Polimorfizam gena za interleukin-6 i monocitni hemoatraktantni protein-1 i nivoi interleukina-17 i interleukina-23 kod obolelih od sistemske skleroze u odnosu na zdrave osobe

Abstract

Sistemska skeroza (SSc) je bolest nepoznatog uzroka. Jedan od mogućih činilaca u nastanku SSc je poremećaj imunskog odgovora na nivou Th17 ćelija i sekrecije interleukina-23 (IL-23/Th17 osovine). Polimorfizmi tipa zamene jednog nukleotida, u proksimalnom promotoru interleukin-6 (IL-6) gena (-174 G>C), kao i gena za monocitni hemoatraktantni protein-1(MCP-1, –2518 A>G), dovode se u vezu sa predispozicijom za nastanak nekolicine bolesti, ali je malo poznato o njihovom značaju u SSc. Ciljevi: Istraživanje je imalo za cilj ispitati razlike u učestalosti promotorskih varijanti rs1800795 (-174 G>C) IL-6 gena i rs1024611 (-2518G/A) MCP-1 gena, kao i razlike u ekspresiji IL-17A, IL-17F, IL-6 i IL-23 u krvi obolelih od SSc sa različitom kliničkom slikom bolesti u odnosu na kontrolnu grupu zdravih ispitanika. Ispitanici i metode: U studiji preseka učestvovala su 102 bolesnika sa SSc i 93 zdravih ispitanika koju su činili kontrolnu grupu. Za genotipizaciju promotorskih varijanti -2518 G/A MCP-1 gena i -174 C/G IL-6 gena korišćen je PCR-RFLP metod. Nivoi ekspresije IL-6, IL-17A, IL-17F i IL-23 mRNA mereni su pomoću qRT-PCR metode. Za merenje serumskih nivoa IL-17 i IL-23 korišćena je ELISA. Procena stepena oštećenja gastro-intestinalnog trakta (GIT-a) u SSc vršena je pomoću UCLA GIT 2.0 upitnika. Rezultati: Uočena je pozitivna korelacija između dužine trajanja bolesti i nivoa IL-17A mRNA (r = 0.41, p<0.05), IL-23mRNA (r = 0.49, p<0.05) i IL-6 mRNA (r = 0.4, p<0.05). Nivoi ekspresije IL-6 bili su značajno viši kod obolelih od SSc u odnosu na kontrolnu grupu (p<0.05). Nosioci C-alela za IL-6 gen imali su viši nivo ekpresije IL-6 (95.8 vs. 41.2, p<0.05) i viši skor nadutosti (1.4±0.9 vs. 0.78±0.8, p=0.05) u poređenju sa G/G homozigotima. Nije primećena udruženost -2518G/A MCP-1 polimorfizma i kliničke slike bolesti...

Introduction: Systemic sclerosis (SSc) is a disease of unknown origin. One of possible factors contributing to SSc development could be immunological disturbance regarding Th17 cells and secretion of interleukin-23 (IL-23/Th17 axis). Single nucleotide polymorphisms in proximal promoter of IL-6 gene (-174 G>C), as well as MCP-1 gene (–2518 A>G), are associated with predisposition of several chronic diseases development, but little is known about their significance in SSc. Aims: Study aimed to asses differences in frequencies of promoter variants rs1800795 (-174 G>C) IL-6 gene and rs1024611 (-2518G/A) MCP-1 gene, as well as differences in IL-17A, IL-17F, IL-6 and IL-23 expression profiles in the blood of SSc patients with different clinical disease manifestations compared to healthy controls. Subjects and methods: In this case-control study, 102 patients with SSc i 93 healthy controls were included. RCR-RFLP method was performed for genotyping promotor variants -2518 G/A of MCP-1 gene and -174 C/G of IL-6 gene. The expression level of IL-6, IL-17A, IL-17F and IL-23 mRNA were determined by qRT-PCR method. Enzyme-Linked Immunosorbent Assay (ELISA) was used for analysis of IL-23 and IL-17 serum protein levels. We used UCLA GIT 2.0 questionnaire to assess gastrointestinal involvement in SSc patients. Results: We found a positive correlation between disease duration and expression levels of IL-17A mRNA (r = 0.41, p<0.05), IL-23mRNA (r = 0.49, p<0.05) and IL-6 mRNA (r = 0.4, p<0.05).The expression level of IL-6 gene was significantly higher in patients with SSc in comparison with healthy controls (p<0.05). Carriers of C allele of IL-6 gene compared to those with G allele, showed higher expression of IL-6 gene ( 95.8 vs. 41.2, p>0.05) and higher distension scale score (1.4±0.9 vs. 0.78±0.8, p=0.05). We found no significant correlation between the -2518G/A MCP-1 polymorphism and disease manifestations...