摘要
目的研究十一方药酒(以下简称“SYF”)对木瓜蛋白酶诱导的兔膝骨关节炎(KOA)的保护作用及机制。方法将35只兔随机分为空白组、模型组、阳性组(双氯芬酸二乙胺乳胶剂200 mg/kg)、SYF高剂量组(386 mg/kg)、SYF低剂量组(97 mg/kg),每组7只(均为雄性4只、雌性3只)。除空白组外,其余各组兔于第1、4、7天从右后肢膝关节腔注入木瓜蛋白酶混合液(含2%木瓜蛋白酶与0.03 mol/L L-半胱氨酸)0.5 mL复制KOA模型,空白组注射等体积生理盐水。从第15天开始于各组兔右后肢膝关节涂抹药物,每天给药2次,连续20 d,同时在第0、8、14、35天,用游标卡尺测量兔右后肢膝关节直径计算其肿胀度。实验结束后,采用酶联免疫吸附测定法检测滑膜组织白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)、前列腺素E(2 PGE2)的水平;制作滑膜组织苏木精-伊红(HE)染色切片,观测病理学形态变化;采用实时荧光定量聚合酶链反应法检测滑膜组织Toll样受体4(TLR4)/髓样分化因子88(MyD88)/细胞核因子(NF-кB)信号通路中TLR4、MyD88、NF-кB p65 mRNA相对表达水平;采用Western blot法检测滑膜组织TLR4、MyD88、NF-кB p65、磷酸化NF-кB p65(p-NF-кB p65)蛋白相对表达水平。结果与空白组比较,模型组兔膝关节肿胀度升高,滑膜组织病理性损伤严重,滑膜组织中IL-1β、TNF-α、PGE2水平均显著升高(P<0.05),TLR4、MyD88、NF-кB p65 mRNA和TLR4、MyD88、p-NF-кB p65蛋白相对表达水平均显著升高(P<0.05)。与模型组比较,各给药组兔右后肢膝关节肿胀度与滑膜组织病理损伤程度均有显著改善,滑膜组织中IL-1β、TNF-α、PGE2水平均显著降低(P<0.05),TLR4、MyD88、NF-кB p65 mRNA相对表达水平和TLR4、MyD88(SYF低剂量组除外)、p-NF-кB p65蛋白相对表达水平均显著降低(P<0.05)。结论SYF对木瓜蛋白酶诱导的兔KOA有保护作用,其作用机制与降低炎症因子IL-1β、TNF-α、PGE2水平及下调TLR4/MyD88/NF-к
OBJECTIVE To study the protective effect and mechanism of Shiyifang medicinal wine(SYF)on knee osteoarthritis(KOA)of rabbit induced by papain.METHODS Thirty-five rabbits were randomly divided into blank group,model group,positive group(Diclofenac diethylamine emulsion 200 mg/kg),SYF high-dose group(386 mg/kg)and SYF low-dose group(97 mg/kg),with 7 rabbits in each group(all had 4 males and 3 females).Except for the blank group,the other groups’rabbits were injected 0.5 mL papain mixture(containing 2%papain and 0.03 mol/L L-cysteine)into the right knee cavity on day 1,4 and 7,to replicate KOA model.Blank group was given constant volume of normal saline.From the 15th day,drugs were applied to right hind knee joints of rabbits in each group,twice a day for 20 days.At the same time,the diameter of right knee joints of rabbits was measured by vernier calipers at day 0,8,14 and 35 to calculate swelling degree.After the experiment,the levels of IL-1β,TNF-αand PGE2 in synovial tissue were determined by enzyme-linked immunosorbent assay.Hematoxylin-eosin(HE)staining was used to prepare the sections of synovial tissue,and the pathological changes were observed.The relative mRNA expressions of TLR4,MyD88 and NF-кB p65 in the TLR4/MyD88/NF-кB signaling pathway were detected by real-time quantitative polymerase chain reaction.The relative protein expressions of TLR4,MyD88,NF-кB p65 and p-NF-кB p65 were detect by Western blot.RESULTS Compared with blank group,the degree of knee swelling could be increased in model group,the pathological damage of synovial tissue was more serious,and the levels of IL-1β,TNF-αand PGE2 were increased significantly in synovial tissue(P<0.05).The relative expression levels of TLR4,MyD88,NF-кB p65 mRNAs and TLR4,MyD88,p-NF-кB p65 proteins were significantly increased(P<0.05).Compared with model group,swelling degree of right hind knee and the pathological injury degree of synovial tissue were significantly improved in each treatment group,while the levels of IL-1β,TNF-αand PGE2 in synov
作者
张文涛
周玲梅
黄宏海
鲍慧慧
毛德文
田慧
刘舒凌
ZHANG Wentao;ZHOU Lingmei;HUANG Honghai;BAO Huihui;MAO Dewen;TIAN Hui;LIU Shuling(Teaching Experiment Training Center,Guangxi University of Chinese Medicine,Nanning 530200,China;College of Pharmacy,Guangxi University of Chinese Medicine,Nanning 530200,China;Dept.of Hepatology,the First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530023,China)
出处
《中国药房》
CAS
北大核心
2022年第18期2224-2229,共6页
China Pharmacy
基金
广西创新驱动发展专项资金项目(No.桂科AA17202-036)
广西壮族自治区中医药管理局中药壮瑶药医院制剂质量提升项目(No.GZZJ202001)。
