表达CD95L的HepG_2细胞自分泌和旁分泌的细胞毒效应

目的阐明乙型肝炎时肝细胞可表达细胞毒效应分子ＣＤ９５Ｌ的效应机制。方法ＨｅｐＧ２细胞诱导表达ＣＤ９５Ｌ,与ＨｅｎＧ２．２．１５细胞共培养;或以其培养上清液加于另一份未经诱导的ＨｅＰＧｚ细胞中,以流式细胞仪检测后者的凋亡率;以单个ＨｅｐＧ２细胞培养试验自体凋亡。结果表达ＣＤ９５Ｌ的ＨｅｐＧ２细胞致表达ＣＤ９５的ＨｅｐＧ２．２．１５细胞的凋亡率为：２４小时时１６．５％,４８小时时４３．０％;培养上清液引起的凋亡率：２４小时时３８．７％,４８小时时７３．３％,单个ＨｅｐＧ２细胞经诱导的自杀率２４小时为４３．８％,均高于各自的对照。结论ＨｅｐＧｅ细胞以ＣＤ９５Ｌ的膜性或可溶性分子、经旁分泌或自分泌发生“自杀”或“同胞相杀”。     

缺血性脑卒中患者外周血淋巴细胞CD95和CD95L的表达及其临床意义

目的　探讨缺血性脑卒中患者外周血淋巴细胞CD95和CD95L表达的动态变化及其与病情严重程度的关系及临床意义。方法　采用流式细胞术动态检测了 36例轻中重型缺血性脑卒中患者在发病后 3d内 (最早发病后 6h)、第 7天和第 2 1天外周血淋巴细胞CD95和CD95L的表达情况 ,并选择 2 2例同期住院病人作为对照。结果　CD95和CD95L在缺血性脑卒中患者急性期是升高的 ,在发病后 6h～ 3d、第 7天、第 2 1天均高于正常 ,且在 6h～ 3d最高 (P值均 <0 .0 5 )。其升高程度与病情的严重程度有关 ,病情越重 ,升高越明显。CD95和CD95L两者之间呈正相关 (P <0 .0 5 )。结论　在缺血性脑卒中患者外周血淋巴细胞CD95和CD95L发生显著变化 ,且在不同的时间有不同的变化 ,其变化程度与临床病情的严重程度有密切关系。     

乙型肝炎慢性肝病中的肝细胞表达CD95L提示能自行凋亡

乙型肝炎慢性肝病中的肝细胞表达ＣＤ９５Ｌ提示能自行凋亡骆抗先，朱幼芙，何海棠，王新生，王晋豫作者单位：５１０５１５广州第一军医大学附属南方医院ＣＤ９５是介导凋亡的细胞表面分子，慢性肝病时肝细胞可有强表达。ＣＤ的是受体，与其配体（ＣＤ９５Ｌ）相互作用从...     

肝脏病中CD95介导的细胞凋亡

一、ＣＤ９５业已证明,ＣＤ９５（Ｆａｓ／ＡＰＯ－１,ＣＤ９５受体）通过与其自然配体ＣＤ９５Ｌ或可溶性ＣＤ９５Ｌ或特异性活性抗体结合而激发细胞凋亡。与肿瘤坏死因子受体（ＴＮＦ－Ｒ１）、ＴＮＦ－Ｒ２、低亲和性神经生长因子（ＮＧＦ）受体、ＣＤ４０和ＣＤ３０等细胞外结构域中存在２～５个半胱氨酸而统归于ＴＮＦ／ＮＧＦ受体超家族。ＣＤ９５还与ＴＮＦ－Ｒ１,肿瘤坏死因子相关凋亡诱导配体（ＴＲＡＩＬ）－Ｒ１,ＴＲＡＩＬ－Ｒ２和死亡受体３（ＤＲ３）在胞内有一段同源区域而被命名为“死亡区域”。ＣＤ９５凋亡路径是通过…     

慢性乙型肝炎患者外周血CD8~+T细胞CD95表达的研究

目的检测慢性乙型肝炎患者外周血CD8+T细胞CD95分子的表达水平,了解CD95分子表达与炎症进展的关系。方法采集37例慢性乙型肝炎患者及健康对照者外周血,分离外周血单个核细胞(PBMCs),用流式细胞术检测外周血CD8+T细胞CD95分子的表达情况。结果高病毒载量组和低病毒载量组CD8+T细胞CD95的表达均显著高于健康对照组(P0.05);CD95的表达及CD8+T细胞频率之间存在负相关关系(r=-0.3486)。结论慢性乙型肝炎患者CD95的高表达可能促进了CD8+T细胞的凋亡,使CD8+T细胞的抗病毒作用减弱,可能与HBV持续感染、慢性乙型肝炎的形成机制有关。     

老年人外周血淋巴细胞的凋亡及CD28和CD95的表达

目的通过对健康老年人与健康成年人外周血淋巴细胞的凋亡率及CD28和CD95表达水平的比较性研究,观察衰老对淋巴细胞凋亡的影响,以及CD28和CD95与淋巴细胞凋亡的关系。方法将实验对象分为2组：成年组（25～59岁）与老年组（60～90岁）各20例,采用免疫荧光标记流式细胞术检测外周血淋巴细胞CD28和CD95的水平;Annexin-v-FITC／PI双染色流式细胞术测定地塞米松诱导的淋巴细胞凋亡。结果老年组：淋巴细胞凋亡率[（14．90±4．12）％]显著高于成年组[（8．12±3．12）％];CD28^＋CD95[（8．80±4．86）％]及CD28^＋[（36．31±10．38）％]均明显低于成年组[（23．09±3．48）％、（52．29±4．90）％],而CD28^-CD95^＋[（53．23±8．28）％]、CD95^＋[（80．25±7．19）％]及CD28^-[（63．69±10．38）％]明显高于成年组[（33．58±4．72）％、（63．18±4．12）％、（47．71±4．90）O];CD28^＋CD95^＋在两组间差异无统计学意义。淋巴细胞凋亡率与CD28（CD28^＋）呈负相关,与CD95（CD95^＋）呈正相关。结论老年人淋巴细胞凋亡率上升;CD28^＋表达下降,CD28^-、CD95^＋表达上升;淋巴细胞的凋亡率与CD28^＋、CD95^＋有相关性。提示老化导致老年人淋巴细胞凋亡增加,CD28和CD95的变化与淋巴细胞凋亡密切相关。     

乳腺癌患者外周血CD40、CD40L的表达及意义

目的探讨乳腺癌患者外周血CD40、CD40L的表达及其与患者病理分期的关系。方法选择30例乳腺癌患者（乳腺癌组）、30例健康体检者（对照组）,采用流式细胞术检测两组外周血B、T淋巴细胞表面的CD40、CD40L表达水平,并分析其与乳腺癌病理分期的关系。结果乳腺癌组CD40[（9.26±2.12）%]、CD40L[（7.48±1.36）%]表达水平均高于对照组[（7.33±1.64）%、（6.25±1.19）%]（P均〈0.01）,且乳腺癌患者CD40、CD40L表达水平均与其病理分期呈正相关（rCD40=0.88、rCD40L=0.71,P均〈0.01）。结论乳腺癌患者外周血B、T淋巴细胞表面的CD40、CD40L水平可作为判断乳腺病变程度、肿瘤转移的指标。     

CD137／CD137L、白细胞介素-2在大肠癌中的表达及其临床意义

目的：检测大肠癌组织中 CD137、CD137L 和白细胞介素（IL）-2的表达,并探讨其对大肠癌发生和发展的作用。方法收集肠镜切除并经病理检查确诊的30例首发原发结直肠癌患者的癌组织、癌旁组织、远离癌肿的正常组织,采用免疫组织化学法检测其 CD137、CD137L 和 IL-2的表达。结果CD137在3种组织中均呈阴性表达。CD137L 在3种组织中的阳性表达率呈下降趋势（分别为80．0％、16．7％和0）,差异有统计学意义（P ＜0．05）,但不同年龄、性别、肿瘤大小、分化程度及肿瘤位置比较,CD137L 的表达差异无统计学意义（P ＞0．05）。CD137L 与 IL-2间无相关性。结论CD137L 在大肠癌组织及癌旁组织中的阳性表达率较高,CD137L 可能与大肠癌发生有关,对大肠癌早期诊断有一定的指导意义。     

肾综合征出血热患者循环内皮细胞CD95表达的临床研究

目的 探讨肾综合征出血热(HFRS)发热期和多尿期患者循环内皮细胞(CEC)中CD141、CD31及CD95的动态表达及临床意义.方法 应用流式细胞术分析不同型HFRS发热期和多尿期患者外周血细胞CD141、CD31及CD95的表达.多组数据的比较采用单因素方差分析.结果HFRS患者发热期和多尿期CD141+CD31+细胞(CEC)占外周血细胞的9.47%±1.98%和8.26%±1.55%.高于健康对照者的7.05%±1.45%(F=8.42;P=0.000,P=0.029),且发热期CD141+CD31+细胞较多尿期增高(P=0.48).HFRS发热期及多尿期患者CEC中CD95+细胞的平均荧光强度(MFI)较健康对照者明显升高(F=19.93;P=0.000,P=0.000),且发热期较多尿期明显增高(P=0.049).发热期各型患者CEC中CD95+细胞的MFI较健康对照者均明显增高(F=7.36;P=0.000),其中以重(危重)型患者最高,与中型和轻型比较差异有统计学意义(P=0.009,P=0.002).结论 CEC所占比例及CD95表达可能与HFRS病期及病情轻重有关.     

CD95基因转导食管癌细胞的体外抑瘤效应

目的 观察CD95基因表达株表达的CD95蛋白对体外培养食管癌细胞的抑制作用。方法 Western blot检测CD95基因转导前后CD95蛋白的表达水平。直接记数法描述转导株的细胞生长曲线;MTT显色法比较转导前后细胞的体外增殖能力;药敏实验观察转导细胞对VCR、5-FU等化疗药物的敏感性;软琼脂集落形成实验观察基因转导前后细胞的克隆形成力。结果 杂交结果表明．转导株CD95蛋白水平的表达明显高于非转导株。转导细胞的细胞倍增时间(2d)、对数生长期(3d～7d)等均体现了比非转导株(0．8d,2d～8d)更为缓慢和处于抑制状态,体外生长速度的增长指数下降,集落形成能力低下(0．9％),而对VCR、5-FU等化疗药物的敏感性明显增加。结论 CD95基因在食管癌细胞中处于低表达状态;通过真核表达载体的介导,CD95基因导入食管癌细胞后．能有效地表达CD95蛋白。CD95基因转导株的表达蛋白能有效地抑制体外培养的食管癌细胞的生长。     

IgE- and IgE+Ag-mediated mast cell migration in an autocrine/paracrine fashion

Mast cells are the major effector cells for immediate hypersensitivity and chronic allergic reactions. These cells accumulate in mucosal tissues of allergic reactions, where immunoglobulin E (IgE) is produced locally. Here we provide evidence that, in addition to antigen that can attract IgE-bound mast cells, the type of IgE molecules that efficiently activate mast cells can promote the migration of mast cells in the absence of antigen. IgE- and IgE+Ag-mediated migration involves an autocrine/paracrine secretion of soluble factors including adenosine, leukotriene B4, and several chemokines. Their secretion depends on 2 tyrosine kinases, Lyn and Syk, and they are agonists of G-protein-coupled receptors and signal through phosphatidylinositol 3-kinase gamma, leading to mast cell migration. In mouse experiments, naive mast cells are attracted to IgE, and IgE-sensitized mast cells are attracted to antigen. Therefore, IgE and antigen are implicated in mast cell accumulation at allergic tissue sites with local high IgE levels.     

Progesterone as an autocrine/paracrine regulator of human granulosa cell proliferation.

The present study examined the role of progesterone in regulating human granulosa cell proliferation. Human granulosa and luteal cells were obtained from follicular aspirates of women undergoing in vitro fertilization. Cells were plated at 5, 10, or 50 x 10(3) cells/mL and cultured for up to 6 days. At specific times, cells were harvested and assessed for cell number and morphology. The medium was assayed for progesterone. Cells plated at 50 x 10(3) cells/mL did not increase in number after 3 or 6 days of culture, but rapidly differentiated, secreting high amounts of progesterone (> or = 320 nmol/L). Conversely, cells plated at 5 x 10(3) or 10 x 10(3) cells/mL doubled in number over the first 3 days of culture and subsequently differentiated. The addition of 100 ng/mL progesterone or more to the medium inhibited proliferation. Aminoglutethamide blocked progesterone secretion and increased the number of cells present after 3 days of culture. The antiproliferative effects of progesterone were not mimicked by estradiol, testosterone, dihydrotestosterone, or dexamethasone and could not be overridden by epidermal growth factor, a potent mitogen. These data suggest that progesterone plays an autocrine/paracrine role in regulating granulosa cell proliferation.     

CD40 and CD95 induce programmed cell death in the human myeloma cell line XG2

We have previously demonstrated that CD40 stimulation induced a cellular growth arrest of the highly CD40-positive myeloma cell line XG2. To further characterize this inhibition of proliferation, we looked for a possible induction of apoptosis. Since no DNA fragmentation could be detected, we used newly described techniques that enable detection of apoptosis independently of DNA degradation, i.e. supravital exposure to propidium iodide (PI) and Annexin V labelling. We demonstrated that CD40 effectively induced programmed cell death. Furthermore, we have shown that CD95 (Fas) stimulation significantly enhanced the CD40-induced apoptosis.     

Control of autocrine and paracrine myocardial signals: an emerging therapeutic strategy in heart failure

A growing body of evidence supports the hypothesis that autocrine and paracrine mechanisms, mediated by factors released by the resident cardiac cells, could play an essential role in the reparative process of the failing heart. Such signals may influence the function of cardiac stem cells via several mechanisms, among which the most extensively studied are cardiomyocyte survival and angiogenesis. Moreover, besides promoting cytoprotection and angiogenesis, paracrine factors released by resident cardiac cells may alter cardiac metabolism and extracellular matrix turnover, resulting in more favorable post-injury remodeling. It is reasonable to believe that critical intracellular signals are activated and modulated in a temporal and spatial manner exerting different effects, overall depending on the microenvironment changes present in the failing myocardium. The recent demonstration that chemically, mechanically or genetically activated cardiac cells may release peptides to protect tissue against ischemic injury provides a potential route to achieve the delivery of specific proteins produced by these cells for innovative pharmacological regenerative therapy of the heart. It is important to keep in mind that therapies currently used to treat heart failure (HF) and leading to improvement of cardiac function fail to induce tissue repair/regeneration. As a matter of facts, if specific autocrine/paracrine cell–derived factors that improve cardiac function will be identified, pharmacological-based therapy might be more easily translated into clinical benefits than cell-based therapy. This review will focus on the recent development of potential pharmacologic targets to promote and drive at molecular level the cardiac repair/regeneration in HF.     

核心蛋白聚糖在HepG2细胞中的表达及作用

目的 构建分泌型核心蛋白聚糖(DCN)真核表达载体,并观察其在肝癌细胞HepG2中的表达和抗肿瘤作用.方法 应用PCR技术扩增DCN全长基因eDNA片段,与pcDNA3.1载体连接,并转化到大肠杆菌中扩增获得重组载体,应用双酶切、PCR以及测序鉴定此重组载体,脂质体介导转染HepG2,经G418筛选建立稳定转染细胞株,采用RT-PCR、免疫组化检测其表达.MTT检测细胞增殖活力,流式细胞仪分析细胞周期.结果 RT-PCR可见转染组细胞mRNA表达明显增多,免疫组化可见转染组细胞DCN蛋白表达明显增高.细胞生长曲线显示转染组细胞生长缓慢,G,期细胞显著增多.结论 本方法可成功建立稳定转染DCN的HepG:细胞株,并证实DCN能够抑制HepG:细胞株的生长.     

Human CD4+CD25+ regulatory T cells selectively express tyrosine hydroxylase and contain endogenous catecholamines subserving an autocrine/paracrine inhibitory functional loop

CD4+CD25+ regulatory T lymphocytes (Tregs) are specialized T cells playing a key role in the control of immune homeostasis. Here, we show that human Tregs constitutively express tyrosine hydroxylase (TH, EC 1.14.16.2), the rate-limiting enzyme in the synthesis of catecholamines, and contain substantial amounts of dopamine, norepinephrine, and epinephrine, which are released upon treatment with reserpine. Catecholamine release results in reduced production of interleukin-10 and transforming growth factor-beta by Tregs, and in down-regulation of Treg-dependent inhibition of effector T-lymphocyte (Teff) proliferation, which occurs without affecting the production of tumor necrosis factor-alpha or interferon-gamma. Tregs and Teffs express on the cell membrane both D1-like and D2-like dopaminergic receptors to a similar extent (12%-29% of the cells). Catecholamine-dependent down-regulation of Tregs is, however, selectively reversed by pharmacological blockade of dopaminergic D1-like receptors, which in Tregs only (and not in Teffs) are also expressed at the level of mRNA and are functionally coupled to intracellular production of cAMP. These findings indicate that in human Tregs endogenous catecholamines subserve an autocrine/paracrine loop involving dopaminergic pathways and resulting in down-regulation of Treg function.