活血解毒润燥方干预干燥综合征小鼠颌下腺细胞凋亡促进因子Smac表达的机制研究  

Research on the mechanism of apoptosis promoting factor Smac expression in submandibular gland cells of Sjogren’s syndrome mice intervened by Huoxue Jiedu Runzao prescription

作　　者：李方凯[1] 谷新怡 朱跃兰[1] 侯秀娟[1] 曹云松[1] LI Fangkai;GU Xinyi;ZHU Yuelan;HOU Xiujuan;CAO Yunsong(Dongfang Hospital of Beijing University of Traditional Chinese Medicine,Beijing 100078,China)

机构地区：[1]北京中医药大学东方医院,北京100078 [2]北京中医药大学东直门医院通州院区,北京101121

出　　处：《陕西中医》2023年第6期693-698,共6页Shaanxi Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(81874438);北京中医药大学自主课题青年教师项目(2019-JYB-JS-104)。

摘　　要：目的:将干燥综合征(SS)模型小鼠颌下腺细胞凋亡促进因子Smac的表达作为靶点,探究活血解毒润燥方对SS模型小鼠颌下腺组织形态、腺体功能、细胞凋亡的影响及其干预机制。方法:实验选取雌性BALB/c57小鼠为正常对照组,自发性雌性非肥胖型糖尿病(NOD/Ltj)小鼠为SS模型并随机分为模型组、白芍总苷胶囊组和活血解毒润燥方低、中、高剂量组,每组各15只。各组小鼠给药量按照人与动物体表面积比计算等效剂量,正常对照组、模型组小鼠灌胃10 ml/kg蒸馏水,白芍总苷胶囊组(0.234 g/kg)和活血解毒润燥方低剂量组(15.6 g/kg)、中剂量组(31.2 g/kg)、高剂量组(62.4 g/kg)分别灌胃给药,每日1次。药物干预8周后,无菌分离颌下腺,观察各组小鼠颌下腺形态、功能的改变,并分别应用免疫组化法(IHC)、蛋白免疫印迹法(Western blot)、逆转录聚合酶链式反应(RT-PCR)测定细胞凋亡促进因子Smac的表达。结果:与正常对照组比较,模型组唾液流量及颌下腺指数均显著降低(P<0.01),颌下腺组织病理学评分显著升高(P<0.01),IHC和Western blot检测Smac表达明显升高(P<0.05),RT-PCR检测Smac表达显著升高(P<0.01)。治疗结束后,与模型组比较,活血解毒润燥方各剂量组和白芍总苷胶囊组唾液流量及颌下腺指数均显著升高(P<0.01),颌下腺组织病理学评分显著降低(P<0.01)。运用IHC和Western blot检测活血解毒润燥方高剂量组和白芍总苷胶囊组Smac表达明显降低(P<0.05),运用RT-PCR检测活血解毒润燥方高剂量组与白芍总苷胶囊组Smac表达显著降低(P<0.01)。结论:活血解毒润燥方可以改善SS模型鼠颌下腺腺泡的分泌功能,提高其颌下腺指数,增加SS模型鼠的唾液分泌量;其作用机制与下调SS模型鼠颌下腺细胞凋亡、促进因子Smac的表达水平有关。Objective:To investigate the effect of Huoxue Jiedu Runzao prescription on the tissue morphology,glandular function and cell apoptosis of submandibular gland in Sjogren’s syndrome(SS)model mice and its intervention mechanism,using the expression of apoptosis promoting factor Smac as the target.Methods:Female BALB/c57 mice were selected as normal control group,and spontaneous female non-obese diabetes mellitus(NOD/Ltj)mice were selected as SS model.The mice were randomly divided into model group,total glucosides of paeony capsule group(TGPC)and Huoxue Jiedu Runzao prescription low-dose,medium-dose and high-dose groups,with 15 mice in each group.The dosage of mice in each group was calculated according to the ratio of body surface area between human and animal.Mice in normal control group and model group were intragastric with 10 ml/kg distilled water.Total glucosides of paeony capsule group(0.234 g/kg),low dose group(15.6 g/kg),medium dose group(31.2 g/kg)and high dose group(62.4 g/kg)of Huoxue Jiedu Runzao prescription were administrated by intragastric administration,once a day.After 8 weeks of drug intervention,submandible gland was separated aseptically,and the morphological and functional changes of submandible gland in each group were observed.The expression of apoptosis promoting factor Smac was detected by IHC,Western blot and RT-PCR,respectively.Results:Compared with the normal control group,the saliva flow and submandibular gland index in the model group were significantly decreased(P<0.01),histopathological scores of submandibular gland were significantly increased(P<0.01),the expression of Smac was significantly increased by IHC and Western blot(P<0.05),the expression of Smac was significantly increased by RT-PCR(P<0.01).After treatment,compared with model group,saliva flow and submandibular gland index of Huoxue Jiedu Runzao prescription and total glucosides of paeony capsule group were significantly increased(P<0.01),the submandibular gland histopathological score was significantly decreased(P<0.01

关 键 词：干燥综合征 活血解毒润燥方 白芍总苷胶囊 颌下腺 细胞凋亡 细胞凋亡促进因子Smac 

分 类 号：R593.2[医药卫生—内科学]