益气养阴祛瘀方对干燥综合征NOD小鼠颌下腺AQP5、M3R表达的影响  

Effects of Yiqi Yangyin Quyu Prescription on the Expressions of AQP5 and M3R in Submandibular Glands of NOD Mice with Sjogren Syndrome

作　　者：任雅婷 刘丹 周东海 王新昌 REN Yating;LIU Dan;ZHOU Donghai;WANG Xinchang(Zhejiang Traditional Chinese Medicine University,Hangzhou 310053 Zhejiang,China;Zhejiang Shimai Pharmaceutical Company,Hangzhou 310053 Zhejiang,China;Affiliated Hospital of Zhejiang Traditional Chinese Medicine University,Hangzhou 310005 Zhejiang,China)

机构地区：[1]浙江中医药大学第二临床医学院,浙江杭州310053 [2]浙江时迈药业有限公司,浙江杭州310053 [3]浙江中医药大学附属第二医院,浙江杭州310005

出　　处：《中药新药与临床药理》2023年第5期613-619,共7页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目(82074341)。

摘　　要：目的研究益气养阴祛瘀方(黄芪、生地黄、玄参、丹参、益母草等)对干燥综合征NOD小鼠颌下腺水通道蛋白5(AQP5)以及毒蕈碱样胆碱能受体3(M3R)表达的影响。方法将30只NOD小鼠随机分为5组:模型组、西药组(硫酸羟氯喹60 mg·kg^(-1))及益气养阴祛瘀方低、中、高剂量组(11.147、22.295、44.590 g·kg^(-1)),每组6只。每日灌胃给药1次,连续给药8周。给药后第0、4、8周连续测量小鼠每日饮水量;给药结束后,分离小鼠双侧颌下腺并称定其质量,计算小鼠颌下腺指数;采用HE染色法进行颌下腺组织病理学观察,计算灶性指数评分;RT-qPCR法检测小鼠颌下腺组织AQP5、M3R mRNA表达水平;Western Blot法检测小鼠颌下腺组织AQP5、M3R蛋白表达水平;免疫组化法检测颌下腺组织AQP5蛋白表达水平;ELISA法检测血清干扰素γ(IFN-γ)、白细胞介素10(IL-10)含量。结果给药前,与模型组比较,益气养阴祛瘀方低、中、高剂量组及西药组NOD小鼠的日饮水量无明显变化(P>0.05);给药第4、8周,与模型组比较,益气养阴祛瘀方低、中、高剂量组及西药组NOD小鼠的日饮水量均显著减少(P<0.01)。模型组小鼠颌下腺淋巴细胞浸润明显并形成较多浸润灶;与模型组比较,益气养阴祛瘀方低、中、高剂量组及西药组NOD小鼠的颌下腺淋巴细胞浸润情况得到明显改善,浸润灶数量明显减少,灶性指数评分显著降低(P<0.01),颌下腺组织AQP5、M3R蛋白表达水平明显升高(P<0.05,P<0.01),血清IL-10水平明显升高(P<0.05,P<0.01);益气养阴祛瘀方高剂量组和西药组NOD小鼠的颌下腺指数明显升高(P<0.05);益气养阴祛瘀方中、高剂量组和西药组NOD小鼠颌下腺组织AQP5、M3R mRNA表达明显上调(P<0.05,P<0.01),血清IFN-γ含量显著降低(P<0.01)。结论益气养阴祛瘀方对NOD小鼠唾液腺损伤具有改善作用,可能通过上调AQP5、M3R表达,抑制颌下腺淋巴细胞浸润,改善唾液分泌功能,从而减缓Objective To study the effects of Yiqi Yangyin Quyu Prescription(composed of Astragali Radix,Rehmanniae Radix,Scrophulariae Radix,Salviae Miltiorrhizae Radix et Rhizoma and Leonuri Herba)on the expressions of submandibular gland aquaporin 5(AQP5)and muscarinic acetylcholine receptor 3(M3R)in NOD mice with sjogren syndrome(SS).Methods Thirty NOD mice were randomly divided into 5 groups:model group,western medicine group(Hydroxychloroquine Sulphate 60 mg·kg^(-1))and low-,medium-and high-dose groups(11.147,22.295 and 44.590 g·kg^(-1))of the Yiqi Yangyin Quyu Prescription,6 mice in each group.The drug was administered once daily by gavage for 8 consecutive weeks.The daily water intake of the mice was measured continuously at week 0,4 and 8 after the administration of the drug.At the end of the administration,the submaxillary glands of the mice were separated bilaterally and weighed,and the submaxillary gland index was calculated.The histopathological observation of submandibular gland was performed by HE staining and the focal index score was calculated.The mRNA expression levels of AQP5 and M3R in mouse submandibular gland tissue were measured by RT-qPCR;the protein expression levels of AQP5 and M3R in mouse submandibular gland tissue were measured by Western Blot;the protein expression level of AQP5 in submandibular gland tissue was measured by immunohistochemistry;the levels of serum interferonγ(IFN-γ)and interleukin 10(IL-10)were measured by ELISA.Results Before the administration,there was no significant change in the daily water intake of NOD mice in the low-,medium-and high-dose groups of Yiqi Yangyin Quyu Prescription and the western medicine group compared with the model group(P>0.05);at the fourth and eighth weeks of administration,the daily water intake of NOD mice in the low-,medium-and high-dose groups of Yiqi Yangyin Quyu Prescription and the western medicine group were significantly decreased compared with the model group(P<0.01).Compared with the model group,the lymphocyte infiltration in the subm

关 键 词：益气养阴祛瘀方 干燥综合征 NOD小鼠 颌下腺 水通道蛋白5 毒蕈碱样胆碱能受体3 

分 类 号：R285.5[医药卫生—中药学]