Effects Of Panax Notoginseng Polysaccharides On The Proliferation And Expression Of Cytokine Activity In Human Skin Fibroblasts In Vitro

Part 1Determination the Optimal Concentration of Panax Notoginseng Polysaccharides for Improving Cell Proliferation in Cultured Human Skin FibrablastsObjective:To investigate for further studies about the panax notoginseng polysaccharides (PNP)、its optimal concentration of cultured human skin fibroblasts (hSFb) were determined in this part.Methods:Cultured human skin fibroblasts (hSFb) were treated by various concentrations of PNP. Cells were cultured in an incubator at 37℃,5%CO2 for 72 hours. The vitality and proliferation potential of cells were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.Result:Compared with the control group, PNPs at the concentration range of 0.1 mg/ml to 1 mg/ml improved cell proliferation of hSFb (P<0.01 or P<0.05). And its optimal concentration of PNP was 0.5 mg/ml (P<0.05).Conclusions:The optimal concentration of PNP to improve cell proliferation of human skin fibroblasts is 0.5 mg/ml.Part 2Effects of Panax Notoginseng Polysaccharides on the functional activity in human skin fibroblasts in vitroObjective:To investigate panax notoginseng polysaccharides from notoginseng extracts on human skin fibroblasts (hSFb) effects of functional activity cultured in vitro. It provide experimental evidence for clinical application on chronic wounds healing.Methods:1) The effect of PNP on hSFb proliferantion was examined by MTT colorimestric cell proliferation-assay.2) At 72 hours, through hydroxyproline kits detect the supernatant liquid collagen content.3) At 72 hours, the relative expressions of promote angiogenesis factor(FGF-2、Ang-1、VEGF、PDGF), wound healing related factors(TGF-β1、CTGF、α-SMA、Col-1α),proinflammatory factor(IL-6) and chemokine(SDF-1) mRNA were detected by real time PCR, and the results were compared with the gene expressions of control group.Result:1) After 24,48,72 and 96 hours,0.5 mg/ml PNP induced significant proliferation of hSFb as determined by the MTT assay (P<0.05).2) PNP interact on hSFb 72 hours, collagen content in the culture supernatant has gone up, but no significant difference compared with the control group (P>0.05).3) Compared with the control group, in addition to the mRNA expressions of extracellular matrix protein (Col-la) were lower (P>0.05), the rest of the growth factor mRNA expression ability all have varying degrees of increase, but not statistically significant (P>0.05). However the expression levels of Ang-1 mRNA were higher than cultured control group (P<0.05). Compared with the control group, the mRNA expression of inflammatory mediators IL-6 is significantly improved (P<0.01).Conclusions:1) Compared with control group, PNP can promote hSFb proliferation in vitro, hSFb expression higher mRNA level of Ang-1 and inflammatory mediators IL-6. It indicates that angiogenesis and inflammation related cytokines expression activity of hSFb may be up-regulated by PNP.2) Compared with control group, PNP can decrease the hSFb expression wounds of matrix components Col-1α mRNA (F>0.05), but the supernatant liquid collagen content were up-regulated. Thus, whether to inhibit scar formation are worth discussing in the further.