The Effects And Significance Of The Overexpression Of MiR-24 On The Formation Of FX In Trauma Induced Coagulopathy

BackgroundThe mechanisms of coagulation factor deficiency in trauma-induced coagulopathy(TIC)are poorly understood.Dysregulation of micro RNAs may contribute to the progression of TIC.We aimed to explore the biological function that miRNA-24-3p(miR-24)might have in coagulation factor deficiency after major trauma and TIC.Methods15 healthy volunteers and 36 severe trauma patients(Injury Severity Score(ISS)?16)including 28 non-TIC and 8 TIC patients were enrolled.TIC was determined as the initial international normalized ratio(INR)>1.5.The plasma samples were collected on emergency arrival and the 3rd day.The miR?24 expression was evaluated by quantitative reverse transcription-polymerase chain reaction.The plasma concentrations of factor X(FX)and factor XII(FXII)were detected by enzyme?linked immunosorbent assay.Multivariate logistic regression analysis and correlation analysis were used to analyze the correlation between data.ResultsThe plasma miR-24 expression was significantly elevated by 3.17 fold(P=0.043)in major trauma patients and reduced after 3 days(P<0.01).The expression level was significantly higher in TIC than in non-TIC patients(P=0.040).Multivariate analysis showed the higher miR-24 expression was associated with TIC.The plasma concentration of FX in TIC patients was significantly lower than in non-TIC ones(P=0.030)and controls(P<0.01).Spearman's correlation indicated a moderate to strong significant negative correlation between FX on arrival and shock index,ISS,INR,partial thromboplastin time,activated partial thromboplastin time and albumin,and a moderate positive correlation with levels of hemoglobin.A negative correlation was observed between miR?24 and FX,while the correlation between miR?24 and FXII was not significant.ConclusionsmiR-24 was overexpressed in major trauma and TIC patients.The level of FX was associated with many clinical indicators.The negative correlation of miR?24 with FX suggested the possibility that miR-24 might inhibit the synthesis of FX during TIC.It provides a new insight into the mechanisms of TIC and a potential new diagnostic and treatment measures for TIC patients.BackgroundTo investigate whether miR-24 can inhibit the formation of FX in vitro.MethodsIn vitro study was conducted on L02 human liver cell line.The L02 cells were transfected with miR-24 by Lipo2000.The expression of miR-24 was detected by q RT-PCR after transfection.The cell proliferation was detected by MTT assay.The changes of HNF-4? m RNA and protein expression in L02 cells were determined by q RT-PCR and Western blot.The concentration of FX in the supernatant of L02 cells was examined by ELISA.Results24 h after miR-24 mimic transfection,the expression of miR-24 in L02 cells increased about 48 times compared with control group and NC group;cells proliferation in the 3 groups were not significantly different(P>0.05).After transfection of miR-24,the expression of HNF-4? m RNA in L02 cells declined 34.3%(P=0.023);HNF-4? protein content dropped 44.2% compared with control(P=0.013);miR-24 transduction significantly reduced FX level in the supernatant of L02 cells,from(55.38±5.78)ng/m L to(43.30±8.49)ng/m L(P=0.030).ConclusionmiR-24 can inhibit the formation of FX in L02 cells.This study further clarified the role of miRNAs in the formation of coagulation factors.BackgroundThe optimal transfusion ratio of fresh frozen plasma(FFP)to red blood cells(RBC)remains controversial in traumatic patients requiring massive transfusion(MT).The objective of this part of study was to determine the effect of 2 transfusion strategies(FFP:RBC ratios of 1:1 and 1:2.5)on the treatment of trauma-hemorrhagic shock(T-HS)rats during resuscitation and their roles on the expression of miR-24 in the plasma and FX m RNA in the liver.MethodsThe ratio of FFP to RBC of the traumatic patients requiring MT in the first part of the research was calculated.26 male Sprague-Dawley(SD)rats were randomly divided into sham operation group(group A),lactate Ringer's fluid resuscitation group(group B),FFP:RBC 1:1 transfusion resuscitation group(group C)and FFP:RBC 1:2.5 transfusion resuscitation group(group D).Another 25 animals were used for blood harvesting.Animals underwent T-HS were treated by resuscitation of fluids preset for each group to maintain the rat mean arterial pressure(MAP)?80mm Hg.The MAP,mortality and the amount of infused fluids were observed and recorded in each group.After resuscitation,all rats were killed after 2 h of observation.Arterial blood samples were obtained for coagulation and blood gas analysis.The plasma level of miR-24 was detected by q RT-PCR.The level of FX m RNA was determined in rat liver tissue by q RT-PCR.Results9 patients requiring MT were studied in our group,the transfusion ratio of FFP:RBC was about 1:2.5.T-HS rat model was successfully established,the initial weights and MAP,were not significantly different between the 4 groups(P>0.05).The bleeding volumes were not significantly different among the 3 model groups(P>0.05).The mortality of rats in the group B was significantly higher than in the 2 component blood transfusion groups(P<0.01).The MAP in group B was significantly lower than group C and D in the T180 and T210 time points after resuscitation(P<0.05).There was no significant difference of MAP between group C and D during the experiment.Blood gas analysis showed acidosis and the decreased Pa O2 dramatically improved in the 2 component blood transfusion groups compared with group B.Blood gas values in the group C and D were not significant different(P>0.05).To maintain the blood pressure of rats over 80 mm Hg,the blood volume used in group C was significantly higher than in group D(P<0.01).At the end of the observation period,rats in group B developed severe coagulopathy.FFP:RBC 1:1 could normalize the coagulation values of the T-HS rats,with no significant difference compared to Sham(P>0.05).Coagulation values in group D were markedly worse than those in group C and group A(P<0.05).The plasma miR-24 expression was considerably elevated by 2.59 fold(P<0.01)in group B.The reduction of plasma miR-24 in group C was more substantial than in group D(P=0.03),with no significant difference compared to Sham(P=0.70).The FX m RNA levels in rat livers of the group B were decreased to 42.5% of the group A(P<0.01),the FX m RNA expression in group C elevated significantly compared to group D(P<0.01),with no significant difference compared to Sham(P=0.67).Pearson's correlation analysis indicated a significant moderate negative correlation between plasma miR-24 and FX m RNA in the rat livers(rho=-0.68,P<0.01).ConclusionThe FFP:RBC 1:1 transfusion strategy had a better resuscitation effect than 1:2.5 on the recovery of traumatic coagulopathy in T-HS rats despite more blood requirement.At the same time,it could reduce the level of miR-24 in circulation and increase the expression of FX m RNA in the liver.