ABSTRACT
OBJECTIVE@#To investigate the effects of Notch signal on hypoxic induction factor (HIF-1α) and autophagy-associated genes Beclin1, LC3I, LC3II in oxygen-glucose deprivation (OGD) induced myocardial cell injury.@*METHODS@#The OGD model was established using hypoxic culture box and hypoglycemic DMEM medium. The cells were divided into normal control group, OGD group, OGD + NC siRNA group, OGD + Notch1 siRNA group and OGD + HIF-1α siRNA group. Western blot was used to detect the interference effects of HIF-1α siRNA and Notch1 siRNA. The effects of Notch1 siRNA and HIF-1α siRNA on the activity of myocardial cells in OGD model were detected by the CCK-8 assay. The effects of Notch1 siRNA and HIF-1α siRNA on autophage-associated genes Beclin1, LC3I and LC3II expression were detected by Western blot.@*RESULTS@#The results of Western blot showed that HIF-1α siRNA could effectively knock down the expression of HIF-1α in myocardial cells in OGD model, and Notch1 siRNA could effectively knock down the expression of Notch1 and HIF-1α in myocardial cells in OGD model. The result of CCK-8 assay showed that Notch1 siRNA and HIF-1α siRNA reduced the activity of myocardial cells in OGD model, and there was no statistical difference between the two groups. Western blot results showed that Notch1 siRNA and HIF-1α siRNA could reduce the expressions of the autophagy-associated genes Beclin1, LC3I and LC3II, and reduce the ratio of LC3II to LC3I at mRNA level.@*CONCLUSION@#Notch1 plays a role in myocardial protection by regulating the expression of HIF-1α to regulate the autophagy in OGD model cells.
Subject(s)
Humans , Autophagy , Beclin-1 , Metabolism , Cell Hypoxia , Cells, Cultured , Glucose , Hypoxia-Inducible Factor 1, alpha Subunit , Metabolism , Microtubule-Associated Proteins , Metabolism , Myocytes, Cardiac , Cell Biology , Pathology , Oxygen , Receptors, Notch , Metabolism , Signal TransductionABSTRACT
Objective To investigate the changes in CD8+T lymphocyte count and effect on immune function in patients with sepsis after continuous renal replacement therapy (CRRT).Methods General data of septic patients who admitted to the emergency department of a hospital between October 2015 and August 2016 were collected,pe-ripheral blood was taken from patients before and after single CRRT,the total CD8+T cell count,interferon-γ(IFN-γ)-secreting CD8+T cell count,the levels of inhibitory molecules and costimulatory molecules as well as IFN-γand tumor necrosis factor-α(TNF-α)produced by CD8+T cells were detected.Results A total of 37 hospitalized septic patients were infected with gram-negative bacteria, pathogens causing infection were Klebsiella pneumoniae (22 strains),Acinetobacter baumannii (1 1 strains),and Enterobacter cloacae (3 strains).After CRRT,the body temperature,heart rate,white blood cell count,urea nitrogen,and serum creatinine levels in septic patients were all lower than those before CRRT (all P＜0.05).After CRRT,the total CD8+T cell count in septic patients didn’t change significantly (P＞0.05),but IFN-γ-secreting CD8+T cell count increased (P＜0.05).Levels of cytotoxic T lymphocyte-associated antigen 4(CTLA-4),programmed death-1 (PD-1),T-cell immunoglobulin and mucin-do-main-containing molecule 3 (TIM-3)after CRRT were all lower than those before CRRT (all P＜0.05 ),while levels of costimulatory molecules CD28 and secreting IFN-γ elevated after CRRT(all P＜0.05).Conclusion CRRT can not only improve the vital signs and renal function of patients with sepsis,but also enhance the immune function of CD8+T cells.