the spleenocytes was detected by flow cytometry and the expression of MDSC in the lung tissue was detected by immunohistochemistry[Result]Compared to the PBS group,OVA and OVA/PBS groups showed the increase of the percent of PMN-MDSC in the spleen,and the number of MDSC in the lung of mice,whereas M-MDSC in the spleen showed no significant changes.Compared to OVA or OVA/PBS group,OVA/SJMHE1 group induced the decrease of the proportion of PMN-MDSC in the spleen,and the number of MDSC in the lung of mice,whereas OVA/SJMHE1 group provided almost no reduction in M-MDSC in the spleen.[Conclusion]Schistosoma japonicum peptide SJMHE1 may suppress airway inflammation in asthmatic mice by reducing the number of MDSC.更多还原