The Research Of Gelsolin In Cardiopulmonary Bypass Induced Acute Lung Injury And TRIM22 In The Regulation Of Inflammation

Part1 The study of plasma gelsolin in cardiopulmonary bypass induced acute lung injury in infants and young childrenBackground and Objective:Cardiopulmonary bypass(CPB)can induce acute lung injury(ALI),which is a common and serious complication after cardiac surgery.And young children and infants are much more sensitive to CPB induced-ALI.The aim of this study was to detect the perioperative changes of plasma gelsolin(pGSN)in patients(≤3 years of age)with cardiac surgeries and CPB,and discuss if pGSN are associated with the occurrence and development of CPB-related ALI.Methods:77 consecutive infants and young children below 3 years of age with congenital heart diseases(CHD)performed on cardiac surgery and CPB were finally enrolled,and assigned to acute lung injury and non-acute lung injury groups,according to the Consensus Criteria of American-European.Plasma gelsolin and total protein concentrations were measured at dedicated eight time points:before(a)and immediately after(b)CPB,two hours(c),six hours(d),twelve hours(e),twenty-four hours(f),forty-eight hours(g)and seventy-two hours(h)after CPB.Results:27(35.1%)patients occurred CPB-induced ALI in our study,including 11(14.3%)patients with acute respiratory distress syndrome.The earliest significant decline of plasma GSN and normalized plasma GSN(pGSNN)of acute lung injury group were both at six hours after cardiopulmonary bypass(p = 0.04 and P<0.01),which was earlier than those of non acute lung injury group(forty-eight hours,P= 0.03 and twenty-four hours,P<0.01);plasma GSN of non acute lung injury group before CPB and six-hour after CPB were both obviously higher than those of acute lung injury group(P<0.01);plasma GSNN of non acute lung injury group before CPB and six hours after CPB were both clearly higher than those of acute lung injury group(P<0.01,P=0.04);plasma GSN before CPB was the only independent risk factor for predicting the occurrence of CPB induced acute lung injury(OR,1.023;95%CI,1.007-1.039;P<0.01)and AUC was 0.753(95%CI,0.626-0.880);the pGSN before CPB optimal cutoff value was 264.2 mg/L,the sensitivity was 58.3%and specificity was 94.7%.Plasma GSN level before CPB was negatively associated with the severity of CPB induced-ALI(r =-0.45,P<0.01).Conclusion:Patients developing cardiopulmonary bypass-induced acute lung injury had lower pGSN level,which showed a more amount and rapid consumption early after CPB.Plasma gelsolin before surgery was a sensitive and early predictor of CPB induced-ALI in patients undergoing cardiac surgery,and was negatively associated with the severity of CPB induced-ALI.Part 2 The study of gelsolin in the development of acute lung injury Background and Objective:Studies have showed that the decline of pGSN level could induce and aggravate lung injury,while exogenous gelsolin supplement could obviously alleviate the pulmonary microcirculation permeability and inflammatory infiltration which induced by acute lung injury.Therefore,the purpose of this study is to research the expression and distribution of gelsolin and its protective role in acute lung injury.Method:The single pulmonary ischemia(60min)-reperfusion(30,60,120min)injury model was performed on young New Zealand rabbit(400-500g),then blood were collected at these time points:before ischemia,reperfusion,reperfusion with 30min,60min and 120min,PaO2 was measured.Gelsolin in BALF and plasma was detected with ELISA.The single pulmonary ischemia-reperfusion(I/R:60/120min)injury model was performed on New Zealand rabbit,fetal bovine gelsolin(7.8mg/kg)was intravenously injected before experiments,PaO2,pulmonary vascular permeability and cytokines level in BALF and plasma,as well as neutrophilic exudation in focal and systemic inflammation was measured to determine the protective role of exogenous gelsolin supplement in lung injury.Results:After the single pulmonary ischemia(60min)-reperfusion,PaO2 significantly decreased at 30min,60min and 120min after reperfusion,and is negatively related to reperfusion time(r=-0.627,P<0.01).Gelsolin level in BALF obviously increased after reperfusion,however,plasma gelsolin level did not change significantly.In the intravenous injection of fetal bovine gelsolin group,PaO2 was significantly increased(P<0.01)and protein exudation(P=0.019)and TNF-a level of BALF/plasma(P<0.01 and P<0.01)was decreased.Pretreatment with gelsolin significantly decreased white blood cell(WBC)and PMN in plasma(P<0.01 and P<0.01)and MPO activity(P<0.01).Conclusion:The function of oxygenation decreased after the single pulmonary ischemia-reperfusion,and is negatively related to reperfusion time.Gelsolin level in BALF obviously increased after reperfusion,however,there was no significant difference in plasma.Exogenous supplement of gelsolin played a protective role in acute lung injury.Part 3 The role and mechanism of TRIM22 in the regulation of inflammation in monocytesBackground and Objective:Cardiopulmonary bypass induces systemic inflammatory response and destroys alveolar epithelial cells and alveolar epithelial cells,which result in lung injury.Monocytes are key member during inflammatory reaction and can produce a high amount of pro-inflammatory and anti-inflammatory cytokines.Therefore,further research in monocytes on inflammation is helpful for understanding CPB-related systemic inflammatory reaction.Tripartite motif-containing 22(TRIM22),could be induced by IFN and belongs to TRIM superfamily.TRIM22 is involved in viruses restriction,cell proliferation and activation.The homologous gene of TRIM22 is TRIM30 in mice,it is reported that negatively regulate inflammation.We have found that TRIM22 level is lower in sepsis patients than that of non-sepsis patients.However,the role of TRIM22 in regulation of monocytes inflammation and the detailed molecular mechanisms are still unclear.This part was designed to research the role of TRIM22 in inflammation and molecular mechanisms,which is important for understanding CPB-related systemic inflammatory reaction and providing a new approach for CPB-induced ALI treatment.Methods:TRIM22 expression was detected by using real time PCR and immunoblot analysis.The adenoviral vectors encoding TRIM22 gene were constructed and transfected into THP-1 cells.After LPS stimulation,the level of cytokines was determined by Q-PCR and ELISA.LC3B and heme oxygenase-1 were assayed using immunoblot.Results:LPS induced TR1IM22 expression in THP-1 monocytes.Compared to control group,overexpression of TRIM22 decreased pro-inflammatory cytokines TNF-a and IL-1βlevels(73.41 ±6.49 vs.34.84±3.92pg/ml,P<0.0001;90.64±8.91 vs.45.24±9.97pg/ml,P<0.0001).LC3II and HO-1 levels increased after LPS stimulation.Overexpression of TRIM22 up-regulated LC3II and HO-1 expression,compared with control group.Inhibition of autophagy increased the level of cytokines.Conclusion:Overexpression of TRIM22 could negatively regulate inflammatory response in monocytes,which is associated with increased autophagy and HO-1 level.