Effects Of Notoginsenoside R1 On Cell Proliferation And Synthesis Of Collage And The Expression Of Matrix Metallo-proteinase-1 In Human Skin Fibroblasts In Vitro

Object Notoginsenoside R1 is one kind of characteristic chemical constituent in Araliaceae Panax pseudo-ginseng. To observe the expression of MMP-1 protein and cell proliferation and synthesis of collage in human dermal fibroblasts in vitro after UVA and UVB irradiation and then investigate the photo-protection of notoginsenoside R1. The experiment will explore the photo-protection of to notoginsenoside R1 of fibroblasts after UVB and UVA irradiation and explore the relative molecular mechanism. The results will contribute to the exploration and application of the natural anti-photoaging medicine.Methods Human skin fibroblasts were gotten by explant culture and enzymatic digestion and fibroblasts did primary culture in vitro. Fibroblasts were irradiated by UVA and UVB according to the designed dose. Notoginsenoside R1 was added before irradiation. To observe the change of growth and morphology of fibroblasts by UV and explored the impact of notoginsenoside R1 on it. Fibroblasts proliferation and cellular viability was record by MTT assay. The hydroxproline amount that was secreted into the culture supernatant was measured by digestive technique and then the hydroxproline amount was mathematically converted to the procollagen amount. The level of MMP-1 was secreted by fibroblasts was measured with ELISA Kits.Results Human skin fibroblast were gotten by explant culture and enzymatic digestion showed good proliferation, normal shape and typic Fusiform or babylon weeping shape in vitro. Growth curve showed latency phase, index number growth phase of fibroblasts, was about 0 to 2 days, about 2-5 days separately, then went into platform phase. The fibroblasts were damaged significantly after UV irradiation, cell were broken and disorder arrangement. It didn't change in the number of cell and the morphology after adding notoginsenoside R1. The activity of proliferation of UV-induced fibroblast was increased when the dose of notoginsenoside R1 was 5μg/ml and 20μg/ml; Notoginsenoside R1 could increase the amount of hydroxyproline and the collagen synthesis in the cultural medium, by adding the dose of notoginsenoside R1 below 50μg/ml, the amount of hydroxyproline and the collagen synthesis were the highest when the dose of notoginsenoside R1 was 20μg/ml. The MMP-1 protein secretion level was suppressed when the dose of notoginsenoside R1 was 5μg/ml and 20μg/ml separately and the significant change was observed at 20μg/ml.Conclusions All the results indicated that UVA and UVB irradiation could cause fibroblast damage, and then decrease the quantity of fibroblasts. Notoginsenoside R1 protects the damage of fibroblast after UV irradiation. The experiment proved notoginsenoside R1 could be used as one kind of protection medicine to UV irradiation when it is used in a certain concentration range. The mechanism of notoginsenoside R1 may be related to below several aspects:1. Notoginsenoside R1 could increase the activity of proliferation of fibroblast after UV irradiation;2. UV irradiation can inhibit the collagen synthesis of fibrolasts, while notogin-senoside R1 could reduce the inhibition by UV;3. UV irradiation couls increase the MMP-1 protein secreation, while notoginsenoside R1 could decrease the increase by UV.