| Background: Kawasaki disease(KD)is an immune-mediated systemic small and medium vascular vasculitis with unknown etiology.Coronary lesions induced by KD have become a major cause of childhood acquired heart disease.IL-35 is most recently identified member of Interleukin-12(IL-12)family,which is an important immunosuppressive/anti-inflammatory cytokine and may have the effect on reducing the progression of inflammatory and autoimmune diseases.The purpose of this study was to investigate whether IL-35 plays a role in the mechanism and pathogenesis of KD.Methods:(1)The all serum samples of KD patients were obtained before IVIG therapy.Levels of IL-35,IL-17 A,MCP-1,IL-10,VEGF and IL-6 were measured by ELISA and patients were divided into four groups:KD with coronary arterial lesions,KD without coronary arteries lesions,the febrile control group(FC)and the normal control group(NC).(2)The study included 88 children with KD and 100 gender-matched healthy subjects.five Interleukin-35 Single-nucleotide Polymorphisms: rs2243115,rs2243123,rs583911,rs353698 and rs2302164 were detected by SNapShotmethod.(3)The animal model of coronary arteritis was induced by intraperitoneally injecting with LCWE in C57BL/6 mice.The serum levels of IL-35,MMP-9,TGF-βand TNF-α were determined by ELISA and quantitative RT-PCR.HE staining was used to observe inflammatory cell infiltration in the coronary arteries of mice.The distribution of IL-35(p35and EBI3)and p-STAT1 were mesured by immunohistochemistry.(4)The animal model of coronary arteritis was intervened by IL-35,ELISA and Real-time PCR were used to detect the changes of MMP-9,TNF-αand TGF-β.Immunohistochemistry and Western-blot method for detection of p-Stat1 changes.Results:(1)Levels of IL-35,RBC and hemoglobin significantly decreased,and IL-6,IL-17 A,IL-10,MCP-1 and VEGF were signifcantly elevated in the KD group compared with febrile and control groups(P<0.05);Serum levels of IL-35 were decreased in patients with KD in CALs group,and ESR,IL-6,MCP-1 and VEGF were elevated(P <0.05).Serum levels of IL-35 in KD patients were negatively associated with WBC,CRP,IL-6,IL-17 A,IL-10,MCP-1 and VEGF in children with KD(P<0.05).(2)The IL-35 gene polymorphism was not significantly different between the KD group and the control group;The IL-35 rs2243115 was significantly different between KD with CALs group and KD without CALs group(P<0.05);The IL-35 rs353698 was significantly different between complete KD group and incomplete KD group(P<0.05).(3)IL-35 can bedetected on the coronary arteries in the mouse model of coronary artery.The levels of IL-35,MMP-9,TNF-α and TGF-β in mice injected with LCWE were significantly elevated(P<0.05).Expression of p-STAT1 and STAT1 proteins was higher in the LCWE-injected mice than in the PBS group.(4)In the IL-35 intervented LCWE-injected mice,The levels of TNF-α and MMP-9 were significantly decreased and the level of TGF-βwas significantly increased.The expression of p-STAT1 protein was lower than the group of LCWE-injected mice.Conclusions:(1)IL-35 may participate in the pathogenesis of KD by inhibiting the secretion of inflammatory cytokines.(2)IL-35 rs2243115 may be associated with CALs in KD.IL-35 rs353698 may be associated with incomplete KD.(3)IL-35 may be involved in the pathological process of CALs in KD through inhibiting the pro-inflammatory cytokines secretion and promoting the anti-inflammatory cytokines secretion.(4)IL-35 may inhibit the release of inflammatory factors in LCWE-injected mice by inhibiting phosphorylation of the STAT1 signaling pathway. |
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