The Study Of Effects And Mechanism Of Ulinastatin On The Treatment Of Patients With Sepsis

BackgroundSepsis is a serious syndrome that given priority to with systemic inflammatory response caused by kind of infection factors. Severe sepsis is a leading cause of death among critically ill patients in intensive care unit. In recent years on the pathogenesis of sepsis and physiological mechanism, as well as the clinical treatment has a lot of related research. However, the morbidity and mortality of sepsis patients is still high. The pathogenesis of sepsis is complex and oxidative stress is one of the key features of sepsis. Ulinastatin (UTI) is a kind of protease inhibitors (extraction) in the urine, can inhibit a variety of protein and the activity of hydrolytic enzymes, antioxidant, anti-inflammatory and anti apoptosis and signal adjustment. The early clinical trials found that these drugs can improve the prognosis of patients with sepsis, but its role is unclear. However, the potential molecular mechanism is not well understood.ObjectiveTo evaluation the treatment effect of Ulinastatin (UTI)on patients with sepsis, and to study the possible mechanism through the test peroxide material concentration changes in serum. At the cellular level, we aimed to investigate the potential effects of UTI on the (Lipopolysaccharide) LPS-induced oxidative stress in human umbilical vein endothelial cells (HUVECs) and the underlying mechanisms.MethodsCollected clinical data of 40 patients with sepsis,40 patients with sepsis were randomly assigned to Ulinastatin group (n=20), the control group (n=20).And 10 normal physical examination, for this experiment into the normal group. the treatment of control group according to sepsis treatment guidelines, Ulinastatin group patients on the basis of the control group therapy intravenous drip Ulinastatin 100000 U q8h, the treatment for 7 days. Before the treatment and the posttreatment of 4,8 days extraction of venous blood, to detect the change concentration of Malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (gsh-px). In check-up venous blood was collected at normal group, normal test result as the experiment group. Monitor the patient’s heart rate, breathing rate, body temperature and white blood cell count, and statistical mechanical ventilation time, ICU length of hospital stay and 28 d survival rate.On the basis of clinical research. Oxidative stress injury model was constructed by using the lipopolysaccharide (LPS) induce human umbilical vein endothelial cells in vitro. According to the concentration gradient of LPS experiment and time gradient, select 10 ug/ml concentration of drug, the processing time of 24 h as the production of HUVEC oxidative stress model experiment concentration and time. Ulinastatin protection concentration gradient experiment to select 100 U/ml concentration as the experimental concentration. Cell is divided into blank control group, UTI group, the LPS group, UTI+LPS group, the SP600125 plus LPS group, UTI+Y60025+LPS group. The cell survival rate of detection to choose a kind of commonly used methods (determined by MTT method), MDA, GSH-Px kits to test production of MDA, GSH-Px. Using Western blot method to detect SOD, JNK, p-JNK, c-Jun, p-c-Jun the expression of protein. To further investigate whether the UTI on HUVECs injury induced by LPS has a protective effect. Application of LPS induced endothelial cell damage, to discusses UTI through JNK/c-Jun signal pathways against oxidative stress.ResultsExperimental group patients the situation is better than the control group after treatment including heart rate, breathing rate, body temperature and white blood cell count. Two groups of patients before treatment in the serum MDA content compared with normal group increased significantly, and in the serum SOD, gsh-px, significantly lower compared with the normal group. UTI group of patients after treatment than before treatment in the serum MDA decreases, there is significant difference before and after treatment (p< 0.05). SOD, GSH-Px was increased before treatment, but there is no significant difference before and after the treatment (p> 0.05). While the control group after treatment the peroxide index level has no obvious change, there was no significant difference before and after treatment. Before the treatment, Ulinastatin group and the control group there was no statistically significant difference each index (p> 0.05). After treatment4 d Ulinastatin group and control group each indicators have differences, but no statistical significance (p> 0.05). After treatment 8 d the index between two groups was statistically significant difference (p<0.05). UTI group mechanical ventilation time and ICU length of hospital stay is less than the control group, and 28 d survival appear contrary trend.On the cellular level, we found that intracellular MDA content increased, and reduced GSH-Px vigor, SOD protein expression level decreased after LPS stimulation, and LPS significantly increased ROS produced in the cell. UTI after processing can reverse LPS induced endothelial cells to produce this effect. UTI can inhibit LPS of endothelial cells to stimulate the JNK/c-Jun phosphorylation. JNK pathway inhibitor SP600125 can weakened LPS oxidative stress effect obviously. JNK, c-Jun phosphorylation level is abate, the expression of protein is reduced.ConclusionUlinastatin significantly improve the condition of patients with sepsis, shortening the time of mechanical ventilation and ICU length of hospital stay, improve 28 d survival; UTI may protect the vascular endothelium against serious disease induced by oxidative stress such as sepsis.We demonstrated that the anti-oxidant of UTI observed in HUVECs induced by LPS repress through JNK/c-Jun signaling pathway. The data presented here support JNK/c-Jun is a potential therapeutic target for the treatment of sepsis and that inhibiting oxidative stress with UTI administration.