ABSTRACT
Objective:To explored the effect of 78c in treating collagen-induced arthritis (CIA) mice and to investigate its mechanism of effects.Methods:CIA mice model and CD38 +NK cells were treated with 78c. Cytokine concentrations and lymphocyte subtypes were measured in the mice peripheral blood and culture medium using flow cytometry. Mikenyi cell isolation kit was used to isolate CD4 + T cells and NK cells from peripheral blood mononuclear cells of healthy volunteers. CD38 + NK cells were enriched using the Miltenyi CD38 microbeads from the extracted NK cells. CD38 + NK cells with 78c pretreatment or not were cocultured with CD4 +T cells in transwells. The least significant difference (LSD) method was used for comparison between the two groups, and one-way analysis of variance was used for multi-group significance. Pearson correlation analysis was used for correlation analysis. Results:78c treatment significantly suppressed joint inflammation, inhibited the toe thickness of CIA mice, and reduced the number of while cell, neutrophils, platelets, and concentrations of IFN-γ, IL-6 and TNF-α ( t=6.10, P<0.001; t=4.00, P=0.002; t=3.09, P=0.012; t=2.31, P=0.043; t=3.58, P=0.005; t=2.68, P=0.002) in the CIA mice. The proportion of CD38 +NK cells decreased from (3.9±0.9)% to (2.4±0.3)% ( t=2.49, P=0.032), the proportion of regulatory T cell (Treg) increased from (0.81±0.33)% to (1.41±0.26)% ( t=2.74, P=0.021), and the concentration of IL-10 also increased from (99±37) pg/ml to (199±9) pg/ml( t=2.76 , P=0.020). The proportion of Treg in CD4 +T cells cocultured with 78c-pretreated CD38 +NK cells increased from (0.52±0.04)% to (0.69±0.08)% ( t=3.33, P=0.029) , the T helper cells (Th)17/Treg ratio decreased from (4.44±0.26) to (2.59±0.64) ( t=4.76 , P=0.009), and the Th1/Th2 ratio decreased from (14.8±1.6) to (8.1±1.3)( t=5.70 , P=0.005). Conclusion:78c can reduce the proportion of CD38 +NK cells, thereby reducing the inhibition of CD38 +NK cells on CD4 +T cell differentiation into Treg cells, leading to the restoration of immune balance. The results of this study suggest that 78c is a potential therapeutic agent for rheumatoid arthritis.
ABSTRACT
Objective To screen the proteins with decreased expression in the synovial tissues of rheumatoid arthritis (RA) patients by comparing their expression profiles with that of osteoarthritis (OA) and ankylosing spondylitis (AS) patients by a proteomic approach,and to explore the association of reduced expression with disease susceptibility by a single nucleotide polymorphism (SNP) analysis.Methods Proteins extracted from the synovial membranes (n=10 for each disease) were separated by 2-D electrophoresis.The proteins with significantly decreased expression in the RA samples were subjected to MALDI-TOF-MS.The results were verified using Western blotting.Tag SNPs located in the targeted gene were assessed using the Taqman assay in a cohort of 267 Chinese patients with RA and 160 healthy controls.The genotyping results were confirmed in a large cohort of 389 patients with RA and 371 healthy controls.SPSS 11.5 software package was used for one way ANOVA and Fisher's exact test.Results The expression of vitamin D-binding protein (VDBP) in the synovial membranes from patients with RA was significantly decreased when examined by proteomic approach.This result was confirmed by Western blotting analysis.The rs2282679 was significantly associated with RA (P=0.026 794).This result was confirmed in a large cohort of RA( OR=0.678 639,95％CI 0.54l 113-0.851 118,P=0.000 776).Conclusion Compared with samples from patients with OA and AS,RA patients' synovial tissues have low VDBP expression when examined by the proteomic method.The tag SNP rs2282679 located in VDBP is significantly associated with RA.The decreased expression and the genetic effect of VDBP in RA suggest that a novel pathogenic pathway,in which vitamin D contributes,may be involved in the arthritis process of RA.
ABSTRACT
Objective We aim to investigate the association between peptidylarginine deiminase type 4 (PADl4) and colorectal cancer by genotyping threetag single nucleotide polymorphisms (tagSNP) genetic maker, to explore the role of polymorphism of PAD14 gene in development of colorectal cancer. Methods A case-control study was conducted using TaqMan-PCR methods to analyze the genotypes of three TagSNPs such as rs1886302, rs2477131 and rs1635561 for 515 patients with colorectal cancer and 549 health controls. Results There is significantly different in allelic frequencies and genotype frequencies of rs1886302 between cases and controls (P=0.039 and 0.040,respectively). The OR value of A allele and AA genotype is 0.796 and 0.731,respectively,and A allele may reduce the probability of incidence of colorectal cancer. No association Was found among SNPs of rs2477131 and rs1635561 within intronl and intron 15 in terms of coloretal cancer. Conclusion Rs1886302 on PADI4 might be a susceptible SNP to coloretal cancer.