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@#Objective To investigate the mechanism of anti-IL-17A monoclonal antibody(secukinumab) regulating autophagy and inflammation in gout.Methods The peripheral venous blood samples from 57 patients with acute gout(AG),57patients with intermittent gout(IG) and 82 healthy volunteers were collected and measured for the mRNA transcription levels of autophagy-related genes(ATGs) ATG4B,ATG7, A TG16L1,Beclin-1 and LC3B by RT-qPCR.The model of AG inflammation was established by adding monosodium urate(MSU) crystals into the peripheral venous blood samples of healthy volunteers,and the transcription and protein expression of IL-1β were detected by RT-qPCR and ELISA at 0,1,2,4,6 and8 h and different concentrations(0,100,200 and 400 μmol/L) of secukinumab.The peripheral blood samples of healthy volunteers were divided into control(without MSU treatment),MSU(100 μg/mL),MSU+colchicine(100 μg/mL+30 μg/mL) and MSU+secukinumab(100 μg/mL+400 μmol/L) groups,which were detected for the mRNA transcription and protein expression of IL-1 β and ATGs by RT-qPCR and Western blot,and for the expression of IL-1β,IL-12 and IL-35 by ELISA.Results The mRNA expression levels of ATG4B, Beclin-1 and LC3B in AG,IG and healthy control groups were significantly different(F=3.896,11.78 and 3.856,respectively,each P <0.05),among which the mRNA levels in AG were lower than those in IG and HC groups(t=2.692,3.234,2.231 and 2.085,4.795,2.748,respectively,each P <0.05);the expression levels of ATG16L1 mRNA were significantly different in the three groups(F=7.949,P <0.001),and was significantly lower in AG group than HC group(t=3.860,P <0.001).In AG inflammation model,the mRNA and protein expression of IL-1 β reached their peak in 2—4 h,and the anti-inflammation effect of secukinumab was the strongest at the concentration of 400 μmol/L.Compared with MSU group,the mRNA levels of ATG16L1 and LC3B(t=2.343 and 2.916,respectively,each P <0.05) as well as the expression levels of ATG4B,ATG7,Beclin-1,ATG16L1 and LC3B-Ⅱ proteins(t=28.84,11.6,8.402,4.124 and 2.458,respectively,each P <0.05) in MSU+secukinumab group decreased significantly.The expression levels of IL-12 and IL-35 in the control,MSU,MSU+colchicine and MSU+secukinumab groups showed significant difference(F=7.009 and 6.518,respectively,each P <0.01).Compared with MSU group,the expression level of IL-12 significantly decreased(t=2.604,P <0.05)in MSU+secukinumab group,and the expression level of IL-35 also decreased,while with no significant difference(t=1.928,P> 0.05).Conclusion Secukinumab can regulate the mRNA and protein expression of ATGs,reduce the levels of pro-inflammatory cytokines,and inhibit gout inflammation,which provides a reference for the treatment of gout.
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Objective To investigate the effect of human umbilical cord mesenchymal stem cells (HUC-MSCs) on CD4+T cells in liver after hepatic ischemia-reperfusion injury (HIRI) in mice. Methods Two hundred and twenty-five mice were randomly divided into sham group, control group and MSC group, with 75 mice in each group. HIRI model mice were used in MSC group and control group. HUC-MSCs were injected in MSC group through inferior vena cava. Normal saline was injected in control group through inferior vena cava. Only laparotomy and abdominal closure were performed in sham group without blood vessel clipping. At 6, 12 and 24 h after operation, 15 mice of each group were randomly selected to sample eyeball blood and liver tissues, and the 30 mice left in each group were used to extract intrahepatic mononuclear cells. The number of intrahepatic mononuclear cells, percentage, number and positive rate of CD4+T cells in the mice of various groups at different time points were compared. The content of interleukin (IL)-17 in serum and liver tissue as well as expression levels of costimulatory molecules B7-1 and B7-2 messenger RNA (mRNA) in liver tissues of the mice at different time points were compared. Results At 12 and 24 h after operation, the number of intrahepatic mononuclear cells of control group was significantly higher than that of sham group, while the number of intrahepatic mononuclear cells of MSC group was significantly lower than that of control group (P<0.01-0.05). At 6, 12 and 24 h after operation, the percentage, number and positive rate of CD4+T cells of control group were significantly higher than those of sham group (all P<0.01), while the percentage of CD4+T cells of MSC group was significantly lower than that of control group (P<0.01-0.05). At 12 and 24 h after operation, the number and positive rate of CD4+T cells of MSC group were significantly lower than those of control group (P<0.01-0.05). At 6, 12 and 24 h after operation, the IL-17 contents in serum and liver tissues of control group were higher than those of sham group (all P<0.01), while the IL-17 contents in serum and liver tissues of MSC group were lower than those of control group (all P<0.01). At 6 h after operation, the mRNA expression level of B7-2 of control group was higher than that of sham group (P<0.05). At 12 and 24 h after operation, the mRNA expression levels of B7-1 and B7-2 of control group were higher than those of sham group (all P<0.01), while the mRNA expression levels of B7-1 and B7-2 of MSC group were lower than those of control group (all P<0.01). Conclusions HUC-MSCs inhibits the number of CD4+T cells and the secretion of IL-17 in liver after HIRI, as well as decreases the number of intrahepatic mononuclear cells and the mRNA expression of B7-1 and B7-2, thereby alleviating HIRI.
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Objective To detect the levels of helper T cells (Th) 17 and Th9 cells in the peripheral blood of patients with pedi-atric bronchial asthma and the expression of interleukin 17 (IL-17) ,IL-9 and total IgE in serum ,and to discuss their effect and clini-cal significance in pathogenesis of pediatric asthma .Methods A total of 46 children with bronchial asthma ,including asthma attack group (n=26) and asthma remission group (n=20) were selected .Healthy children were selected as healthy control group (n=20) .Flow cytometry was used to detect the percentages of Th17 and Th9 cells in the peripheral blood .The levels of IL-17 and IL-9 in serum were measured by enzyme-linked immunosorbent assay (ELISA) .Serum total IgE levels were measured by the specific protein analyzer .Results The percentages of Th17 and Th9 cells in the peripheral blood were significantly higher in asthmatic group compared with remission group and healthy control group (P<0 .05) .The percentages of Th17 and Th9 cells in the periph-eral blood was significantly higher in remission group compared with healthy control group (P<0 .05) .The levels of IL-17 ,IL-9 and total IgE in serum were significantly higher in asthmatic group compared with remission group and healthy control group (P<0 .05) .The levels of IL-17 ,IL-9 and total IgE in serum were significantly higher in remission group compared with healthy control group (P<0 .05) .The levels of IL-17 and IL-9 in serum of asthmatic children were positively correlated with total IgE levels (P<0 .05) .The correlation analysis results showed that the levels of IL-17 ,IL-9 and total IgE in serum of patients with pediatric bron-chial asthma have a positive correlation .(r=0 .717 ,0 .491 ,0 .786 ,P<0 .05) .Conclusion Th17 ,Th9 cells and their cytokines in pe-ripheral blood of patients with pediatric bronchial asthma play important roles in the pathogenesis of asthma and are positively cor-related with the severity of asthma .