1.Changes of suppressors of cytokine signaling-3 and sterol regulatory element binding proteins-1c pathway in steatosis HepG2/HepG2.2.15 cells
Yan WANG ; Longfeng ZHAO ; Rongrong WANG ; Shengwen ZHI
Chinese Journal of Infectious Diseases 2017;35(6):326-331
Objective To investigate the effects of HepG2 and HepG2.2.15 cells steatosis on the mRNA and protein expressions of suppressors of cytokine signaling-3(SOCS-3) and sterol regulatory element binding proteins (SREBP-1c).Methods The cell model of chronic hepatitis B (CHB) combined with nonalcoholic fatty liver disease (NAFLD) was successfully constructed using an oleic acid-induced HepG2 and HepG2.2.15 cells steatosis.Cells were divided into HepG2 cell control group (HepG2 cell control group), HepG2.2.15 cell control group (HepG2.2.15 cell control group), HepG2 cell steatosis group (HepG2 cell steatosis group) and HepG2.2.15 cell steatosis group (HepG2.2.15 cell steatosis group).The expression levels of SOCS-3 and SREBP-1c mRNA were detected by real-time quantitative polymerase chain reaction (PCR).Changes in protein expressions of SOCS-3 and SREBP-1c were measured by western blot.Results SOCS-3 mRNA expression level in HepG2.2.15 cell control group was significantly lower than that in HepG2 cell control group (P<0.01).The level in HepG2 cell steatosis group was also significantly lower than that in HepG2 cell control group (P<0.01).However, the level of SOCS-3 mRNA in HepG2.2.15 cell steatosis group was lower than HepG2.2.15 cell control group with no statistical significance (P=0.173).There was interaction between cells and steatosis (F=25.547, P<0.01).The expression of SREBP-1c mRNA in HepG2.2.15 cell control group was significantly lower than that in HepG2 cell control group (P<0.01), and was significantly higher in HepG2.2.15 cell steatosis group than that in HepG2.2.15 cell control group (P<0.01).There was no significant difference between HepG2 cell steatosis group and HepG2 cell control group (P=1.000).There was interaction between cells and steatosis (F=5.04, P<0.05).Western blot analysis showed that protein levels of SOCS-3 and SREBP-1c in steatosis cells at 48 h and 72 h were significantly higher than those in non-alcoholic steatosis cells.Conclusions Protein expressions of SOCS-3 and SREBP-1c are up-regulated in both steatosis groups.Factorial analysis shows that there is interaction between cells and steatosis.HBV gene could inhibit SOCS-3 mRNA expression and promote the expression of SREBP-1c mRNA in steatosis cells.
2. Effect of transforming growth factor-β1 on HBV replication and antigen synthesis in HepG2.2.15 cells with steatosis
Yan WANG ; Longfeng ZHAO ; Rongrong WANG ; Shengwen ZHI
Chinese Journal of Hepatology 2017;25(10):732-737
Objective:
To investigate the effect of transforming growth factor-β1 (TGF-β1) on HBV replication and protein expression in HepG2.2.15 cells with steatosis, as well as the association of TGF-β1 with suppressor of cytokine signaling-3 (SOCS-3) mRNA and sterol regulatory element-binding protein-1c (SREBP-1c) mRNA during the steatosis of HepG2.2.15 cells.
Methods:
The cells were divided into HepG2/HepG2.2.15 cell control groups (C1/C2 groups) and HepG2/HepG2.2.15 cell steatosis groups (F1/F2 groups). 5 ng/ml TGF-β1 was added to the two cell systems for intervention to establish TGF-β1 intervention groups (T1/T2 groups) and steatosis+TGF-β1 intervention groups (TF1/TF2 groups). A time-resolved fluorescence analyzer was used to measure HBsAg and HBeAg, and quantitative real-time PCR was used to measure HBV DNA, SOCS-3 mRNA, and SREBP-1 mRNA. A one-way analysis of variance and a factorial analysis were used for the statistical analysis of data.
Results:
TGF-β1 significantly reduced the level of HBeAg in C2 group (
3.Clinical value of myocardial enzymes and umbilical artery blood gas analysis in diagnosis and treatment of neonatal asphyxia
Shengwen ZHI ; Qingxin BAI ; Min CHENG ; Jie CUI
Chinese Journal of Primary Medicine and Pharmacy 2018;25(7):847-850
Objective To investigate the clinical value of myocardial enzymes and umbilical artery blood gas analysis in the diagnosis and treatment of neonatal asphyxia (HIE).Methods Eighty-seven neonates with HIE and 94 healthy neonates(control group) were selected and compared.The myocardial enzymes [aspartate aminotransferase (AST),creatine phosphokinase(CK),creatine phosphokinase isoenzyme (CK-MB),hydroxybutyrate dehydrogenase (HB-DH),lactate dehydrogenase (LDH)] and umbilical artery blood gas analysis [pH,alkali residue (BE),arterial oxygen pressure(PaO2),arterial carbon dioxide partial pressure (PaCO2)] of the two groups were detected and compared.Results The levels of AST,CK,CK-MB,HB-DH and LDH in the observation group were (123.54 ±36.57) U/L,(1 786.83 ± 542.37) U/L,(584.63 ± 164.54) U/L,(652.31 ± 187.38) U/L,(956.38 ± 257.64) U/L,respectively,which were significantly higher than those in the control group [(65.38 ± 20.34) U/L,(675.48 ± 240.32) U/L,(48.61 ± 12.15) U/L,(248.36 ± 51.69) U/L,(581.36 ± 102.67) U/L] (t =13.35,17.75,31.50,20.10,13.04,all P <0.05).The pH,BE,PaO2,PaCO2 of the observation group were (7.02 ±0.13),(9.56 ± 1.74) mmol/L,(3.26 ± 0.26) kPa,(6.88 ± 1.24) kPa,respectively,which of the control group were (7.20 ± 0.16),(8.96 ± 1.52) mmol/L,(3.21 ±0.33) kPa,(6.58 ± 1.84) kPa,respectively.the pH value in the observation group was significantly lower than that in the control group (t =8.27,P < 0.05).The BE,PaO2 and PaCO2 between the two groups had no statistically significant differences (all P > 0.05).In the observation group,the levels of AST,CK,CK-MB,HB-DH and LDH of the neonates with ideal prognosis were (59.68 ±-13.52) U/L,(542.36 ± 103.65) U/L,(49.37 ± 14.25) U/L,(275.36 ± 64.51) U/L,(567.35 ± 115.24) U/L,respectively,which were significantly lower than those of the neonates with poor prognosis [(81.32 ± 36.24) U/L,(1 265.38 ± 362.74) U/L,(168.35 ±50.01)U/L,(602.31 ± 205.34)U/L,(853.64 ± 212.54)U/L],and the pH value of the observation group with ideal prognosis was significantly higher than that of the neonates with poor prognosis [(7.19 ± 0.21) vs.(7.01 ±0.18)],the differences were statistically significant (t =14.67,14.42,17.22,11.14,7.14,2.91,all P < 0.05).Conclusion The myocardial enzymes and umbilical artery blood gas analysis can be used in the early diagnosis of HIE and evaluation of the prognosis,and it has high clinical value.