摘要
探讨金天格胶囊含药血清(JTG-CS)对人类风湿关节炎成纤维样滑膜细胞(FLS)的作用及机制。制备SD大鼠空白血清及低、中、高剂量JTG-CS。超高效液相色谱-质谱联用技术检测血清内氨基酸成分和含量;肿瘤坏死因子-α(TNF-α)刺激原代FLS建立FLS炎症模型,CCK-8检测FLS增殖,伤口愈合实验和Transwell实验分析FLS的迁移和侵袭,Western blot分析相关蛋白的表达。JTG-CS能显著抑制FLS的增殖和迁移,抑制FLS的伤口愈合和侵袭能力,抑制黏附相关蛋白细胞间黏附分子-1(ICAM-1)、血管细胞黏附分子-1(VCAM-1)、基质金属蛋白酶(MMP)2和MMP3的表达,降低炎性细胞因子TNF-α、白细胞介素(IL)-6和IL-1β的水平,抑制PI3K/Akt通路、NF-κB通路和JAK2/STAT3通路的激活。PI3K激动剂可逆转JTG-CS对FLS炎性细胞因子和细胞侵袭的抑制作用。与空白大鼠血清相比,JTG-CS中46种氨基酸含量上升,20种氨基酸含量下降。JTG-CS通过抑制PI3K/Akt通路的激活,抑制TNF-α刺激的FLS增殖、迁移、侵袭和炎症反应。
This study aims to explore the effects and mechanism of Jintiange Capsules-containing serum(JTG-CS)on fibroblast-like synoviocytes(FLS)in human rheumatoid arthritis.The blank serum from SD rats and low,medium,and high-dose JTG-CS samples were prepared.Ultra-high performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS)was employed to determine the amino acid composition and content in the blank serum and JTG-CS.Tumor necrosis factor-α(TNF-α)was used to stimulate FLS for the modeling of inflammation.The proliferation of FLS was determined by the CCK-8 assay.The migration and invasion of FLS were examined by wound healing and Transwell assays.The protein expression of FLS was analyzed by Western blot.JTG-CS significantly inhibited the proliferation,migration,wound healing,and invasion of FLS stimulated by TNF-α,down-regulated the expression of intercellular adhesion molecule-1(ICAM-1),vascular cell adhesion molecule-1(VCAM-1),matrix metalloproteinase(MMP)2,and MMP3,lowered the levels of TNF-α,interleukin(IL)-6,and IL-1β,and inhibited the activation of phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt),nuclear factor(NF)-κB,and Janus kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3)pathways.The treatment with the PI3K agonist reversed the inhibitory effects of JTG-CS on inflammatory cytokines and invasion of FLS stimulated by TNF-α.Compared with the blank serum,JTG-CS showed increased content of 46 amino acids and decreased content of 20 amino acids.JTG-CS inhibited the proliferation,migration,invasion,and inflammation of FLS stimulated by TNF-αby inhibiting the activation of the PI3K/Akt pathway.
作者
叶欣园
沈燚
鲍荣华
刘玉玲
任秋儒
张泉龙
秦路平
张巧艳
YE Xin-yuan;SHEN Yi;BAO Rong-hua;LIU Yu-ling;REN Qiu-ru;ZHANG Quan-long;QIN Lu-ping;ZHANG Qiao-yan(School of Pharmaceutical Science,Zhejiang Chinese Medical University,Hangzhou 310053,China;Hangzou Fuyang Hospital of TCM Orthopedics and Traumatology,Hangzhou 311400,China)
出处
《中国中药杂志》
北大核心
2025年第20期5811-5819,共9页
China Journal of Chinese Materia Medica
基金
浙江省自然科学基金重点项目(LZ22H28002)
国家自然科学基金项目(82370499)。
