LFA-1共刺激诱导狼疮性肾炎外周血单个核细胞IL-10表达的研究

目的：探讨淋巴细胞功能相关抗原－１（ＬＦＡ－１）对活动性狼疮肾炎（ＬＮ）外周血单个核细胞（ＰＢＭＣ）白介素 １０（ＩＬ－１０）的调节作用。方法：利用半定量 ＲＴ－ＰＣＲ技术观察 ＬＦＡ－ １共刺激对活动性 ＬＮ患者 ＰＢＭＣ ＩＬ－１０ ｍＲＮＡ表达的影响。结果：单独抗ＣＤ３抗体（３０ ｎｇ／ｍｌ）能诱导ＬＮ 患者ＰＢＭＣ ＩＬ－１０ ｍＲＮＡ轻微表达（０．４３＋０．０３ｖｓ ０．５５±０．４４,Ｐ＜０．０５）而在抗ＣＤ３抗体（３０ ｎｇ／ｍｌ）存在的情况下,抗ＬＦＡ－１抗体（２ μｇ／ｍｌ,相当于ＬＦＡ－１配体）共刺激时明显增加了ＰＢＭＣ ＩＬ—１０ ｍＲＮＡ表达（１．３１±０．０８ｖｓ ０．５５＋０,０４,Ｐ＜０．０１）。ＬＦＡ－１中和抗体明显抑制了这种共刺激诱导的ＩＬ—１０ ｍＲＮＡ表达（１．３１±０．０８ｖｓ ０．６４ ±０．０３,Ｐ＜０．０１）。抗ＬＦＡ－１抗体单独刺激时并不引起 ＩＬ－１０ ｍＲＮＡ表达变化（Ｐ＞０．０５）。结论：ＬＦＡ－１作为共刺激分子诱导 ＩＬ－ １０ ｍＲＮＡ表达可能是其参与 ＬＮ发病的机制之一。     

狼疮性肾炎患者血清IL-1 0水平与狼疮疾病活动的关系

目的：探讨狼疮肾炎（ＬＮ）患者血清白细胞介素－１０（ｓＩＬ—１０）水平改变与狼疮疾病活动的关系及意义。方法：应用酶联免疫法（ＥＬＩＳＴ）测定３８例ＬＮ患者和２２例健康志愿者ｓＩＬ－１０水平,同时应用狼疮疾病活动指数（ＳＬＥＤＡＩ）对 ＬＮ患者进行疾病活动评分,并对两者进行相关分析。结果： ＬＮ患者 ｓＩＬ－ １０水平明显较正常健康对照组高（１９．７±７．２ ｐｇ／ｍｌ ｖｓ １２．０±２,９ ｐｇ／ｍｌ,Ｐ＜０．０５）,对ＳＬＥＤＡＩ与ｓＩＬ－１０水平进行Ｐｅａｒｓｏｎ直线相关分析,其相关系数ｒ＝０．１０５,Ｐ＝０．６０９（Ｐ＞０．０５）,提示ＬＮ患者ＳＩＬ－１０水平与疾病活动指数无相关性。结论：ＬＮ患者血清白细胞介素－１０异常升高可能是一种内源性的异常而与狼疮疾病活动无明显相关性。     

凋亡相关基因产物Fas/APO-1和bcl-2蛋白在肾癌组织中的表达及意义

目的研究凋亡相关基因Ｆａｓ／ＡＰＯ－１和ｂｃｌ－２在肾癌发生发展中的作用。方法采用免疫组织化学法对３５例肾癌组织和２６例远离肾癌的正常肾组织Ｆａｓ／ＡＰＯ－１和ｂｃｌ－２蛋白的表达进行检测。结果肾癌组织Ｆａｓ／ＡＰＯ－１蛋白表达率为５７．１４％，明显低于正常肾组织中的表达率（８４．６２％，Ｐ＜０．０５），且表达强度也明显低下；而ｂｃｌ－２蛋白表达率为８０．００％，明显高于正常肾组织中的表达率（５３．８５％，Ｐ＜０．０５）。结论Ｆａｓ／ＡＰＯ－１与ｂｃｌ－２基因共同参与了肾癌的发生和发展。     

Fas/FasL表达和细胞凋亡在人类同种异体移植肾急性排斥中的作用

目的探讨移植肾急性排斥（ＡＲ）时细胞凋亡与Ｆａｓ和Ｆａｓ配体（ＦａｓＬ）表达的作用及其临床意义。方法分别用原位末端标记技术（ＴＵＮＥＬ法）和免疫组织化学方法检测２６例移植肾ＡＲ标本中细胞凋亡和Ｆａｓ／ＦａｓＬ表达情况。结果细胞凋亡和Ｆａｓ／ＦａｓＬ表达主要在ＡＲ移植肾小管上皮发生,且凋亡指数和Ｆａｓ／ＦａｓＬ表达与肾组织病理损伤程度平行,与正常肾对照组和移植肾功能稳定组比较差异显著（Ｐ＜０．０１）。结论肾小管上皮细胞凋亡在ＡＲ所致的移植肾损伤中起重要作用,Ｆａｓ／ＦａｓＬ系统可能参与移植肾ＡＲ,是造成肾小管上皮细胞凋亡的重要因素。ＴＵＮＥＬ法检测细胞凋亡可作为判断移植肾病理变化和预后的重要指标     

辽宁绝经妇女骨密度与绝经年限、体重关系研究

目的　研究绝经年限、体重对辽宁地区绝经后女性不同部位骨密度的影响。方法　测定共９６ 例绝经后妇女腰椎（Ｌ２～４）、股骨颈（Ｎｅｃｋ）、大转子（Ｔｒｏｃｈ）、Ｗａｒｄ＇ｓ 区的ＢＭＤ,同时测定了血ＡＬＰ、血尿钙、肌肝（Ｃｒ）等指标,分析其相互关系。结果　１． 绝经后妇女各部位的ＢＭＤ 不同。２．Ｌ２～４的ＢＭＤ与体重（Ｗ ）、血小板、尿Ｃａ／Ｃｒ呈正相关（Ｐ＜ ０．０１、Ｐ＜ ０．０５、Ｐ＜ ０．０１）。与绝经年限的自然对数（ＰＦＮＬ）、ＡＬＰ呈负相关（Ｐ＜ ０．０１、Ｐ＜ ０．０１）。３．Ｎｅｃｋ 区ＢＭＤ 与体重、血小板呈正相关（Ｐ＜ ０．０１、Ｐ＜ ０．０５）,与ＰＦＮＬ呈负相关（Ｐ＜ ０．０５）。４．Ｗａｒｄ＇ｓ 区ＢＭＤ 与体重、血小板、尿Ｃａ／Ｃｒ 呈正相关（Ｐ＜ ０．０１、Ｐ＜ ０．０５、Ｐ＜ ０．０５）,与ＰＦＮＬ及ＡＬＰ呈负相关（Ｐ＜ ０．０１、Ｐ＜ ０．０５）。５． 大转子区骨密度与体重、血小板、转氨酶呈正相关（Ｐ＜ ０．０１、Ｐ＜ ０．０１、Ｐ＜ ０．０５）,与ＡＬＰ呈负相关（Ｐ＜ ０．０１）。结论　体重、绝经年限、尿Ｃａ／Ｃｒ、血小板及血ＡＬＰ能影响骨密度。     

地塞米松和川芎嗪对狼疮肾炎外周血单个核细胞白细胞介素12表达的影响

目的　探讨地塞米松、川芎嗪对活动期、静止期狼疮肾炎（ Ｌ Ｎ） 外周血单个核细胞（ Ｐ Ｂ Ｍ Ｃ） 白细胞介素１２（ Ｉ Ｌ１２） 表达的影响。方法　应用地塞米松（１０ － ５ ｍｏｌ／ Ｌ） ,川芎嗪（３３ μｇ／ｍｌ） 对 Ｌ Ｎ 患者 Ｐ Ｂ Ｍ Ｃ 培养,以正常人为对照,分别采用 Ｅ Ｌ Ｉ Ｓ Ａ 法和半定量 Ｒ Ｔ Ｐ Ｃ Ｒ 法检测细胞上清液及细胞 Ｉ Ｌ１２ 和 Ｉ Ｌ１２ Ｐ４０ ｍ Ｒ Ｎ Ａ 表达。结果 Ｌ Ｎ 活动期、静止期较正常对照组 Ｉ Ｌ１２ 蛋白、基因水平明显增高（ Ｐ ＜ ０ ．０１） 。地塞米松明显抑制狼疮肾炎 Ｐ Ｂ Ｍ Ｃ Ｉ Ｌ１２ 表达（ Ｐ ＜ ０ ．０１） ,而川芎嗪仅对活动期狼疮肾炎 Ｐ Ｂ Ｍ Ｃ Ｉ Ｌ１２ 表达抑制明显（ Ｐ ＜ ０ ．０５） ,两种药物对正常人对照 Ｉ Ｌ１２ 表达无明显抑制作用（ Ｐ ＞ ０ ．０５） 。结论　狼疮肾炎 Ｉ Ｌ１２ 表达水平增高,这与 Ｐ Ｂ Ｍ Ｃ 处于异常活化状态有关;地塞米松、川芎嗪下调狼疮肾炎 Ｉ Ｌ１２ 表达,提示它们通过免疫调节直接抑制了活化的狼疮肾炎 Ｐ Ｂ Ｍ Ｃ 表达 Ｉ Ｌ１２     

心脏手术中小剂量抑肽酶对中性粒细胞CD_(11b)/CD_(18)表达的影响

目的 探讨体外循环术（ＣＰＢ）中小剂量抑肽酶对全身炎性反应的抑制作用。方法２８例首次行心脏瓣膜置换术患者随机分为对照组和抑肽酶组,各１４例,于麻醉诱导前、ＣＰＢ前、ＣＰＢ结束后１ｈ及２４ｈ用免疫流式细胞仪分别测定中性粒细胞表面粘附分子ＣＤ１１ｂ和ＣＤ１８的表达。结果ＣＤ１１ｂ／ＣＤ１８的表达与ＣＰＢ时间呈正相关（相关系数γ分别为０．６４４、０．５３８,Ｐ＜０．０５）,ＣＰＢ结束后１ｈ及２４ｈ对照组ＣＤ１１ｂ的表达较麻醉诱导前及同时点的抑肽酶组明显上调（Ｐ＜０．０５）;ＣＤ１８的表达仅在ＣＰＢ结束后１ｈ明显上调并高于抑肽酶组（Ｐ＜０．０５）,ＣＰＢ结束后２４ｈ表达下凋有所缓解,但仍高于麻醉前基础值（Ｐ＜０．０５）。结论 ＣＰＢ所致的炎性反应可能与转机时间有关,小剂量抑肽酶对ＣＰＢ术后ＣＤ１１ｂ／ＣＤ１８表达增加具有抑制作用。     

实验性肾间质纤维化大鼠模型中Fas表达与细胞凋亡

目的　探讨肾间质纤维化形成过程中的肾间质细胞凋亡及其与Ｆａｓ 表达的关系。方法　建立大鼠单侧输尿管梗阻模型,分别于模型３ 天、７ 天、１４ 天、２８ 天、５６ 天和８４ 天处死大鼠。除光镜、电镜观察外,免疫组织化学染色检测胶原Ⅲ和Ｆａｓ 蛋白表达,末端转移酶介导的ｄＵＴＰ缺口末端标记法检测凋亡细胞。用病理分析软件对肾间质Ｆａｓ 表达和细胞凋亡进行定量分析。结果　该模型显示进行性的肾间质纤维化。模型１４ 天后,肾间质Ｆａｓ 表达阳性率递增,肾间质凋亡细胞亦逐渐增加,且２ 者成正相关（ ｒ ＝０－９４,Ｐ＜０－０５）。结论　细胞凋亡可能为该模型中肾间质细胞丢失的原因,且Ｆａｓ／ＦａｓＬ系统可能介导了该细胞凋亡。     

人外周血淋巴细胞Fas的表达及细胞凋亡的诱导

目的 研究Ｆａｓ系统对外周血淋巴细胞的影响。方法 将分离、纯化后的淋巴细胞进行单向淋合淋巴细胞培养,用流式细胞术动态检测新鲜分离和激活后的Ｔ淋巴细胞表面Ｆａｓ表达水平,并用ＤＮＡ电泳和原位末端标记技术分别从定性和定量的角度观察Ｆａｓ系统激活与否对混合培养后淋巴细胞凋亡率的影响。结果 Ｔ淋巴细胞在被激活后Ｆａｓ表达水平升高,加抗Ｆａｓ单克隆抗体组的细胞凋亡率高于末加抗体的对照组（Ｐ〈０．０５）。结论     

凋亡相关基因产物Fas／APO—1和bcl—2蛋白在肾癌组织中的表达及 …

目的 研究凋亡相关基因Ｆａｓ／ＡＰＯ－１和ｂｃｌ－２在肾癌发生发展中的作用。方法 采用免疫组织化学法对３５例肾癌组织和２６例远离肾癌的正常肾组织Ｆａｓ／ＡＰＯ－１和ｂｃｌ－２蛋白的表达进行检测。结果 肾癌组织Ｆａｓ／ＡＰＯ－１蛋白表达率５７．１４％,明显低于正常肾组织中的表达率（８４．６２％,Ｐ〈０．０５）,且表达强度也明显低下;而ｂｃｌ－２蛋白表达率为８０．０％,明显高于正常肾组织中的表达率（５     

大黄素对全身炎性反应综合征病人外周血中性粒细胞Fas/FasL、Caspase-3表达的影响

目的 探讨全身炎性反应综合征(SIRS)病人外周血中性粒细胞(PMN)凋亡信号Fas/FasL、Caspase-3表达的动态变化及大黄素对SIRS时PMN凋亡的影响及其作用机制.方法 采集6例健康志愿者及8例SIRS病人(均为急性胰腺炎病人)外周血液并从中分离PMN进行体外培养.分为3个实验组:正常组,SIRS组,大黄素干预组.观察各组PMN的凋亡情况;检测各组PMN Fas/FasL和Caspasc-3表达水平的变化.结果 SIRS组PMN凋亡百分率明显低于正常组(P＜0.05),大黄素干预后SIRS病人PMN凋亡百分率明显增加(P＜0.05);SIRS组PMN Fas和Caspase-3的表达水平明显低于正常组(P＜0.05),大黄素能诱导SIRS病人PMN Fas和Caspase-3的表达(P＜0.05).各组PMN在体外培养24 h后,经Western Blotting没有检测到FasL的表达.结论 SIRS病人外周血PMN凋亡存在异常,且与Fas和Caspase-3的表达降低有关;大黄素能通过诱导Fas和Caspase-3的表达,对SIRS时PMN凋亡延迟具有抑制作用.     

Fas介导细胞凋亡通路在慢性排斥反应所致移植物血管病变中的作用

目的 通过建立小鼠心脏及主动脉移植模型探讨Fas途径介导细胞凋亡在慢性排斥反应所致移植物血管病变形成中的作用。方法 (1)正常B6及B6．MRL(Fas-／-)→B10．2R小鼠腹腔异位心脏移植。B10．2R→B6小鼠胸主动脉异位移植。定期获取移植物。观察移植物存活率与组织形态学改变；TUNEL及FITC Annexin V／PI双染色方法检测移植物中凋亡细胞形态及分布；B6小鼠主动脉体外培养。人重组的可溶性FasL诱导平滑肌细胞凋亡。结果 B6．MRL(Fas-／-)供体来源移植物存活期显著延长，其血管平滑肌细胞凋亡明显减少。新生血管内膜中的平滑肌细胞具有抗Fas介导凋亡的特性。结论 在慢性排斥反应所致移植物血管病变形成中Fas途径及平滑肌细胞对Fas介导凋亡的敏感性具有重要作用。     

Effects of uraemia and haemodialysis on neutrophil apoptosis and expression of apoptosis-related proteins.

BACKGROUND: In haemodialysis (HD) patients, it is unclear whether increased apoptosis of neutrophils is due to uraemia or HD itself. The purpose of the current study was to assess the effect of uraemia and HD on the rate of apoptosis and apoptosis-related protein expression in whole blood neutrophils. METHODS: We employed a whole-blood micromethod to test spontaneous apoptosis and expression of apoptosis-regulating proteins in cultured neutrophils from uraemic patients (pre-HD), HD patients and healthy controls. Blood samples were drawn before, after 20 min and after 4 h of haemodialysis, and were then cultured for 20 h. We evaluated the rate of apoptosis from annexin V and propidium iodide staining, and examined bcl-2, Fas/Apo-1 and p53 expression in the cultured neutrophils. RESULTS: Fas/APO-1 expression and total percentage of apoptotic whole blood neutrophils of pre-HD and HD patients before HD were significantly higher than controls. There was a transient but significant decrease in the percentage of apoptotic neutrophils and Fas/APO-1 expression after 20 min of dialysis. The expression of bcl-2 protein was significantly lower from neutrophils in HD patients compared with controls, and HD significantly downregulated bcl-2 expression. The p53 protein content in HD patients before HD was significantly higher than in pre-HD patients. CONCLUSIONS: These findings suggest that uraemia accelerates neutrophil apoptosis by increasing Fas/Apo-1, and that HD does not affect neutrophil apoptosis more than uraemia. In addition, HD produces only in a transient sequestration of potentially apoptotic neutrophils.     

Fas and Fas ligand expression in cystic fibrosis airway epithelium

BACKGROUND: Cystic fibrosis (CF) is a genetic disease caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene and defective expression of CFTR protein in epithelial cells. The main cause of mortality in CF is linked to chronic inflammatory and infectious airway processes. Recent studies have suggested perturbations in the apoptotic process in CF cell lines and enterocytes. A study was undertaken to investigate the expression of Fas and Fas ligand (FasL) in CF bronchial epithelium and CF tracheal cell lines. METHODS: Immunohistochemical staining for Fas (alkaline phosphatase anti-alkaline phosphatase) and FasL (immunoperoxidase) was performed in eight CF bronchial epithelial samples and four controls and immunohistochemical DNA fragmentation (TUNEL) was carried out in four CF patients and four controls. Immunofluorescence staining and flow cytometric analysis of Fas and FasL expression was performed in two human tracheal epithelial cell lines (HTEC) with normal and CF genotype. The dosage of serum soluble FasL was examined in 21 patients with CF and 14 healthy volunteers. RESULTS: FasL expression was markedly increased in patients with CF in both the ciliated and submucosal glandular bronchial epithelium compared with controls; Fas was similarly expressed in bronchial samples from controls and CF patients in both the ciliated epithelium and submucosal glands. High levels of DNA fragmentation were observed in CF but with some epithelial cell alterations. Serum concentrations of soluble FasL were frequently undetectable in patients with CF. In vitro, HTEC expressed Fas and FasL in both genotypes. A higher mean fluorescence intensity for FasL expression was noted in CF genotype HTEC with median (range) for six experiments of 74 (25-101) for CF cells and 42 (21-70) for non-CF cells. CONCLUSION: Fas/FasL interaction is probably implicated in the human CF airway apoptotic pathway. The mechanisms of induction of FasL expression and its role in inducing tissue damage or remodelling or in controlling local inflammatory cell apoptosis remain to be determined.     

B lymphopenia in uremia is related to an accelerated in vitro apoptosis and dysregulation of Bcl-2.

BACKGROUND: Lymphopenia has been described in patients with chronic renal failure (CRF). It is postulated that the decline in lymphocytes is due to accelerated apoptosis. We investigated whether dysregulation of programmed cell death plays a role in the immunodeficiency described in CRF. METHODS: Peripheral blood lymphocytes (PBL) from pre-dialysis uraemic patients (nHD) and haemodialysed patients (HD) were cultured with no stimulus for 96 h. Apoptosis of lymphocytes was measured by propidium iodide staining and flow cytometry. Expression of Fas and Bcl-2 was also analysed by flow cytometry. RESULTS: Peripheral blood B cells were significantly lower in pre-dialysis and haemodialysis uraemic patients compared to control. Lymphocytes from both groups of patients had a higher rate of apoptosis in vitro than those from healthy controls. This effect was more pronounced in B lymphocytes and a significant correlation between the B lymphopenia and the percentage of apoptotic B cells after 48 h of culture without stimulus was observed. The increased lymphocyte apoptosis in CRF was accompanied by a significantly lower in vitro Bcl-2 expression. However, Fas did not seem to play a role in spontaneous lymphocyte apoptosis in end-stage renal disease. CONCLUSIONS: Our data indicate that B lymphopenia in CRF may be partially attributed to an increased susceptibility to cell death by apoptosis that is associated with a decreased expression of Bcl-2.     

滋肾化毒饮联合环磷酰胺对狼疮性肾炎患者外周血淋巴细胞Fas、FasL、Bcl-2表达的影响及临床疗效观察

目的:观察滋肾化毒饮联合间断性环磷酰胺(CTX)静脉冲击疗法对活动期狼疮性肾炎(LN)患者外周血淋巴细胞(PBL)凋亡调控因子及临床指标的影响.方法:将活动性LN患者40例,随机分为治疗组和对照组,每组各20例.治疗组予滋肾化毒饮联合间断性CTX静脉冲击疗法,对照组予间断性CTX静脉冲击疗法.观察治疗3个月前后的PBL的Fas、FasL、Bcl-2表达及临床指标变化,Fas、FasL、Bcl-2用免疫组化法测定.结果:治疗后两组Fas、FasL及治疗组Bcl-2表达下调(P＜0.05),而Fas、FasL表达下调幅度大于对照组(P＜0.05).治疗组SLEDAI、24 h尿蛋白、Hgb、Ccr、C3指标的改善优于同期对照组(P＜0.05).治疗组肝功能受损、白细胞减少、舌苔厚腻、失眠、痤疮发生率明显少于对照组(P＜0.05).结论:调控Fas、FasL介导的凋亡可能是CTX、激素、滋肾化毒饮有效治疗活动狼疮性肾炎机制之一.     

Apoptotic markers on lymphocytes and monocytes are unchanged during single hemodialysis sessions using either regenerated cellulose or polysulfone membranes

BACKGROUND: There is an increased rate of apoptosis of peripheral blood mononuclear cells (PBMCs) in patients undergoing hemodialysis (HD), but little is known about how different dialysis membranes may contribute to the process. We, therefore, studied the influence of two different dialysis membranes on apoptotic markers during HD. METHODS: 8 healthy controls and 8 patients on regular HD 3 times per week were enrolled in this cross-controlled study. Patients received HD using polysulfone and then regenerated cellulose dialysis membranes for one week each, sequentially. Serum was collected for C-reactive protein (CRP) detection; flow cytometry with dual antibody staining was used to measure the apoptotic markers Fas (CD95), FasL (CD 178) and TNF-R2 (CD120b) in T cells (CD3+), B cells (CD19+), and monocytes (CD14+) at 0, 15, 120 and 240 min after starting HD. We also measured total leukocyte numbers and differential white cell counts. RESULTS: Hemodialysis patients revealed lymphocytopenia, monocytopenia, higher CRP levels and higher Fas and TNF-R2 expression on lymphocytes and monocytes at baseline when compared with normal controls. Leukocyte numbers, including neutrophils, lymphocytes and monocytes, dropped significantly after 15 min of dialysis. There were no significant differences in Fas levels during hemodialysis on T and B lymphocytes or on monocytes. T lymphocyte FasL (CD 178) levels remained unchanged throughout the process. There was a significantly lower overall level of CD120b at 15 min of HD, whereas this marker was higher on monocytes after dialysis. There were no significant differences in the levels of apoptotic markers between the two membranes. CONCLUSION: Our results suggest that uremia itself contributes to PBMC apoptosis. The two different dialysis membranes used in this study did not influence apoptotic markers on PBMCs significantly, but increased TNF-R2 expression on monocytes during a single dialysis session.     

Prognostic Significance of Fas and Fas Ligand System-Associated Apoptosis in Gastric Cancer

Background: Previous studies indicate that gastric carcinomas express Fas ligand and downregulate Fas to escape from the host immune attack; however, the prognostic importance of Fas/FasL expression in this tumor is yet to be evaluated.Methods: Specimens from 87 gastric carcinoma patients of different stages treated in a defined period with curative intent were evaluated for apoptosis, Fas, FasL, and CD8 expression using an immunohistochemical method.Results: The percentage of terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL)-positive apoptotic cells expressed as apoptotic index (AI) was higher in 43 patients when the cut-off value was set at the median value. There were no significant correlations between AI and clinicopathologic parameters. Thirty-nine patients showed a high number of CD81 cells within cancer nests. Positive FasL and Fas expression was seen in 53 and 72 patients, respectively. CD8 and FasL expressions were related only to patients age. Fas expression had significant correlations with tumor invasion and Lauren classification. There were significant direct correlations between AI and number of nest CD81 cells and between AI and grade of Fas expression. Apoptotic index, pT stage, CD8 expression, and Fas expression were identified as independent prognostic factors.Conclusions: Spontaneous apoptosis in gastric carcinoma may be an independent prognosticator for survival and is significantly influenced by tumor Fas expression and number of nest CD81 cells.     

不同浓度地塞米松对人骨关节炎软骨细胞凋亡及Fas/FasL基因表达的影响

目的糖皮质激素会破坏软骨,通过观察地塞米松对人骨关节炎软骨细胞(human articular chondrocytes,HACs)凋亡及Fas/FasL基因表达的影响,探讨其促进HACs凋亡的作用机制。方法取行人工膝关节置换的膝骨关节炎患者自愿捐赠软骨组织,体外分离培养HACs,取第2代细胞进行实验。将HACs分别置于含浓度为0.125、1.25、12.5、25及50μg/mL地塞米松培养液中,培养48 h后采用MTT法选择地塞米松最佳工作浓度进行后续实验。将HACs分别采用最佳工作浓度地塞米松(实验组)及不含地塞米松培养液(对照组)培养,0、24、48 h后行TMRE/Hoechst/Annexin V-FITC/7-AAD四重染色检测细胞凋亡,48 h后行实时定量PCR检测Fas/FasL mRNA表达,0、24、48 h后行免疫组织化学染色检测Fas/FasL蛋白表达。结果 25μg/mL浓度组细胞抑制率显著高于50μg/mL浓度组,差异有统计学意义(P<0.05);与其他浓度组比较,差异亦有统计学意义(P<0.05);选择25μg/mL浓度作为后续实验工作浓度。培养0、24、48 h,实验组HACs凋亡细胞百分比分别为5.8%±0.3%、27.0%±2.6%、36.0%±3.1%,呈明显时间依赖性(P<0.05)。培养48 h后实验组及对照组Fas mRNA相对表达量分别为(8.93±1.12)×10—3及(3.31±0.37)×10—3,FasLmRNA相对表达量分别为(5.92±0.66)×10—3及(2.31±0.35)×10—3,两组比较差异均有统计学意义(P<0.05)。随培养时间延长,实验组Fas及FasL蛋白表达逐渐增加,且各时间点均显著高于对照组,差异均有统计学意义(P<0.05)。结论地塞米松可促进HACs细胞凋亡及上调凋亡基因Fas/FasL表达,为进一步探讨Fas/FasL信号通路在地塞米松促HACs凋亡中的作用提供了实验依据。