阿托伐他汀联合葛根素对激素性股骨头缺血性坏死的作用机制研究  

Mechanism of combined application of atorvastatin and puerarin on SANFH rats

作　　者：胡彦婷 汪瑜[1] 熊德建 黄歆[1] 杨云戟 罗梅懿 张霞[2] HU Yanting;WANG Yu;XIONG Dejian;HUANG Xin;YANG Yunji;LUO Meiyi;ZHANG Xia(Orthopedics Department,Chengdu Seventh People’s Hospital,Chengdu,Sichuan 610041,China;Nursing Department,Chengdu Seventh People’s Hospital,Chengdu,Sichuan 610041,China;VIP Clinic,Sichuan Academy of Medical Sciences,Sichuan People’s Hospital,Chengdu,Sichuan 610072,China)

机构地区：[1]成都市第七人民医院骨科,四川成都610041 [2]成都市第七人民医院护理部,四川成都610041 [3]四川省医学科学院、四川省人民医院特需门诊,四川成都610072

出　　处：《安徽医药》2020年第6期1070-1074,I0001,共6页Anhui Medical and Pharmaceutical Journal

摘　　要：目的研究阿托伐他汀和葛根素对股骨头缺血性坏死大鼠的治疗作用及作用机制。方法50只SD大鼠采用随机数字表法分为空白组、模型组、阿托伐他汀组、葛根素组、联合用药组,每组10只。除空白组外,其余各组用脂多糖联合甲强龙建立大鼠激素性股骨头缺血性坏死模型。各组大鼠用对应药物灌胃给药,连续8周。酶联免疫吸附剂测定(ELISA)检测大鼠血清中碱性磷酸酶(ALP)和转化生长因子β1(TGF-β1)的含量;苏木精-伊红(HE)染色观察病理变化及DNA断裂的原位末端标记法(TUNEL法)观察细胞凋亡;实时荧光定量PCR(qPCR)检测核因子-κB受体活化因子(RANK)和核因子-κB受体活化因子配基(RANKL)mRNA,并用蛋白质印迹法(Western Blot)检测骨保护蛋白(OPG)、RANK、RANKL和肿瘤坏死因子受体相关因子6(TRAF-6)蛋白表达。结果空白组大鼠血清中ALP和TGF-β1含量分别为(25.23±1.24)IU/L和(47.12±2.41)ng/mL,而模型组大鼠血清中ALP含量升高,TGF-β1含量降低,分别为(31.86±1.23)IU/L和(37.74±2.68)ng/mL;与模型组相比,联合用药组能够降低ALP和升高TGF-β1含量(P<0.01),分别为(26.96±1.54)IU/L和(44.12±2.42)ng/mL;HE染色结果显示模型组大鼠股骨头组织被破坏,血管减少;联合用药组的软骨细胞排列整齐,骨髓腔内有大量造血细胞,脂肪细胞分布均匀。RT-PCR和蛋白质印迹法结果显示模型组OPG、RANK、RANKL和TRAF-6表达量升高。与模型组相比,联合用药组能提高OPG、RANK和降低RANKL、TRAF-6表达量(P<0.05)。结论阿托伐他汀联合葛根素可有效治疗激素性股骨头缺血性坏死,作用机制可能与OPG/RANKL/RANK信号通路有关。Objective To explore the effects and mechanism of combined application of atorvastatin and puerarin on steroid-induced avascular necrosis of femoral head(SANFH)rats.Methods Fifty SD rats were randomly assigned into five groups(n=10 for each group):blank group,model group,atorvastatin group,puerarin group and combined treatment group.SANFH rat models were established by lipopolysaccharide(LPS)combined with methylprednisolone in all the groups except for the blank group.All animals were treated with appropriate drugs for 8 weeks.The alkaline phosphatase(ALP)and transforming growth factor β1(TGF-β1)contents were measured by enzyme linked immunosorbent assay(ELISA).The changes and apoptosis of femoral head were observed through haematoxylin-eosin(HE)staining and TdT-mediated dUTP nick end labeling(TUNEL)of DNA fragmentation,respectively.Receptor activator of NF-κB(RANK)and receptor activator of NF-κB ligand(RANKL)mRNA were detected by Realtime fluorescent quantitative polymerase chain reaction(RT-PCR)and expressions of bone protection protein(OPG),RANK,RANKL and tumor necrosis factor receptor-related factor 6(TRAF-6)protein were measured by Western blotting.Results The serum ALP and TGF-β1 contents in the blank group were(25.23±1.24)IU/L and(47.12±2.41)ng/mL,respectively,while the serum ALP content in the model group was increased and TGF-β1 content was decreased,which were(31.86±1.23)IU/L and(37.74±2.68)ng/mL,respectively.Compared with the model group,the combined treatment group could reduce ALP and increase TGF-β1 content(P<0.01),which were(26.96±1.54)IU/L and(44.12±2.42)ng/mL,respectively.HE staining results showed that the femoral head tissue of rats of the model group was damaged and the blood vessels were decreased.Chondrocytes were arranged orderly,large number of hematopoietic cells were found in the bone marrow cavity and fat cells are evenly distributed in the combined treatment group.RT-PCR and Western blotting results showed that the expressions of OPG,RANK,RANKL and TRAF-6 were significantl

关 键 词：股骨头坏死/药物疗法 核因子ΚB受体活化因子 RANK配体 骨保护素 碱性磷酸酶 肿瘤坏死因子受体相关因子-6(TRAF-6) 阿托伐他汀 葛根素 

分 类 号：R965[医药卫生—药理学]