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NF-κB-dependent Luciferase Activation and Quantification of Gene Expression in Salmonella Infected Tissue Culture Cells
Published on: January 12, 2020
[Hypercoagulable state is associated with NF-kappa B activation and increased inflammatory factors in patients with
Pingheng Zhang1, Jian Liu2, Bing Tan1
1Anhui University of Chinese Medicine, Hefei 230038, China.
Objective:
To investigate the mechanism of hypercoagulable state based on nuclear factor κB (NF-κB) pathway in patients with rheumatoid arthritis (RA).
Methods:
Thirty-five RA patients were enrolled as well as 20 healthy volunteers as a control group. Interleukin-10 (IL-10), IL-6, IL-4, IL-17, NF-κB activator 1 (Act1), p50, p65, IκBα, platelet activating factor (PAF), PAF-acetylhydrolase (PAF-AH) and anti-cyclic citrullinated peptide (CCP) were detected using ELISA. The number of platelet (PLT) was detected using Sysmex XT-2000i automated hematology analyzer. The levels of D-dimer (D-D), fibrinogen (FBG), thrombin time (TT), prothrombin time (PT), and partial thromboplastin time (APTT) were detected using Sysmex CA-1500 automatic coagulation analyzer. Erythrocyte sedimentation rate (ESR) was detected using Westergren method. C-reactive protein and rheumatoid factor (RF) were detected using Hitachi 7060 automatic biochemical analyzer. Meanwhile, the mRNA expressions of Act1, p65, p50, IκBα and IκB kinase α (IKKα) were detected using semi-quantitative reverse transcription PCR. The expressions of p65, p50 and IκBα proteins were examined using Western blotting. The correlations of the above indexes were analyzed by Spearman correlation test.
Results:
Compared with the normal group, the levels of DD, FBG, PLT significantly increased in the peripheral blood of RA patients, TT decreased, while APTT and PT were not significantly changed. IL-4, IL-10 and PAF-AH were significantly reduced in the sera of RA patients, while IL-6, IL-17, Act1, p50, p65, IκBα, IKKα and PAF were significantly elevated. Spearman correlation analysis showed that coagulant and fibrinolytic indexes were significantly correlated with cytokines, NF-κB, activity indexes and clinical symptoms and signs.
Conclusion:
The hypercoagulable state is common in the peripheral blood of RA patients, and it is closely related to inflammatory factors, activity indexes and abnormal activation of NF-κB.
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