七氟醚通过影响外周血miR-340水平逆转脓毒症大鼠巨噬细胞吞噬功能抑制状态的机制

Mechanism of Sevoflurane Reversing the Inhibition of Macrophage Phagocytosis in Septic Rats by Influencing the Level of Peripheral Blood miR-340

目的:探究七氟醚通过影响外周血miR-340水平逆转脓毒症大鼠巨噬细胞吞噬功能抑制状态的机制。方法:60只雄性Sprague-Dawley大鼠根据研究目的将大鼠分为对照组、脓毒症组和七氟醚组。通过RT-PCR分析各组大鼠外周血miR-340以及促炎因子IL-1β、IL-6和TNF-α的m RNA表达水平。统计各实验组大鼠的存活率。通过血琼脂平板对各组大鼠腹腔液和血液中的细菌进行计数。通过使用荧光显微镜测量吞噬率和吞噬指数。通过蛋白印迹分析p65和NF-κB的蛋白表达。结果:脓毒症组miR-340水平较对照组升高(P<0.05),七氟醚组miR-340水平较脓毒症组降低(P<0.05)。三组不同时间点存活率相比差异无统计学意义(P>0.05);第4 d、8 d脓毒症组大鼠存活率较对照组大鼠降低(P<0.05),七氟醚组大鼠存活率较脓毒症组升高(P<0.05)。脓毒症组IL-1β、IL-6和TNF-α的m RNA表达较对照组升高(P<0.05),七氟醚组IL-1β、IL-6和TNF-α的m RNA表达较脓毒症组降低(P<0.05)。脓毒症组腹腔液和血液中的细菌数量较对照组升高(P<0.05),七氟醚组腹腔液和血液中的细菌数量较脓毒症组减少(P<0.05)。血琼脂平板对各组大鼠腹腔液和血液中的细菌进行计数,脓毒症组腹腔液和血液中的细菌数量较对照组升高(P<0.05),七氟醚组腹腔液和血液中的细菌数量较脓毒症组减少(P<0.05)。脓毒症组p65和NF-κB的蛋白表达较对照组升高(P<0.05),七氟醚组p65和NF-κB的蛋白表达较脓毒症组降低(P<0.05)。结论:miR-340参与了脓毒症大鼠巨噬细胞吞噬功能调节,七氟醚通过降低外周血miR-340水平减少内毒素诱导的大鼠促炎细胞因子的释放,并恢复巨噬细胞吞噬功能。

Objective:To explore the mechanism by which sevoflurane can reverse the inhibition of macrophage phagocytosis in septic rats by affecting the level of miR-340 in peripheral blood.Methods:60 male Sprague-Dawley were divided into control group,sepsis group and sevoflurane group according to research purposes.The mRNA expression levels of peripheral blood miR-340 and pro-inflammatory factors IL-1β,IL-6 and TNF-αwere analyzed by RT-PCR.Statistics of the survival rate of rats in each experimental group.Bacteria in the peritoneal fluid and blood of each group of rats were counted by blood agar plate.The phagocytic rate and phagocytic index were measured by using a fluorescence microscope.The protein expression of p65 and NF-kB was analyzed by Western blot.Results:The level of miR-340 in the sepsis group was higher than that in the control group(P<0.05),and the level of miR-340 in the sevoflurane group was lower than that in the sepsis group(P<0.05).There was no significant difference in survival rate among the three groups at different time points(P>0.05);on the 4th and 8th day,the survival rate of rats in the sepsis group was lower than that in the control group(P<0.05),and the survival rate of rats in the sevoflurane group was higher than that in the sepsis group(P<0.05).The mRNA expression of IL-1β,IL-6 and TNF-αin the sepsis group was higher than that in the control group(P<0.05),and the mRNA expression of IL-1β,IL-6 and TNF-αin the sevoflurane group was higher than that in the sepsis group The symptom group decreased(P<0.05).The number of bacteria in peritoneal fluid and blood in the sepsis group was higher than that in the control group(P<0.05),and the number of bacteria in the peritoneal fluid and blood in the sevoflurane group was lower than that in the sepsis group(P<0.05).The blood agar plate counts the bacteria in the peritoneal fluid and blood of the rats in each group.The number of bacteria in the peritoneal fluid and blood of the sepsis group was higher than that in the control group(P<0.05),and the number of peritoneal fluid and blood in the sevoflurane group The number of bacteria was lower than that in the sepsis group(P<0.05).The protein expression of p65 and NF-kB in the sepsis group was higher than that in the control group(P<0.05),and the protein expression of p65 and NF-kB in the sevoflurane group was lower than that in the sepsis group(P<0.05).Conclusion:miR-340 is involved in the regulation of macrophage phagocytosis in septic rats.Sevoflurane reduces the release of endotoxin-induced rat pro-inflammatory cytokines by reducing the level of peripheral blood miR-340,and restores macrophage phagocytosis Features.