Effect of CCK-8 on expression of MMPs/TIMP-1 in TNF-α-induced RSC-364 / 中国药理学通报

Aim To observe the influence of CCK-8 on expression of MMPs/TIMP-1 in TNF-α-induced rat fibroblast-like synovial cell line RSC-364.Methods The secretion levels of MMP-1, MMP-3, MMP-9 and TIMP-1 were determined using ELISA;MMP-3 and MMP-9 mRNA expressions were detected by RT-PCR.Results MMP-3 and MMP-9 could not be examined in RSC-364 incubated with CCK-8 and unstimulated RSC-364, which was able to product a little MMP-1, TIMP-1 and express even less MMP-3,-9 mRNA.CCK-8 inhibited the increase in MMP-1, MMP-3, MMP-9 secretion and MMP-3,-9 mRNA expression in TNF-α-induced RSC-364.TIMP-1 production was also increased in TNF-α-induced RSC-364.CCK-8 had no effect on TIMP-1 production in TNF-α-induced RSC-364, but was able to reduce the ratios of MMP-1, MMP-3, MMP-9 to TIMP-1.Conclusion The inhibitory effect of CCK-8 on MMPs activity may be related to the decrease of MMPs mRNA expression, MMPs secretion and the ratios of MMPs to TIMP-1 in TNF-α-induced RSC-364, which indicates that CCK-8 might be a possible regulator in the pathogenesis of rheumatoid arthritis.

Clinical validation of the 2012 classification criteria for early rheumatoid arthritisin: a domestic multi-center cohort / 中华风湿病学杂志

Objective To evaluate the value of 2012 classification criteria for early rheumatoid arthritis (ERA),2010 American College of Rheumatology/European League Against Rheumatism (ACR/EULAR) classification criteria,and 1987 ACR classification criteria in the diagnosis of early rheumatoid arthritis (RA).Methods Patients who had at least one swollen and tender joint with disease duration no more than 2 years,and age more than 16 years were enrolled.The patients were diagnosed as RA or other non-RA by 2 experienced rheumatologists.The clinical and laboratory parameters were recorded.The sensitivity and specificity of three RA classification criteria were compared by McNemar test,The areas under the receiver operating characteristic curve (ROC) curve (AUC) of each RA classification criteria were analyzed using MedCalc software.Results Atotal of 310 patients were enrolled in this study,including 182ERA and 128 non-RA.The sensitivity(88.5％) of ERA criteria was much higher than that of the 1987 ACR criteria (45.6％,x2=75.013,P＜0.05),and not significantly different with the 2010 ACR/EULAR criteria (91.8％,X2=1.042,P＞0.05).The specificity of ERA criteria (91.4％) of 2010 ACR/EULAR criteria (87.5％,x2=1.8,P＞0.05) was similar to that of the 1987 ACR criteria (96.1％,x2=3.1,P＞0.05).The AUC of ERA criteria was 0.962 [95％CI(0.934,0.980)],which was slightly better than that of the 2010 ACR/EULAR criteria 0.959 [95％CI(0.931,0.978)],Z=0.380,P=0.7038,and much higher than that of the 1987 ACR criteria 0.885 [95％CI (0.845,0.919)],Z=4.517,P＜0.01.Conclusion Overall evaluation,the diagnostic value of ERA criteria is better than 1987 ACR and 2010 ACR/EULAR criteria in early rheumatoid arthritis.Compared to 2010 ACR/EULAR classification criteria,ERA criteria is more simple and practical.

Correlation among ST-2,adiponectin,growth differentiation factor-15 and senile heart failure / 实用医学杂志

Objective To explore the correlation among ST-2,adiponectin(APN),positive growth dif-ferentiation factor -15(GDF-15)and senile heart failure,and to investigate the diagnostic values of these indica-tors. Methods Totally 129 patients(15 patients of NYHA Ⅰ,60 of NYHA Ⅱ,28 of NYHA Ⅲ and 26 of NYHAⅣin study group)with heart failure and 30 control subjects(control group)were enrolled in the study.Se-rum levels of ST-2,APN and GDF-15 were determined by ELISA,and their correlation with senile heart failure was analyzed.Results Compared with those in control group,serum levels of ST-2,APN and GDF-15 in study group were significantly increased(P<0.05).There were significant differences in serum levels of ST-2,APN and GDF-15 among patients of different classifications in study group(P<0.05)and the higher level of cardiac func-tion,the higher serum ST-2,APN and GDF-15. There was a positive correlation among serum ST-2,APN and GDF-1. Conclusions The serum levels of ST-2,APN and GDF-1 are positively related to the severity of senile heart failure,thus it could be served as the index of diagnosis,curative effect monitoring and prognosis of patients with heart failure.

Analysis of etiological changes of atrial fibrillation in Zhanjiang / 中国基层医药

Objective To analyze the etiological changes of atrial fibrillation(AF) in Zhanjiang. Methods The etiology of 592 AF cases during 1990～1997 were analyzed, and 610 cases during 2000～2007 were analyzed as comparison. Results Rheumatic heart disease(36.8%) was main etiology of AF during 1990～1997. But coronary artery disease(33.1%) has surpassed rheumatic heart disease recently, and the hyperthyroidism and undetermined-e-tiology of atrial fibrillation were decreasing. The etiology of atrial fibrillation in different age groups was significantly different(P <0.05 or P <0.01). Conclusion The etiology of atrial fibrillation changes every year,and age is a pre-dictable factor to the etiology of atrial fibrillation.

Effects of cholecystokinin octapeptide on TNF- α- induced IL- 6 expression and its possible molecular mechanismin rat synovial cell strain RSC-364 / 中国病理生理杂志

AIM: To investigate the effect of sulfated cholecystokinin octapeptide (CCK -8 ) on TNF -α induced IL - 6 mRNA expression, NF - κB activation in the rat fibroblast - like synovial cell strain RSC - 364 and its possible receptor mechanisms. METHODS: RSC -364 cells were stimulated with TNF - α( 10 μg/L) in the presence or absence of sCCK- 8( 10-8 - 10-6 mol/L) or/and CCK receptor antagonist proglumide(2 mg/L). IL -6 and CCK receptor A/B (CCK- AR/CCK/BR) mRNA expression were assayed by reverse transcription polymerase chain reaction (RT- PCR) at 3 h after stimulation, and nuclear factor - κB (NF - κB) binding activity was analyzed by electrophoretic mobility shift assay (EMSA) at lh after stimulation. At 30 min of stimulation the IκB protein level in cytoplasma was measured by Western blotting. RESULTS: Both CCK - AR and CCK - BR were constitutively expressed on RSC - 364. sCCK - 8, at concentrations from 10-8 mol/L to 10 -6 mol/L, significantly increased IL - 6 mRNA expression, CCK - AR and CCK - BR mRNA expression, NF - κB binding activity and IκB protein degradation. The effects of sCCK - 8 on NF - κB activity and IκB degradation level were attenuated by CCK receptor antagonist proglumide. CONCLUSION: sCCK - 8 upregulats TNF - α- induced IL - 6 mRNA expression by NF - κB pathway through its receptor on rat synoviocytes, suggesting its possible regulatory role in the pathogenesis of rheumatoid arthritis.

CCK-8 decreases RSC-364 proliferation and p38 MAPK activation induced by TNF-? in rats / 基础医学与临床

Objective To study the effect of cholecystokinin-octapeptide(CCK-8) on the proliferation of fibroblast-like synovial cell line RSC-364 and p38 MAPK activity induced by TNF-? in rat.Methods The proliferation of RSC-364 cells was measured by monotetrazolium(MTT) colourmetric assay and the level of activation of p38 MAPK was deteced by Western blot.Results An increase in p38 MAPK phosphorylation was detected 5 min after TNF-?((50 ?g/L))addition,and reached a plateau at(15 min),finally returned to the basic level at(2 h).TNF-?(10,25,(50 ?g/L)) increased p38 MAPK phosphorylation in a dose dependent manner at 15 min.CCK-8((10~(-10))～(10~(-6)mol/L))could inhibit the proliferation and the level of phosphorylation of p38 MAPK in a dose dependent manner.Moreover the inhibitory effects were partly reversed by CCK-A receptor specific antagonist CR1409 or CCK-B receptor specific antagonist CR2945.SB203580 inhibited TNF-?-stimulated RSC-364 proliferation.Conclusion CCK-8 inhibited TNF-?-stimulated proliferation by decreasing p38 MAPK phosphorylation in RSC-364 cells,which was mediated through CCK-A receptor or CCK-B receptor.

Effect of CCK-8 on IL-1?-induced RSC-364 proliferation / 中国药理学通报

Aim To investigate the effects and mechanisms of CCK-8 on IL-1? induced proliferation of RSC-364, a rat fibroblast-like synovial cell line. Methods MTT colorimetric assay and Western blot were used to measure cell proliferation and p38MAPK phosphorylation level to elucidate the mechanism of CCK-8 in IL-1? induced RSC-364 proliferation. Results CCK-8 significantly inhibited IL-1?-induced RSC-364 proliferation at 10 -12 , 10 -10 , 10 -8 , 10 -6 mol ? L -1 , and IL-1?-activated p38MAPK activity at 10 -10 , 10 -8 , 10 -6 mol?L -1 in a dose-dependent manner. The effect of CCK-8 was blocked by CR1409 (a CCKA-receptor antagonist) and CR2945 (a CCKB-receptor antagonist). Conclusion CCK-8 inhibits IL-1?-induced RSC-364 proliferation, probably by reducing p38MAPK activity through CCKA and CCKB receptors.

Effects of cholecystokinin octapeptide on TNF-?-induced IL-6 expression and its possible molecular mechanism in rat synovial cell strain RSC-364 / 中国病理生理杂志

AIM:To investigate the effect of sulfated cholecystokinin octapeptide(CCK-8) on TNF-? induced IL-6 mRNA expression,NF-?B activation in the rat fibroblast-like synovial cell strain RSC-364 and its possible receptor mechanisms.METHODS:RSC-364 cells were stimulated with TNF-?(10 ?g/L) in the presence or absence of sCCK-8(10-8-10-6 mol/L) or/and CCK receptor antagonist proglumide(2 mg/L).IL-6 and CCK receptor A/B(CCK-AR/CCK/BR) mRNA expression were assayed by reverse transcription polymerase chain reaction(RT-PCR) at 3 h after stimulation,and nuclear factor-?B(NF-?B) binding activity was analyzed by electrophoretic mobility shift assay(EMSA) at 1h after stimulation.At 30 min of stimulation the I?B protein level in cytoplasma was measured by Western blotting.RESULTS:Both CCK-AR and CCK-BR were constitutively expressed on RSC-364.sCCK-8,at concentrations from 10-8 mol/L to 10-6 mol/L,significantly increased IL-6 mRNA expression,CCK-AR and CCK-BR mRNA expression,NF-?B binding activity and I?B protein degradation.The effects of sCCK-8 on NF-?B activity and I?B degradation level were attenuated by CCK receptor antagonist proglumide.CONCLUSION:sCCK-8 upregulats TNF-?-induced IL-6 mRNA expression by NF-?B pathway through its receptor on rat synoviocytes,suggesting its possible regulatory role in the pathogenesis of rheumatoid arthritis.

Inhibitory effects of CCK-8 on NF-?B activities stimulated by LPS in rat PIMs / 中国病理生理杂志

AIM: To investigate the inhibitory effects of cholecystokinin octapeptide(CCK-8) on nuclear factor-?B(NF-?B) activities stimulated by lipopolysaccharide(LPS) by using forskolin,the activator of adenylate cyclase,and PKA inhibitor H-89 in rat pulmonary interstitial macrophages(PIMs).METHODS: PIMs were isolated and purified.EMDA was applied to detect NF-?B activities and Western blotting was used to analyze the I?B-? protein level in rat PIMs.RESULTS: The NF-?B activity was not detected in normal control rat PIMs.The NF-?B activity in LPS-treated rat PIMs was obviously higher than that in control group(P0.05).The NF-?B activity in CCK+LPS group and LPS+Fsk group were obviously lower than that in LPS group(P

Effects of cholecystokinin octapeptide on expression of IL-1?,IL-6,IL-4 and IL-10 in lipopolysaccharide-attacked mice / 中国病理生理杂志

AIM:To observe the effects of cholecystokinin octapeptide(CCK-8) on the expression of proinflammatory cytokines IL-1?,IL-6 and anti-inflammatory cytokines IL-10,IL-4 in LPS-attacked mice.METHODS: Kunming mice were randomly assigned and injected intraperitoneally with LPS alone or/and CCK-8 at different time points.The expression of IL-1?,IL-6,IL-10 and IL-4 in the serum and lung tissues were assayed by ELISA and RT-PCR.RESULTS: The expression of IL-1?,IL-6,IL-10 and IL-4 were upregulated in LPS-attacked mice.Pre-treatment of CCK-8 decreased both IL-1? and IL-6 expression and augmented IL-10 and IL-4 expression in LPS-attacked mice.CONCLUSIONS: CCK-8 exerts an anti-inflammatory effect by inhibiting the expression of IL-1?,IL-6 and increasing the expression of IL-10,IL-4 in LPS-attacked mice,which could alleviate the inflammatory response in lung tissue.