实验动物与比较医学 ›› 2026, Vol. 46 ›› Issue (2): 178-190.DOI: 10.12300/j.issn.1674-5817.2025.175

• 人类疾病动物模型 • 上一篇    下一篇

抗原类型对诱导型干燥综合征小鼠建模的影响

荣文双1,2,3, 牛源菲4, 刘美婷2,3, 杨梦园5, 崔爽2,3, 马丽娜6, 富尧1, 王连嵋2,3()(), 曹俊岭1,6()()   

  1. 1.北京中医药大学东直门医院, 北京 100700
    2.中国中医科学院中药研究所中草药中精神活性物质发现与防控北京市重点实验室, 中药安全用药风险评控与转化重点实验室, 北京 100700
    3.中国中医科学院中药研究所道地药材品质保障与资源持续利用重点实验室, 北京 100700
    4.北京中医药大学中药学院, 北京 102488
    5.中国中医科学院中药资源中心, 北京 100700
    6.北京中医药大学东方医院, 北京 100078
  • 收稿日期:2025-10-24 修回日期:2026-02-05 出版日期:2026-04-25 发布日期:2026-04-20
  • 通讯作者:
    王连嵋(1981—),女,博士,副研究员,研究方向:中药毒理与安全用药研究。E-mail: lmwang@icmm.ac.cn。ORCID:0000-0001-5328-6223
    曹俊岭(1972—),男,博士,博士生导师,博士后合作导师,研究方向:中药合理使用研究。E-mail: caojunling72@163.com。ORCID:0000-0001-7640-8713
  • 作者简介:荣文双(1996—),女,硕士研究生,研究方向:中医优势病种的中医药治疗。E-mail: rongwenshuang123@163.com
  • 基金资助:
    国家中医药管理局项目“特色饮片品种挖掘应用-柴胡”(GZY-KJ5-2022-057);中国中医科学院中药研究所中央级公益性科研院所基本科研业务费专项资金资助项目“中药材拟境栽培与品质保障重点实验室”(ZXKT25043);“中国中医科学院中药安全用药风险评控与转化重点实验室”(ZXKT25036)

Influence of Antigen Type on the Establishment of an Induced Sjögren Syndrome Mouse Model

RONG Wenshuang1,2,3, NIU Yuanfei4, LIU Meiting2,3, YANG Mengyuan5, CUI Shuang2,3, MA Lina6, FU Yao1, WANG Lianmei2,3()(), CAO Junling1,6()()   

  1. 1.Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing 100700, China
    2.Beijing Key Laboratory of Psychoactive Substances Discovery and Control in Chinese Herbal Medicines, Key Laboratory of TCM Safety Risk Assessment and Translation, Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China
    3.State Key Laboratory for Quality Assurance and Sustainable Use of Dao-di Herbs, Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China
    4.School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 102488, China
    5.National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China
    6.Dongfang Hospital, Beijing University of Chinese Medicine, Beijing 100078, China
  • Received:2025-10-24 Revised:2026-02-05 Published:2026-04-25 Online:2026-04-20
  • Contact: CAO Junling (ORCID: 0000-0001-7640-8713), E-mail: caojunling72@163.com;
    WANG Lianmei (ORCID: 0000-0001-5328-6223), E-mail: lmwang@icmm.ac.cn

摘要:

目的 观察比较颌下腺抗原和唾液腺抗原用于诱导干燥综合征(Sj?gren syndrome,SS)小鼠模型的建模效果,并与自发性SS模型NOD/LtJ小鼠进行比较,明确两种腺体抗原诱导的SS小鼠模型的表型与免疫特征。 方法 选用成年C57BL/6J小鼠(雌雄各半),分别采用颌下腺抗原和唾液腺抗原联合弗氏佐剂免疫小鼠,诱导建立SS模型;以PBS联合弗氏佐剂免疫小鼠作为对照。在第1天和第7天时采用抗原联合弗氏完全佐剂(Freund’s complete adjuvant,FCA)经小鼠背部皮下多点注射进行免疫诱导,第14天采用抗原联合弗氏不完全佐剂(Freund’s incomplete adjuvant,FIA)经小鼠背部皮下多点注射进行加强免疫诱导。同时设置雌性NOD/LtJ小鼠为自发性SS模型组,以其背景品系ICR小鼠为对照,进行对比分析。实验周期为4周,动态监测小鼠体重、饮水量及唾液流量的变化。在实验终点时收集小鼠各脏器组织与血清样本,称量颌下腺、胸腺和脾脏重量,计算脏器指数;对颌下腺和脾脏组织进行病理形态学分析;采用ELISA法检测血清中白细胞介素-17(interleukin-17,IL-17)含量;采用实时荧光定量PCR法检测颌下腺组织中SS抗原A(SS type A,SSA)和SS抗原B(SS type B,SSB)的mRNA表达水平。 结果 与雌性的PBS联合弗氏佐剂对照组相比,颌下腺抗原雌性组小鼠的饮水量明显增加(P<0.05),唾液分泌量明显减少(P<0.05),颌下腺指数、胸腺指数和脾脏指数均无明显差异(P>0.05),颌下腺出现灶性淋巴细胞浸润、脾脏边缘区扩大,血清IL-17水平明显升高(P<0.05),颌下腺组织中SSA/SSB表达水平无明显差异(P>0.05)。与雌性的PBS联合弗氏佐剂对照组相比,唾液腺抗原雌性组小鼠饮水量、唾液分泌量、颌下腺指数以及脾脏指数均无明显差异(P>0.05),但胸腺指数明显降低(P<0.01);颌下腺出现轻微炎性细胞浸润和腺体萎缩,脾脏白髓及边缘区略有扩大;血清IL-17水平与颌下腺组织中SSB表达水平均明显升高(P<0.01),颌下腺组织中SSA表达水平无明显变化(P>0.05)。与雄性的PBS联合弗氏佐剂对照组相比,除颌下腺抗原雄性组小鼠出现轻微颌下腺腺体萎缩外,各造模组的其余成模指标均没有明显差异(P>0.05)。与ICR对照组相比,NOD/LtJ模型组小鼠饮水量明显增加(P<0.05),唾液流量明显减少(P<0.01),颌下腺指数和脾脏指数均无明显差异(P>0.05),但胸腺指数明显增大(P<0.05);颌下腺病理灶性浸润明显,脾脏边缘区明显扩大,血清IL-17水平和颌下腺SSA/SSB表达水平均明显升高(P<0.05)。 结论 颌下腺抗原和唾液腺抗原均能诱导雌性C57BL/6J小鼠表现出SS相关特征,而颌下腺抗原诱导的SS相关表型较唾液腺抗原诱导更为明显,但弱于自发性雌性NOD/LtJ小鼠。用颌下腺抗原和唾液腺抗原接种雄性C57BL/6J小鼠未能诱导出明显的SS表型。

关键词: 干燥综合征, 颌下腺抗原, 唾液腺抗原, C57BL/6J 小鼠, NOD/LtJ小鼠

Abstract:

Objective This study aims to compare the modeling effects of submaxillary gland antigen and salivary gland antigen in the establishment of Sj?gren syndrome (SS) mouse models, and to characterize the phenotypic and immunological features of these models in comparison with spontaneous SS-prone non-obese diabetic (NOD)/LtJ mice. Methods Adult C57BL/6J mice (equal numbers of males and females) were immunized with submaxillary gland antigen or salivary gland antigen, respectively, combined with Freund's adjuvant to induce SS models. Mice immunized with phosphate-buffered saline (PBS) combined with Freund's adjuvant served as the control group. Immunization was induced via multiple subcutaneous injections in the back with antigen combined with Freund's complete adjuvant (FCA) on Days 1 and 7. A booster immunization was administered via multiple subcutaneous injections in the back with antigen combined with Freund's incomplete adjuvant (FIA) on Day 14. Female NOD/LtJ mice were used as the spontaneous SS model group, with ICR mice as the corresponding control strain for comparative analysis. Body weight, water intake, and salivary flow rate of mice were dynamically monitored for 4 weeks. At the end of the experiment, tissue and serum samples were collected, the weights of submaxillary glands, thymus, and spleen were measured, and organ indices (organ-to-body weight ratios) were calculated. Pathological morphological analysis of the submaxillary gland and spleen was performed with hematoxylin and eosin (HE) staining. Serum interleukin-17 (IL-17) level was detected using enzyme-linked immunosorbent assay (ELISA). Real-time quantitative polymerase chain reaction was used to detect the mRNA expression levels of SS type A (SSA) and SS type B (SSB) in submaxillary gland tissues. Results Female mice in the submaxillary gland antigen group exhibited significantly increased water intake (P<0.05) and reduced salivary flow rate (P<0.05) compared with the female control group. No statistically significant differences were observed in the submaxillary gland index, thymus index and spleen index (P>0.05). Focal lymphocytic infiltration was observed in the submaxillary glands, and the splenic marginal zone was enlarged. Serum IL-17 levels were significantly increased (P<0.05). There was no significant difference in submaxillary gland SSA/SSB expression levels (P>0.05). Compared with the female control group, female mice in the salivary gland antigen group showed no statistically significant differences in water intake, salivary flow rate, submaxillary gland index, and spleen index (P>0.05), whereas the thymus index was significantly reduced (P<0.01). Mild inflammatory cell infiltration and glandular atrophy were observed in the submaxillary glands, and the splenic white pulp and marginal zone were slightly enlarged. Serum IL-17 levels and submaxillary gland SSB mRNA expression levels were significantly increased (P<0.01), whereas no significant change was observed in submaxillary gland SSA expression levels (P>0.05). Compared with the male control group, mild submaxillary gland atrophy was observed in male mice in the submaxillary gland antigen group, whereas no obvious changes were found in other modeling-related indicators (P>0.05). Compared with the ICR control group, NOD/LtJ model mice exhibited elevated water intake (P<0.05), significantly reduced salivary flow rate (P<0.01), no significant differences in the submaxillary gland index or spleen index (P>0.05), but a significantly increased thymus index (P<0.05). Marked focal infiltration was observed in the submaxillary glands, the splenic marginal zone was obviously enlarged, and serum IL-17 concentrations as well as submaxillary gland SSA/SSB expression levels were significantly increased (P<0.05). Conclusion Submaxillary gland antigen and salivary gland antigen can induce SS-related features in female C57BL/6J mice. The SS-related phenotype is more pronounced in the submaxillary gland antigen group than in the salivary gland antigen group, but weaker than that in spontaneously SS-prone female NOD/LtJ mice. Immunization of male C57BL/6J mice with submaxillary or salivary gland antigens fails to induce an obvious SS phenotype.

Key words: Sj?gren syndrome, Submaxillary gland antigen, Salivary gland antigen, C57BL/6J mice, NOD/LtJ mice

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