1.Transcription factor ETS-1 mediates the expression of miRNA-21 induced by arsenic in human hepatic L-02 cells
Caihua QI ; Aihua ZHANG ; Xiong CHEN ; Baofei SUN
Chinese Journal of Endemiology 2017;36(1):26-31
Objective To investigate the effect of sodium arsenite (NaAsO2) on the expression of miRNA-21 (miR-21) mediated by transcription factor ETS-1 in human normal hepatocytes (L-02).Methods Dose-effect study:The L-02 cells were treated with different doses of NaAsO2 [0.0 (control),2.5,5.0,10.0,20.0,40.0 μmol/L] for 24 h.Time-effect study:L-02 cells were exposed to 0 (control) and 20 μmol/L NaAsO2 for 12,24,36 and 48 h (n =6).ETS-1 and miR-21 were treated with ETS-1 shRNA and miR-21 inhibitor,respectively.The cells treated with ETS-1 shRNA (100 nmol/L) were divided into 4 groups:①ETS-1 shRNA NC treatment alone (control group);②ETS-1 shRNA NC combined with NaAsO2 (20 μ,mol/L) treatment group;③ETS-1 shRNA treatment alone group;④Treatment with ETS-1 shRNA and NaAsO2 (20 μmol/L) group.The MiR-21 inhibitor (100 nmol/L) treated cells were also divided into 4 groups:① miR-21 inhibitor NC treatment (control group);② miR-21 inhibitor NC combined with NaAsO2 (20 μmol/L);③miR-21 inhibitor group;④miR-21 inhibitor combined with NaAsO2 (20 μ mol/L) treatment group.The expression of ETS-1 mRNA and miR-21 were detected by quantitative real-time PCR (qRT-PCR);the protein expression of ETS-1 was detected by Western blotting.Results Dose-effect study:The expression of ETS-1 mRNA in the groups of 0.0 (control),2.5,5.0,10.0,20.0 and 40.0 μmol/L was 1.008 ± 0.028,1.552 ± 0.029,1.697 ± 0.050,1.842 ± 0.077,2.233 ± 0.096 and 2.235 ± 0.092;miR-21 expression was 1.025 ± 0.094,1.552 ± 0.072,1.683 ± 0.066,1.915 ± 0.171,2.337 ± 0.195 and 2.592 ± 0.177;the expression of ETS-1 protein was 1.060 ± 0.045,1.267 ± 0.160,1.386 ± 0.087,1.723 ± 0.196,2.208 ± 0.122 and 2.284 ± 0.224,respectively,and the differences were statistically significant (F =47.797,8.959,65.748,all P < 0.05),the NaAsO2 dose groups were significantly higher than those of the control group (all P < 0.05),and there was a dose-effect relationship.Time-effect study:The expression of ETS-1 mRNA in L-02 cells was 1.253 ± 0.175,1.623 ± 0.220,1.771 ± 0.324 and 1.913 ± 0.251,respectively at 12,24,36 and 48 h;the expression of miR-21 was 1.502 ± 0.111,1.716 ± 0.113,1.979 ± 0.186 and 2.452 ± 0.304;the expression of ETS-1 protein was 1.196 ± 0.105,1.502 ± 0.076,1.651 ± 0.074 and 1.839 ± 0.139,respectively,there were significant differences between the groups (F =14.936,39.180,39.441,all P < 0.05).The expression of various time points of exposure to NaAsO2 was significantly higher than those in the control group (1.044 ± 0.115,1.044 ± 0.124,1.108 ± 0.088,1.053 ± 0.061;1.092 ± 0.061,1.096 ± 0.169,1.024 ± 0.111,1.057 ± 0.146;1.020 ± 0.017,1.049 ± 0.121,1.024 ± 0.089,1.031 ± 0.124,all P< 0.05),and there was a time-effect relationship.ETS-1 shRNA and miR-21 inhibitor treatment:compared with ETS-1 shRNA NC combined with NaAsO2 (20 μmol/L),ETS-1 shRNA and NaAsO2 (20 μmol/L) could significantly inhibit the expression of ETS-1 (0.912 ± 0.238 vs 1.641 ± 0.225,P < 0.05),and down-regulated the expression of miR-21 (1.313 ± 0.334 vs 2.363 ± 0.252,P < 0.05).There was no significant difference of ETS-1 mRNA expression between miR-21 inhibitor and NaAsO2 (20.μmol/L) group (1.580 ± 0.077 vs 1.576 ± 0.065,P > 0.05) compared with miR-21 inhibitor NC and NaAsO2 (20 μmol/L).Conclusions The expression of ETS-1 and miR-21 in L-02 cells is significantly higher than those in control.The high expression of ETS-1 mediates NaAsO2-induced miR-21 overexpression,which may be an important molecular mechanism of arsenic-induced expression dysregulation of human hepatic miRNAs and liver damage.
2.Effects of a single general anesthesia on neurodevelopment in preschool children
Bing LIU ; Xin QI ; Yaqiu ZHANG ; Caihua FENG ; Hui ZHANG
Chinese Journal of Anesthesiology 2022;42(6):666-670
Objective:To evaluate the effects of a single general anesthesia on neurodevelopment in preschool children.Methods:American Society of Anesthesiologists physical status Ⅰ-Ⅱchildren of either sex, aged 3-6 yr, who were undergoing dental treatment under general anesthesia or non-general anesthesia, were enrolled in the study.Children in general anesthesia group received dental treatment under general anesthesia, while children in control group received pit and fissure sealant.In general anesthesia group, the children entered the operating room accompanied by their parents and inhaled 50% nitrous oxide and 8% sevoflurane, propofol 2 mg/kg, fentanyl 2 μg/kg, and cisatracurium 0.1 mg/kg and dexamethasone 0.2 mg/kg were intravenously injected, mechanical ventilation was performed after nasotracheal intubation under the guidance of a video laryngoscope, and anesthesia was maintained through inhalation of 2.5%-3.0% sevoflurane.The outcomes of neurodevelopment were assessed and scored using Wechsler Preschool and Primary Scale of Intelligence, Behavior Rating Inventory of Executive Function, and Adaptive Behavior Assessment System.Results:Forty-four children were included in general anesthesia group and 19 children in non-general anesthesia group.There were no significant differences between the two groups in the changes in the scores of Wechsler Preschool and Primary Scale of Intelligence, Behavior Rating Inventory of Executive Function, and Adaptive Behavior Assessment System before anesthesia and at 6 months after operation ( P>0.05). Conclusions:A single general anesthesia exerts no significant effect on neurodevelopment in preschool children.