Study on expression of ASC and Caspase-1 in peripheral blood of patients with primary biliary cirrhosis / 重庆医学

Objective To explore the influence of apoptosis-associated speck-like protein containing a caspase recruitment do-main(ASC)and Caspase-1 on the pathogenesis of primary biliary cirrhosis(PBC).Methods The real-time PCR,Western blot,py-rophosphate sequencing and ELISA were adopted to respectively detect the relative expressions of mRNA and protein of peripheral blood mononuclear cells(PBMS)Caspase-1 and ASC as well as the methylation status of ASC promoter region and plasma Caspase-1 and IL-18 expression levels in 30 cases of PBC and healthy controls.Results The mRNA and protein expressions of PBMC Caspase-1 and ASC in the PBC group were significantly higher than those in the control group(P<0.05).The methylation rate of ASC promoter Island1(ISI)was significantly lower than that of the control group(P<0.05),which of Island 2(IS2)was smaller than the background value and had no methylation occurrence.The levels of plasma Caspase-1 and IL-18 in the PBC group were sig-nificantly higher than those in the control group(P<0.05).The ASC mRNA in the PBC group was significantly correlated with the Caspase-1 mRNA expression(P<0.05);the methylation rates at loci 1,2,4,5 of ASC promoter region CpG island were nega-tively correlated with ASC mRNA expression(P< 0.05),and which at loci 3,6 had no correlation with their expressions(P>0.05);plasma Caspase-1 and IL-18 levels showed the obviously positive correlation.Conclusion ASC and Caspase-1 are involved in the immune inflammatory response in PBC.

Correlation analysis of NF-κB signaling pathway activated by IL-18 in CD4+ T cells and the pathogenesis of PBC / 重庆医学

Objective To explore the correlation between NF-κB signaling pathways activated by IL-18 in CD4+ T cells and the pathogenesis of PBC.Methods We detected the expression of IL-18 mRNA in PBMCs,IL-18 level in plasma,receptor IL-18R on surface of CD4+ T cell,proliferation rate of CD4+T cell and its NF-κB signaling pathway protein IκBα and NF-κB p65 by qRT-PCR,ELISA,flow cytometry,MACS and Western blot on 32 cases of patients with PBC (PBC group) and 32 healthy people (control group) in Guizhou provincial people′s hospital.Results The level of IL-18 in PBC group was significantly higher than that in control group (P<0.05).The relative expression of IL-18 mRNA in PBC group was significantly higher than that in control group (P<0.05).The percentage of CD4+T cells expressing IL-18Rα in PBC group was higher than that in control group (P<0.05).The proliferation rate of CD4+T cells stimulated by IL-18 in PBC group was significantly higher than that in healthy control group (P<0.01).The relative expression levels of NF-κB p65 protein were up-regulated in IL-18,and the expression of IκBα protein in each group was significantly increased,especially in PBC group (P<0.01).Conclusion IL-18 can activate NF-κB signal pathway in CD4+ T cells and participate in the pathogenesis of primary biliary cirrhosis.

Study on associations between type Ⅰ interferon pathway IRF5 gene polymorp-hisms and SLE / 中国免疫学杂志

Objective:To investigate the IRF5 rs2004640,rs10954213,rs4728142 loci gene polymorphisms in the progress of Systemic lupus erythematosus(SLE),and explore the influence of the IRF5 gene polymorphisms to the SLE.Methods:218 patients with SLE and 200 health controls were analyzed by using TaqMan-PCR.And all allele frequencies were calculated.The risk factors were compared between cases and controls.At the same time,antinuclear antibodies and double-stranded DNA antibody of 218 SLE cases were analysed with indirect immunefluorescence method,specific autoantibodies in plasma spectrometry were determined using linear immunoassay in plasma.Results:T allele frequency of IRF5 rs2004640 loci in SLE was higher than the controls,allele frequency of the G/T distribution differences in two groups were statistically significant(χ2=6.809,P=0.009).The GG/TT genotype frequencies between the control and the SLE were statistically significant(χ2=5.111,5.035;P=0.024,0.025).Compared control group with SLE group,G allele frequency of IRF5 rs10954213 loci in SLE group was higher than the controls,the difference was statistically significant(χ2=4.332,P=0.037).And GG genotype distribution had significant difference in the two groups(χ2=5.805,P=0.016).SLE group AA/AG/GG genotype distribution of IRF5 rs4728142 loci,compared with control group,there were no statistically difference(χ2=1.273,0.902,1.853;P=0.259,0.342,0.173).And allele frequency A/G also were not statistically differences in two groups(χ2=2.651,P=0.104).The T allele frequency of IRF5 rs2004640 and autoantibodies(anti-Sm,anti-Rib-P)significant associations were found with SLE patient.There were no statistically significant difference between IRF5 rs10954213 with autoantibodies.Compared with SLE remission patients,ANA,ds-DNA increased obviously.Anti-NUC,anti-His,anti-Rib-P were higher,there were statistically significant differences betwen two groups.Conclusion:The GG/GT/TT polymorphism of IRF5 rs2004640,GG/GA/AA polymorphism of rs10954213 were related to SLE.But the GG/GA/AA polymorphism of IRF5 rs4728142 were not significant difference.IRF5 rs2004640 T comtributed to the anti-Sm,anti-Rib-P.In active SLE patients,anti-ds-DNA,anti-NUC,anti-His,anti-Rib-P were higher than the other group.

Study on the relationship between polymorphism site rs13146124 of IRF2 and systemic lupus erythematosus patients﹡ / 重庆医学

Objective To investigate the correlation between single nucleotide polymorphism rs13146124 of interferon regu‐latory factor2(IRF2) on type Ⅰ interferon pathway and systemic lupus erythematosus(SLE) in a population from Guizhou Prov‐ince .Methods The polymorphism IRF2(rs13146124) was detected by using Taqman‐PCR in 366 cases of patients with SLE and 218 healthy controls .The genotype and allele frequencies were calculated and analyzed .Results The genotype frequencies of AA , AG and GG in IRF2 rs13142164 site in patients with SLE were 0 .011 ,0 .246 and 0 .743 respectively ,compared with the control group ,the difference was not statistically significant (χ2 =0 .093 ,0 .205 ,0 .136 ;P=0 .761 ,0 .651 ,0 .712) .The allele frequencies of A and G in IRF2 rs13146124 site in SLE patients were 0 .13 ,0 .87 respectively ,compared with the control group ,the difference was not statistically significant(χ2 =0 .071 ,P=0 .790) .There was no significant difference between the allele frequencies of A and G in IRF2 rs13146124 site in SLE and ANA ,dsDNA and other specific antibodies .There was no correlation between the allele frequen‐cies of A and G in IRF2 rs13146124 site in SLE and clinical features such as arthritis ,kidney damage ,etc .Conclusion The poly‐morphism of rs13146124 in IRF2 may not be associated with SLE in the population from Guizhou Province .