2023 年第 1 期 第 18 卷

生长分化因子15对巨噬细胞吞噬杀菌功能影响的初步实验探究

Preliminary experimental study on the effect of growth differentiation factor 15 on phagocytosis and bactericidal function of macrophages

作者：王历李欢汤冬玲张平安

英文作者：Wang Li Li Huan Tang Dongling Zhang Ping′an

单位：武汉大学人民医院检验科，武汉430060

英文单位：Department of Clinical Laboratory Renmin Hospital of Wuhan University Wuhan 430060 China

关键词：生长分化因子15；巨噬细胞；吞噬作用；杀菌功能

英文关键词：Growthdifferentiationfactor15;Macrophages;Phagocytosis;Bactericidalfunction

• 摘要：

• 目的  探究生长分化因子15（GDF15）对巨噬细胞吞噬杀菌功能的影响。方法  将小鼠巨噬细胞RAW264.7和人单核巨噬细胞THP-1，分别分为对照组和GDF15组，各设3个复孔。GDF15组每孔加入用磷酸盐缓冲液稀释的不同浓度（50、100、150、200 μg/L）的GDF15 10 μl，其中RAW264.7细胞采用小鼠GDF15重组蛋白处理、THP-1细胞（加入佛波酯刺激分化为巨噬细胞）采用人GDF15重组蛋白处理；对照组用磷酸盐缓冲液代替GDF15。采用细胞计数盒8（CCK-8）法检测GDF15对2种细胞活性的影响。采用大肠埃希菌作为实验菌株，选择合适的GDF15浓度，采用菌落计数法分析GDF15对巨噬细胞吞噬杀菌功能的影响。结果  CCK-8实验结果显示，在RAW264.7细胞中，50、100、150、200 μg/L GDF15组与对照组吸光度值比较差异无统计学意义（P＞0.05），THP-1来源巨噬细胞中，200 μg/L GDF15组吸光度值明显低于对照组（P=0.026），因此选择150 μg/L作为GDF15对2种巨噬细胞的共同作用浓度进行菌落计数检测。在RAW264.7细胞中，庆大霉素处理0.5 h后GDF15组菌落计数高于对照组［（8 767±351）CFU/ml比（7 267±473）CFU/ml］，处理6 h后GDF15组菌落计数低于对照组［（1 333±379）CFU/ml比（2 267±115）CFU/ml］，GDF15组杀菌比例高于对照组［（0.85±0.05）比(0.69±0.06）］（均P＜0.05）。在THP-1来源巨噬细胞中，庆大霉素处理0.5 h后GDF15组菌落计数高于对照组［（2 167±404）CFU/ml比（1 267±208）CFU/ml］，处理6 h后GDF15组菌落计数低于对照组［（200±100）CFU/ml比（567±153）CFU/ml］，GDF15组杀菌比例高于对照组［（0.88±0.68）比（0.58±0.16）］（均P＜0.05）。结论  GDF15可以增强巨噬细胞的吞噬杀菌功能。

• Objective  To explore the effect of growth differentiation factor 15 (GDF15) on the phagocytosis and bactericidal function of macrophages. Methods  Mouse macrophage RAW264.7 and human monocyte macrophage THP-1 were divided into control group and GDF15 group respectively, each setting up 3 replicates. The macrophages in GDF15 group were intervened by different concentrations of 10 μl GDF15 (50, 100, 150, 200 μg/L) diluted by phosphate buffer (PBS), of which RAW264.7 was treated with mouse GDF15 recombinant protein, and THP-1 (stimulated to differentiate into macrophages by phorbol esters) was treated with human GDF15 recombinant protein. The control group was treated with PBS instead of GDF15. Cell counting box-8 (CCK-8) method was used to detect effect of GDF15 on the two kinds of macrophages. Escherichia coli was used as experimental strain. The appropriate concentration of GDF15 was selected, and the colony counting method was used to anlyze effect of GDF15 on the phagocytosis and bactericidal function of macrophages. Results  CCK-8 trail showed that in RAW264.7, there was no significant differences in absorbance value among 50, 100, 150, 200 μg/L GDF15 groups and control group (P>0.05). In THP-1 derived macrophages, absorbance value in 200 μg/L GDF15 group was lower than that in control group (P=0.026). Finally, 150 μg/L was selected as the concertration of GDF15 for the two kinds of macrophages for colony count detection. In RAW264.7, the colony count in GDF15 group was higher than that in control group 0.5 h after gentamicin intervention ［（8 767±351）CFU/ml vs （7 267±473）CFU/ml］, the colony count in GDF15 group was lower than that in control group 6 h after gentamicin intervention ［（1 333±379）CFU/ml vs （2 267±115）CFU/ml］, and the bactericidal ratio in GDF15 group was higher than that in control group ［（0.85±0.05） vs (0.69±0.06）］(all P<0.05). In THP-1 derived macrophages, the colony count in GDF15 group was higher than that in control group 0.5 h after gentamicin intervention ［（2 167±404）CFU/ml vs （1 267±208）CFU/ml］, the colony count in GDF15 group was lower than that in control group 6 h after gentamicin intervention ［（200±100）CFU/ml vs （567±153）CFU/ml］, and the bactericidal ratio in GDF15 group was higher than that in control group ［（0.88±0.68） vs （0.58±0.16）］(all P<0.05). Conclusion  GDF15 can enhance the phagocytosis and bactericidal function of macrophages.