润燥活血汤对干燥综合征模型小鼠血清及颌下腺M3R表达的影响  

Effect of Runzao Huoxue decoction on muscarinic acetylcholine 3 expressions in blood and submandibular gland tissue of Sjogren's Syndrome mice

作　　者：张丹娜[1] 叶丽红[1] 刘喜德[1] ZHANG Dan-na;YE Li-hong;LIU Xi-de(Department of Athropathy,The Hospital of Integrated Traditional and Western Medicine of Zhejiang Province,Hangzhou,Zhejiang,310003,China)

机构地区：[1]浙江省中西医结合医院老年病科,杭州310003

出　　处：《浙江中西医结合杂志》2021年第7期597-602,共6页Zhejiang Journal of Integrated Traditional Chinese and Western Medicine

基　　金：浙江省中西医结合医院院内课题(No.hhyn201904)支持

摘　　要：目的观察润燥活血汤对干燥综合征(SS)模型小鼠血清及颌下腺毒蕈碱样乙酰胆碱3型受体(M3R)表达的影响。方法8周龄雄性BALB/c小鼠47只,按区组随机选出5只处死摘取颌下腺,剩余的小鼠随机分为正常组10只,模型组32只。参照文献的方法,采用背部皮内5个部位多点注射抗原的方法进行免疫,建立SS小鼠模型。在造模后第5周随机选取2只造模小鼠处死,通过病理形态学观察判定造模成功。另30只小鼠随机分为模型组10只,润燥活血汤组10只和羟氯喹(HCQ)组10只。润燥活血汤组小鼠按11.4g·kg^(-1)·d^(-1)剂量灌胃润燥活血汤,模型组小鼠灌胃等量生理盐水,HCQ组小鼠按0.06g·kg^(-1)·d^(-1)剂量灌胃HCQ混悬液(HCQ与生理盐水混合液),每天1次;正常组小鼠常规饲养;同时观测各组小鼠唾液量、饮水量。连续用药30天后眼眶取血后处死小鼠,摘取颌下腺、脾脏。采用苏木素-伊红(HE)染色评价颌下腺淋巴细胞浸润情况,采用ELISA法测定血清M3R水平,免疫组化法检测颌下腺M3R表达水平。结果与正常组比较,模型组小鼠血清、颌下腺M3R表达增加[血清M3R:(12.23±10.10)pg/mL比(2.72±1.21)pg/mL,P<0.05;颌下腺M3R:(2.23±0.11)%比(1.21±0.11)%,P<0.01];颌下腺淋巴细胞浸润程度增加,病理评分明显增高[(2.42±0.22)分比(0.18±0.05)分,P<0.01];与模型组比较,润燥活血汤组、HCQ组小鼠血清、颌下腺M3R表达降低[血清M3R:(3.30±6.25)pg/mL、(3.17±0.87)pg/mL比(12.23±10.10)pg/mL,P均<0.05;颌下腺M3R:(1.62±0.40)%、(1.53±0.06)%比(2.23±0.11)%,P均<0.05];颌下腺淋巴细胞浸润浸润程度减轻,病理评分明显降低[(1.21±0.33)分、(1.08±0.09)分比(2.42±0.22)分,P均<0.05];润燥活血汤组和HCQ组之间比较差异无统计学意义(P>0.05)。与模型组比较,润燥活血汤组小鼠唾液量增加[(25.53±4.27)mg比(22.42±2.52)mg,P<0.05];润燥活血汤组小鼠唾液量与HCQ组比较,差异无统计学意义(P>0.05)。结论润燥活血Objective To investigate the effect of Runzao Huoxue(RH)decoction on the expressions of muscarinic acetylcholine 3(M3R)in blood and submandibular gland tissue of Sjogren's syndrome mice.Methods Forty-seven eight-week-old male BALB/c mice were used.Five mice were randomly selected to have submandibular glands detached.The remaining mice were randomly divided into normal(10 mice)and model(32 mice)groups.The model group(Sjogren's Syndrome,SS)was established by injecting antigens intravenously into 5 sites at the back of mice.At the 5th week of model establishment,two mice were randomly selected for SS confirmation judged by relevant tissue pathomorphological observation prior to further experiments.The remainders of 30 mice were randomly divided into control(n=10),RH decoction(n=10)and hydroxychloroquine(HCQ)groups(n=10).Mice in the normal group were fed regularly.Mice in the RH and HCQ groups were given Runzao Huoxue decoction at a dose of 11.4g/kg per day or HCQ suspension(HCQ mixed with normal saline)at a dose of 0.06g/kg per day,respectively,while in the control group were given the same amount of normal saline.Quantities of saliva production and drinking water consumption were recorded.After 30 days of continuous administration,all mice were sacrificed and submandibular glands were harvested.Submandibular gland was sampled to be stained with hematoxylin-eosin(HE),and infiltration was evaluated.Serum or submandibular gland M3R was analyzed by ELISA or immunohistochemistry,respectively.Results Compared with the normal group,the M3R expressions were significantly increased in the blood(12.23±10.10 vs.2.72±1.21pg/mL;P<0.05)and submandibular gland(2.23±0.1 vs.1.21±0.1%;P<0.01)in the model groups,and infiltration degree of submandibular gland was increased in the model group as pathological score was significantly higher(model vs.normal,2.42±0.22 vs.0.18±0.05;P<0.01).Compared with the model group,M3R expressions in blood and submandibular gland in the RH decoction group and HCQ group(RH vs.HCQ vs.model)were dec

关 键 词：小鼠 润燥活血汤 干燥综合征 毒蕈碱样乙酰胆碱3型 血清 颌下腺 

分 类 号：R285.5[医药卫生—中药学]