白细胞介素-4对肠道期旋毛形线虫感染的保护作用  

Protective effect of IL-4 to Trichinella spiralis infection in the intestinalphase

作　　者：骆泽妮 吴安琪 汪志锴 潘晋[1] 孙希萌[1] LUO Zeni;WU Anqi;WANG Zhikai;PAN Jin;SUN Ximeng(Department of Medical Microbiology and Parasitology,School of Basic Medical Sciences,Capital Medical University,Beijing 100069,China;Grade 2021 Class Medical Laboratory Testing,School of Basic Medical Sciences,Capital Medical University,Beijing 100069,China)

机构地区：[1]首都医科大学基础医学院病原生物学系,北京100069 [2]首都医科大学基础医学院,北京100069

出　　处：《中国寄生虫学与寄生虫病杂志》2024年第3期354-359,366,共7页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：北京市自然科学基金(7222007);国家自然科学基金(81772213)。

摘　　要：目的 探讨小鼠白细胞介素-4 (IL-4)对肠道期旋毛形线虫感染的保护作用。方法 消化旋毛虫保种小鼠肌肉,收集肌幼虫,超声破碎后离心,取上清制备旋毛虫抗原。将10只野生型BALB/c小鼠随机分成未感染组和野生感染组,每组5只;另取4只IL-4敲除(IL-4KO) BALB/c小鼠作为IL-4KO感染组。野生感染组和IL-4KO感染组每鼠灌胃旋毛虫肌幼虫400条,未感染组灌胃等体积PBS。感染旋毛虫后8 d,采集各组小鼠眼眶静脉血,离心收集血清,ELISA检测单核细胞趋化因子1 (MCP-1)含量。剖取十二指肠和近端空肠,制备石蜡切片后进行苏木精-伊红(HE)染色,使用Aperio ImageScope 12.4.3测量测量十二指肠、空肠的肠绒毛和隐窝的长度与杯状细胞的数量和大小,计算肠绒毛长度/隐窝长度的比值(V/C)和杯状细胞数/绒毛长度的比值(GC/V)。分离肠系膜淋巴结中的淋巴细胞,与旋毛虫抗原(10μg/ml)共培养72 h,收集细胞上清,高通量液相蛋白芯片(Luminex)技术检测IL-1β、IL-12p70、IL-2、IL-5、IL-6、干扰素γ (IFN-γ)、肿瘤坏死因子α (TNF-α)的含量。使用SPSS 26.0统计软件对数据进行分析,两两比较采用独立样本t检验,多样本比较采用单因素方差分析。结果 ELISA结果显示,未感染组、野生感染组、 IL-4KO感染组小鼠血清中MCP-1含量分别为(344.90±21.80)、(350.50±38.30)、(467.94±190.01) pg/ml,IL-4KO感染组高于野生感染组(t=0.681,P <0.05)。HE染色结果显示,未感染组小鼠的十二指肠和空肠肠黏膜完整、绒毛结构正常,无炎症改变;野生感染组、IL-4KO感染组小鼠的十二指肠和空肠肠黏膜均出现空泡状的杯状细胞、绒毛长度变短,炎症明显,IL-4KO感染组炎症更为严重。IL-4KO感染组小鼠十二指肠和空肠V/C分别为2.62±0.12、2.78±0.25,均低于野生感染组的3.46±0.05、3.65±0.12 (F=24.09、20.46,P <0.01、0.05);未感染组空肠V/C为4.69±0.16,高于野生感染组(F=25.43,P <0.01)。IL-4KO�Objective To investigate the protective effects of interleukin 4(IL-4)to Trichinella spiralis infection in the intestinal phase in mice.Methods The muscle samples of T.spiralis reservation mice were digested for collection of muscle larvae,which were sonicated and centrifuged to collect supernatant for preparing Trichinella antigen.Ten wild-type BALB/c mice were randomly divided into the non-infection group and wild-type infection group,with 5 mice each group,while additional 4 IL-4 knockout mice(IL-4KO)were assigned to IL-4KO infection group.The mice of wild-type infection group and IL-4KO infection group were given with 400 muscle larvae each by gavage respectively,while the non-infection group received the same volume of PBS.Eight days after infection,orbital blood samples were obtained to collect sera by centrifugation,and the content of monocyte chemotactic protein-1(MCP-1)was detected by ELISA.The mice of all groups were dissected to collect the duodenum and proximal jejunum for preparation of paraffin sections.The sections were stained with Hematoxylin and eosin(HE).Measure the length of villi and crypts as well as the number and size of goblet cells useing Aperio ImageScope 12.4.3,and calculate the vllus length/crypt length(V/C)and the ratio of the number of goblet cell/villus length(GC/V).Lymphocytes isolated from the mesenteric lymph nodes were cultured with Trichinella antigen(10μg/ml)for 72 hours,and the supernatant was collected.A Luminex assay was performed to measure the levels of cytokines including IL-1β,IL-12p70,IL-2,IL-5,IL-6,interferon-(IFN-)and tumor necrosis factor-α(TNF-α).The data was analyzed using SPSS 26.0 software with independent sample t-tests for pairwise comparisons and one-way ANOVA for multiple sample comparisons.Results ELISA results showed that serum MCP-1 levels were(344.90±21.80),(350.50±38.30)and(467.94±190.01)pg/ml in the non-infection,wildtype infection and IL-4KO infection group,respectively,the IL-4KO infection group was higher than the wild-type infection group(t

关 键 词：旋毛形线虫 白细胞介素-4 肠道期 感染 抵抗作用 

分 类 号：R532.14[医药卫生—内科学]