丹紫康膝冲剂通过抑制IKK/IκB/NF-κB信号通路改善膝骨关节炎大鼠关节软骨、血瘀状态的作用研究  

Effect of Danzi Kangxi Gradule(丹紫康膝冲剂)on Articular Cartilage and Blood Stasis Status in Knee Osteoarthritis Rats by Regulating IKK/IκB/NF-κB Pathway

作　　者：何花[1] 董大立[1] HE Hua;DONG Da-li(The Second Affiliated Hospital of Hunan University of Chinese Medicine,Changsha Hunan 410005,China)

机构地区：[1]湖南中医药大学第二附属医院,湖南长沙410005

出　　处：《中医药导报》2021年第10期42-48,共7页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：湖南省教育厅科学研究项目(18C0396);湖南省中医药科研计划项目(2020077)。

摘　　要：目的:探究丹紫康膝冲剂通过抑制IKK/IκB/NF-κB信号通路对膝骨关节炎大鼠关节软骨及血瘀状态的改善作用。方法:将70只SD大鼠随机分为假手术组、模型组、双氯芬酸钠组、丹紫康膝冲剂低剂量组、丹紫康膝冲剂中剂量组、丹紫康膝冲剂高剂量组和PDTC组,每组10只。采用Hulth法建立KOA模型,假手术组不切断膝前交叉韧带。各药物组大鼠灌胃给予相应药物,假手术组和模型组大鼠均灌胃给予等体积生理盐水,1次/d,连续28 d。HE染色和番红-O-固定绿染色观察软骨病理损伤情况,免疫组织化学法检测软骨组织NF-κB蛋白表达情况;蛋白免疫印记法检测NF-κB p65、p-IκBα、IκBα、p-IKKα、IKKα、p-IKKβ、IKKβ、NLRP3蛋白表达;酶联免疫吸附实验检测外周血IL-1β、IL-18、TNF-α、一氧化氮(NO)、低氧诱导因子-1ɑ(HIF-1ɑ)、前列腺素E2(PGE2)、抗Ⅱ型胶原抗体亚型(anti-ColⅡ-IgG、anti-COLⅡ-IG1、anti-ColⅡ-IgG2a)、血栓素B2(TXB2)、6-酮-前列腺素F1α(6-keto-PGF1α)水平。结果:与模型组比较,双氯芬酸钠组,PDTC组和丹紫康膝冲剂低、中、高剂量组大鼠软骨损伤和纤维增生减轻;胞核和胞浆NF-κB蛋白发生核转移的数量减少,p-IκBα、p-IKKα、p-IKKβ、NLRP3蛋白表达降低;外周血浆中IL-1β、IL-18、TNF-α、anti-ColⅡ-IgG、anti-ColⅡ-IgG2a、NO、PGE2、HIF-1ɑ水平降低;血浆中TXB2水平、TXB2/6-keto-PGF1α比值降低,6-keto-PGF1α升高,差异均有统计学意义(P<0.05);丹紫康膝冲剂高剂量组大鼠软骨组织p-IKKβ、NLRP3蛋白表达及外周血清中TXB2水平、6-keto-PGF1α水平、TXB2/6-keto-PGF1α比值与双氯芬酸钠组比较,差异均无统计学意义(P>0.05),而p-IκBα、p-IKKα蛋白表达均低于双氯芬酸钠组(P<0.05)。结论:丹紫康膝冲剂能通过抑制IKK/IκB/NF-κB信号通路的活化,进而降低大鼠关节软骨炎症因子的表达,有效改善大鼠的血瘀状态,以减轻软骨组织病理性损伤,而且随�Objective:To discuss the effect of Danzi Kangxi Gradule on articular cartilage and blood stasis status in knee osteoarthritis(KOA)rats by regulating IKK/IκB/NF-κB pathway.Methods:The 70 SD rats were randomly divided into sham-operation group,model group,Diclofenac Sodium group,Danzi Kangxi Gradule low-dose group,Danzi Kangxi Gradule medium-dose group,Danzi Kangxi Gradule high-dose group and PDTC(NF-κB inhibitor)group,10 rats in each group.KOA model was established by Hull method,while the anterior cruciate ligament was not transected in rats of sham-operation group.The rats were given the corresponding drugs by gavage respectively in each administration group,and the sham operation group and model group were given normal saline by gavage,once a day for 28 days.The pathological change of joint tissue was examined by HE staining or Safranin-O-fast Green staining.Immunohistochemical method was used to detect NF-κB protein expression in cartilage tissue.Western blotting was used for the detection of NF-κB p65,p-IκBα,IκBα,p-IKKα,IKKα,p-IKKβ,IKKβand NLRP3 proteins,and ELISA was used to detect serum IL-1β,IL-18,TNF-α,NO,Hypoxia inducible factor-1ɑ(HIF-1ɑ),Prostaglandin E2(PGE2),anti-ColⅡ-IgG,anti-ColⅡ-IgG1,anti-ColⅡ-IgG2a,thromboxane B2(TXB2)and 6-keto-PGF1αlevels.Results:Compared with model group,the chondrocyte damage and fibroplasia gradually reduced in Diclofenac Sodium group,PDTC group,Danzi Kangxi Gradule low-dose group and Danzi Kangxi Gradule medium-dose group and Danzi Kangxi Gradule high-dose group;NF-κB protein was mainly expressed in nucleus,but few in cytoplasm in those groups,and the levels of p-IκBα,p-IKKα,p-IKKβand NLRP3 proteins decreased in Diclofenac Sodium group,PDTC group,Danzi Kangxi Gradule low-dose group,Danzi Kangxi Gradule medium-dose group and Danzi Kangxi Gradule high-dose group;The levels of IL-1β,IL-18,TNF-α,anti-ColⅡ-IgG,anti-ColⅡ-IgG2a,NO,PGE2,HIF-1ɑ,TXB2 and TXB2/6-keto-PGF1αdecreased,and the level of 6-keto-PGF1αincreased in Diclofenac Sodium group,

关 键 词：膝骨关节炎 丹紫康膝冲剂 关节软骨 血瘀状态 核因子B信号通路 大鼠 

分 类 号：R285.5[医药卫生—中药学]