东莨菪碱法和高渗盐水法复制小鼠干燥综合征模型的比较

Comparison of mice models of Sjögren's syndrome with scopolamine method and hypertonic saline method

目的:比较东莨菪碱法和高渗盐水法复制小鼠干燥综合征(SS)模型的特点。方法:将6~8周龄SPF级雄性BALB/c小鼠随机分为3组:正常组、高渗盐水组和东莨菪碱组(n=10)。除正常组给予生理盐水外,东莨菪碱组小鼠经皮下注射1 mg/ml氢溴酸东莨菪碱0.2 mg/次/20 g体重(qid),高渗盐水组经鼠眼缓滴无菌2.8 mg/ml高渗氯化钠溶液[100μl/眼(qid)],连续造模10 d。期间检测各组小鼠体质量、饮水量消耗和泪液结晶变化。实验结束后利用ELISA方法检测各组小鼠血清中自身抗体SSA和SSB、抗核抗体ANA、BAFFR蛋白和炎症因子TNF-α表达量,计算脾脏系数;HE染色分析脾脏病理变化,Chisholm-Mason组织学评分标准评价外分泌腺组织中淋巴细胞浸润程度。结果:与高渗盐水组相比,东莨菪碱组小鼠体质量下降,饮水量消耗显著增加(P<0.05或P<0.01),泪液分泌减少且无蕨形结晶分布散乱(Ⅳ型);血清中抗体SSA、ANA、TNF-α表达量、脾脏系数均显著升高(P<0.05);HE染色结果表明,东莨菪碱组小鼠脾脏白髓中淋巴细胞增殖明显,脾小体萎缩,达到3级,而高渗盐水组小鼠脾小体结构较为整齐,仅有淋巴细胞增殖,为1级;东莨菪碱组小鼠颌下腺、腮腺组织中淋巴细胞浸润评分升高(P<0.05)。结论:东莨菪碱法复制小鼠模型更接近SS病理特征,优于高渗盐水滴眼法。

Objective:To compare the characteristics between Sjögren's syndrome(SS)mice models replicated respectively by scopolamine method and hypertonic saline method.Methods:SPF grade 6~8 weeks old male BALB/c mice were randomly divided into three groups:normal group,hypertonic saline group and scopolamine group(n=10).Except for normal group was given saline,mice in scopolamine group were injected subcutaneously with 1 mg/ml scopolamine hydrobromide solution 0.2 mg/time/20 g body weight(qid),and mice in hypertonic saline group were slowly dropped into mice eyes with sterile 2.8 mg/ml hypertonic sodium chloride solution[100μl/eye(qid)],processing continuously for 10 days.Body weight,water consumption and tear crystallization of mice in each group were measured in the duration.After the end,expressions of SSA and SSB autoantibodies,antinuclear antibody ANA,BAFFR protein and inflammatory factor TNF-αin serum were detected by ELISA method,indexes of spleen were calculated;pathological changes of spleen were analyzed by HE staining,and degrees of lymphocytes infiltration in exocrine glands were evaluated by Chisholm-Mason histological scoring criteria.Results:Compared with hypertonic saline group,body weight was decreased,water consumption was increased(P<0.05 or P<0.01),tear secretion,scattered and not-fern shaped tear crystals(typeⅣ)were decreased in scopolamine group.Expressions of SSA antibody,antinuclear antibody ANA and TNF-αin serum and index of spleen were significantly increased(P<0.05);HE staining showed that lymphocytes proliferated significantly in white pulp of spleens and spleen corpuscles shranked in scopolamine group(grade 3),while structure of spleen corpuscles in hypertonic saline group was neat,less lymphocytes proliferated(grade 1).In scopolamine group,score of lymphocytes infiltration in submandibular gland and parotid gland were significantly increased(P<0.05).Conclusion:Mice model of scopolamine method is better than that of hypertonic saline eye drop method,and its indexes are closer to the pathological characteristics of SS.