丹皮酚对强直性脊柱炎模型小鼠Wnt和BMP/Smad信号转导通路的影响  

Effects of Paeonol on Wnt and BMP/Smad Signal Transduction Pathway in the Ankylosing Spondylitis Model Mice

作　　者：吴琪 吴倩[2] 周晓红 吴玲慧 WU Qi;WU Qian;ZHOU Xiaohong;WU Linghui(Teaching and Demonstration Center of Clinical Skill,Hubei University of TCM,Wuhan 430061,China;Hemodialysis Room,South Branch of Shanghai East Hospital,Shanghai 200123,China;College of Acupuncture and Orthopaedics-Traumatology,Hubei University of TCM,Wuhan 430061,China;Dept. of Immunology,Child Health Center,Xianning Center for Maternal and Child Health Care,Hubei Xianning 437000,China)

机构地区：[1]湖北中医药大学临床技能实训中心,武汉430061 [2]上海市东方医院南院血液透析室,上海200123 [3]湖北中医药大学针灸骨伤学院,武汉430061 [4]咸宁市妇幼保健院儿童保健中心计划免疫科,湖北咸宁437000

出　　处：《中国药房》2018年第11期1500-1504,共5页China Pharmacy

基　　金：湖北省教育厅科学研究计划资助项目(No.Q20162011)

摘　　要：目的:研究丹皮酚对强直性脊柱炎(AS)模型小鼠分泌型糖蛋白(Wnt)和骨形态发生蛋白(BMP)/细胞信号转导分子(Smad)通路的影响,探讨丹皮酚防治AS的机制。方法:将40只小鼠随机分为正常组、模型组、柳氮磺吡啶组(阳性对照,9 mg/kg)和丹皮酚组(3 mg/kg),每组10只。除正常组外的其余各组小鼠均采用完全弗氏佐剂+蛋白聚糖腹腔注射法复制AS模型。各给药组小鼠在成模后灌胃相应药物,正常组和模型组小鼠灌胃等体积蒸馏水,每天给药1次,连续20 d。末次给药后处死小鼠,透射电镜下观察各组小鼠骶髂关节滑膜细胞超微病理结构变化,采用酶联免疫吸附法检测小鼠血清中肿瘤坏死因子α(TNF-α)、Wnt信号通路病理性骨化相关因子(DKK-1)含量,实时荧光定量聚合酶链式反应法检测小鼠滑膜组织中骨形态发生蛋白2(BMP-2)、细胞内核心结合因子α1(Cbfα1)、Smad1 m RNA表达。结果:与正常组比较,模型组小鼠血清中TNF-α含量明显增加、DKK-1含量明显减少,滑膜组织中BMP-2、Cbfα1和Smad1 m RNA表达水平明显升高,差异均有统计学意义(P<0.05或P<0.01);电镜下可见模型组小鼠滑膜细胞增生,排列紊乱,分泌活性细胞器分泌亢进,细胞间隙增宽。与模型组比较,柳氮磺吡啶组和丹皮酚组小鼠血清中TNF-α含量均明显减少,丹皮酚组小鼠血清中DKK-1含量明显增加,滑膜组织中BMP-2、Cbfα1和Smad1 m RNA表达水平明显降低,差异均有统计学意义(P<0.05或P<0.01);电镜下可见丹皮酚组小鼠滑膜细胞线粒体、溶酶体、粗面内质网的形态明显改善。结论:丹皮酚防治AS的机制可能与降低血清中TNF-α含量、升高血清中DKK-1含量,下调滑膜细胞中BMP-2、Cbfα1和Smad1 m RNA表达,抑制Wnt和BMP/Smad骨化相关信号转导通路逆转滑膜细胞成骨分化有关。OBJECTIVE：To study the effects of paeonol on Wnt and BMP/Smad pathway in ankylosing spondylitis（AS）model mice,and to investigate prevention and treatment mechanism of paeonol for AS. METHODS：40 mice were randomly divided into normal group,model group,sulfasalazine group（positive control,9 mg/kg）and paeonol group（3 mg/kg）,with 10 mice in each group. In addition to the normal group,Freund＇s adjuvant and proteoglycan were used to establish AS model in each group. After modeling,mice in each administration group were given relevant medicine intragastrically;normal group and model group were given constant volume of distilled water intragastrically,once a day,for consecutive 20 d. After last administration,the mice were killed,TEM was used to observe the ultrastructural pathologic change of synovial cells in articulationes sacroiliaca.The serum contents of TNF-α and DKK-1 were determined by ELISA,and the m RNA expressions of BMP-2,Cbfα1 and Smad1 in synovial tissue were detected by real-time fluorescent quantitative PCR. RESULTS：Compared with normal group,serum content of TNF-α in model group was increased significantly,while serum content of DKK-1 was decreased significantly;m RNA expressions of BMP-2,Cbf α 1 and Smad1 in synovial tissue were increased significantly,with statistical significance（P〈0.05 or P〈0.01）.Under the electron microscope,the synoviocytes of mice in model group were proliferated and arranged in disorder,and the secretory activity of active organelles were hyperactivity and the gap among cells became widened. Compared with model group,serum content of TNF-α in sulfasalazine group and paeonol group were decreased significantly;serum content of DKK-1 was increased significantly and m RNA expressions of BMP-2,Cbf α 1 and Smad1 in synovial tissue were decreased significantly in paeonol group,with statistical significance（P〈0.05 or P〈0.01）. Electron microscopy showed that mitochondria,lysosome and rough endoplasmic reticulum structure of synovial cell were impr

关 键 词：丹皮酚 强直性脊柱炎 分泌型糖蛋白 骨形态发生蛋白/细胞信号转导分子 小鼠 

分 类 号：R285[医药卫生—中药学]