胍丁胺对脓毒症大鼠免疫细胞凋亡的影响

The effects of agmatine on the apoptosis of immune cells in septic rats

目的通过建立内毒素诱导的脓毒症大鼠模型,探讨胍丁胺对脓毒症大鼠脾细胞与树突状细胞凋亡的影响。方法取健康清洁级雄性SD大鼠90只,随机(随机数字法)分为对照组、内毒素组、胍丁胺组,每组30只;对照组经股静脉注射生理盐水(10 mL/kg)、内毒素组经股静脉注射脂多糖(10 mg/kg)、胍丁胺组经股静脉注射脂多糖(10 mg/kg)并同时腹腔注射胍丁胺(200 mg/kg);三组大鼠于建模后0h、12h、24h分别随机抽取10只(标记为0h、12h、24h亚组)麻醉处死;称取脾脏重量;HE染色观察脾脏病理;流式细胞术检测脾细胞与树突状细胞的总凋亡率;结果运用SPSS 17.0软件进行统计分析,采用独立样本t检验进行分析,以P<0.05为差异有统计学意义。结果内毒素组中12 h(1.2633±0.0652)、24 h亚组(1.5576±0.0711)的脾脏重量(g)与对照组中相应亚组[(0.8876±0.0361)、(0.9079±0.0425)]相比明显升高(P<0.05),胍丁胺组中12 h(1.1052±0.0585)、24 h亚组(1.3262±0.0682)的脾脏重量与内毒素组中相应亚组相比明显降低(P<0.05);HE染色见内毒素组睥脏组织炎症反应与对照组相比明显加重(P<0.05),胍丁胺组与内毒素组相比明显减轻(P<0.05);内毒素组中12h、24h亚组的脾细胞[(13.31±1.26)、(19.53±1.68)]及树突状细胞[(19.5±1.52)、(16.09±1.15)]的总凋亡率与对照组中相应亚组[(6.27±0.71)、(6.01±0.67)及(4.99±0.51)、(5.30±0.66)]相比均明显升高(P<0.05),胍丁胺组中12 h[(9.19±0.95)、(12.19±1.25)]、24 h亚组[(12.71±1.19)、(10.76±1.09)]的上述检测指标与内毒素组中相应亚组相比均明显降低(P<0.05);内毒素组及胍丁胺组中24 h亚组的脾细胞凋亡率与同组中12 h亚组相比较明显升高(P<0.05),而树突状细胞凋亡率与同组中12 h亚组相比较明显降低(P<0.05)。结论脾细胞、树突状细胞的凋亡与脓毒症的发生发展密切相关,胍丁胺可抑制脓毒症大鼠脾细胞、树突状细胞的凋亡。

Objective By establishing the rats model of sepsis induced by endotoxin,exploring the effects of agmatine on the apoptosis of splenocyte and dendritic cells in the septic rats.Methods SD rats(healthy and clean,male,90)were randomly(random number)divided into control groups,endotoxin groups and agmatine groups,30 rats per group.The control groups were injected with normal saline via femoral vein(10 mL/kg),endotoxin groups were injected with lipopolysaccharide via femoral vein(10 mg/kg),agmatine groups were injected with lipopolysaccharide via femoral vein(10 mL/kg)and intraperitoneal injected of agmatine(200 mg/kg).The three groups were randomly selected 10 rats separately at 0 h,12 h,24 h(marked as 0 h,12 h,24 h subgroups,n=10),anesthetized to death.Weigh the spleen;HE staining was used to observe the pathological changes of the spleen;The cell apoptosis of splenocyte and dendritic cells were detected by flow cytometry.The results were statistically analyzed using SPSS 17.0 software,analyzed by independent sample t test,p<0.05 was considered statistically significant.Results The weights(g)of spleen in 12 h(1.2633±0.0652),24 h(1.5576±0.0711)subgroups of the endotoxin groups were increased significantly than the corresponding subgrou ps[(0.8876±0.0361),(0.9079±0.0425)]ofthe control groups(P<0.05),the 12 h(1.1052±0.0585),24 h(1.3262±0.0682)subgroups of the agmatine groups were decreased significantly than the corresponding subgroups of the endotoxin groups(P<0.05).HE staining showed that the spleen tissue inflammatory reaction in the endotoxin groups were worse than the control group(P<0.05),the agmatine groups were better than the endotoxin group(P<0.05).The apoptosis rates of the splenocyte[(13.31±1.26)、(19.53±1.68)]and dendritic cells[(19.5±1.52)、(16.09±1.15)]in the spleen from 12 h,24 h subgroups of the endotoxin groups were significantly higher than the corresponding subgroups[(6.27±0.71),(6.01±0.67)and(4.99±0.51)、(5.30±0.66)]of the control groups(P<0.05),the 12 h[(9.19±0.95),(12.19±1.25)],24 h[(12.71±1.19),(10.76±1.09)]subgroups of the agmatine groups were significantly lower than the corresponding subgroups of the endotoxin groups(P<0.05);The apoptosis rates of the splenocyte in the 24 h subgroups of the endotoxin groups and the agmatine groups were significantly higher than the 12 h subgroups of the same group(P<0.05),but the apoptosis rates of the dendritic cells were significantly lower than the 12h subgroups of the same group(P<0.05).Conclusion The apoptosis of splenocyte and dendritic cells were closely related to the occurrence and development of sepsis,Agmatine could inhibit the apoptosis of splenocyte and dendritic cells the rats with sepsis.