益气养阴祛瘀方含药血清对干燥综合征人颌下腺细胞AQP5和M3R表达的影响  

Effects of Yiqi Yangyin Quyu Recipe Containing Serum on AQP5 and M3R of Human Submandibular Gland Cells with Sjögren's Syndrome

作　　者：刘丹 秦源 谢志敏 刘东洋 王静 朱秋爽 王新昌[2] LIU Dan;QIN Yuan;XIE Zhi-min;LIU Dong-yang;WANG Jing;ZHU Qiu-shuang;WANG Xin-chang(Second Clinical Medical College,Zhejiang Chinese Medical University,Hangzhou ,310053;Department of Rheumatism Immunity,Second Hospital Affiliated to Zhejiang Chinese Medical University,Hangzhou,310005)

机构地区：[1]浙江中医药大学第二临床医学院,杭州310053 [2]浙江中医药大学第二附属医院风湿免疫科,杭州310005

出　　处：《中国中西医结合杂志》2019年第3期330-334,共5页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助项目(No.81473646)

摘　　要：目的观察益气养阴祛瘀方含药血清对干燥综合征(■;gren′s syndrome, SS)人颌下腺细胞(human submandibular gland cells, HSG cells)中水通道蛋白5(aquaporin protein-5, AQP5)和毒蕈碱样胆碱能受体3(type 3 muscarinic acetylcholine receptors, M3R)表达的影响。方法制备益气养阴祛瘀方含药血清,将对数生长期的HSG细胞随机分为空白组(RPMI 1640培养液)、空白血清组(10%的空白血清+RPMI 1640培养液)、含药血清组(10%的益气养阴祛瘀方含药血清+RPMI 1640培养液)、IFN-γ组(IFN-γ+RPMI 1640培养液)、IFN-γ+IFN-γ拮抗剂组(IFN-γ+IFN-γ拮抗剂+RPMI 1640培养液)、IFN-γ+含药血清组(IFN-γ+10%益气养阴祛瘀方含药血清+RPMI 1640培养液),共6组。培养24 h后,Western blot法检测HSG细胞中AQP5和M3R蛋白表达;RT-PCR法检测HSG细胞AQP5 mRNA和M3R mRNA表达;免疫荧光法观察HSG细胞AQP5蛋白通道表达变化。结果与空白血清组比较,含药血清组AQP5、M3R蛋白及AQP5 mRNA和M3R mRNA表达升高(P<0.05)。与IFN-γ组比较,空白组、空白血清组、含药血清组、IFN-γ+IFN-γ拮抗剂组AQP5、M3R蛋白及mRNA表达均升高(P<0.05)。结论益气养阴祛瘀方具有与IFN-γ拮抗剂类似功效,能上调主管水运的AQP5、M3R表达,增加唾液量的分泌。Objective To observe effect of Yiqi Yangyin Quyu Recipe(YYQR) containing serum on aquaporin protein-5(AQP5) and type 3 muscarinic acetylcholine receptors(M3R) of human submandibular gland cells with Sj?gren′s syndrome(SS). Methods YYQR containing serum was prepared. HSG cells in logarithmic growth phase were randomly divided into the blank group(RPMI 1640 medium), the blank serum group(10% blank serum + RPMI 1640 medium), the medicated serum group(10%YYQR containing serum + RPMI 1640 medium), the IFN-γ group(IFN-γ+ RPMI 1640 medium), the IFN-γ+ IFN-γ antagonist group(IFN-γ+ IFN-γ antagonist +RPMI 1640 medium), and the IFN-γ+ medicated serum group(IFN-γ+ 10%YYQR containing serum + RPMI 1640 medium). After 24 hours of culture, protein and mRNA were extracted. Protein expressions of AQP5 and M3R in HSG cells were detected by Western blot. AQP5 and M3R mRNA expressions in HSG cells were detected by RT-PCR. Protein channel expression changes of AQP5 in HSG cells was observed by laser confocal microscope. Results Compared with the blank serum group, protein and mRNA expressions of AQP5 and M3R in the medicated serum group increased(P<0.05). Compared with the IFN-γ group, protein and mRNA expressions of AQP5 and M3R in the remaining five groups increased(P<0.05). Conclusion YYQR has similar efficacy with IFN-γ antagonist, it can increase the AQP5 and M3R expressions which were in charge of water transportation, and increase the secretion of saliva.

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

分 类 号：R285[医药卫生—中药学]