The Expression Of T-bet And GATA-3mRNA In Peripheral Blood Mononuclear Cells Of Youg Children Suffered From VSD Combined With Different Level Of PAH

Objective: Ventricular septal defect could also to be abbreviated forVSD, which is one of the most frequent disease of congenital heartdisease(CHD), and accounts for23to30percent. Because of the openning ofthe pulmonary circulation after the child's birth, the pulmonary arterialpressure is decreased suddenly, and the right ventricular pressure grows downaccordingly, and resulted in significant pressure difference between the leftventricle and the right ventricle. When the interventricular septum appeareddefection, some blood could flow from the left ventricle to the right ventriclethrough the defect. When this state continued for a long time, the tunica mediaof arteriola in lung would become accrementition, and endomembrane becomethickening, and pulmonary vascular resistance increased further, and finalyformed pulmonary artery hypertension. If not treated in time, the pathologicalchanges of arteriolopathy in lung would aggravate, and formed inconvertibleangiemphraxis in lung. When the pulmonary vascular resistance exceeds theresistance of genetal circulation, would resulted in right-to-left shunt, and thepatients will appear cyanosis, which is called Eisenmenger Syndrome.Pulmonary arterial hypertension (PAH) is one of the most frequentcomplication of ventricular septal defect, the complicated pathogenesy ofwhich has not been understood clearly up to now. Besides, the youg childrenwith VSD might experience the danger condition of PAH in around theoperation time, which was one of the most causes of death at the operation.And there was a big gap between the internal and the international in childrenwith VSD and combined PAH, which was one of the main challenges ofpedo-cardiac surgery in our country in the future.Some studies reported that there were low and disorder immune functions in children with CHD, in other words, they existed the drift betweeen T helpertype1(Th1) and T helper type2(Th2), and children in pulmonary congestionwas worse than the children in pulmonary oligemia. Th1, Th2and the balancebetween them were important in the immune response. T-bet and GATA-3were respectively expressed in Th1and Th2specificly. The T-bet and GATA-3are the key and specific transcription factors who decide the differentiation ofthe Th0cell to Thl cell or to Th2cell.The real time fluorescence quantitative reverse transcription-polymerasechain reaction (RT-PCR) is a kind of new technology which is based ontraditional PCR detection method. It effectively solves the limitation that thetraditional PCR only detects quantity at end-point. The RT-PCR posses thecharacteristics of high specificity, reliability, the stability of experiment results,repeatability etc.We applied the real time fluorescence quantitative reversetranscription-polymerase chain reaction (RT-PCR) to detect the expression ofT-bet mRNA and GATA-3mRNA in the peripheral blood of patients withventricular septal defect and combined with pulmonary arterial hypertension,to explore the relationship between the key transcription factors controlling ofTh1cells and Th2cells differentiation, to know VSD combined with PAHpatients existing Th1and Th2cells balance or not, and to learn Th1and Th2cells in the pathogenesis of VSD combined with PAH at the transcriptionallevel.Method: Forty young children with VSD combined PAH were involvedin the study, who were from the department of cardiac surgery in Children'sHospital of He Bei Province.All of them were younger than1year old and thebody weight was less than10Kg. Inclusion criteria:(1) all the patients werediagnosed by medical examination, auxiliary examination and the ultrasonicinspection.(2)there were4groups, and each group had10patients wereinvolved in the study. And each group were respectively with lower grade,middle grade, and high grade PAH.(3)Not accompanied with grave disease ofother organs, autoimmune diseases or other serious systemic diseases.(4) primary pulmonary hypertension were excluded.Peripheral venous blood was obtained from the patients. The expressionsof T-bet mRNA and GATA-3mRNA in peripheral blood mononuclear cells(PBMC) were detected by the real time fluorescence quantitative reversetranscription-polymerase chain reaction (RT-PCR). All of the data wasanalyzed by the statistical package of the SPSS13.0. Student's t test waschosen between the two groups, ANOVA or Kruskal-Wallis H test was chosenamong the groups. P <0.05was statistically significant.Results:1The results of Children suffered from VSD combined with different levelof PAH were as follows:1.1Gene T-bet: the level of T-bet in children with mid-grade PAH group andhigh-grade PAH group were lower than the no PAH group,and the values ofΔCt respectively were5.74±2.15,6.93±1.45,7.51±1.96,7.43±1.31, and theresults showed statistical significances, the value P respectively were0.024and0.033.1.2Gene GATA-3: the level of GATA-3in children with low-grade PAH,mid-grade PAH group and high-grade PAH group were lower than the no PAHgroup, and the values of Δ Ct respectively were3.79±2.10,5.43±1.29,5.36±1.36,5.35±1.20, and the results showed statistical significances, and Pvalue respectively were0.025,0.018and0.020.2The results of children suffered from VSD combined with different levelof PAH surrounding the operation were as follows:2.1Gene T-bet: the levels of T-bet in children at three days after operationand seven days after operation were lower compared with the children atperoperative, and the values ofΔCt respectively were5.97±1.49,6.74±2.29,7.30±1.22,7.17±1.27, and the P value respectively were0.045and0.002. Oneday after the operation was lower than the level of seven days after theoperation.2.2Gene GATA-3: there were no statistical significances in each group. Conclusions: At the transcriptional level, the results showed that therewere low and disorder immune functions in children suffered from VSDcombined with different level of PAH. The level of T-bet in Children withmid-grade PAH group and high-grade PAH group were higher than the noPAH group. And this means the quantity of Th1cells in children withmid-grade PAH and high-grade PAH were reduced. The level of GATA-3inchildren with low-grade PAH, mid-grade PAH group and high-grade PAHgroup were higher than the no PAH group, and this hints that the quantity ofTh2cells were reduced too. The levels of T-bet in children at one day afteroperation, three days after operation and seven days after operation werehigher compared with the children at peroperative, and showed that thequantity of Th1cells were reduced after the operation. The humoral immunitywas no difference before operation and after operation.