| Abstract [eng] |
Research of Novel Therapeutic Targets for Sepsis in Plasma from Septic Patients Sepsis is one of the deadliest known diseases all over the world with the case fatality rate exceeding 40 %. In recent years there has been growing evidence, that current strategies of sepsis therapy, which in fact did not undergo any significant changes since the World War II, are not efficient enough. This makes it clear that there is a huge need for novel targets of therapy to be found. One of the major mechanisms for sepsis caused death is higly related to endothelial barrier disruption and increased permeability that is very common during sepsis. We have tried to investigate the amount of high mobility group box 1 protein (HMGB1), as a possible target for sepsis therapy, in plasma from septic patients (n=98) as well as its effect on endothelial cells. An enzyme linked immunosorbent assay (ELISA) based system for quantitative detection of HMGB1 was created. It was shown that during sepsis HMGB1 concentration increases hundred of times in comparison to healthy (n=30) individuals (384,26±122,28 ng/ml and 2,7±1,4 ng/ml respectively; p=0,007). It was also shown that there is a statistically significant difference between HMGB1 concentrations in plasma from sepsis survivors in comparison to lethal cases of sepsis (256,17±72,74 ng/ml and 427,33±68,92 ng/ml respectively; p=0,04), as well as between those who developed multiple organ dysfunction syndrome (MODS) and those who did not (560±65,68 ng/ml in comparison to 324,6±104,12 respectively; p=0,02). It suggests that HMGB1 might be used as sepsis biomarker with ability to distinguish both between septic and non-septic individuals and between those septic patients who are at a high risk of MODS and death and those who are not. Another part of this project was based on the investigation of HMGB1 cytotoxicity on HUVEC primary endothelial cells. While using calorimetric LDH assay, it was shown that HMGB1 in septic plasma is cytotoxic to the endothelium (150 ng/ml, 12 h, 18,4±6,2 %; 200 ng/ml 12 h, 31,7±3,9 %; 400 ng/ml, 12 h, 54,1±11,3 %). The same experiments were repeated with respective concentrations of recombinant HMGB1. Although it was shown that recombinant HMGB1 also has a similar cytotoxic effect on HUVEC, statistical analysis did not show any significant differences from septic plasma mixture with respective amount of HMGB1 (p=0,06). Further experiments need to be done in the future in order to find out if HMGB1 alone is responsible for endothelial barrier disruption during sepsis and might become a novel target for sepsis therapy. |
