Effects Of Viaminate On The Proliferation And Differentiation Of Keratinocytes And Photoproduct Analysis By LC-MS

ObjectiveTo investigate the effects of viaminate on the proliferation anddifferentiation of human keratinocytes ( HaCaT cell line )in vitro, andanalyze the photoproduct of viaminate by LC-MS. To further identify themechanism of viaminate adjust the growth of keratinocytes andinvestigate the stability of viaminate under light.Methods1. The changes of proliferation level of the HaCaT cells at 24, 48and 72 h after the treatment with viaminate were detected by MTT assay;2. The changes of the cycle of HaCaT cells were measured byflow cytometry;3. The changes of keratin 10 and involucrin mRNA expressionwere semi-quantitatively analyzed by reverse transcriptionpolymerasechain reaction (RT-PCR) technique;4. The photoproduct of viaminate was analysed by LC-MS.Results1. Viaminate could significantly inhibit the proliferation ofHaCaT cells with a dose-dependent manner under the same duration ofco-culture (after 48 and 72 hours), p＜0.05;2. The DNA flow cytometric analysis showed that the G₁ phaserate of cell was significant increased in 48 hours compared with thecontrol, but the rates of S and G₂ phase were decreased. The cells werearrested in the G₁/G₂ phase by viaminate;3. The mRNA expression of involucrin on HaCaT cells was upregulated, and was related to the concentration of viaminate, but themRNA expression of keratin 10 was down-regulated under the samecondition;4. LC-MS detected a photoproduct of viaminate under light, themolecular weight was 448 equal with viaminate. Condusion1. Viaminate plays an inhibitory role in the keratinocytesproliferation and induces the cell differentiation;2. The photoproduct of viaminate under light with the equalmolecular weight might be the photoisomer of viaminate, with whichviaminate was stable under the condition protected from light.