解毒化瘀生津方对干燥综合征模型小鼠颌下腺AQP5表达的影响

Effect of Jiedu Huayu Shengjin Decoction on AQP5 Expression in Submandibular Gland of Sjogren Syndrome Model Mice

目的:观察解毒化瘀生津方对干燥综合征模型小鼠颌下腺AQP5表达的影响,探讨其作用机制。方法:C57BL/6小鼠随机分为正常组,模型组,硫酸羟氯喹组,解毒化瘀生津方低剂量组、中剂量组、高剂量组。采用免疫诱导法建立SS动物模型,造模的同时即开始灌胃给药,正常组、模型组,给予生理盐水10 mL·kg^(-1);硫酸羟氯喹组予硫酸羟氯喹片60 mg·kg^(-1);解毒化瘀生津方低剂量组10.4 g·kg^(-1),中剂量组20.8 g·kg^(-1),高剂量组41.6 g·kg^(-1),28 d后采用酶联免疫吸附法检测小鼠颌下腺AQP5水平表达、光镜下观察其病理改变,并进行免疫组织化学法检测及荧光染色图片的观察。结果:对小鼠颌下腺AQP5免疫学及平均吸光度方面影响,解毒化瘀生津方中剂量组、高剂量组与硫酸羟氯喹组3组疗效相似,优于中药低剂量组(P<0.05);病理组织HE染色,在光学显微镜下观察,在抑制颌下腺破坏方面,中药高剂量组优于其他各组,并且病理组织切片的淋巴细胞浸润的阳性率最低(P<0.05);免疫荧光染色图片上可观察到高剂量组的变化优于其他治疗组。结论:解毒化瘀生津方能够明显抑制腺体破坏,上调颌下腺AQP5的表达,增加唾液分泌量,这可能是该药能够治疗干燥综合征的疗效机制之一。

Objective： To observe the effect of Jiedu Huayu Shengjin Decoction on the expression of AQP5 in submandibular gland of mice with Sjogren＇s syndrome model and to explore its mechanism. Methods： C57 BL/6 mice were randomly divided into normal group,model group,hydroxychloroquine sulfate group,low dose group,middle dose group and high dose group. The model of SS animal was established with immunization induction method. At the same time,the rats were given 10 mL·kg^-1 saline in normal group and model group. The hydroxychloroquine sulfate group was given 60 mg·kg^-1 hydroxychloroquine sulfate. Give Jiedu Huayu Shengjin Decoction of 10. 4 g·kg^-1 in the low dose group and 20. 8 g·kg^-1 in the middle dose group and 41. 6 g·kg^-1 in the high dose group. The expression of AQP5 level in the submandibular gland of mice was detected by enzyme-linked immune-sorbent assay. The pathological changes were observed under light microscope,detect with immunohistochemical method and observe the fluorescence staining. Results： The effect on the AQP5 immunoreactivity and the average absorbance of the submandibular gland in mice of hydroxychloroquine sulfate group,TCM middle dose group and high dose group were similar,which were better than that of the low dose group（ P〈0. 05）. The inhibition of submandibular gland damage was best in the high dose group,and the positive rate of lymphocyte infiltration was the lowest（ P〈0. 05）. The significant changes of immunofluorescence staining were observed in the high-dose group and other treatment groups. Conclusion： Jiedu Huayu Shengjin Decoction can obviously inhibit glandular destruction,increase the expression of AQP5 in submandibular gland and increase the amount of saliva secretion,which may be a therapeutic way to treat Sjogren＇s syndrome.