丁酸盐通过调控酸性鞘磷脂酶缓解烫伤诱导的T细胞损失的小鼠实验研究  

Butyrate relieves scald-induced T cells loss of mouse experimental study by regulating acid sphingomyelinase

作　　者：张雷[1] 武斌[1] 高东东[1] 杨喜明[1] ZHANG Lei;WU Bin;GAO Dong-dong(Department of Burn Plastic Surgery,Yan'an University Affiliated Hospital,Yan'an Shaanxi 716000,China)

机构地区：[1]延安大学附属医院烧伤整形手外科,陕西延安716000

出　　处：《临床和实验医学杂志》2018年第24期2581-2584,共4页Journal of Clinical and Experimental Medicine

基　　金：陕西省自然科学基础研究计划项目(编号:2014JM4199)

摘　　要：目的探讨丁酸盐调控烫伤诱导的T细胞变化的作用机制。方法将小鼠随机分为3组:对照组(腹腔注射灭菌的生理盐水1. 5 ml)、烫伤组(烫伤模型)和丁酸组(烫伤模型+腹腔注射1 g/kg的丁酸注射)。采用高效液相色谱于烫伤模型建立后第1、2、3、4、5、6、7和8天检测对照组和烫伤组小鼠粪便中丁酸盐浓度。平板培养法检测3组中肠道菌群,试剂盒法检测酸性鞘磷脂酶(Asm)活性,流式细胞术检测T淋巴细胞各细胞亚型的表达。采用Ficoll-Paque梯度离心法从小鼠脾脏中分离T淋巴细胞。将细胞分为3组:对照组(不处理)、Asm抑制组(10μM的Asm抑制剂地昔帕明)和Asm抑制+丁酸组(Asm抑制剂+100μM的丁酸),检测各组细胞中Asm活性及T淋巴细胞各细胞亚型的表达。结果与对照组相比,烫伤后第3、5、6、7和8天小鼠粪便中丁酸浓度均显著降低(P <0. 05)。丁酸盐可显著增加烫伤后小鼠粪便中乳酸杆菌属、双歧杆菌属、普拉梭菌和罗氏菌属的含量(P <0. 05),降低硫酸盐还原菌的含量(P <0. 05)。丁酸盐可促进烫伤后Asm激活,增加烫伤后脾脏CD4 T细胞、CD4初始T细胞、CD8 T细胞及CD8初始T细胞的数量(P <0. 05)。Asm抑制组中Asm的活性显著低于对照组,而Asm抑制+丁酸组高于Asm抑制组(P <0. 05)。Asm抑制组CD4 T细胞、CD4初始T细胞、CD8 T细胞及CD8初始T细胞的数量均低于对照组,而Asm抑制+丁酸组高于Asm抑制组(P <0. 05)。结论丁酸盐可通过激活Asm修复烫伤诱导的肠道菌群失衡,缓解烫伤诱导的T细胞丢失。Objective To explore the mechanism of butyrate to regulate scald-induced T cell changes.Methods Mice were randomly divided into 3 groups:control group,scald group(scalding model)and butyric acid group(scalding model+10 g/kg of butyric acid injection).The concentrations of butyrate in the stool of the control group and the scald group were measured by high performance liquid chromatography on the 1st,2nd,3rd,4th,5th,6th,7th and 8th day after the establishment of the scald model.The intestinal microbiota of the 3 groups was detected by plate culture method.The activity of acid sphingomyelinase(Asm)was detected by kit method,and the expression of subtypes of T lymphocytes was detected by flow cytometry.T lymphocytes were isolated from mouse spleens using Ficoll-Paque gradient centrifugation.The cells were divided into 3 groups:control group(no treatment),Asm inhibitor group(10μM Asm inhibitor desipramine)and Asm inhibitor+butyric acid group(Asm inhibitor+100μM butyric acid).The expression of Asm and the expressions of each subtype of T lymphocytes in each group of cells were examined.Results Compared with the control group,the concentration of butyric acid in the stool of mice was significantly decreased on the 3rd,5th,6th,7th and 8th day after scald(P<0.05).Butyrate could significantly increase the numbers of Lactobacillus,Bifidobacterium,Faecalibacterium prausnitzii and Roseburia in the stool of mice(P<0.05),and reduce the numbers of sulfate-reducing bacteria(P<0.05)after scald.Butyrate promoted the activation of Asm after scald,and increased the numbers of spleen CD4 T cells,na ve CD4 T cells,CD8 T cells and na ve CD8 T cells after scald(P<0.05).Asm inhibitors could reduce the numbers of the T cells.The activity of Asm in Asm inhibitor group was significantly lower than that in the control group,while the Asm inhibitor+butyric acid group was higher than the Asm inhibition group(P<0.05).The numbers of CD4 T cells,na ve CD4 T cells,CD8 T cells and na ve CD8 T cells in Asm inhibitor group were lower than those in

关 键 词：小鼠 烫伤 丁酸盐 酸性鞘磷脂酶 肠道菌群 T细胞 

分 类 号：R644[医药卫生—外科学]