多房棘球蚴感染小鼠外周血中TREM-1对Treg细胞及其相关因子变化的影响  

The effects of TREM-1 from peripheral blood of Echinococcosis multilocularis infected mice on the Treg cells and its related factors

作　　者：马海梅 张峰波[2] 李玉娇[3] 沙桐 赵骁 陈志强 丁剑冰[4] MA Haimei;ZHANG Fengbo;LI Yujiao;SHA Tong;ZHAO Xiao;CHEN Zhiqiang;DING Jianbing(Department of Microbiology,College of Basic Medical,Xinjiang Medical University,Urumqi 830011,China;Clinical Laboratory,College of Basic Medical,Xinjiang Medical University,Urumqi 830011,China;Department of Blood Transfusion,the First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,China;Department of Immunology, College of Basic Medical, Xinjiang Medical University, Urumqi 830011, China)

机构地区：[1]新疆医科大学基础医学院微生物学教研室,乌鲁木齐830011 [2]新疆医科大学第一附属医院医学检验中心,乌鲁木齐830011 [3]新疆医科大学第一附属医院输血科,乌鲁木齐830011 [4]新疆医科大学基础医学院免疫教研室,乌鲁木齐830011

出　　处：《新疆医科大学学报》2020年第7期841-847,共7页Journal of Xinjiang Medical University

基　　金：国家自然科学基金(81960373,81660343,8460307)。

摘　　要：目的探讨多房棘球蚴感染小鼠外周血中髓系细胞触发受体-1(Triggering recepetor expressed on myeloid cells 1,TREM-1)对调节性T细胞及其相关因子表达的影响及意义。方法选用雌性BALB/c小鼠,采用腹腔接种多房棘球蚴原头蚴的方式建立小鼠模型后,分别给予抗TREM-1单克隆抗体(250μg/kg),TREM-1抑制剂(17个氨基酸组成的多肽—LP17合成短肽)(3.5 mg//kg)、生理盐水尾静脉注射,分为单抗处理组、LP17处理组、模型组,同时设立正常对照组。观察各组小鼠在感染早期(8 d)和感染晚期(180 d)大体改变及囊泡湿重;采用酶联免疫吸附法(ELISA)检测外周血TREM-1、白细胞介素-10(interleukin-10,IL-10)、转化生长因子-β(transforming growth factor-β,TGF-β)含量;实时荧光定量PCR(qRT-PCR)法检测外周血TREM-1、叉头样转录因子P3(fork-head transcription factor P3,FoxP3)、白细胞介素-10(IL-10)、转化生长因子-βmRNA水平;流式细胞术检测CD4^+CD25^+FoxP3^+Treg细胞(regulatory T cell,Treg)比例,并分析TREM-1与CD4^+CD25^+FoxP3^+Treg细胞及关联因子的相关性。结果在感染晚期,单抗处理组与LP17处理组间囊泡湿重抑制率比较差异无统计学意义(P=0.071),但囊泡湿重较模型组有所减少;在感染早期和晚期,小鼠外周血中TREM-1、Foxp3、TGF-β、IL-10的含量及mRNA水平和CD4^+CD25^+FoxP3^+Treg细胞百分比均较正常组有明显升高,且感染早期升高的幅度较晚期大,差异有统计学意义(P<0.05);TREM-1与CD4^+CD25^+FoxP3^+Treg细胞及其特征转录因子Foxp3、细胞因子TGF-β、IL-10均呈正相关(P<0.05)。结论在感染多房棘球蚴后,激活TREM-1可通过上调CD4^+CD25^+FoxP3^+Treg细胞参与早期的炎症反应;而阻断TREM-1可使促炎因子小幅度下降,上调抗炎因子,来介导抗炎作用;随着病程的延长,TREM-1可能通过募集IL-10、TGF-β等来调节Treg细胞的动态平衡、肝脏纤维化和肝损伤的发生,但具体机制有待进一步探讨。Objective To explore the effect and significance that the triggering receptor expressed on myeloid cells 1(TREM-1)influence the fuction of Treg cells and its related factors in peripheral blood of Echinococcosis multilocularis infected mice.Methods E.multilocularis infected mouse model was established by using SPF female BALB/c mice,and treated with anti-TREM-1 monoclonal antibody(250μg/kg),TREM-1 inhibitors LP1(3.5 mg/kg)and normal saline,respectively.The healthy mice were normal control group.At the early stage of infection(8 d)and the late stage of infection(180 d),the concentration of TREM-1,IL-10,TGF-βwere determined by ELISA;the percentage of CD4^+CD25^+FoxP3^+Treg cells was detected by flow cytometry;the mRNA expression of TREM-1,FoxP3,IL-10 and TGF-βwere detected by qRT-PCR;The correlation between TREM-1 and CD4^+CD25^+FoxP3^+Treg cells and its related factors were analyzed by statistic software.Results At the late stage of infection,there was no statistically significant difference in the inhibition rate of vesicle wet weight between the monoclonal antibody treatment group and the LP17 treatment group(P>0.05),but the vesicle wet weight was lower than that of the model group.At the early and late stage of infection,the mRNA expression and the concentration of TREM-1,IL-10,TGF-βand even the percentage of CD+4CD+25Foxp3+Treg cells in peripheral blood of the model group was increased significantly compared with the normal group,and the increasing amplitude at the early stage of infection was significantly greater than that at the late stage of infection(P<0.05).TREM-1 was positively correlated with the number of CD4^+CD25^+FoxP3^+Treg cell and FoxP3,TGF-βand IL-10(P<0.05).Conclusion After infection of E.multilocularis,the activation of TREM-1 can up-regulate the proportion of CD4^+CD25^+FoxP3^+Treg cell and the expression of FoxP3,which is involved in early inflammatory response.However,blocking TREM-1 can slightly up-regulate the proportion of CD4^+CD25^+FoxP3^+Treg cells and the expression of FoxP3,th

关 键 词：多房棘球蚴 TREM-1 CD4^+CD25^+FoxP3^+Treg细胞 IL-10 TGF-Β 

分 类 号：R383.33[医药卫生—医学寄生虫学]