High Glucose On Human Fibroblasts Wnt/Beta-catenin Signaling Pathway

Objective:Wound healing is a complicated biological process, which consists of inflammatory cells,repairing cells,extracellular matrix,cell factors, et al.Those factors participate in the process of wound healing in great harmony and coordination. Fibroblast play an important role in wound healing.Diabetic skin is vulnerable to injury, delayed wound healing or nonunion occur after injury.In recent years,it has become clear that high glucose on rat skin fibroblasts significantly inhibited its proliferation.But the specific mechanism is not clear.Studies indicate that the proliferation of human fibroblasts increased in vitro after activation of Wnt signaling pathway.Activation of Wnt signaling pathway can promote the healing of diabetic ulcers, also can promote skin wound healing.Previous animal experiments show that increased expression of Wnt signaling pathway allows to speed up wound healing in diabetic rats.We speculate that high glucose may be caused by Wnt signaling pathway to reduce the proliferation of human fibroblasts, thus affecting wound healing.And the relationship between the human fibroblast proliferation and the Wnt signaling pathway has not been reported, in order to confirm our speculation, We have designed this experiment to study high glucose on human fibroblast proliferation and Wnt signaling pathway, and to explore the relationship between the human fibroblast proliferation and the Wnt signaling pathway.Methods:1 fibroblast culture, each discarded cell culture supernatant of the bottle, with 10% FBS high glucose DMEM,4-5ml, placed in 37℃,5% CO2 incubator to cultivate.1-2 days, according to cell growth medium was changed once,3-5 days cell culture bottle covered, after the passage Asian fusion. In the training process on the morphology of human fibroblasts were observed.Experimental group,Control group, (medium containing 25 mmolPL glucose), high glucose group:high glucose group 1 (medium containing 40 mmolPL glucose) high glucose group 2 (medium containing 55 mmolPL glucose) high glucose group 3 (medium containing 70 mmolPL glucose) high glucose group 4 (medium containing 85 mmolPL glucose) Mannitol group:isotonic glucose with mannitol mannitol group 1 is 40 mmolPL isotonic mannitol (medium containing 15mmolPL mannitol) mannitol group 2 is 55 mmolPL isotonic mannitol (medium containing 30mmolPL mannitol) mannitol group 3 is 70 mmolPL isotonic mannitol (medium containing 45mmolPL mannitol) mannitol group 4 is 85 mmolPL isotonic mannitol (medium containing 60mmolPL mannitol).The role of each group were in vitro cultured human fibroblasts.2 Determine reproductive activity of human fibroblasts by MTT method. Human fibroblast proliferation under Different Factors in 12h,24h,48h each time is been detected.3 Expression of beta-catenin protein was detected by Western blot.Results:High glucose can inhibit fibroblast proliferation, mtt assay detect high glucose on human fibroblast proliferation inhibition, Western blot detect high glucose on human fibroblasts wnt/beta-catenin signaling pathway inhibition.Conclusion:The high glucose on human fibroblast proliferation is associated with the Wnt signaling pathway.