高迁移率族蛋白B1对磷酸钙诱导巨噬细胞释放炎症因子的协同作用

背景：研究表明，巨噬细胞及其炎症反应参与了肾结石的发生发展。前期实验发现结石晶体可刺激巨噬细胞释放高迁移率族蛋白B1。 目的：观察高迁移率族蛋白B1对磷酸钙诱导巨噬细胞释放白细胞介素1β、白细胞介素6、肿瘤坏死因子α、单核细胞趋化因子1的协同作用。 方法：实验分两部分：①将成功诱导为巨噬细胞的U937细胞分为空白组、100 mg/L磷酸钙组、100 μg/L高迁移率族蛋白B1组、100 mg/L磷酸钙+100 μg/L高迁移率族蛋白B1组，干预1，2，4 h后收集细胞上清液。②将已成功诱导为巨噬细胞的U937细胞分为100 mg/L磷酸钙组、磷酸钙+10 μg/L高迁移率族蛋白B1组、磷酸钙+50 μg/L高迁移率族蛋白B1组、磷酸钙+100 μg/L高迁移率族蛋白B1组，干预4 h后收集细胞上清液。Elisa法检测白细胞介素1β、白细胞介素6、肿瘤坏死因子α、单核细胞趋化因子1水平。 结果与结论：ELISA结果显示，磷酸钙组，100 μg/L高迁移率族蛋白B1组上清液白细胞介素1β、白细胞介素6、肿瘤坏死因子α、单核细胞趋化因子1质量浓度均高于空白组，磷酸钙+100 μg/L高迁移率族蛋白B1组上清液上述因子质量浓度均显著高于其他3组(P < 0.05)，且呈时间依赖性。不同质量浓度高迁移率族蛋白B1+磷酸钙组细胞上清液白细胞介素1β、白细胞介素6、肿瘤坏死因子α、单核细胞趋化因子1水平均显著高于磷酸钙组(P < 0.05)，且呈浓度依赖性。结果表明，磷酸钙及高迁移率族蛋白B1均可诱导巨噬细胞释放白细胞介素1β、白细胞介素6、肿瘤坏死因子α、单核细胞趋化因子1；高迁移率族蛋白B1可协同磷酸钙诱导巨噬细胞释放白细胞介素1β、白细胞介素6、肿瘤坏死因子α、单核细胞趋化因子1。 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程全文链接：

Synergistic effect of high mobility group protein B1 on calcium phosphate-induced release of inflammatory cytokines from macrophages

BACKGROUND:More and more evidence suggests that macrophages and inflammation reactions are involved in the formation and development of nephrolithiasis. Previous studies have found that calculi crystals can stimulate macrophages to release high mobility group protein B1. OBJECTIVE:To investigate the synergistic effect of high mobility group protein B1 in calcium phosphate induced release of interleukin-1β, interleukin-6, tumor necrosis factor α and monocyte chemotactic factor 1 from human macrophages.         METHODS:(1) The induced U937 cells were respectively stimulated with RPMI (blank), 100 mg/L calcium phosphate, 100 μg/L high mobility group protein B1 and 100 mg/L calcium phosphate+100 μg/L high mobility group protein B1 for 1, 2 and 4 hours to collect cell supernatant. (2) The induced U937 cells were respectively stimulated with 100 mg/L calcium phosphate, 100 mg/L calcium phosphate+10 μg/L high mobility group protein  B1, 100 mg/L calcium phosphate+50 μg/L high mobility group protein B1, 100 mg/L calcium phosphate+100 μg/L high mobility group protein B1 for 4 hours to collect cell supernatant. Levels of interleukin-1β, interleukin-6, tumor necrosis factor α and monocyte chemotactic factor 1 were determined by ELISA. RESULTS AND CONCLUSION:The levels of interleukin-1β, interleukin-6, tumor necrosis factor α and monocyte chemotactic factor 1 in the cell culture supernatant of 100 mg/L calcium phosphate group and 100 μg/L high mobility group protein B1 group were both higher than those in the blank group in a time-dependent manner (P < 0.05). The levels of interleukin-1β, interleukin-6, tumor necrosis factor α and monocyte chemotactic factor 1 in the cell culture supernatant of different concentrations of high mobility group protein B1 groups were all higher than those in the 100 mg/L calcium phosphate group in a concentration-dependent manner (P < 0.05). The results suggest that both calcium phosphate and high mobility group protein B1 can induce the release of interleukin-1β, interleukin-6, tumor necrosis factor α and monocyte chemotactic factor 1 from human macrophages and the high mobility group protein B1 has the synergistic effect with calcium phosphate to induce interleukin-1β, interleukin-6, tumor necrosis factor α and monocyte chemotactic factor 1 from human macrophages.