ABSTRACT
Objective@#To explore the role of iTr35 cells in the pathogenesis of children with pulmonary artery hypertension (PAH) in children, and the percentage of iTr35 cells and serum interleukin(IL)-35 levels in peripheral blood of patients with PAH were investigated.Their inhibitory effects on the expression of vascular cell adhesion molecule-1 (VCAM-1) on vascular endothelial cells were also analyzed.@*Methods@#After 3 mL peripheral blood of 30 congenital heart disease (CHD) patients with PAH, 22 CHD patients without PAH and 30 age or gender matched healthy controls (HC) were collected, the percentage of iTr35 cells were detected by flow cytometry and the concentrations of serum IL-35 were detected by Luminex, as well as soluble VCAM-1 (sVCAM-1). Human pulmonary artery endothelial cells (HPAECs) were cultured in vitro and divided into control group, tumor necrosis factor (TNF)-α group and IL-35+ TNF-α group.The expression of VCAM-1 and nuclear factor(NF)-κB P65 protein of each group were detected by flow cytometry and Western blot.The adhesion of peripheral blood mononuclear cells (PBMCs) to HPAECs was observed by fluorescence microscope.@*Results@#Compared to CHD patients without PAH, the percentage of iTr35 cells[0.86(0.45-1.63)% vs.1.14(0.46-2.11)%](H=20.52, P<0.05) in peripheral blood and serum IL-35 levels[2.43(1.76-2.85) μg/L vs.3.17(2.92-5.66) μg/L](H=119.56, P<0.05) were significantly decreased in CHD patients with PAH.However, serum sVCAM-1 concentration[923.1(892.6-1 118.7) μg/L vs.776.1(743.5-932.3) μg/L ] in CHD patients with PAH were significant increased (H=65.65, P<0.05). In addition, the concentration of serum IL-35 were negatively correlated with sVCAM-1 in PAH patients (r=-0.374 P=0.042). In vitro, the positive rate of VCAM-1 on HPAECs was significant decreased in IL-35+ TNF-α group as compared to the TNF-α group [(2.07±0.82)% vs.(5.83±1.34)%, F=1 197.18, P<0.05]. In addition, the protein expression of NF-κB P65 in HPAECs was significantly decreased in TNF-α+ IL-35 group as compared to TNF-α group, as well as the adhesion of PBMCs to HPAECs (F=212.04, 2 533.51, all P<0.05).@*Conclusions@#The percentages of iTr35 and levels of IL-35 are reduced in the peripheral blood of patients with PAH.Through in vitro experiments, IL-35 is found to reduce PBMCs adhesion by inhibiting VCAM-1 expression in HPAECs.
ABSTRACT
Objective To explore the role of iTr35 cells in the pathogenesis of children with pulmonary artery hypertension(PAH)in children,and the percentage of iTr35 cells and serum interleukin(IL)_35 levels in peripheral blood of patients with PAH were investigated. Their inhibitory effects on the expression of vascular cell adhesion mole_cule_1(VCAm_1)on vascular endothelial cells were also analyzed. Methods After 3 mL peripheral blood of 30 congenital heart disease( CHD)patients with PAH,22 CHD patients without PAH and 30 age or gender matched healthy controls(HC)were collected,the percentage of iTr35 cells were detected by flow cytometry and the concentra_tions of serum IL_35 were detected by Luminex,as well as soluble VCAm_1(sVCAm_1). Human pulmonary artery endothelial cells(HPAECs)were cultured in υitro and divided into control group,tumor necrosis factor( TNF)_α group and IL_35+TNF_α group. The expression of VCAm_1 and nuclear factor( NF)_κB P65 protein of each group were detected by flow cytometry and Western blot. The adhesion of peripheral blood mononuclear cells(PBmCs) to HPAECs was observed by fluorescence microscope. Results Compared to CHD patients without PAH,the percent_age of iTr35 cells[0. 86(0. 45_1. 63)% υs. 1. 14(0. 46_2. 11)%](H=20. 52,P<0. 05)in peripheral blood and serum IL_35 levels[2. 43(1. 76_2. 85)μg/L υs. 3. 17(2. 92_5. 66)μg/L]( H=119. 56,P<0. 05)were signifi_cantly decreased in CHD patients with PAH. However,serum sVCAm_1 concentration[923. 1(892. 6_1 118. 7)μg/L υs. 776. 1(743. 5_932. 3)μg/L ]in CHD patients with PAH were significant increased( H=65. 65,P<0. 05). In addition,the concentration of serum IL_35 were negatively correlated with sVCAm_1 in PAH patients(r= _0. 374 P=0. 042). In υitro,the positive rate of VCAm_1 on HPAECs was significant decreased in IL_35+TNF_α group as compared to the TNF_α group[(2. 07 ± 0. 82)% υs.(5. 83 ± 1. 34)%,F=1 197. 18,P<0. 05]. In addition,the protein expression of NF_κB P65 in HPAECs was significantly decreased in TNF_α+IL_35 group as compared to TNF_α group,as well as the adhesion of PBmCs to HPAECs(F=212. 04,2 533. 51,all P<0. 05). Conclusions The percentages of iTr35 and levels of IL_35 are reduced in the peripheral blood of patients with PAH. Through in υitro ex_periments,IL_35 is found to reduce PBmCs adhesion by inhibiting VCAm_1 expression in HPAECs.