Effect Of Glucocorticoid On Inflammatory Mediators And MiRNA-155 In Septic Shock

Objective:Sepsis is defined as life-threatening organ dysfunction caused by a dysregulated host response to infection.Septic shock is one of the most severe complications of sepsis,systemic inflammatory response syndrome leading to increased capillary permeability,lack of effective circulating blood volume,persisting hypotension despite adequate volume resuscitation,and high hospital mortality.In the early stage,the patients often died of uncontrolled inflammatory reaction and circulatory failure.In the late stage,they were mainly characterized by immunosuppression,delayed infection,multiple organ dysfunction and multiple organ failure.In this study,the effects of glucocorticoid on inflammatory mediators and miRNA-155 in patients with septic shock were discussed to provide evidence for clinical use of corticosteroids controlling excessive inflammatory response to septic shock.Part One:Effect of glucocorticoid on prognosis of ICU septic shock.323 patients with septic shock were treated in the first hospital of Jlin university From September 2013 to September 2015 were selected as the research subjects.A retrospective analysis was conducted to collect the original and etiological data of patients by a unified questionnaire.The general data included age,sex,ICU time,APACHEII score,sequential organ failure estimation score(Sequential Organ Failure Assessment,SOFA),mean arterial pressure(MAP),temperature(T),heart rate(HR),respiratory frequency(RR),MODS or not.323 patients were divided into glucocorticoid group and control group(the unused glucocorticoid group)according to whether received glucocorticoid.The time of ICU,the time of mechanical ventilation,the mortality of ICU and the death rate of 28 days were collected.During September 2013-September2015,2782 patients were admitted to ICU and 2640 patients met the sepsis diagnostic criteria.323 patients were diagnosed septic shock,account for 12.23%(323/2640)of septic patients.112 patients died of septic shock,with a mortality of 34.67%(112/323).According to the use of glucocorticoid,septic shock patients were divided into glucocorticoid group(211 cases)and control group,that is,no corticosteroid group(112 cases).There was no significant difference in sex and age between the two groups(P>0.05).The primary lesions of ICU septic shock were mainly from respiratory system infection,abdominal infection,urinary infection,blood infection and skin infection,among which respiratory infection was the main infection.A variety of clinical specimens(including sputum specimens,puncture fluid,blood,etc.)were cultured for infected microorganism.The positive results were mainly Gram-positive(G+)coccus(G+),gram-negative(G-)bacilli and fungi,among which Gram-negative bacteria were the main pathogenic bacteria,which accounted for the species of infected bacteria.Most of the patients(about 75% percentage)had no statistically significant difference in the location of infection between the two groups with Glucocorticoid(P>0.05).In addition,there was no statistical difference between the time of mechanical ventilation and the time of ICU hospitalization(P>0.05),but the mortality rate,either ICU or 28 day mortality,showed statistically significant difference(P<0.05).In particular,as a dependent variable,age,sex,SOFA score,APACHEII score,and the use of glucocorticoid as independent variables were used as dependent variables.The unconditional Logistic multifactor analysis was carried out.The results showed that the APACHE II score,SOFA score,and no glucocorticoid were independent risk factors for death in patients with septic shock.Part Two:Effect of glucocorticoid on inflammatory mediators and T lymphocytes in patients with septic shock100 cases of septic shock treated in the intensive care unit in No.1 Hospital of Jilin University were randomly divided into glucocorticoidtreatment group(50 cases)and conventional treatment group(50 cases)during March 2016 to December 2016.Exclusion criteria: previous blood diseases,immune system diseases or immunodeficiency,long-term use of glucocorticoid or immunosuppressive agents,pregnant women or lactating women,malignant tumor,cardiac arrest,hospitalized time less than 3 days.A total of 10 healthy volunteers were selected and blood samples were taken as healthy control group.Routine treatment procedures were followed by a 6-hour cluster treatment process for fluid resuscitation after the onset of septic shock.The main contents include: stable breathing,tracheal intubation,mechanical ventilation,and maintenance of blood oxygen saturation over 95%;Patients with shock retained arterial and central venous catheters to monitor blood pressure and central venous pressure(CVP);Fluid resuscitation,analgesic analgesia on demand,can be used to maintain the mean arterial pressure(MAP)above 65 mmHg,monitor and maintain the urine volume more than 0.5ml/kg/h,and can be treated with blood filtration if necessary.Glucocorticoid treatment group combined with small dose of glucocorticoid treatmenton the basis of conventional treatment,with recommend dose of hydrocortisone,100 mgintravenous infused two times a day,continuous application of 7 d.Collect basic clinical features in patients with(basic diseases,the source of infection,organ function,support treatment technology application time),and the diagnosis of septic shock in the ICU and treatment after 7 days of APACHE II score,SOFA score,total number of white blood cells,neutrophils and lymphocytes level,CD4 +,CD8 + count,fibrinogen,CRP and PCT,albumin,transaminase,total bilirubin,creatinine,and blood lactic acid.3-5ml of peripheral blood was extracted from 1 day,2 days,3 days,5 days and 7 days after the treatment,and the serum detected cytokines TNF-a,Il-6 and Il-10.The results showed that there was no significant difference in the basic data of sex,age and infection sources between the two groups before treatment.The 7 day mortality rate in the glucocorticoid treatment group were lower than those in the conventional treatment group,and the difference was statistically significant.There was no significant difference in28 day mortality rate.There was no significant difference between the levels of serum IL-6,TNF-alpha and IL-10 in the two groups before treatment(P>0.05),but with the duration of treatment,the levels of the three were decreasing.In particular,the levels of IL-6 and TNF-X alpha in the patients with hormone therapy began to decline after receiving the treatment of Day1,while the conventional treatment group began to decline after the treatment of Day2,but both were significantly lower than those before the treatment,and the difference was statistically significant(P<0.05).Especially in the glucocorticoidtreatment group,the levels of IL-6 and TNF-alpha decreased more significantly than those in the conventional treatment group,and the difference was statistically significant(P<0.05).In addition,IL-10 began to decline after the treatment of Day2 in the glucocorticoid treatment group,and the conventional treatment group decreased after the treatment of Day3,which was significantly lower than before the treatment,with a statistically significant difference(P<0.05).The level of IL-10 in the glucocorticoid treatment group was significantly lower than that in the conventional treatment group(P<0.05).The results also showed that there was no significant difference in the level of T cell subgroup in the blood of the two groups(P>0.05).The changes of T cell subgroups in the two groups after 7 days were all higher than those before the treatment,and the number of CD4+T cells and CD8+T cells in the glucocorticoid treatment group was significantly higher than that in the conventional treatment group,and the difference was statistically significant(P <0.05).Part Three:The third part: the effect of glucocorticoid on the level of miRNA-155 in septic shock patients.By collecting GEO database included has publicly sepsis gene expression spectrum data and the miRNA expression spectrum data,through bioinformatics analysis,explore the sepsis specificity of micrornas-gene regulating function module,a systemic analysis of the molecular mechanisms of sepsis development potential.Biostatistics and bioinformatics research method was applied,the GEO database collection has publicly sepsis gene expression profile and integrating the miRNA expression spectrum data analysis,the results show that with miR-146-a and mi R-155 as the core of micrornas-gene regulation and control function module in sepsis play an important regulating role in the development process.Collect basic clinical features in patients with(basic diseases,the source of infection,organ function),and the diagnosis of septic shock in the ICU and treatment after 7 days of APACHE II score,SOFA score,total number of white blood cells,neutrophils and lymphocytes level,CD4 +,CD8 + count,fibrinogen,CRP and PCT,albumin,transaminase,total bilirubin,creatinine,and blood lactic acid.3-5ml of peripheral blood was extracted from 1 day,2 days,3 days,5 days and 7 days after treatment,and serum mi-RNA155 was detected by centrifugation.Patients before treatment the miRNA-155 levels and APACHE ? scores were positively correlated(r = 0.71,P< 0.05),patients before treatment the mi RNA-155 level and the level of CD4 + and CD8 + T cells was negatively correlated(rCD4 + = 0.68,rCD8 + = 0.65,P<0.05).there was no statistically significant difference of microRNA-155 level before treatment in both groups(0 d)(T = 0.263,P> 0.263).With the extension of treatment time,the level of two groups of patients with serum microRNA-155 are declining,glucocorticoid treatment group 3 d and 7 d miRNA levels are lower than 0 d,and 7 days the mi RNA levels below 3 d,differences were statistically significant(t3-0 = 2.218,t7 has 0 = 4.013,t7 has-3 = 2.150,P< 0.05),the glucocorticoid treatment group 3 d and 7 d miRNA levels lower than the conventional treatment group at the same time the miRNA levels,the difference was statistically significant(t3d = 2.137,t7 d = 2.137,P< 0.05);When the conventional treatment group 7 d miRNA-155 levels lower than 0 d,was statistically significant difference(t = 2.857,P< 0.05),two groups of patients before treatment(0 d)CD4 + and CD8 + T cell levels has no statistically significant difference(tCD4 + = 0.299,tCD8 + = 0.299,P> 0.05),with the extension of treatment time,the two groups of patients the level of CD4 + and CD8 + T cells showed a trend of increase,the glucocorticoid treatment group 3 d and 7 d CD4 + and CD8 + T cell levels were higher than 0 d,7 d and CD4 + T cell levels higher than 3 d,differences were statistically significant(tCD4 +(3-0)= 4.526,tCD4 +(7-0)= 5.927,tCD4 + 3(7-)= 2.597,tCD8 +(7-0)= 2.975,P< 0.05).Glucocorticoid treatment group 3 d and 7 d level higher than the same period to CD4 + T cells in the conventional treatment group(t3d = 2.755,t7 d = 2.755,P< 0.05),theglucocorticoid treatment group level of CD8 + T cells,there was no statistically significant difference in each period and the conventional treatment group(t3d = 0.748,t7 d = 0.748,P> 0.05);When the conventional treatment group 7 d CD8 + T cell levels higher than 0 d,the difference was statistically significant(T = 3.788,P< 0.05),two groups of patients before treatment(0 d)CD4 + and CD8 + T cell levels has no statistically significant difference(tCD4 + = 0.299,tCD8 + = 0.299,P> 0.05),with the extension of treatment time,the two groups of patients the level of CD4 + and CD8 + T cells showed a trend of increase,the glucocorticoid treatment group 3 d and 7 d CD4 + and CD8 + T cell levels were higher than 0 d,7 d and CD4 + T cell levels higher than 3 d level,differences were statistically significant(tCD4 +(3-0)= 4.526,tCD4 +(7-0)= 5.927,tCD4 + 3(7-)= 2.597,tCD8 +(7-0)= 2.975,P< 0.05).Glucocorticoid treatment group 3 d and 7 d level higher than the same period to CD4 + T cells in the conventional treatment group(t3d = 2.755,t7 d = 2.755,P<0.05),the glucocorticoid treatment group level of CD8 + T cells,there was no statistically significant difference in each period and the conventional treatment group(t3d = 0.748,t7 d = 0.748,P> 0.05);The difference was statistically significant(T =3.788,P<0.05)when the level of CD8+T cells was higher than 0 d in the conventional treatment group.Conclusions:(1)Glucocorticoid help to improve the mortality of septic shock.(2)Glucocorticoid can decrease the release of inflammatory mediators and reduce the apoptosis of lymphocytes in patients with septic shock.(3)miR-146 a and miR-155 plays an important regulatory role in the development of sepsis.