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Objective: To observe Paeoniflorin (paeoniflorin, Pae) on tumor necrosis factor-α (tumor necrosis factor-α, TNF-α) stimulation of human synovial fibroblast cells (fibroblast-like synoviocytes, FLS) proliferation and secretion of prostate Su E 2 (pros-taglandin E 2 , PGE 2 ), interleukin-1β (interleukin-1β, IL-1β) levels effects; observe human FLS after TNF-α stimulated PGE 2 receptor (E-prostanoid, EP), inhibitory G protein subunit 2 (inhibitory subunit of G protein, Gαi 2 ), activated G protein subunit (stimulatory subunit of G protein, Gαs), cyclic adenosine monophosphate (cyclic adenosine monophosphate, cAMP) levels change, whether through the influence of human FLS Pae on EP receptors, thus affecting human FLS G protein-cAMP signaling pathways exert its inhibition of human FLS abnormal proliferation. Methods: After informed consent, collect line knee arthroplasty in patients with synovial tissue, using tissue culture method of human synovial cells were cultured. Take the first three generations of synovial cells with different concentrations of TNF-α (0.02,0.2,2.0,20 μg · L -1 ) stimulation, observation PGE 2 , IL- 1β, cAMP levels and the Pae role. MTT assay proliferative response of human FLS. Radioimmunoassay detection of IL-1β, enzyme-linked immunosorbent assay PGE 2 and cAMP levels. By indirect immunofluorescence and fluorescence microscopy to detect human FLS Gαs, Gαi 2 , EP subtype EP 2 expression. Results: 1.Pae different concentrations can inhibit TNF-α stimulation servant FLS abnormal proliferation and secretion of PGE 2 , IL-1β levels. Take the first three generations of FLS, in TNF-α (0. 02,0.2,2.0,20 μg · L -1 ) concentrations stimulated proliferation significantly enhanced its secretion IL-1β, PGE 2 levels were significantly elevated; while Pae (10-8-10-4 mol · L -1 ) can inhibit different concentrations of TNF-α stimulation servant abnormal proliferation of FLS and secretion of PGE 2 , IL-1β levels. Tip: Pae by inhibiting TNF-α stimulation servant FLS abnormal proliferation and secretion of PGE 2 , IL-1β levels to exert anti-inflammatory effects. (2) adjusting EP 2 expression, maintaining Gαs / Gαi balance recovery PGE 2 -EP-G protein-cAMP normal signal transduction, may be the effect of suppressing Pae human FLS abnormal proliferation and secretion of hyperthyroidism important molecular mechanisms. Take the first three generations of FLS, in TNF-α (0. 02,0.2,2.0,20 μg · L -1 ) concentrations stimulated cAMP levels were significantly reduced, and Pae (10-8-10 - 4 mol · L -1 ) can significantly restore cAMP levels. Immunofluorescence analysis of test results found, TNF-α (20μg · L -1 ) stimulation can inhibit human FLS EP subtype EP 2 expression, Pae (10-8 mol · L -1 ) to restore EP 2 expression. TNF-α (20μg · L -1 ) stimulation can inhibit the expression of human synovial cells Gαs promote Gαi 2 expression. Pae (10-8 mol · L -1 ) can be restored Gαs, Gαi 2 expression restored Gαs / Gαi balance. Tip: Adjust the EP 2 expression, maintaining Gαs / Gαi balance, restore PGE 2 -EP-G protein-cAMP signal transduction normal, may be inhibited Pae play human FLS abnormal proliferation and secretion of hyperthyroidism important molecular mechanisms. Conclusion: 1.Pae by inhibiting TNF-α stimulation servant FLS abnormal proliferation and secretion of PGE 2 , IL-1β levels to exert anti-inflammatory immune function. 2.Pae by promoting the EP 2 receptor expression, restoring Gas / Gai balance, regulating PGE 2 -EP-G protein-cAMP signal transduction inhibition of human FLS abnormal proliferation and hyperactivity secretion.
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