The Study Of Inflammatory Responses And Ultrastructural Changes Of IL-6 Co-cultured Neurons In Vitro And Hippocampal Neurons In Postoperative Sleep Deprivation Rats

Objective: The sleep deprivation model of rats after splenectomy was established with modified multi-platform method(MMPM),and IL-6 was used to stimulate hippocampal neurons in vitro to explore the inflammatory response and ultrastructural changes of the two models.Methods: Twenty four adult rats(6-7 weeks)weighing 200-220 g were adapted for 7days before the intervention,and then randomly divided into four groups on the 7th day.The groups were expressed as control group(Control group,n=6),surgery group(S group,n=6),sleep deprivation group(SD group,n = 6),and postoperative sleep deprivation group(SSD group,n=6).On the 8th day,the S group and SSD group were anesthetized and underwent splenectomy.On day 9,the SD group and the SSD group were subjected to sleep deprivation with MMPM for 24 hours.On the 5th day after splenectomy,the hippocampus tissues were obtained by craniotomy after cardiac perfusion and brain fixation,which was used for the subsequent observation by transmission electron microscopy and the measurement of IL-6 concentration in the hippocampal tissue by ELISA.IL-6(10ng / ml)was co-cultured with hippocampal neurons for 24 h in vitro,followed by MTT assay,flow cytometry and transmission electron microscopy.After that,we analyzed the changes of IL-6 concentration and ultrastructure in rat hippocampal tissue cells,as well as the proliferation rate,apoptosis and ultrastructure of hippocampal neurons in vitro.Results: Compared with the control group,all groups showed higher levels of IL-6.Compared with the S group,the concentration of IL-6 in the SSD group decrease.Under transmission electron microscope,the hippocampal neurons in the control group had intact cell membrane,homogeneous cytoplasm and no damage to organelles.In the S group,SD group and SSD group,hippocampal tissues neurons showed different degrees of mitochondrial swelling,crest disorder,vacuolization and endoplasmic reticulum expansion,and the degree of neuron injury was more obvious in the S group.Compared with the control group,IL-6 co-cultured with hippocampal neurons in vitro,cell average proliferation rate was 73.53%,and apoptosis or death of neurons increased significantly.Hippocampal neurons with mitochondrial swelling,cristae disorder and rupture,outer membrane rupture and vacuolation,endoplasmic reticulum expansion,secondary lysosomes increasing,chromatin aggregation and nuclear rupture of some neurons were observed under transmission electron microscopy.Conclusion: Acute 24 h sleep deprivation one the basis of surgery can decrease the inflammatory response in the hippocampal tissue.The 10 ng / ml IL-6 co-cultured neurons were consistent with the ultrastructural changes of nerve cells in rat hippocampal tissue caused by 24-hour sleep deprivation after surgery,and the former was more severely damaged.