| Rheumatoid arthritis(RA)is a chronic systemic autoimmune disease that primarily results in inflammation,loss of bodily function,and persistent damage to bones and joints.This disease also has a certain degree of invasiveness,causing the function of external organs to be affected.According to the global burden disease statistics,the global prevalence rate has increased by 7.4% and the incidence rate has increased by8.2% in this century,and women are more prone to get RA.RA has become the most common clinical disease with disability rates in global clinical medicine and has also become the focus of attention for many researchers.At the moment,the etiology of RA is not fully understood,and there are also many gaps in the study of its pathogenesis.However,research has found that vitamin D deficiency and metabolic damage can lead to a decrease in vitamin D levels,which is related to the occurrence and development of RA.Therefore,vitamin D supplements can be used as a treatment strategy,but the molecular mechanism is still unclear.The group-specific component(GC)gene encodes the vitamin D binding protein(VDBP),which is crucial for the transport and activation of vitamin D,and several genetic variations have been found in the GC gene.Therefore,this study aims to analyze the pathogenesis of RA based on population-specific genetic variations and explore the molecular system for the prevention and treatment of rheumatoid arthritis.In this study,the exploration is mainly based on two perspectives: bioinformatics analysis and laboratory testing analysis.Firstly,various bioinformatics software was used to explain the characteristics of VDBP,with Signal P5.0 and TMHMMV.2.0software analyzing single nucleotide polymorphisms(SNP)and amino acid sequence variations;SMART online software analyzes structural domains;Net PPhos 3.1 and Net NGly 1.0 servers determine protein phosphorylation and glycosylation modifications.The changes in the secondary and tertiary structures were determined using PSIPRED and SWISS models,respectively.Then,samples were collected from patients with rheumatoid arthritis and control groups in Peshawar,Khyber Pakhtunkhwa Province,Pakistan from 2021-2022,and biochemical indicators were detected.The genotypes of rs4588 and rs7041 were analyzed by a polymerase chain reaction and restriction fragment length polymorphism,to determine the correlation between the disease mutant genotype and vitamin D level.The results of bioinformatics research indicate that there are certain differences in molecular weight,isoelectric point,instability coefficient,aliphatic amino acid coefficient,and hydrophilicity between normal VDBP proteins and VDBP polymorphic proteins containing rs7041 and rs4588 mutation points.Sequence analysis showed that there was a cleavage site between 16 and 17 sequences,indicating that VDBP protein was a secreted protein with a signal peptide.SMART domain analysis revealed the presence of two albumin domains.Secondary and tertiary structural analysis showed maximum similarity between normal and polymorphic proteins.Overall,no substantial differences were observed between normal and polymorphic proteins in VDBP.The detection results of biochemical indicators show that rheumatoid factors can become a marker for the diagnosis of RA,and cyclic citrullinated peptide(CCP)is an important biomarker for the prognosis and diagnosis of RA.C-reactive protein(CRP)is a biomarker used to evaluate the severity and treatment response of the disease.In this study,the correlation between age,gender,body mass index(BMI),and biochemical parameters of RA patients was analyzed simultaneously.The results indicate that combining demographic data to analyze multiple biochemical parameters has a certain significance,as these two parameters are related to environmental and geographical factors.In this study,a significant increase in the concentration of biochemical parameters was observed in women Expression analysis for respective polymorphism was determined by Polymerase chain reaction(PCR)and restriction fragment length polymorphism(RFLP).The overall analysis of GC gene polymorphism and allele frequency of subjects in the disease group and the control group concluded that homozygous wild-type genotype(CC)was more common in the control group,while homozygous mutant genotype was more common in the disease group.There is a significant difference in the distribution of allele frequency between the diseased group and the control group.Homozygous mutant genotype GG and heterozygous GT are more common in the diseased group,while homozygous wild type genotype(TT)is more widespread in the control group.The rs4588 polymorphism may be related to the development of the disease and can be used as a biomarker for diagnosis and risk assessment.Analyze the correlation between two genotype variants and vitamin D deficiency through enzyme-linked immunoassay(ELISA).In rs4588 and rs7041,the data obtained indicate a significant relationship between these two variables(p<0.001).In the control subjects,the detected amount of vitamin D was mostly insufficient.In the case of RA,a shortage of quantity was observed in rs7041 and insufficient in rs4588.There is a strong correlation between disease and mutant genotypes.In addition,vitamin D deficiency was detected in both sexes of RA subjects and was significantly associated with mutant genotypes,indicating that vitamin D deficiency may be a contributing factor in the development of RA.The above results indicate that the VDBP GC gene mutation caused by the SNP mechanism has the ability to affect vitamin D.This study provides significant information for the development of more effective diagnostic and treatment strategies for RA patients.Moreover,these analyses will help to identify other modifiable risk factors that could be targeted to improve Vitamin D status in RA patients as well. |
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