Influence Of The Aloe Vera Polysaccharide On Both Proliferation And Collagen Synthesis And Secretion In Vitro Cultured Human Fibroblast Cells

Objective:To investigate the effect of the Aloe vera polysaccharides on the proliferation and collagen synthesis and secretion of human fibroblast cells cultured in vitro .Method:1. The fibroblast cells cultured in vitro were randomly divided into polysaccharide (25, 50, 100, 200, 400mg/L) and control groups. The cell proliferation was measured by MTT. The cell cycle was detected by flow cytometry analysis. The cell apoptosis was assayed for the evaluation toxic side effect of the Aloe vera polysaccharides by AO-EB stain method.The leakage rate of lactate dehydrogenase (LDH) was assayed for the evaluation of the cell injury.2. The fibroblast cells DNA synthesis was measured by [3H]-proline;Express mRNA of collagen type I,III in the cultured fibroblast was measured by RT-PCR ;FITC-immunocytochenistry was assayed for the evaluation of collagen protein type I,III in the cultured fibroblast.3. The levels of collagen type I,III,haluronic acid,and hydroxyproline in the supernatants of cultured fibroblast cells in polysaccharide and control groups were measured by ELISA.Result:1. There was no significant difference in the morphology of the fibroblast cells among the groups under inverted phase contrast microscope. The cells treated with polysaccharide were seen to have proliferated actively by MTT measurement.The percentage of cell cycle phase G0/G1 treated with polysaccharide decreased significantly ,but G2/M and S increased significantly compared with that control group (P <0.05). There was no significant difference in the morphology of the fibroblast cells among the groups by AO-EB stain menthod. The leakage rate of the cells treated with polysaccharide was lower than those control group (P <0.05).2. The fibroblast cells treated with polysaccharide synthesize DNA and express mRNA and synthesize collagen type I,type III in the cultured fibroblast higher than that control group.There was significant difference in the synthesis DNA and express mRNA and synthesize collagen type I, III and haluronic acid and human hydroxyproline of the fibroblast cells among the groups than that in control group (P <0.05).3. The levels of collagen type I,III,haluronic acid,and hydroxyproline in the supernatants of cultured fibroblast cells treated with polysaccharide were significantly higher than that control group in a dose- dependent manner (P <0.05);The ratio of collagen type I,III decreased dramatically.Conclusion:These data indicate the Aloe vera polysaccharides may play an important role in the extracellular matrix remoding during the phase of wound healing, through promoting both fibroblast cells proliferation and synthesis and secretion the production of type I,III collagen, and haluronic acid, hydroxyproline ,fastening wound healing.