The Protective Effect And Mechanism Of Panax Notoginseng Saponins On The Nerve And Heart Of Sleep Deprived Mice

Objective:To investigate the neuroprotective and cardiooprotective effect and mechanism of Stem-leaf saponins from Panax notoginseng(SLSP)on mice with sleep deprivation(SD).The neuroprotective effects of Panax notoginseng saponins on C57BL/6 mice after sleep deprivation were tested by Morris Water Maze(MWM)and multi-platform sleep deprivation.Mouse hippocampal neurons HT-22 cells and rat cardiac H9c2 cells were used in this study.Methods:1.The effect of SLSP on the learning and memory ability of mice with sleep deprivation: The Morris water maze was used to train mice’s learning and memory ability and the mice were subjected to 48 hours of sleep deprivation by modified multi-platform method.2.The mechanism of SLSP on improving learning and memory ability in mice with sleep deprivation: Nissl staining,transmission electron microscopy,immunofluorescence and western blot were used to study the protective effect of Panax notoginseng saponins on hippocampal neurons in sleep deprived mice.mechanism.3.Protection of SLSP on HT-22 cells with abnormal autophagy and apoptosis induced by rapamycin: CCK-8 was used to detect the effect of SLSP on cell viability of HT-22 cells pretreated by rapamycin.The effects of SLSP on the autophagy of hippocampal neurons HT-22 cells were observed by transient transfection of tf-LC3,orange staining and flow cytometry.Western blot was used to observe the mechanism of SLSP on rapamycin-induced excessive autophagy in mouse hippocampal neurons HT-22 cells.4.The protective effect and mechanism of SLSP on sleep deprived mice: HE staining,immunofluorescence and western blot were used to study the protective mechanism of Panax notoginseng saponins on the cardioprotection of sleep deprived mice.5.The effects of SLSP on rat cardiac H9c2 cells were observed by transient transfection of GFP-LC3 and tf-LC3,orange staining and flow cytometry.Western blot was used to observe the mechanism of SLSP on rapamycin-induced excessive autophagy and apoptosis on rat cardiac H9C2 cells.1.SLSP can significantly decrease escape latency of mice before SD and improve the frequency of crossing the platform after SD.2.SLSP can alleviate morphological damage of hippocampal neurons and decrease the autophagosome of SD mice.SLSP can also inhibit excessive expression of LC3 B in hippocampus of SD mice.SLSP can inhibit autophagy and apoptosis-related proteins of hippocampus SD mice through modulating PI3K/Akt/m TOR pathway.3.SLSP can supress abnormal autophagy and apoptosis induced by rapamycin of HT-22 cell through PI3K/Akt/m TOR pathway.4.SLSP can inhibit morphological change in heart tissue of SD mice.SLSP can suppress expression of proteins related autophagy and apoptosis of heart in SD mice by modulating PI3K/Akt/m TOR pathway.5.The SLSP can significantly inhibit the excessive autophagy and apoptosis of H9C2 cells induced by rapamycin through modulating PI3K/Akt/m TOR pathway.Conclusion: SLSP can significantly inhibit hippocampal neuronal injury and heart injury in mice with sleep deprivation.The mechanism of SLSP is to inhibit excessive autophagy and apoptosis of hippocampal neurons and cardiomyocytes by regulating PI3K/Akt/m TORt pathway.