多房棘球蚴感染晚期小鼠肝脏中NK1.1的表达对T细胞功能的影响  

Effect of NK1.1 expression on liver T cell function in the advanced stage of mice infected with Echinococcus multilocularis

作　　者：赵涵玥 李锦田 薛俊隆 卡力比夏提·艾木拉江 林仁勇 吐尔干艾力·阿吉[1,3] ZHAO Hanyue;LI Jintian;XUE Junlong;KALIBIXIATI Aimulajiang;LIN Renyong;TUERGANAILI Aji(Department of Hepatobiliary&Hydatid Diseases,Digestive Vascular Surgery Center,the First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,Xinjiang,China;Institute of Clinical Medicine,the First Affiliated Hospital of Xinjiang Medical University,State Key Laboratory of pathogenesis,Prevention and treatment of High Incidence in Central Asia,Urumqi 830054,Xinjiang,China;Xinjiang Laboratory of Hydatid Fundamental Medicine,the First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,Xinjiang,China)

机构地区：[1]新疆医科大学第一附属医院消化血管外科中心肝胆包虫外科,新疆乌鲁木齐830054 [2]新疆医科大学第一附属医院临床医学研究院省部共建中亚高发病因与防治国家重点实验室,新疆乌鲁木齐830054 [3]新疆医科大学第一附属医院新疆包虫病基础医学重点实验室,新疆乌鲁木齐830054

出　　处：《中国寄生虫学与寄生虫病杂志》2024年第5期594-600,607,共8页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：新疆维吾尔自治区自然科学基金重点项目(2022D01D17);国家自然科学基金(81960377);新疆维吾尔自治区科技厅科技创新领军人才(2022TSYCLJ0034)。

摘　　要：目的 观察多房棘球蚴感染晚期小鼠肝T细胞的NK1.1表达变化,分析NK1.1表达对T细胞功能的影响。方法 C57BL/6N小鼠随机分为对照组和感染组。感染组小鼠经肝门静脉注射2 000个多房棘球蚴原头节,对照组注射等体积生理盐水。感染后36周取肝组织,苏木素-伊红(HE)染色观察肝组织病理变化,天狼星红和Masson染色观察肝组织纤维化情况。收集肝淋巴细胞,流式细胞术检测小鼠肝CD4^(+)T细胞和CD8^(+)T细胞占比,并检测不同T细胞的NK1.1、颗粒酶B (GrB)、穿孔素、γ干扰素(IFN-γ)和白细胞介素-17 (IL-17)表达变化。采用SPSS 25.0软件进行统计学分析,两组间比较采用独立样本t检验。结果 感染多房棘球蚴36周后,HE染色显示感染组小鼠肝组织内可见具有生发层结构的病灶,周围出现大量炎性细胞浸润;Masson染色和天狼星红染色显示感染组小鼠肝组织纤维化明显。流式结果显示,感染组小鼠肝脏的CD4^(+)T和CD8^(+)T细胞的占比分别为(32.0±6.0)%、(42.0±13.9)%,与对照组的(36.6±9.0)%、(50.2±5.2)%差异均无统计学意义(t=1.068、1.446,均P> 0.05)。感染组NK1.1^(+)CD4^(+)T细胞占CD4^(+)T细胞的(30.0±14.5)%,低于对照组NK1.1^(+)CD4^(+)T细胞的(69.0±10.7)%(t=5.573,P <0.01)和感染组NK1.1-CD4^(+)T细胞的(51.6±10.8)%(t=2.920,P <0.05)。感染组NK1.1^(+)CD8^(+)T细胞占CD8^(+)T细胞的(22.0±12.9)%,低于对照组NK1.1^(+)CD8^(+)T细胞的(43.6±5.9)%(t=3.999,P <0.01)和感染组NK1.1-CD8^(+)T细胞的(64.0±13.6)%(t=5.496,P <0.01)。感染组小鼠分泌GrB、IFN-γ和IL-17的NK1.1^(+)CD4^(+)T细胞占比分别为(4.8±2.8)%、(6.8±6.0)%和(6.9±5.2)%,均低于对照组的(12.6±4.8)%、(23.6±7.3)%和(22.1±7.1)%(t=3.462、4.171、4.124,均P <0.01);分泌GrB、IFN-γ和IL-17的NK1.1^(+)CD8^(+)T细胞占比分别为(1.5±0.9)%、(5.3±2.7)%和(3.7±2.3)%,均低于对照组的(6.0±2.7)%、(18.1±9.5)%和(16.9±9.3)%(t=3.861、3.196、3.361,P <0.01、0.05、0.01)。感染组分泌穿�ObjectiveTo observe the changes in NK1.1 expression of liver T cells in the advanced stage of mice infected with Echinococcus multilocularis,and analyze the effect of NK1.1 expression on liver T cells' function.MethodsC57BL/6N mice were assigned into the infection group and the control group randomly.Mice in the infection group were injected with 2 000 E.multilocularis protoscoleces through hepatic portal vein,while mice in the control group injected with the same volume of normal saline.The livers were collected at the advanced stage of infection(36 weeks post-infection).The pathological changes of the liver tissues were observed by hematoxylin-eosin(HE)staining,and the degree of live fibrosis was observed by Masson staining and Sirius red staining.The lymphocytes were collected and the flow cytometry was performed to detect the proportions of CD4^(+)T and CD8^(+)T cells.The expression changes of NK1.1,granzyme B(GrB),perforin,interferon-γ(IFN-γ) and interleukin-17(IL-17) in different T cells were also detected.SPSS 25.0 software was used for statistical analysis,and an independent sample t-test was used to compare the mean between the two groups.ResultsHE staining showed that lesions with germinal layer structure were formed in the livers of mice in the infection group 36 weeks post-infection,with infiltration of inflammatory cells in surrounding tissues,and results of Masson staining and Sirius red staining showed the significant hepatic fibrosis.Flow cytometry results showed the proportions of CD4^(+)T and CD8^(+)T cells in the liver of mice in the infection group 36 weeks post-infection were(32.0 ± 6.0)% and(42.0 ± 13.9)%,respectively,which were(36.6 ± 9.0)%and(50.2 ± 5.2)% in the control group,with no significant differences(t = 1.068,1.446,both P > 0.05).The proportion of NK1.1^(+)CD4^(+)T cells in the infection group was(30.0 ± 14.5)%,which was lower than NK1.1^(+)CD4^(+)T cells in the control group [(69.0 ± 10.7)%](t = 5.573,P < 0.01) and NK1.1-CD4^(+)T cells in the infection group [(51.6 ± 10.8

关 键 词：多房棘球蚴病 NK1.1 CD4^(+)T细胞 CD8^(+)T细胞 

分 类 号：R532.32[医药卫生—内科学]