Study On The Mechanism Of Chronic Intermittent Hypoxia Induced Heart And Brain Damage Of Rats And The Effects Of Drug Intervention

Objective: In vitro and in vitro experiments were conducted to explore the potential molecular mechanism of the FOXO1 inhibitor AS1842856 in the treatment of myocardial apoptosis and myocardial hypertrophy induced by CIH,providing a theoretical basis for AS1842856 in the treatment of OSAS myocardial hypertrophy complications.Methods: H9C2 myocardial cells were cultured In vitro.IH model were established in Oxycycler model C21 hypoxic cell incubator.The changes of apoptotic protein were detected by Western Blot.The number of apoptotic cells was detected by Flow cytometry.The effects of FOXO1 and Bim on myocardial apoptosis were observed by using si RNA technology and lentivirus technology.The upstream and downstream relationship between FOXO1 and Bim was detected by immunoprecipitation.In animal experiments,Oxy Cycler A84 low oxygen chamber was used to make CIH environment.40 SD male rats were randomly divided into normoxic oxygen(NC)group;normoxic oxygen +AS1842856(NC+T)group;CIH group;CIH+AS1842856(CIH+T)group.Each group had 10 rats,and the experiment lasted 8 hours per day,with 7 days per week.The the time of building animal model lasted eight weeks.The rats in the NC+T group and the CIH+T group were injected with FOXO1 inhibitor AS1842856,respectively on the28 th day and on the 42 nd day.Immunohistochemical technique and Western Blot technique were used to observe the change of FOXO1 and Bim in myocardial cells.Hematoxylin-Eosin and Mession staining were used to observe the myocardial structureof each group.TUNEL technique was used to observe the apoptosis of myocardial cells in each group.Results: In vitro cell experiment,IH induced the phosphorylation of FOXO1 and improved the expression level of FOXO1.The activation of Bim could lead to the apoptosis of myocardial cells induced by IH.In IH environment,FOXO1 was Bim upstream regulatory factor in H9C2 cardiomyocytes,and they had a direct interaction relations,FOXO1 activated the expression of Bim,which regulated the occurrence of apoptosis,inhibiting the expression of FOXO1 could reduce myocardial cell apoptosis induced by IH.In vivo,the expression of FOXO1 and Bim increased significantly in cardiac hypertrophy tissue induced by CIH,AS1842856 inhibitd the expression of apoptosis factor Bim significantly and improved the myocardial hypertrophy and myocardial cell apoptosis induced by CIH.Conclusions: In H9C2 cell experiment,FOXO1 / Bim signaling pathways regulated the H9C2 myocardial cell apoptosis.In animal experiments,CIH induced myocardial cell apoptosis and myocardial hypertrophy,FOXO1 / Bim signaling pathways regulated the myocardial cell apoptosis induced by CIH,AS1842856 obviously alleviated the myocardial cell apoptosis and myocardial hypertrophy induced by CIH.Objective: This experiment mainly intended to study the mechanism of FOXO1/Bim signaling pathway leading to aortic smooth muscle cell apoptosis in the process of aorta damage induced by chronic intermittent hypoxia in rats,and to research the treatment effect of FOXO1 inhibitor AS1842856 on anti-apoptosis,which may seeking new approaches and therapeutic targets for the treatment of aortic arch damage caused by OSAS.Methods: Using Oxy Cycler A84 low-oxygen chamber to make CIH environment and create Animal model,40 SD male rats were randomly divided into normal oxygen(NC)group,normal oxygen +AS1842856(NC+T)group,CIH group,CIH+AS1842856(CIH+T)group,each group had 10 rats.The CIH group and the CIH+T group were placed in hypoxia chamber during the daytime sleep,and the experiment lasted 8 hours per day,with 7 days per week.The the time of building animal model lasted eight weeks.After the model was finished,the Polygraph System was used to measure aortic systolic blood pressure in rats,the damage of the aortic arch smooth muscle cells was detected by the Hematoxylin-Eosin staining method and the internal-medial membrane thickness of the aortic arch was measured,Immunohistochemical technique was used to detect the expression of FOXO1,Bim and Ki67 in aortic smooth muscle cells in each group,The apoptosis of smooth muscle cells in each group was observed by TUNEL technique.Results: CIH induced intervention increased blood pressure in SD rats,and FOXO1 inhibitor AS1842856 reduced the elevated blood pressure induced by CIH.The CIH induced intervention lead to thickening of the internal-medial membrane of the aorta,and the FOXO1 inhibitor AS1842856 can reverse the thickening of the aortic arch.The CIH induced intervention resulted in the increase of FOXO1 and Bim expression in the aortic arch smooth muscle cells,and the expression of Bim was decreased after the inhibition of FOXO1.CIH induced intervention resulted in the decrease of normal proliferation in the aortic arch smooth muscle cells,but promoted abnormal apoptosis.However,the FOXO1 inhibitor AS1842856 can correct the normal proliferation of the smooth muscle cells of the aortic arch and can reduce the occurrence of abnormal apoptosis.Conclusions: FOXO1 / Bim signaling pathways can mediate the apoptosis process of the aorta smooth muscle cells induced by chronic intermittent hypoxia,AS1842856 can significantly reversed this process,and may be a potential targeted drug for the treatment of OSAS with high blood pressure complication.Objective: This experiment Mainly intended to study the mechanism of FOXO1/Bim signaling pathway leading to hippocampus neuronal cell apoptosis in the process of hippocampus damage induced by chronic intermittent hypoxia in rats,and to research the treatment effect of FOXO1 inhibitor AS1842856 on anti-apoptosis,which may seeking new approaches and therapeutic targets for the treatment of hippocampus damage caused by OSAS.Methods: Using Oxy Cycler A84 low-oxygen chamber to make CIH environment and create Animal model,40 SD male rats were randomly divided into normal oxygen(NC)group,normal oxygen+AS1842856(NC+T)group,CIH group,CIH+AS1842856(CIH+T)group,each group had 10 rats.CIH group and the CIH+T group were placed in hypoxia chamber during the daytime sleep,and the experiment lasted 8 hours per day,with 7 days per week.The time of building animal model lasted eight weeks.Morris water maze experiment was used to detect the cognitive function of rats.HE staining method was applied to detect the damage extent of hippocampal structure.Immunohistochemical technique was used to detect the expression of FOXO1,Bim and Ki67 in hippocampus neuronal cells in each group.The apoptosis of hippocampus neuronal cells in each group was observed by TUNEL technique.Results: CIH induced intervention resulted in the damage of hippocampal neuronal cell,resulting in increased expression of FOXO1 and Bim in hippocampal neuronal cells.After the expression of FOXO1 was inhibited,the expression of Bim decreased,CIH induced intervention resulted in the decrease of normal proliferation of hippocampal neuronal cells,and promoted abnormal apoptosis.However,the FOXO1 inhibitorAS1842856 can correct the normal proliferation of hippocampal neuronal cells and reduce the occurrence of abnormal apoptosis,and AS1842856 can improve the cognitive impairment of SD rats induced by CIH.Conclusions: FOXO1/Bim signaling pathway can mediate the apoptosis process of hippocampal neuronal cells induced by CIH,and the FOXO1 inhibitor AS1842856 can significantly reverse this process,which might be a potential target drug for the treatment of OSAS with hippocampal damage complication.