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The effects of subdiaphragmatic vagotomy on post-septic sleep deprivation-mediated aggravation of systemic inflammation and injuries of lung,liver and kidneyObjective: To explore the effects of subdiaphragmatic vagotomy(SDV)on the systemic inflammation and injuries of lung,liver and kidney induced by short-term sleep deprivation after systemic inflammation.Methods: Adult male C57BL/6J mice were intraperitoneally injected with 5 mg/kg lipopolysaccharide(LPS)followed by 3 consecutive days of sleep deprivation using a multi-platform method.SDV or sham operation was performed 14 days prior to LPS administration,and mice were allowed to recovery for 14 days.Mice were euthanized to record the weight and spleen weight on day 4 after LPS administration.The plasma levels of interleukin(IL)-6,tumour necrosis factor-?(TNF-?)and IL-10 were detected by enzyme-linked immunosorbent assay(ELISA).The pathological damage of the spleen,lung and liver was detected by hematoxylin-eosin(HE)staining,and the glycogen content in the renal tubules was detected by periodic acid-schiff(PAS)staining.The expression of IL-6 in lung,liver and kidney was detected by western blotting and quantitative Real-time PCR(qRT-PCR).Results: Three consecutive days of sleep deprivation after systemic inflammation increased the plasma levels of IL-6 and TNF-? and spleen weight,decreased the plasma levels of anti-inflammatory cytokine IL-10,and exacerbated the pathological injury as well as protein and m RNA expression of IL-6 in lung,liver and kidney,which could be abrogated by SDV 14 days prior to LPS administration.Conclusion: Short-term sleep deprivation after systemic inflammation could aggravate systemic inflammation and multi-organ inflammatory injuries,which could be reversed by SDV 14 days prior to systemic inflammation.Part ? The role of gut microbiota-vagus nerve axis in sleep deprivation-mediated aggravation of systemic inflammation after LPS administrationObjective: To investigate the effects of subdiaphragmatic vagotomy(SDV)on the gut microbial diversity and composition in mice with short-term sleep deprivation after 5mg/kg lipopolysaccharide(LPS)administration,and to study the role of gut microbiota in the sleep deprivation-aggravated systemic inflammation mediated by the subdiaphragmatic vagus nerve after LPS administration.Methods: Adult male C57BL/6J mice were intraperitoneally injected with 5 mg/kg LPS followed by 3 consecutive days of sleep deprivation using a multi-platform method or treated with broad-spectrum antibiotics for 14 consecutive days to construct the post-septic sleep deprivation model and pseudo germ-free(PGF)mouse model,respectively.The PGF mice received fecal suspension from LPS-treated mice with or without sleep deprivation after 14 days of antibiotic treatment.SDV or sham operation was performed 14 days before LPS administration or antibiotic treatment.Fecal samples were collected on day 4 after LPS administration,and the diversity and composition of fecal microbiota were detected with 16 s RNA high-throughput sequencing.The PGF mice weight and spleen weight were recorded on day 14 after fecal microbial transplantation.The plasma levels of interleukin(IL)-6,tumour necrosis factor-?(TNF-?)and IL-10 were detected by enzyme-linked immunosorbent assay(ELISA).The expression of IL-6 m RNA in the lung,liver and kidney was detected by q RT-PCR.Results: SDV did not alter the ?-diversity and ?-diversity of fecal microbiota,but could regulate the alteration of fecal microbial composition in mice with short-term sleep deprivation after systemic inflammation.SDV reversed the increase in the relative abundance of Proteobacteria and its subgroups caused by short-term sleep deprivation after systemic inflammation.Furthermore,the spleen weight,spleen/body weight ratio,plasma levels of IL-6 and TNF-?,as well as the m RNA expression of IL-6 in lung,liver and kidney were significantly increased,whereas the plasma level of IL-10 was significantly decreased in PGF mice transplanted with fecal suspension from LPS-treated mice with sleep deprivation compared with that in PGF mice transplanted with fecal suspension from LPS-treated mice without sleep deprivation,which was abrogated by SDV 14 days prior to antibiotics administration.Conclusion: SDV could regulate the alteration of fecal microbial composition induced by short-term sleep deprivation after systemic inflammation,while the alteration of fecal microbiota induced by sleep deprivation after systemic inflammation might be involved in the sleep deprivation-aggravated systemic inflammation and the inflammatory injuries of lung,liver and kidney through the subdiaphragmatic vagus nerve after LPS administration.Part ? The role of gut microbiota-spleen axis in sleep deprivation-mediated aggravation of systemic inflammation after LPS administrationObjective: To explore the role of gut microbiota in the sleep deprivation-aggravated systemic inflammation mediated by spleen after LPS administration.Methods: Adult male C57BL/6J mice were treated with broad-spectrum antibiotics for 14 consecutive days to construct the pseudo germ-free(PGF)mouse model.The splenectomy or sham operation was performed 14 days before antibiotic treatment.The fecal suspension from LPS-treated mice with or without sleep deprivation was transplanted after 14 days of antibiotic treatment.The plasma levels of interleukin(IL)-6,tumour necrosis factor-?(TNF-?)and IL-10 were detected by enzyme-linked immunosorbent assay(ELISA)on day14 after fecal microbial transplantation.Results: In PGF mice underwent sham-operation,fecal microbial transplantation with fecal suspension from LPS-treated mice with sleep deprivation significantly increased the plasma levels of IL-6 and TNF-? and decreased the plasma IL-10 level compared with that from LPS-treated mice without sleep deprivation,which was abrogated by splenectomy 14 days prior to antibiotics administration.Conclusion: Gut microbiota might be involved in the post-septic sleep deprivation-aggravated systemic inflammation through spleen. |
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