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Objective To determine the inhibitory effect and mechanism of metformin ( MF) on photo-damage of human skin fibroblasts ( HSF) induced by UVA .Methods Human skin fibroblasts were randomly divided into con-trol group, UVA group and UVA+MF group.The proliferation of HSF was detected by CCK-8 assay kit.SA-β-gal staining was performed to evaluate the senescence state .The level of ROS was examined by fluorescence probe DCF-DA staining using flow cytometry .Real-time PCR was used to determine mRNA expression of senescence -asso-ciated signals of MMP 1 and MMP3.The protein expression of MMP 1, MMP3, SOD1 and SOD2 were measured by Western blot .R esults To the proliferation of HSF , 0.01 mmol/L Metformin had no significant effect , but 0.1 and 1 mmol/L Metformin depressed significantly ( P<0.05 ) .Compared with the Control group , it showed that UVA irradiation increased the positive rate of SA-β-gal staining ( P<0.01 ) , the level of ROS ( P<0.05 ) , mRNA and protein expression of MMP1 and MMP3 significantly(P<0.01);Also decreased the expression of SOD1 and SOD2 ( P<0.01) .Compared with the UVA group , it showed that metformin decreased the positive rate of SA-β-gal staining (P<0.05), the level of ROS(P<0.05), mRNA and protein expression of MMP1 and MMP3 significantly(P<0.05);Also increased the expression of SOD 1 ( P<0.01 ) and SOD2.Conclusions Metfomin can inhibit photo-damage of human skin fibroblasts induced by UVA via decreasing ROS and metal matrix protease generation , also the improvement of cellular antioxidant capacity .
ABSTRACT
OBJECTIVE@#To explore the influence of extract of Ginkgo biloba (EGB) on the proliferation, apoptosis of ACC-2 cell and Survivin gene expression in adenoid cystic carcinoma (ACC) of lacrimal gland.@*METHODS@#ACC-2 cell in human with ACC of lacrimal gland was in vitro cultured. MTT method was used for cell proliferation detection. Annexin V/PI double-staining flow cytometer was used to detect cell apoptosis and cell cycle. Survivin gene expression was analyzed by RT-PCR and Western blotting.@*RESULTS@#EGB had inhibitory effect on the proliferation of ACC-2 cell with significant dose-effect relationship, and there was statistical difference when compared with the control group (P<0.01). The inhibitory concentration 50 % (IC(50)) is 88 mg/L. The flow cytometer test indicated that EGB can gradually increase ACC-2 cell in G(0)-G(1) stage and decrease it in G(2)-M and S stage. With the increase of dose, the apoptosis rate of ACC-2 cell was obviously increased (P<0.05 or P<0.01). EGB had certain inhibitory effect on Survivin gene expression of ACC-2 cell, and Survivin gene expression was decreased with the increasing of the EGB concentration (P<0.01).@*CONCLUSIONS@#EGB can effectively inhibit Survivin gene expression of ACC-2 cell in human with ACC of lacrimal gland, induce the apoptosis of ACC-2 cell and inhibit tumor cell proliferation.