在动脉粥样硬化性病变的进展过程中,一个关键的步骤就是易损斑块的形成.而巨噬细胞凋亡可促进斑块的进展,并与易损斑块形成密切相关.动脉粥样硬化性血管疾病是糖尿病主要并发症之一,因此总结与糖尿病相关的脂代谢紊乱、胰岛素抵抗、脂肪因子等巨噬细胞凋亡影响因素,并初步探讨其介导斑块内巨噬细胞凋亡的信号转导途径,对于防治糖尿病患者动脉粥样硬化具有重要意义.

During the progress of atherosclerotic lesions,the formation of the vulnerable plaque is a critical step.Macrophage apoptosis can promote the progress of plaque,and is closely related to vulnerable plaque formation.Since atherosclerotic vascular disease is a major complication of diabetes mellitus,the factors associated with macrophage apoptosis,such as dyslipidemia,insulin resistance and adipokines etc,which are related to diabetes mellitus are summarized,and the signaling pathways that mediate macrophage apoptosis in the atherosclerotic lesions are also discussed.Deep understanding of the mechanism is important for the prevention and treatment of atherosclerosis in patients with diabetes mellitus.

为研究结核分枝杆菌Hsp16.3对感染巨噬细胞凋亡的影响。分别用结核分枝杆菌国际标准强毒株H37Rv株(H37Rv)、结核分枝杆菌国际标准强毒株H37Rv株Hsp16.3基因缺失突变株(△H37Rv)、卡介苗(BCG)和卡介苗Hsp16.3基因缺失突变株(△BCG)感染RAW264.7巨噬细胞株,使用流式细胞技术于感染1、3、6及12h检测各组感染巨噬细胞的凋亡率,并分析其变化。结果显示,巨噬细胞感染结核分枝杆菌后凋亡率升高,其中在感染1、3、12h,BCG组与H37Rv组比较差异具有统计学意义(P0.05);在感染1、3h,△BCG组与△H37Rv组比较差异具有统计学意义(P0.05);在感染3、6、12h,△H37Rv组与H37Rv组比较差异具有统计学意义(P0.05),△BCG组与BCG组比较差异具有统计学意义(P0.05)。由此可知,结核分枝杆菌Hsp16.3可抑制感染巨噬细胞的凋亡。

Objective :To investigate the effect of Mycobacterium tuberculosis Hsp16 .3 on apoptosis of infected macrophages . Methods :RAW264 .7 were infected with H37Rv ,△ H37Rv ,BCG and △BCG and the apoptotic rate of the infected macrophages were detected by flow cytometry at 1h ,3h ,6h and 12h after infection ,respectively .Results :The apoptotic rate of the infected macrophages were increased significantly ,especially the apoptotic rate of macrophages in BCG group were higher than H 37Rv group(P<0 .05)at 1h ,3h and 12h after MTB infection ;the apoptotic rate of macrophages in △BCG group were higher than△H37Rv group(P<0 .05)at 1h and 3h after M TB infection ;at 3h ,6h and 12h after M TB infection ,the apoptotic rate of macro-phages in △ H37Rv group and △BCG group were higher than H37Rv group and BCG group ,ordinally(P<0 .05) .Conclusion :Mycobacterium tuberculosis Hsp16 .3 can restrain the apoptosis of the macrophages infected .

为了探究布鲁氏菌侵染宿主巨噬细胞过程中铁蛋白(FTH1)生物学功能,本研究根据FTH1蛋白核苷酸序列,分别设计4条特异性小干扰RNA(short interfering RNA,siRNA)分子,亚克隆到RNAi-Ready pSIREN-RetroQ ZsGreen载体,并转染RAW264.7巨噬细胞,实时定量PCR筛选干扰效果最优的质粒;布鲁氏菌侵染干扰前、后的RAW264.7巨噬细胞,实时定量PCR检测布鲁氏菌基因16S rRNA和细胞凋亡相关基因的mRNA表达量。结果显示:获得了4条特异性siRNA分子,经测序正确,pSIREN-C siRNA质粒对FTH1干扰效果最高可达98%。干扰后布鲁氏菌侵染抑制率为84%,凋亡相关基因的mRNA表达量5个降低,3个上升。本研究表明:FTH1基因沉默后布鲁氏菌胞内的生存能力下降;布鲁氏菌侵染可以抑制细胞凋亡发生,FTH1基因沉默后细胞凋亡升高。

To explore the biological function of ferritin heavy chain (FTH1) in Brucella infecting mouse macrophage cells RAW264.7,four specific small interfering RNA were designed according to the nucleotide of FTH1 gene and inserted into RNAi-Ready pSIREN-RetroQ ZsGreen vector in this study.The recombinant vectors were transfected into mouse macrophage cells RAW264.7.Quantitative Real-time PCR was used to screen the plasmid with optimal interference effect.We detected the mRNA expression level of 16S rRNA and cell apoptosis-related genes in Brucella before and after infected with/without interference plasmids.Four specific siRNAs were obtained and sequenced correctly.The interference effect of plasmid pSIREN-C targeting FTH1 was up to 98%.The inhibiting rate of Brucella infection was 84% and the mRNA expressions of five apoptosis-related genes were down regulated,and three were up regulated after interference of FTH1.This study showed that inhibiting the expression of FTH1 could affect the abil

目的 探讨氧化应激在石英粉尘致肺泡巨噬细胞内质网应激性凋亡途径中的作用机制.方法 健康成年雄性Wistar大鼠72只,随机分为对照组、染尘组、染尘+N-乙酰半胱氨酸(NAC)组及NAC组,每组18只.采用非暴露式气管内注入法染尘,染尘组及染尘+NAC组一次性注入含5％超细石英粉尘的生理盐水1 ml,对照组及NAC组注入等量的无菌生理盐水,染尘+NAC组及NAC组在大鼠染尘后第2天开始,以灌胃的方式接受每天NAC 100 mg/kg抗氧化治疗,每周连续给药5d,直至实验结束.每组分别于染尘结束后14、28、56 d随机选取6只大鼠,股动脉放血处死、取肺,进行支气管肺泡灌洗,收集支气管肺泡灌洗液,分离纯化巨噬细胞,流式细胞术检测肺泡巨噬细胞凋亡率及巨噬细胞活性氧(ROS)含量,紫外比色法检测巨噬细胞蛋白羰基含量,蛋白免疫印迹(Western blot)法检测肺泡巨噬细胞caspase-12的蛋白表达水平.结果 与对照组比较,染尘组大鼠在染尘后第14天ROS含量、蛋白羰基含量、caspase-12蛋白表达水平及细胞凋亡率开始升高,染毒后第28、56天上述指标持续升高,差异有统计学意义(P＜0.05).与染尘组比较,染尘+NAC组大鼠在染尘后第14、28及56天巨噬细胞ROS含量均降低,差异有统计学意义(P＜0.05).与染尘组比较,染尘+NAC组大鼠染

Objective To investigate the role of oxidative stress in the endoplasmic reticulum stressinduced apoptosis of alveolar macrophages triggered by quartz dust.Methods Seventy-two healthy adult Wistar rats were randomly divided into control group,quartz dust group,quartz dust plus N-acetyl cysteine (NAC) group,and NAC group,with 18 rats in each group.One milliliter of sterile saline (for the control and NAC groups) or l ml of saline with 5％ ultrafine quartz dust (for dust group and dust plus NAC group) was given to each rat by non-exposed endotracheal infusion.From the second day after dust infusion,rats in dust plus NAC group and NAC group received intragastric administration of NAC (100 mg/kg).In each week,the treatment with NAC lasted for 5 consecutive days,followed by 2 days'' interval.For each group,6 rats were randomly selected on the 14th,28th,or 56th day after dust exposure; they were sacrificed by bloodletting from the femoral artery,and the lungs were collected.Broncho

目的：探讨Toll样受体4（ TLR4）对氧化型低密度脂蛋白（ oxidized low density lipopro-tein，ox-LDL）诱导的巨噬细胞凋亡的影响及其机制研究。方法体外培养THP-1细胞，用PMA（丙二醇甲醚醋酸酯）诱导成巨噬细胞，分别设立正常对照组、ox-LDL组、ox-LDL＋LPS组、衣霉素组，用MTT和流式细胞术检测细胞存活率及凋亡情况、油红O染色观察细胞吞噬脂质情况，q-RT-PCR、Western blot检测内质网应激相关基因及葡萄糖调节蛋白78（glucose-regulated protein78， GRP78）和CCAAT／增强子结合蛋白同源蛋白（ CCAAT／enhancer-binding protein homologous protein ， CHOP ）的表达情况，并且用TLR4-siRNA抑制TLR4活性观察其对通路的影响。结果流式及MTT结果显示ox-LDL＋LPS组的凋亡细胞数较ox-LDL组明显增加（P＜0．01）， ox-LDL＋LPS组的内质网应激相关基因和蛋白GRP78、CHOP均较ox-LDL组增加，且两组都明显大于正常对照组（ P＜0．01），抑制TLR4活性后内质网应激程度明显减轻（ P＜0．05）。结论 TLR4加重ox-LDL诱导的巨噬细胞凋亡，其机制是加重内质网应激程度，增加CHOP表达，起到促凋亡作用。

Objective To study the effects of Toll-like receptor 4(TLR4) on oxidized low density lipoprotein ( ox-LDL) induced macrophage apoptosis and its possible mechanism .Methods THP-1 derived macrophages were divided into four groups including untreated control group , ox-LDL treated group , ox-LDL+LPS treated group and tunicamycin treated group .MTT assay and flow cytometry analysis were performed to measure cell vitality and cell apoptosis , respectively .Oil red O staining was used to observe the phagocytosis of lipids by macrophages .The persistent and intense endoplasmic reticulum ( ER) stress markers were de-tected by analyzing the expression of glucose-regulated protein 78 ( GRP78 ) and CCAAT/enhancer-binding protein homologous protein ( CHOP) at mRNA and protein levels by q-RT-PCR and Western blot .Small in-terfering RNA ( siRNA) was used to silence the expression of TLR 4 to further elucidate its possible mecha-nism.Results Flow cytomotry and MTT assay showed that the number of apop