| ObjectiveComparison of IL-2,IL-4,IL-6,IL-10,IL-17 A,TNF-? and IFN-? cytokines and NLRP-3,GAST-5 mRNA expression level in children AA and s-JIA in order to explore theirs' role in the pathogenesis of AA and S-JIA. Methods1.There were 38 cases of children with AA(SAA 26 cases,N-SAA 12 cases),28 cases of children with JIA(S-JIA 13 cases,oligoarticular JIA 15 cases),normal healthy children(20 cases) each 3ml of fasting peripheral blood into the drying tube,after low-speed centrifugation to extract the upper serum was stored in vials,and put into-80 refrigerator to save as backup,application Aimplex? Human Th1 / Th2 / Th17 ?7-plex kit after specimen collection complete,using flow cytometry to detect the serum levels of IL-2,IL-4,IL-6,IL-10,IL-17 A,TNF-? and IFN-? seven kinds of cytokines.2.They were the newly diagnosed AA children,newly diagnosed JIA as the experimental group,the control group of healthy children.While collecting 21 cases of children with AA(SAA 13 cases,N-SAA 7 cases),17 cases of children with JIA(s-JIA 8 cases, oligoarticular JIA 9 cases) of bone marrow fluid each 2ml,as well as children with AA 32 cases(s AA 21 cases, AA 11 cases),28 cases of children with JIA(s-JIA 13 cases,oligoarticular JIA 15 cases),normal healthy children(20 cases)of each fasting peripheral blood 3ml,density gradient centrifugation of bone marrow mononuclear cells,reference manual Trizol reagent to extract bone marrow and peripheral blood mononuclear cells in the total RNA,the purity and integrity of RNA detection;by reverse transcription kit synthesized cDNA;using Real-time PCR assay NLRP-3,GAST-5 mRNA expression. Results 1. Comparison of serum levels of cytokines:(1) In the N-SAA,SAA group and control group comparison:IL-2, IL-4, IL-6, IL-17 A, TNF-?, IFN-? levels of SAA were higher than the normal control group, TNF-?, IFN-?levels in SAA group was higher than N-SAA group, the differences were statistically significant(P <0.05); IL 10-level N-SAA group, SAA group and control group among the three groups showed no significant difference(P> 0.05).(2) In the S-JIA group, JIA oligoarthritis group and control group comparison:IL-6, IL-17 A, TNF-? levels of S-JIA group was higher than the normal control group, TNF-? levels were higher S-JIA JIA oligoarthritis group, IL-10 levels in the S-JIA group was lower than normal the control group, the differenthan ces were statistically significant(P <0.05); IFN-? levels oligoarticular JIA group were higher than S-JIA group and normal control group, the difference was statistically significant(P <0.05); IL- 2, IL-4, IFN-?levels of S-JIA group and normal control group, the difference was not statistically significant(P> 0.05).(3)In SAA group,S-JIA group and control group comparison:IL-2, IL-6, IL-10, IL-17 A, IFN-?levels in SAA group was higher than S-JIA group, IL-4, TNF-? levels in SAA group than S-JIA group, the differences were there was statistically significant(P <0.05). 2.GAST-5, NLRP3 mRNA expression levels of comparison:(1) Expression in bone marrow fluidGAST-5, NLRP-3 mRNA in S-JIA group was higher than N-SAA group, SAA group, GAST-5 mRNA in oligoarticular JIA group was higher than N-SAA group, SAA group, NLRP-3 mRNA in S- JIA group was higher than oligoarticular JIA group, the difference was statistically significant(P <0.05).(2) Compare of the expression in peripheral bloodGAST-5, NLRP-3 mRNA expression in water S-JIA group were higher than group N-SAA, SAA group and normal control, NLRP3 mRNA in S-JIA group than in oligoarticular JIA group, the differences were statistically significance(P <0.05).(3) GAST-5, NLRP-3 mRNA expression levels were compared in the peripheral blood and bone marrow fluidGAST-5 mRNA was lower than in the SAA marrow peripheral blood, NLRP-3 mRNA in S-JIA group, JIA oligoarthritis group of bone marrow fluid was higher than the expression in peripheral blood, the difference was statistically significant(P < 0.05). Conclusion1.IL-6, IL-17, TNF-? and IFN-? were all involved in AA and s-JIA pathogenesis; IL-6, IL-17 and IFN-? expression in AA were higher than S-JIA, suggesting that these three cytokines increases to a certain critical value, it may lead to bone marrow failure; TNF-? was higher than AA in S-JIA, suggesting that TNF-? at different levels of community values, different mechanisms of action, resulting in different clinical which performed.2.IL-4 was one of the adjustment factors AA hematopoiesis; IL-10 could antagonize the S-JIA inflammation, inhibit the inflammatory response.3.Reduced GAST-5 mRNA level of AA was one of the causes bone marrow failure and reduced peripheral triple systems; increased NLRP-3 mRNA level was one of the important factors leading to S-JIA produce inflammatory response. |
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