日本血吸虫源多肽SJMHE1对变应性鼻炎小鼠干预作用分析  

A preliminary study on the intervention effect of Schistosoma japonicum-derived peptide SJMHE1 on allergic rhinitis mice

作　　者：高学荣 郑婷婷 董利阳 徐小卫 汪雪峰[2] 毛朝明[2] GAO Xuerong;ZHENG Tingting;DONG Liyang;XU Xiaowei;WANG Xuefeng;MAO Chaoming(College of Public Health and Management,Jiangsu Vocational College of Medicine,Yancheng,Jiangsu 224005,China;Department of Nuclear Medicine,the Affiliated Hospital of Jiangsu University,Zhenjiang,Jiangsu 212001,China)

机构地区：[1]江苏医药职业学院公共卫生与管理学院,江苏盐城224005 [2]江苏大学附属医院核医学科,江苏镇江212001

出　　处：《热带医学杂志》2024年第5期656-661,F0004,共7页Journal of Tropical Medicine

基　　金：国家自然科学基金(81871243,81900562);镇江市重点社会发展项目(SH2021050)。

摘　　要：目的探究日本血吸虫源多肽SJMHE1对卵清蛋白(OVA)诱导的小鼠变应性鼻炎(AR)的保护作用与机制。方法将15只雌性BALB/c小鼠随机分为正常组、AR组以及AR+SJMHE1组,每组5只。正常组全程未给予任何处理;AR组在第0、7、14天腹腔给予OVA佐剂混悬液致敏,随后在第28~32天鼻腔滴注5%OVA溶液(20μL)进行局部激发,每日1次;AR+SJMHE1组在诱导变应性鼻炎的基础上于第0、14、28天皮下注射SJMHE1乳化液进行干预。末次激发后观察各组小鼠20 min,并进行行为学(挠鼻和打喷嚏)记录;苏木精-伊红染色观察各组小鼠鼻黏膜病理学改变;酶联免疫吸附试验检测各组小鼠血清中白细胞介素(IL)-4、IL-13和IL-10含量;流式细胞术检测小鼠脾脏CD19+细胞中IL-10+细胞比例。另选6只雌性BALB/c小鼠分为空白对照组和异硫氰酸荧光素(FITC)-SJMHE1组,每组3只。空白对照组注射磷酸盐缓冲液(PBS),FITC-SJMHE1组皮下注射FITC标记的SJMHE1(FITC-SJMHE1)。流式细胞技术检测FITC阳性细胞在脾脏中的含量,并观察CD19+细胞与FITC-SJMHE1结合比例。结果与AR组挠鼻次数(33.20±5.07)次/20 min和打喷嚏次数(中位数=8次/20 min)相比,AR+SJMHE 1组小鼠挠鼻次数(17.20±6.02)次/20 min和打喷嚏次数(中位数=1次/20 min)显著下降,差异均有统计学意义(F/H=33.71、10.81,P均<0.01),且鼻粘膜病理损伤得到缓解。与AR组小鼠血清中IL-4(21.55±2.78)pg/mL、IL-13(40.80±3.54)pg/mL和IL-10(61.32±15.30)pg/mL的水平相比,AR+SJMHE1组IL-4(14.39±1.53)pg/mL、IL-13(22.49±3.32)pg/mL水平降低,IL-10(108.65±25.26)pg/mL水平升高,差异均有统计学意义(F=58.92、80.10、17.12,P均<0.01)。与AR组小鼠脾脏CD19+细胞中IL-10+细胞比例(0.79±0.32)%相比,AR+SJMHE1组IL-10+比例(3.35±0.99)%显著上调,差异有统计学意义(F=19.55,P<0.01)。流式细胞术结果显示FITC-SJMHE1可分布于小鼠脾脏且与CD19+细胞结合。结论SJMHE1对OVA诱导的小鼠变应性鼻炎有干预作用,可Objective To explore the protective effect of Schistosoma japonicum-derived peptide SJMHE1 on allergic rhinitis(AR) induced by ovalbumin(OVA) in mice and its mechanism.Methods Fifteen female BALB/c mice were randomly divided into normal group,AR group,and AR+SJMHE1 group,with 5 mice in each group.The normal group did not receive any treatment throughout the entire process.The AR group was sensitized by intraperitoneal administration of OVA adjuvant suspension on days 0,7,and 14,subsequently;from 28 to 32 days,mice were given a daily infusion of 5% OVA for local stimulation.On the basis of inducing allergic rhinitis,the AR+SJMHE1 group received subcutaneous injection of SJMHE1 emulsion at 0,14,and 28 days for intervention.After the last challenge,mice were observed for 20 minutes and their behavior(nasal rubbing movements and sneezes) was recorded.The pathological changes of mice nasal mucosa were observed by hematoxylin-eosin staining.Interleukin-4(IL-4),IL-13 and IL-10 in the serum of each group of mice were measured by enzyme-linked immunosorbent assay.The ratio of IL-10~+ cells in mice spleen CD19~+ cells was measured by flow cytometry.Six female BALB/c mice were divided into blank control group and fluorescein isothiocyanate(FITC)-SJMHE1group,three mice in each group.The blank control group was injected with phosphate buffer solution(PBS),and FITCSJMHE1 group was injected with FITC-labeled SJMHE1(FITC-SJMHE1) subcutaneously.Proportion of FITC-positive cells in spleen and proportion of CD19~+ binding FITC-SJMHE1 were detected by flow cytometry.Results Compared with the AR group,the number of rubbing [(33.20±5.07)times/20 min vs.(17.20±6.02)times/20 min] and sneezing [(median=8 times/20 min) vs.(median=1 time/20 min)] in the AR+SJMHE1 group decreased significantly;the differences were statistically significant(F/H=33.71,10.81;all P<0.01),and the pathological damage to the nasal mucosa was alleviated.Compared with the serum concentrations of IL-4(21.55±2.78) pg/mL,IL-13(40.80±3.54) pg/mL and IL-10(61.32±15.

关 键 词：日本血吸虫 SJMHE1 变应性鼻炎 脾脏 Breg细胞 

分 类 号：R765.21[医药卫生—耳鼻咽喉科]