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Aim To investigate the effects of DAP on human rheumatoid arthritis synovial fibroblasts(RA-FLS)and its relationship with endoplasmic reticulum stress(ERS)PERK/ATF4/CHOP signaling pathway.Methods RA-FLS cells were cultured and identified by immunofluorescence assay for Vimentin and CD68.CCK-8 detected(0,5,10,20,30,40,50,60,70)mg·L-1 DAP on the proliferation activity of RA-FLS cells.According to the proliferation activity,the experiment was divided into blank group(Blank),low dose group(L-DAP),medium dose group(M-DAP),high dose group(H-DAP)and high dose+specific inhibitor 4-PBA group(H-DAP+4-PBA).The levels of TNF-α and IL-6 were detected by ELISA.Cell apoptosis was detected by flow cytometry.The invasion and migration of cells were detected by Transwell and scratches assays,and the expressions of endoplasmic reticulum stress-related proteins GRP78,PERK,P-PERK,ATF4,CHOP,Caspase-12,C-Caspase-12 and Bcl-2 were assessed by Western blot.Results CCK-8 results showed that compared with 0 mg·L-1 DAP group,the proliferation activity of each group in 20-70 mg·L-1 DAP group was significantly different(P<0.05),and the proliferation rate corresponding to 0,20,40 and 60 mg·L-1 DAP group was significantly different(P<0.05).This concentration was used as the basis for blank,low-dose,medium-dose,high-dose groups and high-dose+4-PBA experimental grouping.DAP could inhibit the release of inflammatory cytokines IL-6 and TNF-α from RA-FLS cells in concentration,reduce the invasion ability of cells,promote apoptosis,upregulate the expression of PERK,P-PERK,ATF4,GRP78,CHOP,Caspase-12, C-caspase-12 proteins and decrease the expression level of anti-apoptotic protein Bcl-2.Another set of experiments demonstrated that high dose DAP+4-PBA could up-regulate the expression of IL-6 and TNF-α,as well as the invasion of cells,and inhibit the expression of apoptosis and ERs-related proteins.Conclusions Daphresin regulates the secretion of inflammatory factors by activating the PERK/ATF4/CHOP signaling pathway,inhibits the proliferation and invasion of RA-FLS cells,and induces apoptosis,which is expected to be a potential therapeutic pathway for RA.
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Objective:To investigate the mechanism of sinomenine (SIN) in inducing the immunosuppressive effects of rat-derived dendritic cells (DCs).Methods:The bone marrow-derived precursor cells from Wistar rats were cultured in vitro. The morphological differences between sinomenine treated DCs (sinomenine modified group, SIN group) and conventional induced DCs (conventional induced group, control group) were observed under microscope. The CD phenotype of DCs was detected by flow cytometry. DCs were induced maturation by lipopolysaccharides (LPS) stimulation. The impact of SIN on the expressions of Toll-like receptor (TLR)2, TLR3 and TLR4 on the DCs surface were detected by flow cytometry. Results:In the conventional induction group, the cells showed clusters or suspension growth, with obvious granular sense on the cell surface; while in the sinomenine induction group, the cells were clustered together, with no significant change in cell volume and morphology. The relative expressions of CD80 and CD86 were 70.7% and 71.3% in the conventional induction group, while 51.7% and 49.4% in the SIN group. The relative expression of TLRs on DCs in SIN + LPS group was TLR2 (51.2±0.34)%, TLR3 (50.3±0.14)%, TLR4 (52.1±0.16)%, which were significantly lower than those in LPS group [(94.35±0.16)%, (97.55±0.16)%, (94.6±0.12)%].Conclusions:SIN may induce immune tolerance by inhibiting the maturation of DCs via inhibiting the TLRs signaling pathways.
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<b>Objective::To study the chemical constituents from <italic>n</italic>-butanol extract of <italic>Akebia trifoliata</italic> caulis. <b>Method::The 100 kg caulis of <italic>A</italic>. <italic>trifoliata</italic> was extracted with 75% ethanol (EtOH) for three times by heating reflux. These 3 extracts were decompressed and concentrated, and then dissolved in water. The solvent was successively extracted with dichloromethane, ethyl acetate and <italic>n</italic>-butanol. The chemical constituents from the <italic>n</italic>-butanol fraction were isolated by macroporous, silica gel, sephadex LH-20 and ODS columns, and semi-preparative high performance liquid chromatography, and their chemical structures were determined through MS, NMR analysis (<sup>1</sup>H and <sup>13</sup>C-NMR) and spectroscopic data from literatures. <b>Result::Totally 14 compounds were isolated and identified as mutongsaponin B(<bold>1</bold>), mutongsaponin C(<bold>2</bold>), saponin PH(<bold>3</bold>), begoniifolide A(<bold>4</bold>), 2<italic>α</italic>, 3<italic>β</italic>, 23-trihydroxy-30-noroleana-12, 19-dien-28-oicacid-<italic>O</italic>-<italic>β</italic>-<italic>D</italic>-xylopyranosyl-(1→3)-<italic>α</italic>-<italic>L</italic>-rhamnopyranosyl-(1→4)-<italic>β</italic>-<italic>D</italic>-glucopyranosyl-(1→6)-<italic>β</italic>-<italic>D</italic>-glucopyranosyl ester(<bold>5</bold>), akemisaponins D(<bold>6</bold>), akemisaponins E(<bold>7</bold>), asiaticoside(<bold>8</bold>), saponin PJ1(<bold>9</bold>), scheffoleoside A(<bold>10</bold>), symplocosneolignan A(<bold>11</bold>), kalopanax-saponins D(<bold>12</bold>), leonticin E(<bold>13</bold>), ciwujianoside A<sub>1</sub>(<bold>14</bold>). <b>Conclusion::Compounds <bold>1-4</bold>, <bold>11</bold>, <bold>13, 14</bold> were isolated from this plant for the first time. The discovery of these compounds further enriched the chemical constituents of <italic>A</italic>. <italic>trifoliata</italic>, and provided experimental and scientific basis for the comprehensive development and utilization of <italic>A</italic>. <italic>trifoliata</italic>.
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BACKGROUND@#Clinical outcomes of undifferentiated arthritis (UA) are diverse, and only 40% of patients with UA develop rheumatoid arthritis (RA) after 3 years. Discovering predictive markers at disease onset for further intervention is critical. Therefore, our objective was to analyze the clinical outcomes of UA and ascertain the predictors for RA development.@*METHODS@#We performed a prospective, multi-center study from January 2013 to October 2016 among Chinese patients diagnosed with UA in 22 tertiary-care hospitals. Clinical and serological parameters were obtained at recruitment. Follow-up was undertaken in all patients every 12 weeks for 2 years. Predictive factors of disease progression were identified using multivariate Cox proportional hazards regression.@*RESULTS@#A total of 234 patients were recruited in this study, and 17 (7.3%) patients failed to follow up during the study. Among the 217 patients who completed the study, 83 (38.2%) patients went into remission. UA patients who developed RA had a higher rheumatoid factor (RF)-positivity (42.9% vs. 16.8%, χ = 8.228, P = 0.008), anti-cyclic citrullinated peptide (CCP) antibody-positivity (66.7% vs. 10.7%, χ = 43.897, P < 0.001), and double-positivity rate of RF and anti-CCP antibody (38.1% vs. 4.1%, χ = 32.131, P < 0.001) than those who did not. Anti-CCP antibody but not RF was an independent predictor for RA development (hazard ratio 18.017, 95% confidence interval: 5.803-55.938; P < 0.001).@*CONCLUSION@#As an independent predictor of RA, anti-CCP antibody should be tested at disease onset in all patients with UA.
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Objective To observe the expressions of bone morphogenetic protein-2(BMP-2)and bone morphogenetie protein-7(BMP-7)in the synovial tissue of fluorosis rats and its correlation with pathogenic mechanism of fluorosis arthritis.Methods Thirty-two SD rats were randomly divided into 4 groups:the control group,low,moderate and high-dose fluoride group.The control group ate commou fodder.The low,moderate and high dose fluoride group were fed with fodder composed of 25%.35%and 68%of corn(containing fluorine of 148.00 mg/kg)in chronic endemic fluorosis region in Guizhou Province.After 140 days,the expressions of BMP-2 and BMP-7 protein were determined by immunohistochemistry and assayed the absorbanee by computer image-pattern analysis system.Light microscope was used to observe the synovial tissue by Hematoxin Eosin,and calculated the pathological integral of synovium according to pathological grade standard.Results The expressions of BMP-2 (32.50±2.73)and BMP-7(38.90±2.56)in the control group was spare.Compared with the control group,the expressions of BMP-2(59.43±5.12,79.82±6.41,101.76±7.56)and BMP-7(55.10±4.82,78.42±5.61,98.46± 6.05)in the synovial tissue was up-regulated in each experimental groups(P＜0.05),especially in the moderate dose and the high-dose groups(P＜0.05).Compared with the control group(0.54±0.21).the pathological integral of synovium increased(P＜0.05)in each experimental groups(1.04±0.98,4.69±1.28,8.60±2.07).The expressions of BMP-2 and BMP-7 in the synovial tissue was found to be positively related with the pathological integral of synovium(r=0.98,0.99,P＜0.05).Conclusion The BMP-2 and BMP-7 play an important role in the development of fluorosis arthritis,probably by affecting osteogenesis.