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
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(
)(
)
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:
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:
RONG Wenshuang1,2,3, NIU Yuanfei4, LIU Meiting2,3, YANG Mengyuan5, CUI Shuang2,3, MA Lina6, FU Yao1, WANG Lianmei2,3(
)(
), CAO Junling1,6(
)(
)
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;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。
中图分类号:
荣文双,牛源菲,刘美婷,等. 抗原类型对诱导型干燥综合征小鼠建模的影响[J]. 实验动物与比较医学, 2026, 46(2): 178-190. DOI: 10.12300/j.issn.1674-5817.2025.175.
RONG Wenshuang,NIU Yuanfei,LIU Meiting,et al. Influence of Antigen Type on the Establishment of an Induced Sjögren Syndrome Mouse Model[J]. Laboratory Animal and Comparative Medicine, 2026, 46(2): 178-190. DOI: 10.12300/j.issn.1674-5817.2025.175.
引物名称 Primer name | 引物序列 Primer sequence |
|---|---|
| m-SSA-F | 5'-GCTGGTCTCACAAAGATCTCCTCC-3' |
| m-SSA-R | 5'-TCTCAGCCTCCACAGAAAGCG-3' |
| m-SSB-F | 5'-ATGACTGCTTTGGAGGCCAAAATC-3' |
| m-SSB-R | 5'-TGTCAGCCGGTTTAGCCTGTTGAA-3' |
| m-β-Actin-F | 5'-AGAGGGAAATCGTGCGTGAC-3' |
| m-β-Actin-R | 5'-CAATAGTGATGACCTGGCCGT-3' |
表1 PCR 引物序列
Table 1 PCR primer sequences
引物名称 Primer name | 引物序列 Primer sequence |
|---|---|
| m-SSA-F | 5'-GCTGGTCTCACAAAGATCTCCTCC-3' |
| m-SSA-R | 5'-TCTCAGCCTCCACAGAAAGCG-3' |
| m-SSB-F | 5'-ATGACTGCTTTGGAGGCCAAAATC-3' |
| m-SSB-R | 5'-TGTCAGCCGGTTTAGCCTGTTGAA-3' |
| m-β-Actin-F | 5'-AGAGGGAAATCGTGCGTGAC-3' |
| m-β-Actin-R | 5'-CAATAGTGATGACCTGGCCGT-3' |
图 1 免疫诱导后雌性C57BL/6J小鼠的干燥综合征相关表型
Figure 1 Sj?gren syndrome-related phenotypes in antigen-induced female C57BL/6J mice
组别 Group | 第0天唾液流量/ (mg·min-1) Day 0 salivary flow rate /(mg·min-1) | 第30天唾液流量/ (mg·min-1) Day 30 salivary flow rate /(mg·min-1) | 颌下腺指数 Submaxillary gland index | 胸腺指数 Thymus index | 脾指数 Spleen index |
|---|---|---|---|---|---|
PBS-佐剂雌性对照组 PBS-adjuvanted female control group | 6.60±1.11 | 14.77±1.04 | 0.28±0.03 | 0.26±0.04 | 0.44±0.04 |
唾液腺抗原雌性组 Salivary gland antigen female group | 6.28±1.10 | 13.10±1.05 | 0.29±0.03 | 0.20±0.02∗∗ | 0.51±0.13 |
颌下腺抗原雌性组 Submaxillary gland antigen female group | 6.14±1.16 | 10.52±1.06∗ | 0.30±0.03 | 0.24±0.04 | 0.50±0.06 |
表2 雌性C57BL/6J各组小鼠唾液流量和脏器指数 (
Table 2 Salivary flow rate and organ indices in female C57BL/6J mice of each group
组别 Group | 第0天唾液流量/ (mg·min-1) Day 0 salivary flow rate /(mg·min-1) | 第30天唾液流量/ (mg·min-1) Day 30 salivary flow rate /(mg·min-1) | 颌下腺指数 Submaxillary gland index | 胸腺指数 Thymus index | 脾指数 Spleen index |
|---|---|---|---|---|---|
PBS-佐剂雌性对照组 PBS-adjuvanted female control group | 6.60±1.11 | 14.77±1.04 | 0.28±0.03 | 0.26±0.04 | 0.44±0.04 |
唾液腺抗原雌性组 Salivary gland antigen female group | 6.28±1.10 | 13.10±1.05 | 0.29±0.03 | 0.20±0.02∗∗ | 0.51±0.13 |
颌下腺抗原雌性组 Submaxillary gland antigen female group | 6.14±1.16 | 10.52±1.06∗ | 0.30±0.03 | 0.24±0.04 | 0.50±0.06 |
图 2 免疫诱导后雄性C57BL/6J小鼠的干燥综合征相关表型
Figure 2 Sj?gren syndrome-related phenotypes in antigen-induced male C57BL/6J mice
组别 Group | 第0天唾液流量/ (mg·min-1) Day 0 salivary flow rate /(mg·min-1) | 第30天唾液流量/(mg·min-1) Day 30 salivary flow rate /(mg·min-1) | 颌下腺指数 Submaxillary gland index | 胸腺指数 Thymus index | 脾指数 Spleen index |
|---|---|---|---|---|---|
PBS-佐剂雄性对照组 PBS-adjuvanted male control group | 6.21±0.88 | 14.60±0.90 | 0.38±0.03 | 0.18±0.06 | 0.41±0.07 |
唾液腺抗原雄性组 Salivary gland antigen male group | 6.72±0.73 | 11.17±0.62 | 0.40±0.04 | 0.18±0.05 | 0.37±0.14 |
颌下腺抗原雄性组 Submaxillary gland antigen male group | 6.17±0.90 | 11.54±0.70 | 0.38±0.06 | 0.17±0.03 | 0.34±0.05 |
表3 雄性C57BL/6J各组小鼠的唾液流量和脏器指数 (
Table 3 Salivary flow rate and organ indices in male C57BL/6J mice of each group
组别 Group | 第0天唾液流量/ (mg·min-1) Day 0 salivary flow rate /(mg·min-1) | 第30天唾液流量/(mg·min-1) Day 30 salivary flow rate /(mg·min-1) | 颌下腺指数 Submaxillary gland index | 胸腺指数 Thymus index | 脾指数 Spleen index |
|---|---|---|---|---|---|
PBS-佐剂雄性对照组 PBS-adjuvanted male control group | 6.21±0.88 | 14.60±0.90 | 0.38±0.03 | 0.18±0.06 | 0.41±0.07 |
唾液腺抗原雄性组 Salivary gland antigen male group | 6.72±0.73 | 11.17±0.62 | 0.40±0.04 | 0.18±0.05 | 0.37±0.14 |
颌下腺抗原雄性组 Submaxillary gland antigen male group | 6.17±0.90 | 11.54±0.70 | 0.38±0.06 | 0.17±0.03 | 0.34±0.05 |
图 3 雌性NOD/LtJ模型小鼠与 ICR小鼠的干燥综合征相关表型
Figure 3 Sj?gren syndrome-related phenotypes in female NOD/LtJ model mice and ICR mice
组别 Group | 第0天唾液流量/(mg·min-1) Day 0 salivary flow rate /(mg·min-1) | 第30天唾液流量/(mg·min-1) Day 30 salivary flow rate / (mg·min-1) | 颌下腺指数 Submaxillary gland index | 胸腺指数 Thymus index | 脾指数 Spleen index |
|---|---|---|---|---|---|
| ICR mice | 8.37±0.46 | 15.85±1.25 | 0.39±0.01 | 0.13±0.03 | 0.39±0.08 |
| NOD/LtJ mice | 6.42±0.05∗ | 5.11±0.46∗∗ | 0.40±0.04 | 0.16±0.03∗ | 0.31±0.05 |
表4 雌性NOD/LtJ模型小鼠与ICR小鼠的唾液流量和脏器指数比较 (
Table 4 Salivary flow rate and organ indices in female NOD/LtJ model mice and ICR mice
组别 Group | 第0天唾液流量/(mg·min-1) Day 0 salivary flow rate /(mg·min-1) | 第30天唾液流量/(mg·min-1) Day 30 salivary flow rate / (mg·min-1) | 颌下腺指数 Submaxillary gland index | 胸腺指数 Thymus index | 脾指数 Spleen index |
|---|---|---|---|---|---|
| ICR mice | 8.37±0.46 | 15.85±1.25 | 0.39±0.01 | 0.13±0.03 | 0.39±0.08 |
| NOD/LtJ mice | 6.42±0.05∗ | 5.11±0.46∗∗ | 0.40±0.04 | 0.16±0.03∗ | 0.31±0.05 |
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