髓系细胞触发受体-1对大鼠肠巨噬细胞凋亡的影响

Effect of TREM-1 expression on the apoptosis of intestinal macrophages of rats

目的 探讨髓系细胞触发受体-1 （triggering receptor expressed on myeloid cells-1,TREM-1）的表达对大鼠肠巨噬细胞凋亡的影响.方法 体外培养肠巨噬细胞,实验3分组,每组6孔：对照组、LPS组及LPS＋LP17组.加入药物LPS（1 mg/L）和LP17 （0.1 mg/L）,培养6h后用TUNEL试剂盒及流式细胞仪对大鼠肠巨噬细胞凋亡进行检测,利用SPSS 18.0软件进行统计分析.结果 细胞生长状态良好,分布均匀,细胞活力约为80％～85％,CD14免疫荧光鉴定证实为肠巨噬细胞.经药物处理后,TUNEL检测细胞凋亡情况：LPS组（44.33±7.74）％较对照组（19.17±6.01）％明显增多（P=0.000）,LPS＋ LP17组（28.33±6.53）％的凋亡细胞比LPS组（44.33±7.74）％明显减少（P =0.004）,对照组（19.17 ±6.01）与LPS＋ LP17组（28.33±6.53）％的凋亡细胞差异无统计学意义（P =0.050）.采用流式细胞仪检测细胞凋亡情况：LPS组（16.47±1.66）％较对照组（7.70±1.52）％明显增多（P=0.000）,LPS＋LP17组（11.47±3.12）％的凋亡细胞比LPS组（16.47±1.66）％明显减少（P=0.018）,对照组（7.70±1.52）％与LPS＋ LP17组（11.47±3.12）％的凋亡细胞差异无统计学意义（P =0.061）.结论 LP17能抑制肠巨噬细胞TREM-1的表达,减少肠巨噬细胞凋亡.

Objective To investigate the effect of triggering receptor expression in myeloid cells-1 （TREM-1） on intestinal macrophage apoptosis in rat.Methods In vitro,the achieved rat intestinal macrophages were divided into 3 groups：control group,LPS （Lipopolysaccharides） group and LPS ＋ LP17 group （n =6 holes of culture plate in each）.The concentrations of LPS and LP17 were 1 mg/L and 0.1 mg/L,respectively.The intestinal macrophage apoptosis was measured by using TUNEL kit and flow cytometry after culture for 6 h.All data were statistically analyzed using SPSS 18.0 software.Results The shape and growth of rat intestinal macrophages were quite favorable after culture.The membrane marker of intestinal macrophages,CD14 was clearly observed under immunofluorescence.After macrophage was treated with specific procedure,the cell apoptosis found in LPS group （44.33 ± 7.74）% was significantly higher than that in control group （19.17 ± 6.01） % （P=0.000） measured by TUNEL;the cell apoptosis in LPS ＋LP17 group （28.33 ± 6.53）% apparently reduced compared with LPS group （44.33 ±7.74） % （P =0.004）;there was no significant difference in cell apoptosis between control group （19.17 ± 6.01） % and LPS ＋ LP17 group （28.33 ± 6.53） % （P =0.050）.By flow cytometry,the apoptotic cells in LPS group （16.47 ± 1.66） % was significantly increased compared with control group （7.70 ± 1.52） % （P =0.000）;apoptotic cells in LPS ＋ LP17 group （11.47 ± 3.12） % was significantly reduced in comparison with LPS group （16.47 ± 1.66） % （P =0.018）.There was no significant difference in apoptotic cells between control group （7.70±1.52）% and LPS ＋ LP17 group （11.47±3.12） % （P =0.061）.Conclusion LP17 can inhibit TREM-1 expression in intestinal macrophages and reduce intestinal macrophage apoptosis.