| ObjectiveInvestigate the neuroprotective effect of 3-n-butylphthalide on sepsis mice.MethodsSeventy healthy adult male C57BL/6 mice were divided into three groups:Sham group(n=10),sepsis group(n=30),sepsis + NBP group(n=30).All mice were given Morris water maze(MWM)training every day three days before CLP or sham operation.The hidden platform test and exploration test were conducted on the day before operation;modeling;6 hours after CLP or sham operation,the mice in NBP group were given 70mg/kg/d of 3-n-Butylphthalide,and the same volume vegetable oil was fed by nasal feeding in sham group and CLP group.The number of deaths in each group was recorded for 7 days to calculate the mortality rate;The nerve reflex was evaluated before and after operation,4th day and 7th day after operation;5 surviving mice in each group were perfused and sampled 24 hours after CLP or sham operation,and oxidative stress and inflammatory indexes were detected after the homogenate of hippocampus;the survival mice of each group were tested and explored on the 4th and 7th days after sham operation or CLP.Immediately after MWM,the surviving mice in each group were perfused and hippocampal tissue was taken.HE staining was performed on the left side and apoptotic cells were detected on the right side.Results1.24 hours after CLP or sham operation,there was no death in Shamgroup,5 in CLP Group and 4 in NBP group.The mortality of CLP Group and NBP group was 83.3% vs 86.6%(P=1.00),and there was no significant difference between the two groups.On the 4th day after operation,no mice died in Sham group,15 mice died in CLP Group,and 10 mice died in NBP group(Note: the above dead mice did not include mice used for 24-hour determination of oxidative stress and inflammatory factors).The mortality of mice in CLP Group and NBP group was(60.0% vs 40.0%,P<0.05).On the 7th day after operation,there was no death in Sham group,19 in CLP Group and 10 in NBP group.The mortality of CLP Group and NBP group was 76.0% vs 40%(P<0.05).Kaplan-Meier analysis showed that there were differences between Sham group and CLP Group(P<0.05),between CLP group and NBP group(P<0.05),and between Sham group and NBP group(P<0.05).2.Compared with Sham group,the concentrations of SOD and MDA in CLP Group were significantly increased(P<0.05);the concentrations of TNF-αand IL-6 were significantly increased(P<0.05).Compared with CLP Group,the concentrations of SOD and MDA in NBP group were lower(P<0.05),and the concentrations of TNF-α and IL-6 were lower(P<0.05);but compared with Sham group,the concentrations of SOD and MDA,TNF-α and IL-6 in NBP group were higher(P<0.05).3.24 hours after operation,the nerve reflex evaluation scores of CLP group and NBP group were significantly lower than Sham group,and there were statistical differences between the two groups(P<0.05).Although the score of NBP group was slightly higher than that of CLP group,there was no statistical difference between the two groups(P=0.108).On the 4th day after operation,the nerve reflex scores of CLP group and NBP group were still lower than Sham group,and there were statistical differences between the two groups(P<0.05);the nerve reflex scores of NBP group were higher than those of CLP group,but there was no statistical difference between CLP group and NBP group(P=0.11);on the 7th day after operation,the nerve reflex scores of CLP group and NBP group were still lower than Sham group,and there were still differences between the two groups and Sham group.The score of NBP group was higher than CLP group,and there was statistical difference between the two groups(P<0.05).4.On the day of operation,there was no significant difference in escape latency,proportion of entering target quadrant and times of entering target quadrant among all groups;on the fourth day after operation,compared with sham group,escape latency was significantly prolonged,proportion and times of entering target quadrant were significantly decreased in CLP group and NBP group,and there was significant difference between the two groups and Sham group(P<0.05).Compared with CLP group,NBP group had shorter latency extension,lower proportion and frequency of entering the current quadrant(P<0.05);compared with Sham group,CLP Group had longer escape latency,lower proportion and frequency of entering the target quadrant on the 7th day after operation(P<0.05);however,compared with the 4th day after operation,the performance of CLP group was improved and the survival rate was improved,and the difference was statistically significant(P<0.05).The latency avoidance time,proportion and times of entering target quadrant in NBP group were significantly better than those in Sham group on day 4,and there was no significant difference between NBP group and Sham group(P=0.681).5.Under the light microscope,the size of neurons and glial cells in the hippocampus of Sham group was normal,and the cell structure was clear;in CLP group,the nuclear of neurons were pyknosis and hyperchromatic,the cytoplasm of neurons was edematous,and the stroma was edematous;in NBP group,the neurons were pyknosis and hyperchromatic,and the surrounding tissue was edematous,but the degree was lighter than that in CLP group.6.A small amount of green fluorescence appeared in the blue background of Sham group.The green fluorescence of CLP group was significantly morethan that of Sham group.The green fluorescence of NBP group was more than that of Sham group,but less than that of CLP group.Conclusions3-n-butylphathlide can reduce the level of inflammatory factors and the degree of oxidative stress reaction in the hippocampus of septic mice,reduce the apoptosis of neurons,improve the spatial memory impairment of mice,and reduce the long-term mortality of mice. |
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