中文摘要：

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）-lBmRNA和IL-1β蛋白分泌规律。方法体外培养THP—1单核细胞，分别以5．5mmol／L葡萄糖（LG）和25．0mmol／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2～48h提取单核细胞，检测细胞凋亡、IL-1β蛋白、iNOS和IL-1BmRNA表达。细胞凋亡采用AnnexinV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．timequantitativePCR）法，检测iNOS和IL-1βmRNA表达情况。结果（1）高糖刺激THP-1单核细胞12—48h后儿1β蛋白、iNOS和IL-18mRNA表达较刺激前显著增加（P〈0．05），iNOS和IL-1βmRNA表达在24h达最高值，IL-1β蛋白分泌48h达高峰。细胞凋亡在高糖刺激48h-4d较刺激前显著增加（P〈0．05）。（2）两组细胞加入HKSA后6～48h表达iNOS和L-lβ显著高于刺激前（P〈0．01），24h达高峰，48h表达下降；IL-1β蛋白分泌在48h达最高值。两组细胞凋亡率在加入HKSA后12h、24h、48h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-1BmRNA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-1BmRNA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。

英文摘要：

Objective To investigate the effects of heat-k/lied Staphylococcus aurem（HKSA） on the apoptosis and expression of iNOS and IL-Iβ 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-115 were exm-ined. Apoptosis was analyzed by flow cytometry and expressions of 1L-115 and iNOS were quantitated by real-tlme PCR. Bmults The expression of iNOS and IL-Iβ 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 significandy after 48 h to 4 days. HKSA infection enhanced apoptosis,but inhibited the expression of iNOS and IL- 1β in the presence of higfi concentration of glucose. The expression of iNOS and IL-Iβ increased significantly at 6 h（P〈0. 01 ） ,reaching the highest level at 12 h,but the levels were significantly lower than those in contml groups （ P〈0. 05 ）. Conclusion These data suggest that high concentration of glucose can interfere with the anti-bacterial function of monocvtes by reducing their expression of iNOS and IL-lβ and enhancing theirapoptosis.