益气养阴祛瘀方对干燥综合征颌下腺细胞AQP5及M3R表达的影响Study on the Influence of Yiqiyangyin Quyu Formulae on AQP5 and M3R Expression of Sjogren's Syndrome Submandibular Gland Cells
秦源,林崇泽,孙晗,李志宇,王新昌
摘要(Abstract):
目的观察益气养阴祛瘀方含药血清对干燥综合征(SS)患者颌下腺细胞AQP5及M3R表达的影响。方法制备不同剂量SD大鼠益气养阴祛瘀方含药血清,再分别给予以下处理:空白组(仅含RPMI1640培养液),空白血清组(10%空白血清+RPMI1640培养液),分别予含5%、10%、15%、20%不同浓度的含药血清+RPMI1640培养液组,共6组。培养细胞24 h后,分别以倒置相差显微镜、激光共聚焦显微镜观察细胞,Western blot检测细胞中AQP5及M3R蛋白表达水平。结果益气养阴祛瘀方含药血清作用后,激光共聚焦显微镜观察抗体标记细胞的荧光强度明显增强。Western blot检测AQP5与M3R的蛋白表达均有上调。含药血清组中的AQP5及M3R蛋白的表达明显高于空白血清组,且10%、15%含药血清组高于5%含药血清组(P<0.05),但与20%相差不大(P>0.05)。结论益气养阴祛瘀方可以上调颌下腺细胞中AQP5及M3R蛋白的表达,而且与之成一定的剂量依赖关系。
关键词(KeyWords): 干燥综合征;益气养阴祛瘀方;含药血清;水通道蛋白AQP5;M3R
基金项目(Foundation): 国家自然科学基金资助项目(81473646)
作者(Author): 秦源,林崇泽,孙晗,李志宇,王新昌
参考文献(References):
- [1]王新昌,谢志军,范永升,等.益气养阴祛瘀法治疗原发性SS的疗效观察[J].中国基层医学,2009,16(1):92-95.
- [2]王新昌,温成平,范永升,等.益气养阴祛瘀药对SS患者性激素水平影响的研究[J].浙江中医药大学学报,2009,33(1):48-50.
- [3]王新昌,范永升,曹灵勇,等.益气养阴祛瘀中药对SS自发性模型非肥胖糖尿病小鼠的治疗作用及机制[J].浙江中医药大学学报,2012,36(2):171-174.
- [4]黄臣虎,陆茵,严令耕,等.中药血清药理学研究进展[J].中国实验方剂学杂志,2011,17(10):266-271.
- [5]Itoh T,Rai T,Kuwahara M,et al.Identification of a novel aquaporin,AQP12,expressed in pancreatic acinar cells[J].Bioehem Biophys Res Commun,2005,330:832-838.
- [6]Mitsiasd I,Tzioufasa G,Veiopoulou C,et al.The Th1、Th2 cyto-kine balance changes with the progress of the immunopathological lesionof Sjogren’s syndrome[J].Clinical and Experimental Immunology,2002,128:562.
- [7]Moore M,Ma T,Yang B,et al.Tear secretion by lacrimal glands in transgenic mice lacking water channels AQP1,AQP3,AQP4 and AQP5[J].Eye Res,2002,70(5):557.
- [8]肖林,胡静,杨军,等.AQP5在SS患者唇腺中表达的实验研究[J].临床口腔医学杂志,2004,20(8):457-460.
- [9]李菁,赵岩,杜德顺,等.水分子通道蛋白-5在SS唇腺中的异常表达[J].中华风湿病学杂志,2005,9(8):453-456.
- [10]Sankar V,Brennan MT,Kok MR,et al.Etanercept in Sj?gren′s syndrome:a twelve-week randomized,double-blind,placebo-controlled pilot clinical trial[J].Arthritis Rheum,2004,50(7):2240-2245.
- [11]Berggreen E,Nylkken K,Delaleu N,et al.Impaired vascular responses to parasympathetic nerve stimulation andmuscarinic receptor activation in the submandibular gland in nonobese diabetic mice[J].Arthritis Resther,2009,11(1):1-12.
- [12]Nguyen KH,Brayer J,Cha S,et al.Evidence for antimuscarinic acetylcholine receptor antibody-medi ated secretory dysfunction in nod mice[J].Arthritis Rheum,2000,43(10):2297-2306.
- [13]Bacman S,Berra A,Sterin-Borda L,et al.Muscarinic acetylcholine receptor antibodies as a new marker of dry eye Sjogren’s syndrome[J].Invest 0phthalmol Vis Sci,2001,42(2):321-327.
- [14]Salomonsson S,Jonsson MV,Skarstein K,et al.Cellular basis of ectopic germinal center formation and autoantibody.Production in the target organ of patients with Sj觟gren′s syndrome[J].Arthritis Rheum,2003,48:3187-201.
- [15]王頔,秦臻,周党侠,等.M3受体激动剂诱导大鼠腮腺细胞内AQP5和脂质筏的核转位[J].基础医学与临床,2015,35(2):237-239.
- [16]Wang D,Yuan Z,Inoue N,et al.Abnormal subcellular localization of AQP5 and downregulated AQP5 protein in parotid glands of streptozotocin-induced diabetic rats[J].Biochim Biophys Acta,2011,1810:543-554.