Protective effect of ginsenoside Rg1 on 661W cells exposed to oxygen-glucose deprivation/reperfusion via keap1/nrf2 pathway

AIM:To construct an in vitro model of oxygen-glucose deprivation/reperfusion(OGD/R)induced injury to the optic nerve and to study the oxidative damage mechanism of ischemia-reperfusion(I/R)injury in 661W cells and the protective effect of ginsenoside Rg1.METHODS:The 661W cells were treated with different concentrations of Na2S2O4 to establish OGD/R model in vitro.Apoptosis,intracellular reactive oxygen species(ROS)levels and superoxide dismutase(SOD)levels were measured at different time points during the reperfusion injury process.The injury model was pretreated with graded concentrations of ginsenoside Rg1.Real-time polymerase chain reaction(PCR)was used to measure the expression levels of cytochrome C(cyt C)/B-cell lymphoma-2(Bcl2)/Bcl2 associated protein X(Bax),heme oxygenase-1(HO-1),caspase9,nuclear factor erythroid 2-related factor 2(nrf2),kelch-like ECH-associated protein 1(keap1)and other genes.Western blot was used to detect the expression of nrf2,phosphorylated nrf2(pnrf2)and keap1 protein levels.RESULTS:Compared to the untreated group,the cell activity of 661W cells treated with Na2S2O4 for 6 and 8h decreased(P<0.01).Additionally,the ROS content increased and SOD levels decreased significantly(P<0.01).In contrast,treatment with ginsenoside Rg1 reversed the cell viability and SOD levels in comparison to the Na_(2)S_(2)O_(4)treated group(P<0.01).Moreover,Rg1 reduced the levels of caspase3,caspase9,and cyt C,while increasing the Bcl2/Bax level.These differences were all statistically significant(P<0.05).Western blot analysis showed no significant difference in the protein expression levels of keap1 and nrf2 with Rg1 treatment,however,Rg1 significantly increased the ratio of pnrf2/nrf2 protein expression compared to the Na_(2)S_(2)O_(4)treated group(P<0.001).CONCLUSION:The OGD/R process is induced in 661W cells using Na_(2)S_(2)O_(4).Rg1 inhibits OGD/R-induced oxidative damage and alleviates the extent of apoptosis in 661W cells through the keap1/nrf2 pathway.These results suggest a potential protective effect of Rg1 against retinal I/R injury.

Surface modification of apatite-wollastonite glass ceramic by synthetic coupling agent

In this study, lysine was introduced into the surface of apatite-wollastonite glass ceramic （AW-GC） to improve its cytocompatibility by two steps reaction procedure. Firstly, lysine connected to N-β-（aminoethyl）-y-aminopropyl trimethoxy silane （A-1120） by covalent binding of amide group. Secondly, the lysine-functionalized A-1120 was deposited on the surface of AW-GC through a silanization reaction involving a covalent attachment. FTIR spectra indicated that lysine had been immobilized onto the surface of AW-GC successfully. Bioactivity of the surface modified AW-GC was investigated by simulated body fluid （SBF）, and the in vitro cytocompatibility was evaluated by coculturing with human osteosarcoma cell MG63. The results showed that the process of hydroxyapatite layer formed on the modified material was similar to AW-GC while the mode of hydroxyapatite deposition was changed. The growth of MG63 cells showed that modifying the AW-GC surface with lysine enhances the cell adhesion and proliferation.