Rheumatoid arthritis (RA) belongs to Bi syndrome (arthralgia) in Chinese medicine. Till now there lacks effective therapeutic methods. Recently cyclooxygenases (COXs) inhibitors, having regulator roles for many pro-inflammatory cytokines, have been widely used in RA treatment. But due to existing cardiovascular risks, researches on targeting the downstream specific factors of COXs have been under discussion. Considering the key role of blood stasis syndrome (BSS) in the pathology of RA and the fact that thromboxane A2 (TXA2) plays a pivotal role in BSS, we theoretically explored possible regulatory roles of Compound Danshen, a representative therapy in blood activating stasis removing method in the downstream path of COXs in synovial cells of RA. We proposed a brand new research direction of RA researches.
Arthritis, Rheumatoid ; diagnosis ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Medicine, Chinese Traditional ; methods ; Prostaglandin-Endoperoxide Synthases ; metabolism ; Salvia miltiorrhiza ; chemistry ; Synovial Membrane ; drug effects ; metabolism

AIMTo study the effects of lipopolysaccharide (LPS), the supernatant of U937 cells stimulated with LPS and dexamethasone on interleukin-6 (IL-6) expression in the synoviocyte from patients with rheumatoid arthritis (RA).

METHODSFibroblast-like synoviocytes (FLS) from the joint tissue of patients with rheumatoid arthritis were cultured and incubated for 24 h with LPS (1 mg.L-1) or the supernatant of U937 cells stimulated with LPS (1 mg.L-1) for 24 h. Dexamethasone was added to the supernatant of U937 cells and FLS was incubated for 24 h. The expression of IL-6 protein was detected by radioimmunoassay. The mRNA expression of IL-6 was accessed by RT-PCR.

RESULTSThe growth of FLS was not markedly affected by LPS, and the protein secretion and mRNA expression of IL-6 were not markedly changed in FLS treated with LPS. The IL-6 secretion and IL-6 mRNA expression were significantly increased in FLS cultured with the supernatant from U937 cell treated with LPS. Dexamethasone markedly inhibited the protein secretion and mRNA expression of IL-6 in FLS cultured with the supernatant from U937 cell stimulated with LPS. The inhibitory effects were increased as the concentration of dexamethasone increased.

CONCLUSIONLPS was not shown to directly affect the expression of IL-6 in FLS, but it indirectly causes the increase of the IL-6 expression in FLS by stimulating U937 cell. Dexamethasone can inhibit this increase of the IL-6 expression.

Arthritis, Rheumatoid ; pathology ; Cell Division ; drug effects ; Cells, Cultured ; Dexamethasone ; pharmacology ; Fibroblasts ; drug effects ; metabolism ; Gene Expression ; drug effects ; Humans ; Interleukin-6 ; biosynthesis ; genetics ; Lipopolysaccharides ; pharmacology ; RNA, Messenger ; genetics ; Synovial Membrane ; drug effects ; metabolism ; U937 Cells

OBJECTIVETo study the effect of pulchinenoside (PULC) in modulating SFRP2 expression in fibroblast-like synoviocytes (FLS) of rheumatoid arthritis (RA) model rats.

METHODThe effect of PULC in treating RA rats was evaluated by rat arthritis score and paw swelling score. The inhibitory effect of PULC on FLS proliferation was detected by MTT reagent. The effects of PULC gavage treatment in modulating gene expression of FLS SFRP2, critical gene beta-catenin of Wnt pathway and downstream effector genes C-myc of of Wnt pathway were detected by RT-PCR and Western blotting.

RESULTPULC had a significant effect in treating RA rats and that SFRP2 expression was down-regulated in FLS. After PULC gavage treatment, FLS SFRP2 expression was obviously up-regulated, whereas beta-catenin and C-myc gene expressions were significantly down-regulated.

CONCLUSIONPULC can inhibit abnormal proliferation of synovial membrane by modulating Wnt pathway of RA rats.

Animals ; Arthritis, Rheumatoid ; drug therapy ; metabolism ; Disease Models, Animal ; Gene Expression Regulation ; drug effects ; Male ; Membrane Proteins ; genetics ; Rats ; Rats, Sprague-Dawley ; Saponins ; pharmacology ; Synovial Membrane ; drug effects ; metabolism

OBJECTIVETo observe the effect of Sanshui Baihu Decoction (SBD) containing serum on the proliferation of in vitro cultured fibroblast-like synoviocytes (FLS) derived from rheumatoid arthritis (RA) and osteoarthritis (OA) and its secretion of interleukin-6 (IL-6) and IL-17, and to explore the pharmacological mechanism of SBD.

METHODSThe FLS obtained from cultured RA and OA patients' synovial tissue were cultured and passaged in vitro in a routine way. The cultured medium was changed to DMEM with 20% SBD containing serum and cultured for 72 h after cultured for 3 to 6 generations. The proliferation rate of FLS was detected by MTT assay. Levels of IL-6 and IL-17 in the supernatant were detected by ELISA. Leflunomide and saline containing serum were used as positive and negative control respectively.

RESULTSSBD containing serum significantly inhibited the proliferation of RA-FLS and OA-FLS, and decreased the secretion of IL-17 in RA-FLS. Its inhibition efficiency of SBD was equivalent to that of Leflunomide. No obvious inhibition on the secretion of IL-6 in RA-FLS was observed. It had no significant effect on the secretion of IL-17 and IL-6 in OA-FLS.

CONCLUSIONSBD could inhibit the proliferation of FLS and the secretion of IL-17 in RA-FLS, which might be one of its pharmacological mechanisms for treating RA.

Animals ; Arthritis, Rheumatoid ; metabolism ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Fibroblasts ; drug effects ; secretion ; Humans ; Interleukin-17 ; metabolism ; Interleukin-6 ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Synovial Membrane ; cytology ; drug effects

OBJECTIVETo study the effect and related mechanism of seaweed polysaccharide (SP) on apoptosis of fibroblast-like synoviocytes in patients with rheumatoid arthritis (RA-FLS).

METHODSRA-FLS were in vitro cultured using modified tissue culture method. Effect of SP (0, 15, 20, and 25 mg/mL, respectively) at different time points (0, 3, 4, and 5 days, respectively) on the proliferation and apoptosis of RA-FLS, and protein expressions of Caspase-3, Bax, and Bcl-2 was detected by cell counting kit-8 (CCK-8) assay, Hoechst 33258 staining assay, TUNEL assay, and Western blot, respectively.

RESULTSCompared with 0 mg/mL SP at the same time point, the proliferation of RA-FLS was inhibited, and the apoptosis was promoted 3, 4, and 5 days after intervened by 15, 20, and 25 mg/mL SP, respectively (P<0.01) in time- and dose-dependent manners. RA-FLS Bax protein expression was up-regulated, Bcl-2 protein expression down-regulated, Caspase-3 activated and split by 15, 20, and 25 mg/mL SP, respectively for 4 days (P<0.05, P<0.01). Besides, the changes were in a dose-dependent manner.

CONCLUSIONSSP could inhibit RA-FLS proliferation and induce its apoptosis in dose- and time-dependent manners. Its apoptosis mechanism might be through up-regulating intracellular Bax protein expression and down-regulating Bcl-2 protein expression, thus influencing the mitochondrion signaling pathway, further promoting Caspase-3 activation and split, resulting in the apoptosis of RA-FLS.

Apoptosis ; drug effects ; Arthritis, Rheumatoid ; pathology ; Caspase 3 ; metabolism ; Cells, Cultured ; Fibroblasts ; cytology ; drug effects ; Humans ; Polysaccharides ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Seaweed ; Synovial Membrane ; cytology ; drug effects ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo explore the effects of Bushen Shuji Granule (BSG) on inhibiting the interleukin 6 (IL-6) level in the synovial fluid sample of fibroblast cells from the ankylosing spondylitis (AS) patients.

METHODSUsing serum pharmacologic method, the IL-6 level in the culture fluid sample of fibroblast cells was observed by ELISA method with different concentrations of medicated serum containing BSG. The IL-6 level at the mRNA level was detected using reverse transcriptase-polymerase chain reaction (RT-PCR). The vehicle serum and sulfasalazine (SSZ) serum were taken as controls.

RESULTSResults of ELISA showed the IL-6 level in the AS group was more than that in the vehicle serum group with obvious statistical difference. BSG could obviously inhibited the IL-6 level, showing statistical difference when compared with the vehicle serum group. Besides, obvious dose-dependent correlation existed between BSG and its inhibition on fibroblast proliferation. And the IL-6 level at the mRNA level in the AS group was higher than that in the vehicle serum group, showing statistical difference by semi-quantitative analysis.

CONCLUSIONBSG could play its clinical role of anti-inflammation and anti-fibrosis through inhibiting the IL-6 level in the culture fluid sample of fibroblast cells.

Animals ; Cell Line ; Drugs, Chinese Herbal ; pharmacology ; Female ; Fibroblasts ; drug effects ; secretion ; Humans ; Interleukin-6 ; metabolism ; Male ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Spondylitis, Ankylosing ; Synovial Fluid ; metabolism ; Synovial Membrane

OBJECTIVETo investigate the mechanism of anti-death receptor 5-10 (AD5-10) combined with epirubicin in treating rheumatoid arthritis (RA).

METHODSWe detected the cell viability of the fibroblast-like synoviocytes (FLS) from RA patients with MTT. The expression level of apoptosis signaling pathways protein, p53, and p21 were evaluated with Western blot.

RESULTSWe found that epirubicin, at different doses, could enhance the effect of AD5-10 on FLS, promoting the apoptosis of FLS. The expression levels of caspase-3, -8, -9, c-FLIP, Bcl-2, p53, and p21 in the FLS changed after epirubicin treatment.

CONCLUSIONEpirubicin may coordinate with AD5-10 in inducing FLS apoptosis through affecting the levels of p53, p21, c-FLIP, and Bcl-2.

Antibodies, Monoclonal ; pharmacology ; Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; metabolism ; Arthritis, Rheumatoid ; drug therapy ; metabolism ; pathology ; Cells, Cultured ; Epirubicin ; pharmacology ; Humans ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; immunology ; Synovial Membrane ; cytology ; drug effects ; metabolism

The role of flavonoids of Echinps latifolius (FELT) in Wnt signaling was investigated in adjuvant arthritis (AA) rats. The therapeutic effects of FELT on AA rats were detected by rat arthritis score and MTT. The effect of FELT gavage treatment on the Wnt signaling key gene β-catenin, C-myc and cyclin D1 in synovium from AA rats was detected by Real-time qPCR, and the effects of FELT gavage treatment on the upstream negative regulation gene SFRP 1,2,4,5 in synovium from AA rats were detected by Real-time qPCR. The results showed that FELT gavage treatment significantly inhibited arthritis score and MTT values in AA rats, significantly inhibited the expression of the Wnt signaling gene β-catenin, C-myc and cyclin D1, significantly up-regulated the expression of the up- stream negative regulation gene SFRP 1,2,4. FELT has a better therapeutic effect for AA rats.
Animals ; Arthritis, Experimental ; drug therapy ; genetics ; metabolism ; Asteraceae ; chemistry ; Disease Models, Animal ; Down-Regulation ; drug effects ; Drugs, Chinese Herbal ; administration & dosage ; Flavonoids ; administration & dosage ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; Male ; Membrane Proteins ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Synovial Membrane ; drug effects ; metabolism ; Wnt Signaling Pathway ; drug effects ; beta Catenin ; metabolism

OBJECTIVETo research the effects of Qubi Zhentong Recipe (QZR) on the expressions of interleukin-1beta (IL-1beta), interleukin-8 (IL-8), and vascular endothelial growth factor (VEGF) in the synovial of rats with collagen-inducing arthritis (CIA), and to discuss its mechanisms of action.

METHODSHealthy male Wistar rats were recruited and randomly divided into the model group ( n = 50) and the normal control group (n = 10). Rats of the model group were injected with type II collagen of bovine (BC II) emulsion in the tail and nape to establish the CIA model. After successful modeling, 30 successfully modeled rats were selected and randomly divided into three groups, i.e., the model group (n = 10), the QZR group (n = 10), and the methotrexate (MTX) group (n = 10). Rats in the normal control group and the model group were administered with physiological saline by gastrogavage, while those in the QZR group were administered with QZR at 22.9 g/kg by gastrogavage. All medication was performed once daily. The rats in the MTX group were administered with MTX suspension at 0.78 mg/kg by gastrogavage, once per week. After 30-day treatment, the levels of IL-1beta, IL-8, and VEGF in the synovial were detected by immunohistochemical method. The arthritis index (AI) was scored before and after medication.

RESULTSAfter treatment the AL score of the QZR group and the MTX group was obviously lower than that of the model group (P < 0.01). The AI score of the two drug groups were lower than that before treatment (P < 0.01). Compared with the normal control group, the expression levels of IL-1beta, IL-8, and VEGF obviously increased in the model group (P < 0.01). Compared with the model group, the expression levels of IL-1beta, IL-8, and VEGF were significantly lower in the two drug groups (P < 0.01). But there was no statistical difference between the QZR group and the MTX group (P > 0.05).

CONCLUSIONDecreasing the expression levels of IL-1beta, IL-8, and VEGF in the synovial of CIA rats may be one of the mechanisms for treating CIA by QZR.

Animals ; Arthritis, Experimental ; drug therapy ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Interleukin-1beta ; metabolism ; Interleukin-8 ; metabolism ; Male ; Rats ; Rats, Wistar ; Synovial Membrane ; drug effects ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo study the effect of genistein (Gen) on MAPK signal pathway in the CIA rat fibroblast-like synoviocytes (FLS).

METHODSThe rat model of collagen-induced arthritis (CIA) was established. The cultured FLS of CIA rats were divided using randomized method. The effects of Gen (at the concentration of 50, 100, and 200 micromol/L, respectively) on the proliferation of FLS in CIA rats using methyl thiazolyl tetrazolium (MTT) assay. Effects of Gen (at the concentration of 50, 100, and 200 pmol/L, respectively) on the expressions of extracellular signal-regulated kinase (ERK) and phosphorylated extracellular signal-regulated kinase (p-ERK) in the FLS of CIA rats were detected.

RESULTSGen could inhibit the proliferation of FLS in CIA rats. The FLS proliferation in the high dose Gen group at 72 h was only 1.10+/-0.04, significantly lower than that in the model group (2.12+/-0.03, P<0.01). Besides, after Gen's action on FLS, the expression of p-ERK was down-regulated. It was only 0.34+/-0.02 in the high dose Gen group, significantly lower than that in the model group (2.68+/-0.14, P<0.01). There was no change in the expression of ERK (P>0.05).

CONCLUSIONSGen could inhibit the proliferation of FLS in CIA rats. Its mechanism of action was mainly correlated to down-regulating the tyrosine kinase of MAPK signal transduction pathway and inhibiting phosphorylation of ERK.

Animals ; Arthritis, Experimental ; metabolism ; Cells, Cultured ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Female ; Genistein ; pharmacology ; MAP Kinase Signaling System ; drug effects ; Rats ; Rats, Sprague-Dawley ; Synovial Membrane ; cytology ; drug effects ; metabolism