Protection Of RUWE Against HepG-2Cells Injury Induced By H₂O₂

Objective:To investigate the protective effect of Rhaponticum uniflorum water extract (RUWE) in H2O2-induced HepG-2cell injury on the cellar level, and concluded that the protection of RUWE in human liver injury research and its mechanisms.Methods:1, Establish H2O2induced HepG-2cell damage model, with different concentration at the same time and the same time dealing with different concentration H2O2to cells, using MTT determination of cell proliferation was inhibition, kit detection cells on growing the clear of Lactic dehydrogenase (LDH), glutamic-pyruvic transaminase (ALT) and glutamic-oxaloacetic transaminase (AST) activity, have the best H2O2induction HepG-2concentration cell damage.2、MTT method for detection RUWE, HepG-2cells toxicological effect, get to HepG-2leaks reed cell non-toxic role concentration; Will HepG-2cells into the control group, the model group, RUWE low, medium and high dose group, HepG-2cells to stick wall, RUWE according to50,100,200mg/L dose pretreatment24h, model group and RUWE group with H2O2induction damage, such as growing the volume control group added, the determination of the medium LDH, ALT and AST activity, cell cytoplasm activity of superoxide dismutase (SOD)n glutathione (GSH) and malondialdehyde (MDA) content, get the best protection RUWE concentration.3、In the best protection concentration as the research object, using immunohistochemistry methods (Western blot) detection cells extracellular signal regulating protein kinase (ERK), c-Jun amino-terminal kinase (JNK), P38silk crack the original amp-activated protein kinase (P38) and its phosphorylation form protein expression.Result:In To HepG-2cells was the damage with the increase of the dose effect and the time of the growth and strengthen, in H2O2concentration for300umol/L, effect time2h HepG-2cells on the inhibition of45.78+2.11%, in medium ALT and LDH AST levels are normal group increased significantly (P<0.01), and to extend the time to increase concentration can still make the damage to strengthen, but flattened range.2、 Concentrations RUWE after pretreatment, compared with model group, RUWE can significantly improve the cell survival; Reduce HepG-2cells in the medium, ALT、AST and LDH level (P<0.01), and a dose dependent on; Reduce the cell level MDA (P<0.01), also increase the activity of SOD in cells and the content of GSH (P<0.01). dose the effect is not obvious.3、Western results show that, H2O2make the HepG-2cells’ERK、 JNK、P38、P-ERK、P-JNK and P-P38higher, compared with the model group, RUWE group of ERK, JNK protein expression are lower, P-ERK, P-JNK protein expression significantly reduced (P<0.01), P38and P-P38protein expression no change.Conclusion:RUWE on H2O2HepG-2damage caused by cells apparently had a protective effect, can significantly inhibit H2O2HepG-2cells to the damage effect. Its role may increase and oxidation resistance, restrain ERK, JNK protein activation relevant.