Effects Of IL-35 On Angiogenesis Factors In Osteoblasts

Objective: Rheumatoid arthritis(RA)is a chronic autoimmune disease whose main clinical manifestations are synovitis,destructive polyarthritis and bone destruction.RA's inflammation cause bone loss and damage can not be rebuilt and repaired in time,that will lead the dynamic balance between bone resorption mediated by osteoblast and bone formation by osteoclast mediated destroyed,cause bone mass,bone mineral density and abnormal in bone structure,that will trigger osteoporosis(OP),which often accompanied joint dysfunction and activity restriction.This experiment studying the IL-35 on osteoblast's proliferation,apoptosis and its influence on osteoblast secrete angiogenic factors,as well as the function and the relationship with the Wnt/?-catenin signaling pathways,to explore the influence of IL-35 in bone formation,offers a new treatment direction for RA merger OP.Methods: Using cell culture technology culture osteoblast cell line MC3T3-E1 and stimulate with different concentrations of IL-35(0-100 ng/ml),CCK-8 detecting cell's proliferation,FCM analysis the apoptosis rate of osteoblast,RT-PCR detecting the m RNA expression of FGF-2,VEGF and its receptors in osteoblast.Treadted with Wnt/?-catenin inhibitor DKK-1(0.1ug/ml)and then adding different concentrations of IL-35,the expression of VEGF's m RNA was detected by RT-PCR.Results: After stimulated by different concentrations of IL-35,the light absorption value increased with the level of IL-35 concentration,and the cell proliferation ability was enhanced and the difference was statistically significant(P<0.05).In the role of IL-35,the apoptosis rate of 25 ng/ml(4.25±0.35%),50 ng/ml(3.0±0.14%)and 100ng/ml(1.3±0.28%)groups are significantly lower than the control group's apoptosis rate(6.35±0.64%),the difference is statistically significant(P < 0.05),and the trend is negative correlation with the concentration of IL-35.With the increase of IL-35 concentration,compared with the control group the m RNA expression of FGF-2,VEGF and Flt-1 in osteoblasts was significantly higher(P<0.05),and the expression of Flk-1 was not detected.after treated with DKK-1,compared with the control group,VEGF m RNA was significantly reduced in osteoblasts and negative correlated with the increase of IL-35 concentration,and the difference was statistically significant(P<0.05).Conclusion: Il-35 can promote the proliferation of osteoblast and inhibit its apoptosis.IL-35 can promote the secretion of angiogenic factors FGF-2,VEGF and its receptor Flt-1,this impact maybe has relationship with the Wnt/?-catenin signal pathway.