Babak Afkari1, Zohreh Babaloo2, Sanam Dolati3, Alireza Khabazi4, Farhad Jadidi-Niaragh5, Mahsa Talei5, Dariush Shanehbandi6, Shiva Mahmoudi6, Bita Hazhirkarzar7, Ebrahim Sakhinia8. 1. Connective Tissue Research Center, Tabriz University of Medical Science, Iran; Immunology Research Center, Tabriz University of Medical Sciences, Iran; Drug Applied Research Center, Tabriz University of Medical Sciences, Iran; Student's Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran. 2. Connective Tissue Research Center, Tabriz University of Medical Science, Iran; Immunology Research Center, Tabriz University of Medical Sciences, Iran; Drug Applied Research Center, Tabriz University of Medical Sciences, Iran; Department of Immunology, Faculty of Medicine, Tabriz University of Medical Sciences, Iran. Electronic address: zbabaloo@tbzmed.ac.ir. 3. Connective Tissue Research Center, Tabriz University of Medical Science, Iran; Immunology Research Center, Tabriz University of Medical Sciences, Iran; Department of Immunology, Faculty of Medicine, Tabriz University of Medical Sciences, Iran. 4. Connective Tissue Research Center, Tabriz University of Medical Science, Iran. 5. Immunology Research Center, Tabriz University of Medical Sciences, Iran; Department of Immunology, Faculty of Medicine, Tabriz University of Medical Sciences, Iran. 6. Immunology Research Center, Tabriz University of Medical Sciences, Iran. 7. Research Center for Evidence Based Medicine, Tabriz University of Medical Science, Iran. 8. Connective Tissue Research Center, Tabriz University of Medical Science, Iran; Department of Genetics, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.
Abstract
BACKGROUND: Interleukin 10 (IL-10) is a cytokine with potent anti-inflammatory properties that play a fundamental role in restrictive host immune response to pathogens, by means of that is a crucial importance for chronic inflammatory disease studies. Therefore, the goal of this study was to measure the correlation of the IL-10 gene polymorphisms with the susceptibility to Behçet's disease compared with the control group in the Azeri population and to determine the expression of this gene in the two groups. Also, real-time PCR was performed for evaluate the IL-10 mRNA expression of the associated polymorphisms. METHODS: In this study, blood samples from 47 (1 missed) patients and 58 (3 missed) healthy control were taken, and then mononuclear cells isolated with ficoll protocol. The DNA and RNA were subsequently extracted. They were examined for -592A/C (rs1800872) of IL-10 gene single nucleotide polymorphism (SNP) using RFLP-PCR. Allele and genotype distributions were evaluated among groups using chi-square or Fisher's test. Following this, the extracted RNA was converted to cDNA using the RT-PCR method, after that expression of IL-10 evaluated by Real-time PCR. Serum levels of IL-10 were measured using Enzyme-linked immunosorbent assay (ELISA). RESULTS: Rates of the rs1800872 A allele was statistically lower in the control group compared with BD patients (p = 0.0315 and OR = 1.90 (1.05-3.42)). Also, as we expected, the expression level of the IL-10 gene was seen to significantly decrease in the patient group compared to the control. CONCLUSIONS: Our study showed that the rs1800872 A allele of the IL-10 gene may contribute to the genetic susceptibility of BD by regulating the expression of IL-10. Also as we expected, the expression level of this gene was seen to significantly decrease in the patient group compared to the control.
BACKGROUND:Interleukin 10 (IL-10) is a cytokine with potent anti-inflammatory properties that play a fundamental role in restrictive host immune response to pathogens, by means of that is a crucial importance for chronic inflammatory disease studies. Therefore, the goal of this study was to measure the correlation of the IL-10 gene polymorphisms with the susceptibility to Behçet's disease compared with the control group in the Azeri population and to determine the expression of this gene in the two groups. Also, real-time PCR was performed for evaluate the IL-10 mRNA expression of the associated polymorphisms. METHODS: In this study, blood samples from 47 (1 missed) patients and 58 (3 missed) healthy control were taken, and then mononuclear cells isolated with ficoll protocol. The DNA and RNA were subsequently extracted. They were examined for -592A/C (rs1800872) of IL-10 gene single nucleotide polymorphism (SNP) using RFLP-PCR. Allele and genotype distributions were evaluated among groups using chi-square or Fisher's test. Following this, the extracted RNA was converted to cDNA using the RT-PCR method, after that expression of IL-10 evaluated by Real-time PCR. Serum levels of IL-10 were measured using Enzyme-linked immunosorbent assay (ELISA). RESULTS: Rates of the rs1800872 A allele was statistically lower in the control group compared with BDpatients (p = 0.0315 and OR = 1.90 (1.05-3.42)). Also, as we expected, the expression level of the IL-10 gene was seen to significantly decrease in the patient group compared to the control. CONCLUSIONS: Our study showed that the rs1800872 A allele of the IL-10 gene may contribute to the genetic susceptibility of BD by regulating the expression of IL-10. Also as we expected, the expression level of this gene was seen to significantly decrease in the patient group compared to the control.