Effects Of Ecdysterone In The Oxidative Stress Induced Hepatic Gluconeogenesis

Objective: Diabetes mellitus is a deficiency in insulinsecretion or insulin resistance caused by elevated blood sugar as the mainmanifestations of chronic diseases,its incidence rises year by year,aboutits pathogenesis and treatment is also increasingly in-depty study.Theinsulin resistance mechanism as the main research object in Thepathogenesis of type2diabetes study.In recent years, more and morestudies show that oxidative stress in the occurrence and development ofinsulin resistance plays a crucial role.Its mechanism is still not veryclear,hepatic gluconeogenesis in vivo glucose metabolism as theimportant link, its two key enzymes PEPCK, G-6-Pase expression onblood glucose level is of decisive significance.Ecdysterone in diabetesmellitus and its complications related to aspects of the effects such asincreased glucose consumption,antioxidative stress has also beenconfirmed by experiment, the experiment of normal rat liver cells an invitro rat liver cells injury induced by oxidative stress model,after be theconcentration of ecdysterone （ecdysterone, ECR） damage after theintervention, and with pioglitazone （Pioglitazone, PIG） was used as apositive drug control,quasi from horizontal cells to oxidative stress wasobserved and gluconeogenesis and the relationship between insulinresistance, through to the intracellular gluconeogenesis factor expression level on,Understanding of ecdysteroid on oxidative stress induced bygluconeogenesis, indirect finding ecdysterone insulin resistancemechanism.Methods:This experiment in normal rat liver cells forexperimental cell model,from the laboratory for normal BRL-3A rat livercell line,be in high-glucose medium conventional culture, randomgrouping:（1）Normal cellular environment group:Continue to be culturedin high glucose medium, without any other interveningfactors,（2）Oxidative stress model group:With the concentration of100umol/L H2O2high-glucose medium cultured for4hours; and thenrandomly divided into three groups,a group of stopping cultivation,extraction of protein and degeneration after-20℃, pure hydrogenperoxide group;A group, continue to be ecdysterone intervention, be withthe concentration of ECR1×10^-6mmol/L, ECR1×10^-7mmol/L, ECR1×10^-8mmol/L three concentrations of high glucose cultured for24hours；A group of be positive control drug pioglitazone intervention, bewith concentration of1×10^-5mmol/L pioglitazone continued to train for24hours;Above each cell culture liquid is added with concentration of100nM insulin, to simulate the normal cellular environment.Results:（1）Drug effects on BRL-3A cells: when ecdysterone concentrations inexcess of1x10^-5mol/L concentration cell od began to appear to dropapparently，the tips of this concentration range of ecdysterone couldsignificantly inhibit the cell growth （P＜0.01）， when ecdysterone concentrations below1×10^-9mol/L, cell OD higher, tip is smaller thanthe concentration effect of ecdysterone on small cell,ecdysteroneconcentration of1×10^-6-1×10^-8mol/L, cell OD at around0.7,combined with other experimental data, and the concentration as theconcentration of ecdysterone intervention;2) The experimental group Akt,p-Akt, FoxO1protein expression level changes: through each group ofexperimental statistics contrast is found, the experimental group Aktprotein expression differences do not have statistical significance（p<0.01）；With pure hydrogen peroxide groups compared, follow-up todrug intervention groups Akt phosphorylation levels increased, theexpression of p-Akt protein increased in different degrees, the differencehas statistical significance（p<0.01），the downstream of FoxO1proteinexpression appears different level of reduction.（3） Intracellular FOXO1,PEPCK and G-6-Pase mRNA expression:Compared with the controlgroup, only hydrogen peroxide pre-treated cells PEPCK and G-6-PasemRNA expression with the highest（p<0.01），in contrast, subsequent useof ecdysterone and pioglitazone intervention of cellular PEPCK andG-6-Pase mRNA expression is decreased in different degree（p<0.05），Conclusions:（1） A high concentration of ecdysterone on BRL-3A of ratliver cells cytotoxicity;（2） Hydrogen peroxide induced oxidative stressinjury intervention can make the intracellular FoxO1protein expressionincreased, so that the phosphorylation of Akt reduction,at the same time, gluconeogenesis enzymes PEPCK, G-6-Pase two gene expression isincreased, suggests that oxidative stress may induce gluconeogenesis,increased hepatic glucose output.（3） Ecdysterone intervention after cellp-Akt expression level elevated the tips of ecdysterone could increase theAkt phosphorylation levels, while the downstream of FoxO1proteinexpression levels can also be inhibited, PEPCK, G-6-Pase expression wasalso decreased, suggestive of ecdysterone reduces oxidative stress leadsto excessive gluconeogenesis, the the mechanism may be on insulinsignaling pathway in many aspects。...