Expression Of NF-E2 In Immune Organ Of Burn Septic Rat

Objective To investigate expression pattern of Nrf2 in different immune organ of septic rat and preliminary analysis of Nrf2 in the development of sepsis in the possible mechanismMethod Taking wistar rat as animal model, it is divided into 4 groups. Normal control group,Pure burn group, Burn with Staphylococcus sepsis group, Burn with Pseudomonas sepsis group. according to different time intervals as 2 hours,8 hours, and 24 hours. divided into three sub-group after injection of bacteria. Mouse were killed by breaking the cervical spine and under aseptic measure, laparotomy was done and specimen (spleen, lungand liver) was taken from all group according to the above time interval group. A portion of specimen were preserved in liquid nitrogen, Separately done RT-PCR and Western Blot experiment to study the expression of Nrf2 in all group of tissues in different time interval ,rest portion were fixed in 10% formalin. HE staining was done for all paraffin imbedded slides. Microscopic study was done to observe for the histological changes of tissues.Result1. There is slight expression of Nrf2 in spleen, lung and liver of rat in normal control group. Expression is abundant in lung and spleen, then in liver in this experiment.2. Expression of Nrf2 mRNA in all tissue of pure Burn group is obviously down regulated. But in spleen, its expression is down regulated up to lowest level at 8 hours. As time goes on, its expression is up regulated near to reach normal tissue level at 24 hours.3. In staphylococcus attacked group, Nrf2 mRNA expression is up regulated only in liver tissue; it is down regulated in lung and spleen tissue. In liver tissue up regulated tendency is seen in 2-8 hours. In 2 hour, reached in peak value. In 24 hours tendency of expression is down regulated. Expression of NRf2 mRNA in lung tissue and spleen tissue were almost down regulated in 24 hours. Western blot result shows that there is down regulated tendensy of expression of Nrf2 in spleen,liver and lung in 24 hours.4. In burn with pseudomonas sepsis group, except up regulated expression in liver tissue at 2 hours group, all other group had down regulated expression.Conclusion1. There is slight expression of Nrf2 in lung, liver and spleen of rat in normal control group. Expression is abundant in spleen and lung, then in liver in this experiment. It is consider that the distribution of Nrf2 mainly localize in immune organ2. pure burn group lead direct down regulated exprsion of Nrf2,then gradually increased.It showed that Nrf2 direct take part in post burn immune response.3. In burn sepsis,expression of Nrf2 in lung and spleen is down regulated,but it is up regulated in liver in early stage.It showes that there is activation of innate immune response by large ammount of endotoxins in liver tissue. Simillary there is down regulated expression of Nrf2 in immune organs after severe infection.This revealed that there is significant inhibition of auto immune and oxidative stress reaction caused by severe infection.