摘要
目的探讨养阴活血方对原发性干燥综合征非肥胖糖尿病(NOD)模型小鼠颌下腺组织血管活性肠肽(VIP)/蛋白激酶A(PKA)/水通道蛋白5(AQP5)信号通路的影响。方法采用简单随机抽样法将30只NOD小鼠分为5组,每组各6只,依次为模型组,养阴活血方高剂量组(4 g/kg)、中剂量组(2 g/kg)、低剂量组(1 g/kg),以及阳性药组(羟氯喹,40 mg/kg),模型组灌服等量去离子水。给药6周后取颌下腺组织进行免疫组化检测,观察颌下腺中VIP、AQP5表达情况;Western blotting检测颌下腺VIP、AQP5、PKA蛋白表达水平;qRT-PCR检测VIP、AQP5、PKA基因表达水平。结果免疫组化结果显示,养阴活血方各组NOD小鼠颌下腺组织AQP5、VIP呈高表达,平均光密度值增高(P<0.01)。Western blotting结果显示,与模型组比较,养阴活血方高、中剂量组小鼠颌下腺组织VIP、AQP5、PKA蛋白表达显著升高,差异有统计学意义(P<0.05)。qRT-PCR结果显示,与模型组比较,养阴活血方高、中剂量组小鼠颌下腺组织VIP、AQP5、PKA基因表达显著升高,差异有统计学意义(P<0.05)。结论养阴活血方可能通过上调VIP表达,维持AQP5蛋白高表达,激活VIP/PKA/AQP5信号通路,从而发挥治疗原发性干燥综合征的作用。
Objective To explore the effect of Yangyin Huoxue Prescription on the vasoactive intestinal peptide(VIP)/protein kinase A(PKA)/aquaporin-5(AQP5)signaling pathway in submandibular gland tissues of non-obese diabetic(NOD)mice model with primry Sjogren’s syndrome.Methods A total of 30 NOD mice were divided into 5 groups using the simple random sampling method with 6 mice in each group,which were the model group,the high dose(4 g/kg),medium dose(2 g/kg)and low dose(1 g/kg)groups with Yangyin Huoxue Prescription,and the positive drug group(hydroxychloroquine,40 mg/kg),respectively.The model group was given an equal amount of deionized water.After 6 weeks of drug administration,the submandibular gland tissues were taken for immunohistochemistry to observe the expression of VIP and AQP5 in the submandibular gland.Western blotting was used to detect the protein expression of VIP,AQP5 and PKA in the submandibular gland.qRT-PCR was used to detect the expression level of VIP,AQP5 and PKA genes.Results Immunohistochemistry results showed that AQP5 and VIP were highly expressed in the submandibular gland tissues of NOD mice in all groups with Yangyin Huoxue Prescription,with an increase in average optical density value(P<0.01).The Western blotting results showed that the protein expression of VIP,AQP5,and PKA was significantly higher in the submandibular gland tissues of mice in the high and medium dose groups with Yangyin Huoxue Prescription than that in the model group,with a statistically significant difference(P<0.05).qRT-PCR results showed that the genes expression of VIP,AQP5,and PKA and was significantly higher in the submandibular gland tissues of mice in the high and medium dose groups with Yangyin Huoxue Prescription than that in the model group,with a statistically significant difference(P<0.05).Conclusion The Yangyin Huoxue Prescription may activate the VIP/PKA/AQP5 signaling pathway by up-regulating the expression of VIP and maintaining the high expression of AQP5 protein,thus exerting the therapeutic effect on primry Sjogren’s syndrome.
作者
李进红
童胜脂
吴雨峰
LI Jinhong;TONG Shengzhi;WU Yufeng(Department of Stomatology,Hangzhou Hospital of Traditional Chinese Medicine Affiliated to Zhejiang Chinese Medical University,Zhejiang,Hangzhou 310007,China)
出处
《中国医药科学》
2024年第23期4-7,共4页
China Medicine And Pharmacy
基金
浙江省中医药科技计划项目(2023ZL520)。
关键词
原发性干燥综合征
养阴活血方
水通道蛋白5
血管活性肠肽
Primry Sjogren’s syndrome
Yangyin Huoxue Prescription
Aquaporin-5
Vasoactive intestinal peptide
