摘要目的 观察高糖状态下热灭活金黄色葡萄球菌(HKSA)诱导的人单核细胞株THP-1凋亡特点,以及表达诱导型一氧化氮合酶(iNOS)、白细胞介素(IL)-1β mRNA和IL-1β蛋白分泌规律.方法 体外培养THP-1单核细胞,分别以5.5 mmol/L葡萄糖(LG)和25.0 mmol/L葡萄糖(HG)培养12h～8d.将HG和LG培养6d的单核细胞加入HKSA.分别于HKSA加入前和加入后2～48 h提取单核细胞,检测细胞凋亡、IL-1β蛋白、iNOS和IL-1β mRNA表达.细胞凋亡采用Annexin V与PI双染法,流式细胞技术检测；IL-1β蛋白分泌采用ELISA法检测；提取细胞总RNA,反转录生成cDNA后采用实时定量逆转录聚合酶链反应(real-time quantitative PCR)法,检测iNOS和IL-1β mRNA表达情况.结果 (1)高糖刺激THP-1单核细胞12 ～48 h后IL-1β蛋白、iNOS和IL-1β mRNA表达较刺激前显著增加(P＜0.05),iNOS和IL-1β mRNA表达在24 h达最高值,IL-1β蛋白分泌48 h达高峰.细胞凋亡在高糖刺激48 h～4 d较刺激前显著增加(P＜0.05).(2)两组细胞加入HKSA后6～48 h表达iNOS和L-1β显著高于刺激前(P＜0.01),24h达高峰,48 h表达下降；IL-1β蛋白分泌在48 h达最高值.两组细胞凋亡率在加入HKSA后12h、24h、48 h逐渐增加(P＜0.01).单核细胞加入HKSA前和加入HKSA后12h,高糖组与低糖组比较,高糖组IL-1β蛋白、iNOS和IL-1β mRNA表达均低于低糖组,凋亡率高于低糖组(P＜0.05).结论 高糖和HKSA对单核细胞分泌IL-1β蛋白、表达iNOS和IL-1β mRNA的影响与刺激时间有关；单核细胞处于持续高糖状态时,高糖可以增加细胞凋亡率,降低IL-1β蛋白分泌、iNOS和IL-1β表达.

abstractsObjective To investigate the effects of heat-killed Staphylococcus aureus (HKSA) on the apoptosis and expression of iNOS and IL-1β in THP-1 monocytes in the presence of high concentration of glucose.Methods THP-1 cells were cultured in medium containing 25.0 mmol/L(HG) or 5.5 mmol/L (LG,control) D-glucose for 12 h-8 d.The THP-1 cells cultured for 6 d were extracted on the 0-48 h with or without HKSA,then apoptosis and expression of iNOS and IL-1β were examined.Apoptosis was analyzed by flow cytometry and expressions of IL-1β and iNOS were quantitated by real-time PCR.Results The expression of iNOS and IL-1β in THP-1 monocytes was increased significantly in the presence of high concentration of glucose for 12-48 h(P＜0.05),reaching the highest level at 24 h and returned to baseline after 4 d.The expression was significantly lower than that of control after 4-6 d.Apoptosis rate was also increased significantly after 48 h to 4 days.HKSA infection enhanced apoptosis,but inhibited the expression of iNOS and IL-1 β in the presence of high concentration of glucose.The expression of iNOS and IL-1β increased significantly at 6 h(P＜0.01),reaching the highest level at 12 h,but the levels were significantly lower than those in control groups (P＜0.05).Conclusion These data suggest that high concentration of glucose can interfere with the anti-bacterial function of monocytes by reducing their expression of iNOS and IL-1β and enhancing their apoptosis.