四神煎对膝骨关节炎大鼠滑膜病变的影响及机制  

Effect and Mechanism of Sishenjian on Synovial Lesions in Knee Osteoarthritis Rats

作　　者：胡子旋 黄雪君 姚楠 彭莎 黄丹娥 姜涛 甘海宁 黄晓丹 HU Zixuan;HUANG Xuejun;YAO Nan;PENG Sha;HUANG Dan'e;JIANG Tao;GAN Haining;HUANG Xiaodan(Guangdong Provincial Second Hospital of Traditional Chinese Medicine(TCM),Guangdong Provincial Engineering Technology Research Institute of TCM,The Fifth Clinical College of Guangzhou University of Chinese Medicine,Guangzhou 510095,China;Guangdong Provincial Key Laboratory of Research and Development in TCM,Guangzhou 510095,China)

机构地区：[1]广东省第二中医院,广东省中医药工程技术研究院,广州中医药大学第五临床医学院,广州510095 [2]广东省中医药研究开发重点实验室,广州510095

出　　处：《中国实验方剂学杂志》2024年第11期18-26,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：广东省基础与应用基础研究基金项目(2023A1515010752);广东省中医药局科研项目(20222011);广东省医学科研基金项目(B2022208);广东省第二中医院科研创新基金项目(SEZYY2023A02)。

摘　　要：目的:探讨四神煎对碘乙酸钠诱导的大鼠膝骨关节炎(KOA)滑膜病变的影响及机制。方法:60只雌性SD大鼠随机分为6组,分别为正常组、模型组、塞来昔布组和四神煎高、中、低剂量组。通过关节腔内注射碘乙酸钠复制KOA大鼠模型。根据分组给予塞来昔布(18 mg·kg^(-1))和四神煎(14.4、7.2、3.6 g·kg^(-1))进行灌胃干预。连续给药4周后处死所有大鼠。测量大鼠双侧膝关节的横径并进行膝关节大体观察,苏木素-伊红(HE)染色、天狼猩红染色观察膝关节滑膜组织病理学改变,免疫组化法(IHC)检测滑膜组织血管内皮生长因子A(VEGFA)蛋白表达。酶联免疫吸附测定法(ELISA)检测关节滑液中炎性细胞因子的表达水平。实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白免疫印迹法(Western blot)分别检测膝关节滑膜转化生长因子-β_(1)(TGF-β_(1))/Smad2/3通路相关mRNA和蛋白表达。结果:与正常组比较,模型组大鼠膝关节横径显著增加(P<0.01);与模型组比较,四神煎各剂量组大鼠膝关节横径显著减小(P<0.01)。与正常组比较,模型组大鼠膝关节滑液白细胞介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)表达水平显著升高(P<0.01);与模型组比较,四神煎各剂量组大鼠膝关节滑液IL-1β和TNF-α表达水平显著降低(P<0.01)。与正常组比较,模型组大鼠膝关节滑膜TGF-β_(1)、Smad2/3、磷酸化(p)-Smad2/3、Ⅰ型胶原_(α1)(ColⅠ_(α1))、Ⅲ型胶原_(α1)(ColⅢ_(α1))和VEGFA蛋白表达显著升高,TGF-β_(1)、Smad2/3、ColⅠ_(α1)、ColⅢ_(α1)mRNA表达显著升高(P<0.01);与模型组比较,四神煎各剂量组大鼠膝关节滑膜TGF-β_(1)、Smad2/3、p-Smad2/3、ColⅠ_(α1)、ColⅢ_(α1)和VEGFA蛋白表达明显降低,TGF-β_(1)、Smad2/3、ColⅠ_(α1)、ColⅢ_(α1)mRNA表达明显降低(P<0.05,P<0.01)。结论:四神煎可以抑制滑膜炎和血管新生,并且可以通过调节TGF-β_(1)/Smad2/3通路减少滑膜纤维化,因而有望成�Objective:To explore the effect and mechanism of Sishenjian on synovial lesions induced by monosodium iodoacetate(MIA)in rats with knee osteoarthritis(KOA).Method:Sixty female SpragueDawley(SD)rats were randomly divided into the following six groups:normal group,model group,celecoxib group,and high-,medium-,and low-dose Sishenjian group.The KOA rat model was established by intraarticular injection of MIA.Celecoxib(18 mg·kg^(-1))and Sishenjian(14.4,7.2,3.6 g·kg^(-1))were administered by gavage according to the groups.All rats were euthanized after four weeks of continuous administration.The transverse diameter of the bilateral knee joints of rats was measured,and gross observation of the knee joint was performed.Pathological changes in knee joint synovial tissue were observed by hematoxylin-eosin(HE)staining and picrosirius red staining.Immunohistochemistry(IHC)was used to detect the expression of vascular endothelial growth factor A(VEGFA)in synovial tissue.The levels of inflammatory cytokines in the joint synovial fluid were detected by enzyme-linked immunosorbent assay(ELISA).Real-time quantitative polymerase chain reaction(Real-time PCR)and Western blot were used to detect the expression of mRNA and proteins related to the transforming growth factor-β_(1)(TGF-β_(1))/Smad2/3 pathway in knee joint synovium.Result:Compared with the normal group,the transverse diameter of the knee joint in the model group significantly increased(P<0.01).Compared with the model group,the transverse diameter of the knee joint in rats of each Sishenjian group significantly decreased(P<0.01).Compared with the normal group,the expression levels of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)in the knee joint synovial fluid of model group significantly increased(P<0.01).Compared with the model group,the expression levels of IL-1βand TNF-αin the knee joint synovial fluid of rats in each Sishenjian group significantly decreased(P<0.01).Compared with the normal group,the expression levels of TGF-β_(1),Smad2/3,phosphor

关 键 词：四神煎 滑膜炎 血管新生 滑膜纤维化 

分 类 号：R2-0[医药卫生—中医学] R22R285.5R289R33