Changes And Significance Of SIRT1 Gene Expression In Placenta Of Preeclampsia

Objective To investigate the expression of Sirt1 in placental tissues of PE patients and predictive value of plasma SIRT1 level on the pathogenesis and prognosis of PE,and the effects of up-regulated Sirt1 expression on placental trophoblast apoptosis,inflammatory response and oxidative stress responseMethods A total of 111 PE patients and their placental tissues were collected from January 2019 to January 2021 in our hospital and divided into mild PE group(n=84)and severe PE group(n=27)according to their conditions.According to the treatment prognosis,the patients were divided into poor prognosis group(n=12)and good prognosis group(n=99).A total of 58 healthy pregnant women and their placental tissues delivered in our hospital during the same period were selected as healthy control group.The expression level of Sirt1 gene in placental tissues was determined by fluorescence quantitative PCR,and the expression level of Sirt1 protein in placental tissues was determined by Western blot.Using enzyme-linked immunosorbent method determination of oxidative stress in the placenta grinding fluid index [superoxide dismutase(SOD),catalase(CAT),glutathione peroxidase(GSH-px),reactive oxygen species(ROS),lipid peroxide(LHP),nicotinamide adenine dinucleotide phosphate oxidase4(NOX4)],inflammation index [white blood cells Interleukin(IL-1β),interleukin-6(IL-6),high mobility group protein B1(HMGB1),C-reactive protein(CRP),intercellular adhesion molecule-1(VCAM-1)],endothelial injury factor[hemagglutinin oxidized low density lipoprotein receptor-1(LOX-1),oxidized low density lipoprotein(ox-LDL),human pentaprotein 3(PTX3),adrenomedullin(ADM)].The expression levels of Sirt1,oxidative stress index,inflammation index and endothelial injury factor in PE patients and healthy control group with different conditions and prognosis were analyzed.Pearson test was used to analyze the correlation between Sirt1 expression level in placenta of PE patients and specific disease indicators.Receiver operating characteristic(ROC)curve was used to analyze the predictive value of Sirt1 expression level in plasma of PE patients for poor prognosis.The level of SIRT1 in plasma of patients in each group was measured by enzyme-linked immunosorbent assay,the correlation between the expression level of SIRT1 in plasma and the level of SIRT1 protein in placental tissue was analyzed by Pearson test,and the predictive value of the expression level of SIRT1 in plasma of patients with PE for poor prognosis was analyzed by receiver operating characteristic(ROC)curve.Trophoblast cells HTR-8/SVeno were selected for subculture,and thesubcultured HTR-8/SVeno cells were inoculated in the culture plate and grouped.The mRNA and protein expression levels of Sirt1,and the protein expression levels of acetylated p53,acetylated NF-κB,acetylated Foxo3 a,and cleaved caspase-3 in all groups were compared.The levels of inflammatory indexes tumor necrosis factor α(TNF-α),interleukin-1 β(IL-1β),interleukin-6(IL-6)and oxidative stress indexes reactive oxygen radical(ROS),malondialdehyde(MDA)and 8-hydroxy-deoxyguanosine(8-OHDG)were compared among all groups.Results(I)The expression and prognosis of Sirt1 in the placenta of preeclampsia: The expression of Sirt1 mRNA in the placenta of severe PE group,mild PE group and healthy control group increased sequentially(P<0.05).The expression of Sirt1 protein in placental tissues of severe PE group,mild PE group and healthy control group increased sequentially(P<0.05).The levels of SOD,CAT and GSH-Px in placenta homogenate of severe PE group,mild PE group and healthy control group were increased in turn,while ROS,LHP and NOX4 levels were decreased in turn(P<0.05).The levels of IL-1β,IL-6,HMGB1,CRP and VCAM-1 in placenta homogenate of severe PE group,mild PE group and healthy control group were decreased sequentially(P<0.05).The levels of LOX-1,OX-LDL,PTX3 and ADM in placental homogenates of severe PE group,mild PE group and healthy control group were decreased sequentially(P<0.05).The expression levels of Sirt1 mRNA and Sirt1 protein in placental tissues of patients with poor prognosis were lower than those of patients with good prognosis,(P<0.05).The expression levels of Sirt1 mRNA and Sirt1 protein in placental tissues of PE patients were positively correlated with SOD,CAT and GSH-Px levels,negatively correlated with ROS,LHP and NOX4 levels,and negatively correlated with IL-1β,IL-6,HMGB1,CRP and VCAM-1levels.Lox-1,OX-LDL,PTX3,ADM levels were negatively correlated.ROC curve showed that the expression levels of Sirt1 mRNA and Sirt1 protein inplasma of PE patients had certain predictive value for poor prognosis.(2)There was no significant difference in plasma SIRT1 level among severe PE group,mild PE group and healthy control group(P > 0.05).Based on the existing data,the correlation between the level of SIRT1 in plasma and the expression level of SIRT1 protein in placenta cannot be studied,and the prediction model of PE cannot be established.(3)Expression and significance of Sirt1 in placental trophoblast cells:mRNA and protein expression levels of Sirt1 in HTR-8/Sveno cells in hypoxia group were lower than those in control group(P<0.05);The mRNA and protein expression levels of Sirt1 in HTR-8/Sveno cells in the hypoxia + resveratrol group were higher than those in the hypoxia + resveratrol group(P<0.05).The expression levels of p53,Cleaved caspase-3,NF-κB and Foxo3 a in HTR-8/Sveno cells in the hypoxia group were higher than those in the control group,respectively.The levels of inflammatory factors TNF-α,IL-1β and IL-6in the medium were higher than those in the control group,and the levels of oxidative stress indexes ROS,MDA and 8-OHDG were higher than those in the control group(P<0.05).The expression levels of p53,Cleaved caspase-3,NF-κB,and Foxo3 a in HTR-8/Sveno cells in hypoxia + resveratrol group were lower than those in hypoxia group,respectively.The levels of inflammatory factors TNF-α,IL-1βand IL-6 in the medium were lower than those in the hypoxia group,and the levels of oxidative stress indexes ROS,MDA and 8-OHDG were lower than those in the hypoxia group,with statistical significance(P<0.05).Sirt1 plasmid +hypoxia group,h TR-8 /Sveno cells acetylated p53,cleaved The expression levels of Caspase-3,acetylated NF-κB and acetylated Foxo3 a were lower than those of NC,plasmid + hypoxia group,respectively.The levels of inflammatory cytokines TNF-α,IL-1β and IL-6 in the medium were lower than those in NC plasmid + hypoxia group,respectively.(P<0.05).Conclusions Sirt1 gene and protein are abnormally low in placental tissues of PE patients,and the specific expression level is closely related to oxidative stress,inflammation and vascular endothelial injury.The injury of trophoblast cells under hypoxia and other conditions can lead to the decrease of Sirt1 expression.The indirect increase of Sirt1 expression by resinotrel can alleviate the injury of trophoblast cells under hypoxia.But,it was not found that the level of SIRT1 in plasma of patients with PE decreased significantly,which could not be confirmed to be related to the SIRT1 protein in placenta.Reduced Sirt1 expression may be one of the important mechanisms for the occurrence of preeclampsia,and targeted upregulation of Sirt1 expression may be one of the new approaches for the treatment of preeclampsia.