风湿祛痛胶囊对类风湿关节炎滑膜Akt和MAPK信号通路的影响

Effect of Fengshi Qutong Capsule on Synovial Akt and MAPK Signaling Pathways in Rheumatoid Arthritis

目的:观察风湿祛痛胶囊(FSQTC)对蛋白激酶B(Akt)和丝裂原活化蛋白激酶(MAPK)信号通路的影响,以探讨其抑制类风湿关节炎(RA)滑膜血管新生的机制。方法:采用SD大鼠诱导建立胶原诱导性关节炎(CIA)模型,灌胃给予FSQTC低、中、高剂量(0. 25,0. 5,1 g·kg^(-1)) 19 d后,取膝关节滑膜组织备用;血管内皮细胞生长因子(VEGF,20μg·L^(-1))诱导人脐静脉内皮细胞(HUVEC),肿瘤坏死因子-α(TNF-α,20μg·L^(-1))体外诱导人RA成纤维样滑膜细胞系MH7A,分别加入不同质量浓度FSQTC(0. 02,0. 1,0. 5μg·L^(-1))作用后,取细胞备用。分别提取滑膜组织以及MH7A和HUVEC两种细胞中的蛋白,采用蛋白免疫印迹法(Western blot)检测磷酸化Akt(p-Akt),p-p38 MAPK,p-细胞外信号调节激酶(ERK),p-氨基末端激酶(JNK)的蛋白表达水平。结果:CIA模型组大鼠关节滑膜中p-Akt,p-p38 MAPK,p-ERK和p-JNK的表达水平均较正常组显著升高(P <0. 01),经0. 25,0. 5,1 g·kg^(-1)·d^(-1)的FSQTC治疗后降低(P <0. 05,P <0. 01);与空白组比较,VEGF或TNF-α能分别诱导p-Akt,p-p38 MAPK,p-ERK和p-JNK在MH7A或HUVEC细胞中的异常升高(P <0. 01),0. 02,0. 1,0. 5μg·L^(-1)的FSQTC能明显降低p-Akt,p-p38 MAPK,p-ERK,p-JNK的表达水平(P <0. 05,P <0. 01)。结论:FSQTC能负调节Akt和MAPK信号通路在CIA大鼠滑膜组织、体外培养的血管内皮细胞和成纤维滑膜细胞中的异常活化,这可能与其对RA滑膜血管新生的抑制作用有关。

Objective: To observe the effect of Fengshi Qutong capsule( FSQTC) on protein kinase B( Akt) and mitogen-activated protein kinase( MAPK) signaling pathways of rheumatoid arthritis( RA). Method:Collagen-induced arthritis( CIA) was induced in SD rats,and the synovial membranes of the knee joints were prepared after 19 days of oral administration of 0. 25,0. 5,1 g·kg^-1 FSQTC. MH7 A cells were induced by tumor necrosis factor-α( TNF-α,20 μg·L^-1) in vitro,and human umbilical vein endothelial cells( HUVEC) were induced by vascular endothelial growth factor( VEGF). FSQTC( 0. 02,0. 1,0. 5 μg·L^-1) were added to MH7 A/HUVEC cells,and then the cells were collected. Proteins of synovial tissue,MH7 A and HUVEC cells were extracted,and then were detected the expresstion of p-Akt,p-p38 MAPK,p-extracellular signal-regulated kinase( ERK) and p-Jun n-terminal kinase( JNK) by Western blot. Result: The expression levels of p-Akt,p-p38 MAPK,p-ERK and p-JNK in the synovial membrane of CIA model were significantly increased compared with normal group( P<0. 01),and the treatmend of 0. 25,0. 5 and 1 g·kg^-1·d^-1 FSQTC significantly decreased their expression levels( P<0. 05,P 0. 01). To compared with control group,the expression levels of p-Akt,p-p38 MAPK,p-ERK and p-JNK in MH7 A or HUVEC cells induced by TNF-α or VEGF were increased( P<0. 01),respectively,and these factors are significantly reduced by 0. 02,0. 1,0. 5 μg·L^-1 FSQTC( P<0. 05,P 0. 01). Conclusion: FSQTC can down-regulate the abnormal activation of Akt and MAPK signaling pathways in the synovial membrane of CIA rats,fibroblast synovial cells and vascular endothelial cells,which is related to the inhibition of synovial angiogenesis in the treatment of RA.