血液透析患者外周血中Bcl-2水平检测及临床意义

目的 :探讨血液透析病人外周血中Bcl 2的变化及临床意义。方法 :用ELISA法检测 30例血透病人和 2 0例正常人Bcl 2水平 ,并与sFas、IL 6和TNF α进行比较。结果 :血透病人Bcl 2值 (7 43± 2 14U ml)显著低于正常对照组 (9 14±2 84)U ml,P <0 0 5 ;透析后Bcl 2水平比透析前进一步降低 (6 0 3± 2 0 1)U ml ,P <0 0 5。血液透析病人Bcl 2水平与sFas、IL 6、Cr水平呈明显负相关 ,r值分别为 - 0 482、- 0 375、- 0 36 3,P <0 0 5。结论 :血液透析病人外周血中Bcl 2水平显著降低 ,可作为衡量血透疗效的一项指标。     

IL-13对小鼠哮喘模型气道黏蛋白基因Muc5ac及Bcl-2、Bax表达的作用

目的研究白细胞介素-13(IL-13)处理小鼠支气管哮喘(哮喘)模型前后肺组织黏蛋白基因Muc5ac、凋亡相关蛋白Bcl-2和Bax表达的作用,探讨气道黏液过度分泌的机制.方法45只雄性BALB/c小鼠随机分为对照组、哮喘组和IL-13组,每组15只.用逆转录-聚合酶链反应(RT-PCR)方法和免疫组化法分别检测Muc5acmRNA、Muc5ac蛋白、Bcl-2蛋白以及Bax蛋白在肺组织的表达.结果哮喘组和对照组肺组织Muc5acmRNA分别为(0.1552±0.0057)和(0.0633±0.0013),Muc5ac蛋白分别为(0.8849±0.0257)和(0.1166±0.0064),两组比较差异均有统计学意义(P<0.01);IL-13组肺组织Muc5acmRNA和蛋白分别为(0.2807±0.0027)和(1.6138±0.0483),与哮喘组、对照组比较差异也均有统计学意义(P均<0.01).与对照组Bcl-2蛋白(0.3279±0.0136)、Bax蛋白(1.7284±0.0263)相比,哮喘组分别增加和降低(分别为0.8383±0.0310和0.8987±0.0106),两组差异均有统计学意义(P均<0.01);IL-13处理后可分别促进Bcl-2和Bax蛋白增加和降低(分别为1.6934±0.0229和0.3522±0.0152),其和哮喘组的差异均有统计学意义(P均<0.01);哮喘组和IL-13组小鼠肺组织Muc5acmRNA、蛋白表达与Bcl-2蛋白表达均呈直线正相关(P均<0.05),而与Bax蛋白表达则均呈直线负相关(P均<0.05).结论IL-13是引起哮喘气道黏液过度分泌的重要细胞因子,它可能通过改变Bcl-2和Bax的表达导致了上述病变.     

Interleukin-2 receptor common γ-chain signaling cytokines regulate activated T cell apoptosis in response to growth factor withdrawal: Selective induction of anti-apoptotic (bcl-2, bcl-xL) but not pro-apoptotic (bax,bcl-xS) gene expression

Cytokine deprivation from activated T cells leads to apoptosis associated with down-regulation of the bcl-2 gene product. It is not clear, however, how cytokines other than interleukin-2 (IL-2) may affect this process and regulate the involvement of other apoptosis-modulating genes. We show that a group of cytokines including IL-2, IL-4, IL-7 and IL-15, which can all signal through the γ chain of the IL-2R (IL-2Rγ), prevent the apoptosis of IL-2-deprived activated T cells. This rescue involves the induction of the anti-apoptosis genes (bcl-2 and bcl-x_L), but causes little change in expression of bax and bcl-x_S, which promote apoptosis. Furthermore, the prevention of apoptosis and induction of proliferation by the common γ chain cytokines can be dissociated. Thus, when proliferation is blocked, the common γ chain cytokines still induce up-regulation of bcl-2 relative to bax and retard apoptosis. These cytokines can thus regulate the persistence or removal of effector T cells by coordinating the balance between genes which promote and those which inhibit apoptosis, events which are probably mediated at least in part by signals through the common γ chain. These data also implicate inappropriate T cell apoptosis resulting from a dysfunctional common γ-chain as part of the pathophysiological defect in patients with X-linked severe-combined immunodeficiency (SCID).     

Bcl-2、bax基因在功能失调性子宫出血中的表达及意义

Bcl-2和Bax是一对重要的凋亡调节基因,其调节过程广泛存在于全身各组织器官.研究发现子宫内膜的周期性脱落依赖于bcl-2和bax基因的调节,bcl-2主要在增生期子宫内膜表达,排卵后表达迅速降低,bax在整个月经周期均有表达,增生期表达弱阳性,分泌期表达明显增强.功能失调性子宫出血(DUB)患者中子宫内膜组织的Bc...     

帕金森病与ATP13A2基因

ATP13A2基因是帕金森病(PD)的致病基因,早发型帕金森病(EOPD)和Kufor-Rakeb综合征(KRS)患者中均发现ATP13A2基因突变。基因突变类型与疾病的严重程度和发病年龄相关,PD患者中黑质多巴胺能神经元也存在ATP13A2基因mRNA表达升高。因此,对ATP13A2基因的研究将有助于该病的基因诊断、病理生理学机制的阐明和治疗。     

大鼠创伤性脑损伤后海马细胞凋亡及bcl-2、NGF-R阳性细胞的变化

目的：探讨大鼠创伤性脑损伤后不同时相海马细胞凋亡及。bcl-2、NGF—R阳性细胞的变化规律。方法：采用大鼠自由落体脑损伤模型，伤后1、2、3、4、5、7、10d取脑切片，作：Nissl染色，用TUNEL法检测细胞凋亡，免疫组化染色观察bcl-2、NGF-R阳性细胞。结果：Nissl染色可见损伤侧海马CA2、CA3区锥体细胞层细胞消失或排列紊乱。损伤侧海马凋亡细胞伤后5d达到高峰，正常侧海马各时相点均未见到凋亡细胞。损伤侧海马bcl-2阳性细胞数量下降，伤后3d至最低点；NGF—R阳性细胞数量增加，伤后5d达到高峰。结论：大鼠创伤性脑损伤后损伤侧海马细胞凋亡数量的变化与伤后时程有关。伤后细胞bcl-2表达下降、NGF-R表达增加是导致细胞凋亡的因素。     

Developmental patterns of Ki-67, bcl-2 and caspase-3 proteins expression in the human upper jaw

The distribution of the Ki-67, bcl-2 and caspase-3 proteins was immunohistochemically analyzed in the developing human upper jaw (5th-10th gestational weeks).During this period, proliferative activity gradually decreased from higher levels at the earliest stages (50-52%) to lower levels, both in the jaw ectomesenchyme and in the epithelium. The highest expression of bcl-2 protein was found in the epithelium and ectomesenchyme of areas displaying lower rates of cell proliferation. High levels of caspase-3 protein were detected during the earliest stages of jaw development, indicating an important role for apoptosis in morphogenesis of early derivatives of the maxillary prominences. The number of Ki-67, bcl-2 and caspase-3 positive cells changed in a temporally and spatially restricted manner, coincidently with upper jaw differentiation. While apoptosis might control cell number, bcl-2 could act in suppression of apoptosis and enhancement of cell differentiation. A fine balance between cell proliferation (Ki-67), death (caspase-3) and cell survival (bcl-2) characterized early human upper jaw development. A rise in the number of apoptotic cells always temporally coincided with the decrease in number of surviving bcl-2 positive cells within the palatal region. Therefore, the upper jaw development seems to be controlled by the precisely defined expression of genes for proliferation, apoptosis and cell survival.     

miR-222通过调控BCL2L13基因促进HBx-HepG2细胞生长

目的:探讨HBx-Hep G2细胞中微小RNA-222(mi R-222)对BCL2L13基因表达的调控作用以及对细胞生长和凋亡的影响,并研究其潜在的分子作用机制。方法:利用实时荧光定量PCR检测mi R-222的表达水平;MTT和集落形成实验检测细胞的生长;流式细胞术检测细胞周期和凋亡;构建BCL2L13 3’UTR双萤光素酶报告载体,通过采用双萤光素酶报告实验验证mi R-222的靶基因。结果:与正常的肝细胞L02相比,mi R-222在HBx-Hep G2细胞过量表达(P0.05)。mi R-222过表达可促进HBx-Hep G2细胞的生长,改变细胞周期,降低细胞的凋亡率;mi R-222表达下调可抑制HBx-Hep G2细胞的生长,改变细胞周期,增加细胞的凋亡率,和对照组相比差异有统计学显著性(P0.05)。与正常肝细胞L02相比,BCL2L13在HBx-Hep G2细胞表达下调(P0.05);mi R-222表达下调可促进BCL2L13的表达(P0.05)。双萤光素酶报告实验和pc DNA3.1-BCL2L13转染实验结果提示mi R-222可以通过作用于BCL2L13的3’UTR区,负向调控其表达,从而促进细胞的生长。结论:mi R-222可以通过靶向调控BCL2L13基因进而促进HBx-HepG2细胞的生长。     

Chronic application of nonylphenol-induced apoptosis via suppression of bcl-2 transcription and up-regulation of active caspase-3 in mouse brain

Nonylphenol (NP) is an endocrine disruptor, which has been reported to have adverse effects on reproductive and immune systems. However, the influence of NP on the central nervous system (CNS) has not been extensively explored. The present study was performed to investigate the effects of chronic administration of NP on the apoptosis-related protein expression in mouse brain by in situ hybridization, RT-PCR and immunoblotting assays. The expression of bcl-2 mRNA was down-regulated by NP at the doses of 100 and 200mg/(kg day) (p<0.05), whereas the expression of bax mRNA was not affected in NP treated mice (p>0.05). Furthermore, as the main executor of apoptosis, the expression of active caspase-3 was up-regulated by 100 and 200mg/(kg day) NP (p<0.01), which is in accord with the results of terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling (TUNEL) (p<0.05). These results suggest that chronic application of NP could sensitize the mice brain to apoptosis.     

APO-1 mediated apoptosis or proliferation in human chronic B lymphocytic leukemia: Correlation with bcl-2 oncogene expression

We studied induction of apoptosis in several human malignant B cells from chronic lymphocytic leukemias (BCLL) by triggering the APO-1 antigen. The APO-1 antigen was found to be expressed on the surface of malignant B cells. In BCLL cells from most patients, APO-1 antigen expression increased following in vitro activation by Staphylococcus aureus Cowan I (SAC) or interleukin-2 (IL-2). In certain cases of BCLL co-stimulation with SAC plus IL-2 resulted in a synergistic up-regulation of the APO-1 antigen on the cell surface and prepared BCLL cells for monoclonal antibody anti-APO-1 mediated apoptosis. Interestingly, bcl-2 mRNA expression decreased upon stimulation with SAC plus IL-2, whereas SAC or IL-2 alone did not affect the level of bcl-2 expression. Thus, in these BCLL cells induction of anti-APO-1-mediated apoptosis appeared to be correlated with bcl-2 mRNA down-regulation. One informative BCLL, however, with a similar pattern of APO-1-antigen expression, did not show SAC plus IL-2-dependent bcl-2 down-regulation. Surprisingly, these cells proliferated in response to anti-APO-1 only when cells were co-stimulated with SAC plus IL-2. Our data suggest that down-regulation of bcl-2 prepares BCLL cells for induction of APO-1-mediated apoptosis. In addition they demonstrate that triggering of the APO-1 antigen may also lead to the induction of proliferation in special cases of BCLL.     

bcl-2基因表达在genistein抑制裸鼠肝癌生长中的作用

目的：探讨bcl-2基因表达变化在genistein治疗裸鼠移植人肝癌中的作用。方法:以裸鼠移植人肝癌为对照, 对照组腹腔注入含0.04%DMSO RPMI-1640培养基0.05 L/kg，genistein治疗组腹腔注入genistein 1 mg kg-1·d-1，3周后观察肝癌增长情况，并应用同位素试剂盒检测肝癌组织IP3 含量,RT-PCR分析癌组织bcl-2 mRNA表达，Western blotting 分析肝癌组织Bcl-2蛋白表达。结果:治疗组肝癌体积和重量均显著低于对照组［体积:（42.7±27.8)mm3 vs (52.3±26.5)mm3;重量:(42.7±27.8)mg vs (91.3±31.4)mg］，IP3含量显著低于对照组［(13.4±1.4)nmol/g protein vs （35.3±6.6）nmol/g protein］，bcl-2 mRNA表达显著低于对照组［RI（灰度与面积之积的相对强度）0.48±0.02 vs 0.56±0.15］, P<0.01，Bcl-2蛋白表达显著低于对照组［RI（灰度与面积之积的相对强度）1.69±0.52 vs 1.37±0.48］。结论:Genistein能减少IP3 生成,下调肝癌组织bcl-2基因表达,抑制裸鼠移植肝癌的生长。     

二苯乙烯苷对同型半胱氨酸诱导血管内皮细胞凋亡及bcl-2、bax、caspase-3表达的影响

 目的：研究何首乌二苯乙烯苷（TSG）对同型半胱氨酸（Hcy）诱导人脐静脉内皮细胞（HUVECs）凋亡及bcl-2、bax和caspase-3 mRNA表达的影响。方法：建立Hcy （3 mmol/L）所致培养HUVECs核损伤模型,TSG（1和10 μmol/L）提前2 h预孵育,然后再以Hcy处理,作为TSG保护组。用Hoechst 33342核染色法观察HUVECs细胞核损伤状态,以流式细胞术检测细胞凋亡情况,用实时荧光定量RT-PCR法检bcl-2、bax和caspase-3 mRNA的表达。结果：经3 mmol/L Hcy处理后,与正常培养的细胞相比,HUVECs核损伤加重,凋亡细胞比例升高,bcl-2表达降低（P＜0.01）,bax和caspase-3表达增加（P＜0.01）。TSG 1 μmol/L和10 μmol/L预孵育后再经3 mmol/L Hcy处理,与单经3 mmol/L Hcy损伤模型组相比,HUVECs细胞核损伤降低,凋亡率下降,bcl-2的表达增加（P＜0.05）,bax和caspase-3表达降低（P＜0.05）。结论：TSG具有降低Hcy所致HUVECs的细胞损伤和抑制凋亡的作用,其机制可能与影响bcl-2、bax和caspase-3的表达有关。     

Enforced expression of human bcl-2 in CD4+ T cells enhances human herpesvirus 7 replication and induction of cytopathic effects

The cytopathic effects (CPE) resulting from the infection of CD4⁺ T cells by human herpes-virus 7 (HHV-7) comprises two major mechanisms: generation of large polyploid cells, which eventually undergo necrotic lysis, and apoptosis, predominantly occurring in small mononucleated cells. To dissect the relative contribution of these two phenomena to the overall cytopathicity of HHV-7 in vitro, we have investigated the effect of acute HHV-7 infection on SupT1 CD4⁺ T cell lines stably transfected either with the bcl-2 anti-apoptotic gene or with the control vector. Overexpression of Bcl-2 protein by these cells was associated with a progressive decline of the total number of viable cells, and a relative increase of enlarged polyploid cell. Of note, the size of polyploid cells was significantly greater in SupT1 cells over expressing bcl-2 than in cells transfected with the control vector. In addition, bcl-2 expression accelerated the kinetics of an acute spreading of HHV-7 infection, as determined by HHV-7-specific indirect immunostaining revealed by either fluorescence microscopy or flow cytometry. Our results indicate that inhibition of apoptosis in HHV-7-infected cultures greatly favors the process of polyploidization and represents a major mechanism to maximize viral transmission.     

Induction of Bax-α precedes apoptosis in a human B lymphoma cell line: potential role of the bcl-2 gene family in surface IgM-mediated apoptosi

The members of the bcl-2 gene family are major regulators of programmed cell death, but their role in sIg-triggered apoptosis remains unclear. Using sensitive and resistant variants of the human B cell line BL-41, we studied the expression of the bcl-2 gene family during surface IgM-mediated apoptosis. We found constitutive Bcl-2 and Bcl-x expression, which remained unaltered after sIg cross-linking, in both resistant and sensitive cells. This and other experiments suggest that constitutive expression of Bcl-2 or Bcl-x alone is not sufficient to protect from activation-induced cell death in B cells. We therefore investigated Bax-α, the death-promoting splice variant of Bax, and found strong induction of both mRNA and protein upon sIg stimulation in sensitive cells. However, resistant subclones showed only weak expression, which was not inducible by sIg cross-linking. We provide evidence that up-regulation of Bax-α and the resulting imbalance of Bcl-2/Bax might be a major regulator of sIg-mediated apoptosis. Additionally, we found strong constitutive expression of Bcl-x_s, the death promoting variant of Bcl-x, in sensitive cells, whereas resistant cells showed only weak Bcl-x_s expression. Thus, we observed a much stronger expression of the death-promoting proteins Bax-α (inducible) and Bcl-x_s (constitutive) in sensitive cells than in resistant cells. We therefore propose a potential role of the novel bcl-2 gene family members bcl-x and bax in surface IgM-triggered apoptosis.     

哮喘豚鼠嗜酸细胞Bcl-2和Bax mRNA表达及意义

目的:观察Bcl-2及Bax mRNA表达与嗜酸粒细胞（Eos）凋亡的关系,探讨其在哮喘发病中的作用。方法:健康豚鼠随机分为正常组、哮喘组,卵蛋白致敏激发制作哮喘模型,检测支气管灌洗液（BALF）中低密度EoS（HEos）及正常密度Eos（NEos）细胞凋亡,原位杂交及RT-PCR法检测Bcl-2及Bax mRNA表达。结果:哮喘组EoS显著高于正常组,以HEos为著（P<0.01）;Eos凋亡率明显低于正常组（P<0.01）。哮喘组不同密度Eos表达Bcl-2 mRNA明显增加,而Eos表达Bax mRNA明显减少。结论:哮喘时Eos存在凋亡抑制,这是哮喘时Eos增多的原因之一,哮喘豚鼠BALF Eos表达Bcl-2增加,表达Bax减少,表明Bcl-2及Bax可通过调节Eos凋亡参与哮喘发病。     

Apoptosis-related factors p53, bcl-2 and the defects of force transmission in dilated cardiomyopathy

The etiology of heart failure in dilated cardiomyopathy involves multiple agents. The purpose of this study was to investigate the presence of apoptosis-related proteins p53, bcl-2, and the defects of force transmission in end-stage dilated cardiomyopathy.     

Regulation of Fas(Apo-1/CD95)- and perforinmediated lytic pathways of primary cytotoxic T lymphocytes by the protooncogene bcl-2

Cytotoxic T cells (CTL) induce cell death of their target cells either by the surface interaction between Fas ligand and Fas or by the release of perforin and granzymes. Both lytic pathways induce apoptosis yet it is not known whether identical or distinct apoptotic pathways are activated. The protooncogene bcl-2 is known to protect various hematopoietic cells from apoptosis induced by diverse agents. Here we show that overexpression of the Bcl-2 protein in the murine mastocytoma line P815 or in concanavalin A-activated splenocytes suppresses apoptotic cell death induced by allospecific primary cytotoxic T lymphocytes (CTL) in which only the Fas lytic pathway was functional. Bcl-2 also reduced target cell killing induced by CTL whose lytic activity was dependent on the perforin/granzyme pathway only. These data provide evidence that, in the target cells studied here, both perforin/granzyme and Fas apoptotic pathways are modulated by Bcl-2 and suggest that these two pathways converge at a step prior to Bcl-2 inhibition.     

胶质瘤bcd—2基因表达水平与其细胞增殖和凋亡关系的研究

目的 探讨胶质瘤细胞ｂｃｌ－２基因表达水平与肿瘤恶性程度、细胞增殖活性及凋亡程度的关系。方法 以６９例不同级别的人胶质瘤组织为研究对象,用原位杂交及免疫组化染色ＡＢＣ法分别检测ｂｃｌ－２ｍＲＮＡ、ｂｃｌ－２蛋白和增殖细胞核抗原（细胞增殖活性标记物）的表达,并用３’末标记法做原位细胞凋亡检测。结果 ６４例（９２．８％）表达ｂｃｌ－２ｍＲＮＡ,６０例（８７．０％）表达ｂｃｌ－２蛋白,两者的表达水平呈正     

Gene rearrangement of bcl-1 and bcl-2 is confined to distinct subgroups of high-grade malignant B-cell lymphomas

The occurrence of bcl-1 and bcl-2 gene rearrangements was investigated in 37 cases of high-grade B-cell lymphomas. Bcl-2 rearrangement was detectable only in single cases of primary centroblastic lymphoma with a follicular growth pattern, whereas secondary centroblastic lymphomas evolving from a centroblastic–centrocytic lymphoma were positive in up to 60 per cent of the cases analysed. Bcl-1 rearrangement was found only in one case of immunoblastic B-cell lymphoma with a history of a pre-existing lymphoplasmacytoid immunocytoma. It is concluded that there may be a subgroup of centroblastic lymphomas with a biology similar to that of centroblastic–centrocytic lymphomas. The detection of bcl-1 rearrangement in high-grade lymphomas may indicate a secondary high-grade lymphoma.     

Amyloid precursor protein, heat-shock proteins, and Bcl-2 form a complex in mitochondria and modulate mitochondria function and apoptosis in N2a cells

Neurons that degenerate in the brains of persons with Alzheimer's disease accumulate mitochondrial amyloid precursor protein (APP), which is thought to negatively affect mitochondrial function and cellular homeostasis. Because proteins that enter mitochondria require assistance from chaperone proteins, we hypothesized that heat-shock proteins (HSPs) help accumulate APP in mitochondria. We found that APP overexpression in N2a cells (APP cells) did not elicit mitochondrial dysfunction. Because cerebral hypoperfusion-associated energy deficiency is an important etiology for Alzheimer's disease, we also challenged the cells with serum starvation. APP/HSP/Bcl-2 complexes formed within the mitochondria of serum-starved APP cells, but not control cells. Mitochondria containing APP/HSP/Bcl-2 complexes induced apoptosis. We hypothesize that APP/HSP/Bcl-2 complexes diminish the functional capacities of HSPs and Bcl-2, which leads to mitochondrial injury and apoptosis.