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Objective To explore the effects of ring finger protein 43 (RNF43) on fibroblast-like synoviocytes (FLS) from patients with rheumatoid arthritis (RA).Methods Synovial tissues from patients with RA treated by knee arthroplasty were used to isolate FLSs by type 2 collagenase.RNF43 lentivirus overexpressing plasmid was constructed and transfected in to RA-FLS.After successful transfection,RNA and super natant of RA-FLS were extracted.QRT-polymerase chain reaction (PCR) and enzyme linked immunosorbent assay (ELISA) were used to detect the mRNA and protein expression levels of matrix metalloproteinase (MMP)-1,MMP-3 and MMP-13.Data were analyzed with Student's t test.Results Transfection efficiency could meet the test requirements when the multiplicity of infection was 40 and was in conjunction with appropriate concentration of polybrene.The mRNA of RNF43 increased for 26158-fold than the control group.In vitro,compared with the control group,RNF43 could significantly inhibit the mRNA of MMP-1,MMP-3 and MMP-13 and MMP-13 [(0.19±0.06),t=28.314,P＜0.05;(0.28±0.07),t=23.413,P＜0.05;(0.21±0.09),t=18.365,P＜0.05]and the protein of MMP-1,MMP-3 and MMP-13 and MMP-13 [(31.0±9.4) pg/ml,(17.1±2.1) pg/ml,t=3.198,P=0.029],MMP-3 [(38.7±8.1) pg/ml,(24.9±3.5) pg/ml,t=3.514,P=0.015],MMP-13 [(35.9±5.4) pg/ml,(20.6±2.9) pg/ml,t=5.632,P=0.001].Conclusion The results of study suggest that RNF43 could inhibit the secretion of MMPs in RA-FLS by suppressing the activity of Wnt signal pathway.
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Objective To study the proliferation and inflammatory phenotypes of human fibroblast-like synoviocytes (FLS) induced by tumor necrosis factor-α(TNF-α). Methods The rheumatoid arthritis (RA)-FLS were cultured in vitro, then treated with different concentrations of diallyl trisulfide (DATS). The proliferation activity was detected by CCK-8 method. Then TNF-α was used to stimulate the RA-FLS, mRNA and protein expression of interleukin (IL)-6, matrix metalloproteinases (MMP)-1 and vascular endothelial growth factor (VEGF) were detected by quantitative real time polymerase chain reaction (qPCR) and enzyme linked immunosorbent assay (ELISA). Differences among groups were determined by one-way analysis of variance (ANOVA), LSD-t test was used for comparison between 2 groups. Results RA-FLS was successfully is-olated and cultured in vitro. The positive rate of CD90 and CD29 in the RA-FLS was more than 90％. The proli-feration activity of RA-FLS treated with 100 μmoL/L, 200 μmol/L and 300 μmol/L DATS was (98.92 ± 0.40)％, (95.91±0.32)％, (94.05±0.24)％, respectively. As Compared with the normal control group, the pro-liferation activity of RA-FLS was lower, and the statistically significant difference is between normal control group 200 μmol/L and 300 μmol/L DATS (t=-4.46, P<0.05; t=-7.98, P<0.05). After TNF-α stimulation, the expression of IL-6's mRNA in experiment group (100μmol/L DATS) is higher compared with the model control group (t=5.74, P<0.05), but the change of IL-6's protein is no significant difference (t=-0.49, P=0.627). The differences of mRNA and protein expression levels of MMP-1 and VEGF between the experiment group (100μmol/L DATS) and the model control group were not statistically significant. The relative mRNA level [(0.42 ± 0.06), t=-23.47, P<0.05;(0.14±0.039), t=-36.59, P<0.05;(0.36±0.09), t=-13.1, P<0.05)] and the protein levels [(108.0±4.7) ng/L, t=-63.79, P<0.05, (26.0±1.0) ng/L, t=-9.68, P<0.05;(57.9±0.7), t=-34.59, P<0.05] of IL-6, MMP-1, VEGF in experiment group (200μmol/L DATS) were significantly decreased. And the relative mRNA level [(0.041 ±0.027), t=-38.48, P<0.05; (0.027 ±0.027), t=-41.22, P<0.05; (0.131 ±0.047), t=-17.74, P<0.05] and the protein levels [(24.2 ±2.3) ng/L , t=-88.69, P<0.05; (22.7 ±1.0) ng/L , t=-14.13, P<0.05; (34.5 ±1.7), t=-48.45, P<0.05] of IL-6, MMP-1, VEGF in the experiment group (300 μmol/L DATS) were also significantly decreased. The difference between the two groups was significant (t=-24.89, P<0.05; t=-4.45, P<0.05; t=-13.87, P<0.05). Conclusion DATS can inhibit the proliferation and the effect of TNF-αinduced secretion of IL-6, MMP-1 and VEGF in RA-FLS. The effect is dose-dependent.
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Fructus psoralea is a tonic traditional Chinese herb commonly used in clinic.The chemical constituents of Fructus psoralea are complicated,mainly containing coumarins,flavonoids and monoterpene phenols with various pharmacological effects.Since the increasing number of reports on the toxicity of Fructus psoralea in clinic,the side effects including toxicity on the liver and kidney,as well as skin allergies have gradually attracted attention.The toxicity of Fructus psoralea is produced from ADME (absorption,distribution,metabolism and excretion) in vivo.In this paper,we collected and clarified the studies of ADME of Fructus psoralea in vivo,and summarized recent adverse clinical events and research over its toxicity.We propose to make a thorough study on the toxic material basis of Fructus psoralea and the toxicological mechanism of its extraction,fractions and compounds.The review provided a possible reference and the direction of research for the safe clinical use of Fructus psoralea.
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Objective:To analyse the biological function of anti-IL-6Rβ(gp130) monoclonal antibody and its regulatory effect on IL-6 signaling.Methods:Biological characteristics of anti-IL-6Rβ(gp130) mAb were assessed by Western blot analysis, capture ELISA and peptide ELISA .The phosphorylation of STAT 3 was tested by Western blot analysis in IL-6-stimulated U266/RA-FLS/RA-PBMC with or without anti-IL-6Rβ(gp130) mAb treatment.Results:3 strains of mouse anti-human gp130 mAb were with high affini-ty and different binding epitopes , the kaff of 10A1 was 2.62E-10.In U266, RA-PBMC and RA-SFMC, IL-6 signaling highly activated STAT3 which could be inhibited by anti-gp130 mAb.Conclusion: Anti-IL-6Rβ( gp130 ) mAb might have different binding epitopes and could affect IL-6 stimulated phosphorylation of STAT3, which provides a preliminary experiment for analyse the correlation of IL-6 signaling and RA .
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Rheumatoid arthritis (RA) is a kind of chronic autoimmune disease and osteoporosis is one of its complications.
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Objective This study is aimed to explore the effect of rhTNFR:Fc and methotrexate (MTX)-rhTN FR:Fc on joint destruction of collagen-induced arthritis ( CIA ) rat by establishing CIA rat model which imitates pathogenic factors of rheumatoid arthritis (RA).Methods CIA rat model were developed by subcutaneous injection of bovine type Ⅱ collagen.The rats with inflammation scores of two or above were randomly divided into four groups:the sterilized water treatment group (0.4 ml/w,intra-peritoneal injection),the MTX treatment group (1 mg/w,intra-peritoneal injection),the rhTNFR:Fc treatment group(0.8 mg Biw,intra-peritoneal injection),the MTX + rhTNFR:Fc treatment group (MTX 1 mg/w and rhTNFR:Fc 0.8 mg Biw,intraperitoneal injection).After treatment for 8 weeks,the rats were sacrificed and took the ankle radiography.Micro-CT scan of proximal tibia was performed and hard-tissue slices were made,and then the ankle's bone damage of each group was observed in order to evaluate trabecular variation and bone quantity changes of proximal tibia.Statisstical analysis was conducted with ANK-q test.Results After treatment for 8 weeks,the percentage of trabecular area and the trabecular number of the rhTNFR:Fc treatment group and the MTX-rhTNFR:Fc treatment were [(29.1±0.3)％,(26.7±0.6)％,(4.4±0.5)/mm,(4.0±0.6)/mm] (P＜0.01),which were evidently higher than the sterilized water treatment group and MTX treatment group (P＜0.01).The trabecular separation of Etanercept treatment group and MTX-rhTNFR:Fc group was obviously less than the sterilized water treatment group and MTX treatment group [(12.9±0.5)％,(13.2±0.4)％ vs (2.0±0.3)/mm,(2.2t0.2)/mm] (P＜0.01).Conclusion rhTNFR:Fc and MTX-rhTNFR:Fc can remarkably inhibit joint destruction of CIA rat.And their effect on inhibiting of inflammation and increasing peri-articular bone quantity.In addition,they are effective on inhibiting the reduction of local trabecular structure and increase of trabecular separation.
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Objective To evaluate the outcome of primary total knee arthroplasty (TKA) in treatment of stiff knee, and discuss the key points of operative technique and rehabilitation. Methods From February 2005 to April 2009, 23 patients with 34 stiff knees were treated with primary TKA. The study included 3 males (4 knees) and 20 females (30 knees), with the mean age of 56.9 years. Primary disease of the patients included rheumatoid arthritis (26 knees in 15 cases) and osteoarthritis (8 knees in 8 cases). Varus deformity was found in 10 knees (5°-15°), and valgus was found in 5 knees (5°-10°). Evaluations included preoperative and postoperative range of motion (ROM) measurement, hospital for special surgery knee score (HSS), blood loss, operative time and assessment of postoperative complications. Results All patients were followed up. The mean follow-up time was 32.2 (range, 24 to 40) months. At the final follow-up visit, the HSS score increased from 42.9±5.2 preoperatively to 85.7±4.3, the range of motion increased from 42.6°±5.7° preoperatively to 89.2°±10.5°. Sixteen knees in 12 cases underwent manipulation at 3 to 8 weeks postoperatively for unsatisfied ROM, but ROM was still less than 90° in 8 knees at the last follow-up. The average blood loss were (632.4±180.2) ml in first 24 hours (450-850 ml) and the operative time were (98.1±18.6) min (80-150 min). Deep venous thrombosis was found in 3 patients. All the symptoms relieved after anticoagulant therapy. Postoperative varus deformity was seen in one patient, but the function of knee was good. No revision was needed. Conclusion Primary total knee arthroplasty is reliable method in treatment of stiff knees. Sufficient soft tissue release during the operation, postoperative muscle strength exercise and essential manipulation are key points for satisfactory outcomes.
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OBJECTIVE:To study the effect and thepossible mechanism of low molecular weight heparin(LMWH)on immunocytes' adhesion to fibroblast-like synovocytes(FLS)isolated from patients with rheumatoid arthritis(RA)and to study LMWH'S possible anti-inflammatory effect on RA.METHODS:LMWH's interference on the adhesion of peripheral blood monouclear cells(PBMC)isolated from healthy volunteers to FLS of RA patients was determined by quantitative counting using flow cytometry.The expression of CYR61 in the in vitro cultured FLS of RA patients was detected using real-time PCR technology.RESULTS:When FLS culture system was added with PBMC,PBMC were obviously found to adhere to FLS,but the number of adhered PBMC decreased after LMWH treatment,which manifested as increase of deciduous PBMC,increased more with the increase of LMWH dose.There was a high expression of CYR61 in synovium tissue in RA patients.CONCLUSION:LMWH inhibited the adheresion of PBMC to FLS from RA patients in a dose-dependent manner,which might be attributed to its competitive combination with heparin sulfate sites on CYR61.