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Objective To study the photodynamic therapy (PDT) mediated by a novel porphyrin-typed photosensitizer on human hepatic carcinoma HepG2 cell and the mechanisms.Methods Experiments were derided into four groups:control group,PDT group,photosensitizer group and photosensitizer+PDT group.The photostability of novel photosensitizer upon repetitive illumination was evaluated by bleaching method,and cell survival rate was determined by MTT assay.Cellular uptake of novel photosensitizer was measured with luminescence spectrometer,and cellular localization ofphotosensitizer was observed by laser scanning confocal microscopy (LSCM).Furthermore,apoptotic cell was detected with Hoechst 333342 staining.Results Novel photosensitizer was stable after repetitive light irradiation,and PDT or photosensitizer alone showed no dark cytotoxicity toward HepG2 cell (P＞0.05),but intense killing was observed in photosensitizer+PDT group (P＜0.05).The IC50 is 1.21 μmol/L.Cellular uptake of novel photosensitizer was concentration-dependent and the highest uptake is at concentration of 12.5 μmol/L.Novel photosensitizer localizes in lysosomes of HepG2 cell,and the death mode of HepG2 cell was mainly apoptosis.Conclusions Novel photosensitizer exerts profound cytotoxic effects on HepG2 cell mainly through the initiation of secondary cell apoptosis by lysosome destruction.
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OBJECTIVE@#To observe the influence of ropivacaine on the proliferation and migration of rat bone marrow mesenchymal stem cells (BMSCs) and provide basis for the clinical application of BMSCs.@*METHODS@#Rat BMSCs were isolated and cultured by adherence method. Surface markers of BMSCs were examined by flow cytometry. Multipotent differentiation of BMSCs was detected by induced adipogenesis, osteogenesis and muscular differentiation. Proliferation of BMSCs was examined by CCK-8 and Brdu incorporation after ropivacaine treatment at different concentrations. Migration of BMSCs was tested by cell scratch assay and Millicell experiment.@*RESULTS@#Cultured cells had representative appearance and surface markers of BMSC, and they had potential multiple differentiation. Ropivacaine treatment at 50 and 100 μmol/L significantly reduced the proliferation rate of BMSCs and Brdu incorporation rate. There was significant difference compared with the control group (P<0.05). Cellular scratch assay and migration experiment indicated that ropivacaine significantly reduced the migration of BMSCs. There was significant difference compared with the control group (P<0.05). All these mentioned effects of ropivacaine on BMSCs were dose-dependent. There was significant difference between groups (P<0.05).@*CONCLUSION@#Ropivacaine can significantly reduce the proliferation and migration of rat BMSCs, suggesting that the influence of local anesthetics on BMSCs has to be taken into account when BMSCs are used in clinical practice.