摘要
目的探索化湿润燥方调控Hippo-Yes相关蛋白(YAP)/具有PDZ结合基序的转录共激活因子(TAZ)通路对干燥综合征(SS)模型小鼠(NOD/Ltj)颌下腺细胞凋亡和唾液分泌功能的影响。方法选取8周龄雌性BALB/c小鼠为空白组,8周龄NOD/Ltj雌性小鼠简单随机抽样分为模型组、羟氯喹组和化湿润燥低、中、高剂量组,给药8周后测量各组小鼠饮水量、唾液流率,采用苏木素-伊红染色检测各组小鼠颌下腺组织病理损伤,免疫组织化学法及蛋白质印迹法检测颌下腺组织中水通道蛋白5(AQP5)、B细胞淋巴瘤/白血病-2蛋白(Bcl-2)、Bcl-2相关X蛋白(Bax)、苄氯素1蛋白(beclin-1)、YAP、磷酸化YAP(p-YAP)、TAZ蛋白表达,TUNEL染色检测各组颌下腺细胞凋亡率。结果与空白组比较,模型组小鼠颌下腺组织中Hippo-YAP/TAZ通路被抑制,淋巴细胞浸润增多、饮水量增加、唾液流率降低、AQP5蛋白表达减少、唾液腺细胞凋亡率增加(均P<0.05)。与其他给药组比较,化湿润燥中剂量能够明显激活Hippo-YAP/TAZ通路,改善颌下腺组织淋巴细胞浸润、减少小鼠饮水量、升高唾液流率、增加AQP5蛋白表达、减少唾液腺细胞的凋亡率(均P<0.05)。结论化湿润燥方能减轻SS模型小鼠颌下腺细胞病理损伤和凋亡,改善颌下腺组织的整体分泌功能,这可能与其激活Hippo通路、下调YAP/TAZ水平相关。
Objective To explore the effects of modulating the Hippo-YAP/TAZ pathway using Huashi Runzao Prescription(HRP)on the apoptosis and salivary secretion function of submandibular gland cells in a naive non-obese diabetic(NOD/Ltj)mouse model of Sjögren′s syndrome(SS).Methods Eightweek-old female BALB/c mice were selected as the blank group.Eight-week-old female NOD/Ltj were randomly divided into the model,HRP-L,HRP-M,HRP-H,and hydroxychloroquine sulphate(HCQ)groups.After eight weeks of drug administration,the water consumption and salivary flow rate of each group were recorded.The histopathological damage of the submandibular gland in each group was determined using hematoxylin and eosin staining,the apoptosis rate of the submandibular gland was determined using TUNEL staining,and AQP5,Bax,Bcl-2,beclin-1,YAP,p-YAP,and TAZ expressions in submandibular gland tissues were determined using immunohistochemistry and Western blotting.Results Compared with the blank group,the Hippo-YAP/TAZ pathway was inhibited in the submandibular gland tissues of the model group,lymphocyte infiltration increased,water consumption increased,salivary flow rate decreased,AQP5 expression decreased,and submandibular gland cells apoptosis rate increased(P<0.05).Compared with all other administered groups,in the HRP-M group,the Hippo-YAZ/TAZ pathway was significantly activated,lymphocyte infiltration in submandibular gland tissues was reduced,water consumption was reduced,the salivary flow rate and AQP5 expression were increased,and the apoptosis rate of submandibular gland cells was reduced(P<0.05).Conclusion HRP reduced the pathological damage and apoptosis of submandibular gland cells in SS model mice.It improved the overall secretory function of submandibular gland tissues,which may be related to Hippo pathway activation and downregulating YAP/TAZ expression.
作者
黄子玮
陶庆文
廖佳禾
余新波
贺倩
唐博杰
罗静
HUANG Ziwei;TAO Qingwen;LIAO Jiahe;YU Xinbo;HE Qian;TANG Bojie;LUO Jing(Graduate School,Beijing University of Chinese Medicine,Beijing 100029,China;Traditional Chinese Medicine Department of Rheumatism,China-Japan Friendship Hospital,Beijing Key Laboratory of Immune Inflammatory Disease,Beijing 100029,China)
出处
《北京中医药大学学报》
CAS
CSCD
北大核心
2024年第11期1527-1540,共14页
Journal of Beijing University of Traditional Chinese Medicine
基金
国家自然科学基金项目(No.81804042)
首都卫生发展科研专项(No.2024-1-4065)
中央高水平医院临床科研业务费专项成果转化项目(No.2023-NHLHCRF-YXHZ-ZRMS-07)
北京中医药科技发展基金项目(No.BJZYQN-2023-04)
中日友好医院“菁英计划”人才培育工程(No.ZRJY2024-GG08)
中华中医药学会风湿病分会青年培英计划项目(No.202327-007)。
