Research The Effect Of Deacetylase(SIRT1) Agonist Resveratrol On The Relationship Between SIRT1 And Reactive Oxygen Species After Oxygen Therapy In Premature Infants In Vivo

Objective: To explore the relationship between silent mating-type information regulation 2 homolog 1（sirtuin 1/SIRT1） and reactive oxygen species（ROS）after oxygen therapy in the peripheral blood mononuclear cells（PBMCs） of the premature infants. To explore the effect of resveratrol on the relationship SIRT1 and reactive oxygen species（ROS）after oxygen therapy in the peripheral blood mononuclear cells（PBMCs） of the premature infants and search a new therapeutic targets for reducing oxidtive stress injury in premature infants.Methods :The experiment was divided into two parts.In the first part,according to the fraction of inspiration O₂（Fi O₂）, 30 cases of premature infants with the diagnosis of respiratory distress syndrome（NRDS） and oxygen requisition（gestational age <32 weeks） during the period from March 2014 to March 2015 at the department of neonatology of the affiliated hospital of Southwest Medical University, were divided into three groups: low dosage oxygen group（Fi O₂<300 m L/L）, moderate dosage oxygen group（Fi O₂ 300 m L/L ⁴00 m L/L）, high dosage oxygen group(（Fi O₂>400 m L/L）. Each group 10 cases, 17 males and 13 females,birth weight（1479 ± 121） g.At the same period,10 cases of prematureinfants as the control group whose gestational age <32 weeks and no oxygen, 5 males and 5 females, birth weight（1473 ± 132） g.The differences of sex, gestational age and birth weight quality were not statistically significant. After 48 hours of oxygen treatment, PBMCs and serum were collected from the peripheral blood. Then the intracellular reactive oxygen species（ ROS） level was detected by laser scanning confocal microscopy and the malondialdehyde（MDA） content in the serum was determined by the whole spectrum spectrophotometer and the SIRT1 localization was observed by immunofluorescence staining and the SIRT1 level in PBMCs were examined by western boltting.In the second part, collected the peripheral blood and isolated PBMCs from 40 cases of premature infants whose gestational age less than 32 weeks and no oxygen after birth during the period from March 2014 to March 2015 at the department of neonatology of the affiliated hospital of Southwest Medical University, each group 10 cases, 19 males and 21 females, birth weight（1460 ± 118） g.The differences of sex, gestational age and birth weight quality were not statistically significant.They were randomly divided into four group: control group,air + Res group, hyperoxia group,hyperoxia + Res group,modeling and cultured in vitro for 48 hours,then the intracellular ROS level was detected by laser scanning confocal microscopy and the MDA content in the medium was determined by the whole spectrum spectrophotometer and the SIRT1 localization wasobserved by immunofluorescence staining and the SIRT1 level in PBMCs were examined by western boltting. Results:With the increase of Fi O₂,compared with the control group, the ROS,MDA content and the rate of SIRT1 nucleocytoplasmic shuttling of PBMCs gradually increased（P<0.05） and SIRT1 protein expression was significantly lowered（P<0.05） and the high dosage oxygen group is the most obvious（P<0.05）in the first part. In the second part, compared with the control group, the ROS, MDA content and the rate of SIRT1 nucleocytoplasmic shuttling of PBMCs in the air + Res were no significant change（P>0.05）, the expression level of SIRT1 increased significantly（P<0.05）. Compared with the control group, the ROS, MDA content and the rate of SIRT1 nucleocytoplasmic shuttling of PBMCs in hyperoxia group increased significantly（P<0.05）. Compared with the hyperoxia group, the ROS,MDA content and the rate of SIRT1 nucleocytoplasmic shuttling of PBMCs in the hyperoxia + Res decreased significantly（P<0.05）, but did not reach the level of the control group. The expression level of SIRT1 in the hyperoxia + Res increased significantly（P<0.05）, but did not reach the level of the control group. Conclusions: Hyperoxia-induced produced large amounts of reactive oxygen species in premature infants,promoted SIRT1 translate from nucleus to cytoplasm, inhibited resistance ability of SIRT1 to oxidative stress. In Hyperoxia-induced resveratrol could improve SIRT1 expression and inhibit SIRT1 shuttle from nucleusto cytoplasm in order to increase the ability of antioxidative stress in PBMCs of the premature infants, thereby reduced the oxidative stress injury of premature infants.