自然流产患者绒毛滋养层细胞PCNA和caspase-3的表达

目的从细胞增殖与细胞凋亡的角度探讨自然流产的发生机制。方法取正常早孕流产和自然流产病例各20例的绒毛组织,光镜观察绒毛形态学变化;免疫组织化学SP法检测增殖细胞核抗原(PCNA)及凋亡蛋白酶caspase-3在滋养层细胞中的表达。结果自然流产绒毛组织均出现不同程度的退行性改变,细胞滋养层的增殖指数较对照组降低,而细胞滋养层、合体滋养层的凋亡指数却明显升高。结论凋亡蛋白酶caspase-3可能参与自然流产的发生机制。     

人早孕绒毛滋养层细胞的分离纯化及马鞭草抗早孕机理的初步研究

目的：探讨马鞭草抗早孕的细胞学作用机理。方法;正常妊娠６－８周早期胎盘Ｐｅｒｃｏｌｌ梯度离心分离出滋养层细胞进行体外培养,观察马鞭草对培养的滋养层细胞形态及绒毛膜促性腺激素分泌功能的影响。结果：２５,５０ｍｇ／ｍｌ马鞭草乙醇提取液对滋养层细胞生长及绒毛膜促性腺激素分泌有明显的抑制作用。结论;一定浓度的马鞭草可直接杀伤滋养层细胞,抑制绒毛膜促性腺激素的分泌而中止早孕。     

Bcl—2和caspase—3在创伤性脑损伤后细胞凋亡中的作用

Bcl-2和caspase-3分别是Bcl-2基因族和caspase-3基因族中具有代表性的基因，它们参与对细胞凋亡的调控．研究表明创伤性脑损伤后存在细胞凋亡及凋亡相关基因的表达．从基因水平探讨创伤性脑损伤后细胞调亡调控机制是近年研究的一个热点．本文仅综述Bcl-2和caspase-3基因在创伤性脑损伤研究中的最新进展．     

丙酸睾丸酮,米非司酮对早孕蜕膜,绒毛组织超微结构的…

使用透射电镜观察丙酸睾丸酮（丙睾），米非司酮，或两者合用对早孕蜕膜，绒毛组织超微结构的影响，结果表明：丙睾组出现蜕膜细胞缩小，结构不清，核固缩，合体滋养细胞核密度增高，核膜不清等严重改变，米非司酮组则合体滋养层细胞内质网扩大，溶酶体增多，微绒毛减少，细胞核异染色质边聚；朗罕细胞线粒体肿胀，空泡变，细胞核异染色介质边聚等严重损害改变，而两药合用组出现较其中任何一组更严重的损害作用（Ｐ〈０．０１）。提     

H2O2通过ERK通路抑制绒毛外细胞滋养层细胞的侵袭能力

目的研究活性氧过氧化氢（H2O2）对绒毛外细胞滋养层细胞（EVCT）侵袭行为的影响及其信号传导机制。方法建立EVCT体外培养模型,利用Transwell细胞侵入系统检测EVCT的体外侵袭作用,应用Western blot法评价细胞外信号调节蛋白激酶（ERK1/2）的活性变化,使用CCK-8测定细胞的生长状况。结果与对照组比较,15μmol/LH2O2和50 μmol/L PD98059均可显著抑制EVCT的侵入指数（分别为43.3±4.7,49.3±7.0）（P〈0.01）,同时降低EVCT的ERK1/2磷酸化水平（P〈0.01）,但并不影响EVCT的细胞活力（P〉0.05）。结论 H2O2可能通过抑制ERK1/2的激活,进而影响EVCT的侵袭行为,参与子痫前期的发病机制。     

早孕绒毛滋养细胞FasL表达异常与自然流产的关系

通过比较正常早孕与自然流产者滋养细胞表面FasL表达,进一步从分子免疫学角度探讨自然流产的发病机理。方法：用特异性的FasL抗体进行免疫组化染色,并通过高清晰彩色病理免疫组化测量系统对其定量分析,SSPS对两组进行比较。结果：自然流产组滋养细胞表面FasL表达面积及强度均明显低于正常早孕组,两者间差异有显著性意义。结论：滋养细胞表面FasL表达减少,引起母胎间的免疫耐受的破坏,是导致自然流产的一重量免疫病因,诱导FasL的产生或调节母胎间的免疫耐受将为临床治疗自然流产提供新的方向。     

Bcl-2和caspase-3在创伤性脑损伤后细胞凋亡中的作用

bcl-2和caspase-3分别是bcl-2基因族和caspase基因族中具有代表性的基因,它们参与对细胞凋亡的调控.研究表明创伤性脑损伤后存在细胞凋亡及凋亡相关基因的表达.从基因水平探讨创伤性脑损伤后细胞调亡调控机制是近年研究的一个热点.本文仅综述bcl-2和caspase-3基因在创伤性脑损伤研究中的最新进展.     

Bcl-2表达与小鼠腹腔巨噬细胞凋亡的关系

利用免疫荧光技术和激光扫描共聚焦显微镜（共焦镜）技术检测了地塞米松介导小鼠腹腔巨噬细胞凋亡时调亡抑制基因Ｂｃｌ－２表达的时空变化。结果显示,免疫荧光技术和共焦镜技术可对细胞内的蛋白表达作定量和定位检测,凋巨噬细胞内Ｂｃｌ－２逐渐减少,核中Ｂｃｌ－２相对量逐渐减少,胞浆中Ｂｃｌ－２相对量逐渐增多,Ｂｃｌ－２表达和地塞米松介导的巨噬细胞凋亡呈极显著负相关。结果表明,地塞米松介导巨噬细胞凋亡时,凋亡抑制     

Bcl-2和Bax蛋白在人早孕胎盘绒毛和蜕膜组织中的表达

目的探讨Bcl-2和Bax蛋白在人早孕绒毛和蜕膜组织中的表达及其意义。方法应用免疫组织化学ABC法检测Bcl-2和Bax蛋白在孕5~7周绒毛和蜕膜组织细胞中的表达,用真彩色病理图像分析系统4.0图像分析软件测定Bcl-2和Bax蛋白表达的积分吸光度值。结果Bc1-2蛋白主要分布于孕5~7周绒毛合体滋养层细胞、蜕膜细胞的细胞质和细胞核中,蛋白表达积分吸光度值依次降低,其差异无统计学意义(P>0.05)。Bax蛋白在孕5~7周绒毛细胞滋养层细胞、合体滋养层细胞中均未见表达;孕6周,Bax蛋白表达于极少部分蜕膜细胞的细胞质中。结论Bc1-2和Bax蛋白在人早孕过程中参与了绒毛滋养层细胞增殖和分化、子宫内膜蜕膜化的过程,在绒毛的发生、发育、胎盘形成和组织结构改建及功能完善等方面发挥着重要作用。     

脑缺血再灌注损伤的海马神经元对Bax、Bcl-2和Caspase-3的表达

目的：凋亡调控基因在脑缺血再灌后海马神经元的表达。方法：采用免疫组织化学的方法，观察 昆明小鼠双侧颈总动脉结扎7min后不同再灌时间组(24h组、48h组、72h组、7d组、14d组)海马CAl区神经元Bax、Bcl-2和Caspase-3的活性形式CM1的免疫反应活性。结果：Bax和CM1阳性神经元数在48h组最多，与其他各组相比差异有显著性(P＜0．01)，72h组明显下降，14d组完全消失；而Bcl-2阳性神经元数在48h组增多(与24h组相比，P＜0．01)，72h组下降，7d组再次上升(与72h组相比，P＜0．01)，14d组最多(与48h组相比，P＜0．01)。在24h、48h、72h、7d组，Bax阳性神经元多于Bcl-2阳性神经元(P＜0．05)，14d组则相反。结论：Bax和caspase-3在脑缺血再灌早期表达增强，然后下降以至消失，Bcl-2于再灌后期表达增强。Bax表达上调可能与Caspase-3激活相关。     

胰岛素对滋养层细胞凋亡的调节作用及机制

目的 :探讨胰岛素对培养的滋养细胞凋亡的影响及可能的机制。方法 :将培养的妊娠早期滋养层细胞 ,分为正常对照组(细胞 +培养液 )、H2 O2 组 (细胞 +培养液 +H2 O2 )和胰岛素 +H2 O2 组 (细胞 +H2 O2 +胰岛素 )。采用透射电镜观察及流式细胞术 ,观察H2 O2 诱导的细胞凋亡及胰岛素对H2 O2 诱导的细胞凋亡的抑制作用。并检测胰岛素对滋养细胞caspase 3的活性及Bcl 2蛋白表达的影响。结果 :H2 O2 可诱导培养的滋养细胞凋亡 ,透射电镜下可见特征性的细胞核改变。胰岛素可显著抑制H2 O2 诱导的细胞凋亡 ,流式细胞仪检测其凋亡率较H2 O2 组显著下降 (P <0 .0 1)。H2 O2 组滋养细胞中caspase 3的活性较对照组显著增高 (P <0 .0 1) ,而Bcl 2蛋白的表达则较对照组显著下降 (P <0 .0 1)。结论 :胰岛素可明显抑制H2 O2诱导的滋养细胞凋亡 ,其机制可能与降低caspase 3的活性和促进Bcl 2蛋白的表达有关     

The effects of mifepristone (RU486) on prostaglandin dehydrogenase in decidual and chorionic tissue in early pregnancy

Prostaglandin dehydrogenase is the main inactivating enzymefor prostaglandins and therefore controls local levels of prostaglandins.Since there is some evidence that the expression of this enzymeis under progesterone control it is reasonable that one of theeffects of antiprogestin is to reduce the concentration of thisenzyme and thus increase the effective concentration of prostaglandinwithin tissue. We have investigated the amount of enzyme activitywithin decidua and chorionic villi from women receiving theantigestagen mifepristone (RU486) 12, 24 and 36 h prior to surgicalabortion, and examined the effect on tissue concentrations ofprostaglandin dehydrogenase. Women receiving mifepristone inall groups had a significant reduction in concentration of prostaglandindehydrogenase enzyme in decidual tissue. There was also a markedreduction in prostaglandin dehydrogenase in decidual cells followingRU486, as demonstrated by immunochemical methods. At this stageof pregnancy, prostaglandin dehydrogenase was present in abundancein cytotrophoblast cells of chorionic villi but virtually absentfrom syncytiotrophoblast. In chorionic villi after RU486 administrationin vivo, there were no obvious differences in prostaglandindehydrogenase distribution or reactivity in the majority ofcases.     

抗HER2单链抗体融合凋亡蛋白对SKOV3细胞的靶向杀伤

目的：探讨分泌表达的抗HER2单链抗体与重构型人caspase—3融合蛋白对HER2抗原阳性肿瘤细胞的靶向杀伤作用。方法：将重构型人caspase—3基因亚克隆人pCMV—eDscFv—PEII—PEIII的相应位点，构建重组真核表达载体pCMV—e23scFv—PEII—revcasP3，并转染人T淋巴瘤细胞系Jurkat，筛选并建系。用ELISA检测培养上清中融合蛋白的分泌表达。通过共培养实验观察含有该融合蛋白的培养上清对人卵巢癌细胞SKOV3生长的抑制作用。结果：融合蛋白基因可在Jurkat细胞中，分泌表达并杀伤SKOV3细胞。结论：分泌表达的抗erbB2单链抗体与重构型人caspase3融合蛋白能够靶向诱导SKOV3细胞死亡。     

Effect of IL-5, glucocorticoid, and Fas ligation on Bcl-2 homologue expression and caspase activation in circulating human eosinophils

IL-5 is a potent eosinophil viability-enhancing factor that has been strongly implicated in the pathogenesis of IgE-mediated inflammation in vivo. Recently published data have suggested that IL-5 (and related cytokines) may act by altering the expression of the anti-apoptotic regulator Bcl-2 or its homologues, but this is controversial. The behaviour of the recently described pro-apoptotic cysteine proteases (caspases) in eosinophils after IL-5 treatment has not been explored. We examined the effect of IL-5 on the expression of four major Bcl-2 homologues, as well as on the expression/activation of key members of the caspase cell death cascade in cultured circulating human eosinophils. The effect of relevant inducers of eosinophil apoptosis (glucocorticoid and Fas ligation) on these regulatory proteins was also examined. We observed baseline expression of the anti-apoptotic Mcl-1 and pro-apoptotic Bax proteins in immunoblots of eosinophil lysates, but not Bcl-x, Bcl-2. IL-5 treatment had the effect of maintaining this basal level of expression over time without altering the balance of Bcl-2 homologues. The (upstream) caspase 8 and (downstream) caspase 3 proenzymes were detected in eosinophils at baseline, and were processed during spontaneous and stimulated eosinophil death. IL-5 completely blocked caspase processing in spontaneous and dexamethasone-induced cell death, and significantly slowed processing during Fas ligation. Our data do not support the theory that IL-5 acts by altering the balance of anti-apoptotic and pro-apoptotic Bcl-2 homologues, but suggest that it may act by regulating activation of the caspase cell death cascade.     

Endogenous association of Bim BH3-only protein with Mcl-1, Bcl-xL and Bcl-2 on mitochondria in human B cells

Bim is an essential regulator of lymphoid system homeostasis and appears essential for B cell apoptosis induction. The mechanism by which Bim isoforms are held in an inactive form remains poorly documented in normal B cells. In the current study, we demonstrated that in normal tonsil B cells the three major Bim isoforms are strongly associated with the anti-apoptotic Bcl-2 family members Mcl-1, Bcl-2 and Bcl-x(L). On the other hand, only a weak association of BimEL and L with the dynein LC8 chain has been found. In addition, there is no free Bim in normal B cells. Moreover, subcellular fractionation demonstrated that Bim and the anti-apoptotic counterparts are localized preferentially in the mitochondria-rich fraction. The fact that most Bim was found in this fraction supports the hypothesis that it is sequestered by anti-apoptotic molecules in mitochondria where its pro-apoptotic activity is controlled. Of interest, BimS is essentially complexed to Mcl-1 and the Mcl-1/Bim complex is the most abundant among the three types of complexes. This supports the idea that this complex is critical for the control of B cell death. In conclusion, these results favor a model in which Bim release from anti-apoptotic proteins is a critical event for initiation of apoptosis.     

自然流产中的早孕期蜕膜细胞Bcl-2 /Bax 比例异常

探讨早孕期蜕膜细胞细胞凋亡异常与妊娠失败的关系。应用TUNEL法测定凋亡的发生及凋亡细胞的定位，免疫组化法检测Bcl-2、Bax蛋白的表达和相互关系。结果发现：（1）正常早孕40d蜕膜组织细胞大量凋亡，Bcl-2蛋白表达量较低，Bax蛋白有较强表达。（2）正常早孕50d，凋亡细胞明显减少，Bcl-2的表达显著增强，Bax蛋白表达减弱。（3）早孕50d自然流产组，蜕膜组织大量凋亡，与同时期正常蜕膜组织相比，P＜0．01。Bcl-2蛋白表达明显降低，Bax蛋白表达明显增强，Bcl-2/Bax比例降低。早孕期蜕膜组织凋亡异常可能是自然流产的机制之一，Bcl-2/Bax途径可能是诱导早孕期蜕膜细胞凋亡的重要因素。     

凋亡调节蛋白Bcl-2、p53和Caspase-3在乳腺癌组织中的表达及其相互关系

目的通过观察人乳腺癌组织中凋亡调节蛋白p53、bcl-2和caspase-3表达,探讨bcl-2、p53和caspase-3在乳腺癌发生、发展过程中的作用及其相互关系,为乳腺癌的生物治疗提供实验依据。方法收集手术切除的人乳腺癌组织和癌旁相对正常乳腺组织,用免疫组织化学方法和图像分析技术对21例乳腺癌标本进行检测。结果乳腺癌组织中bcl-2蛋白阳性表达率(52.4%,11/21)明显高于相对正常乳腺组织(19.0%,4/21);乳腺癌组织中p53蛋白阳性表达率(57.1%,12/21)也明显高于相对正常乳腺组织(0%);而乳腺癌组织中caspase-3蛋白阳性表达率(38.0%,8/21)明显低于相对正常乳腺组织(76.1%,16/21)。结论p53、bcl-2和caspase-3蛋白在乳腺癌发生、发展过程中发挥着既独立又协同的作用。     

PLCgamma2 regulates Bcl-2 levels and is required for survival rather than differentiation of marginal zone and follicular B cells

B cells from phospholipase C (PLC)gamma2-deficient mice express reduced levels of the pro-survival protein Bcl-2 and show a defect in the development of transitional T3 and marginal zone (MZ) B cells that reflects reduced B cell survival. Introduction of a bcl-2 transgene restored the numbers of MZ, T3 and follicular B cells in PLCgamma2(-/-) mice. Restricting the B cell repertoire in PLCgamma2-deficient mice by the introduction of a BCR transgene resulted in a striking reduction in the number of IgM-positive B cells and a paucity of IgD-expressing cells in the spleen which was also rescued by the bcl-2 transgene. BCR-stimulated ERK and IkappaBalpha phosphorylation were PLCgamma2 dependent, while calcium flux was reduced, but not abrogated, in the absence of PLCgamma2, suggesting an ancillary role for PLCgamma1. The bcl-2 transgene rescued development of PLCgamma2(-/-) B cells and serum IgM levels but did not restore BCR-mediated signaling, proliferation or serum IgG3 levels. These data suggest that PLCgamma2 performs a critical role in B cell development through regulation of survival rather than differentiation.     

Pregnancy: The effect of dose of mifepristone and gestation on the efficacy of medical abortion with mifepristone and misoprostol

Although it has been demonstrated that a combination of mifepristoneand a prostaglandin is an effective method of inducing abortionin early pregnancy, the optimum dose of the antigestogen isunknown. Women (n = 220) requesting abortion in early pregnancy(63 days amenorrhoea) were randomized to receive a single doseof either 600 or 200 mg mifepristone followed 48 h later bya single dose of 600 µg misoprostol by mouth. The percentageof women who had a complete abortion (93.6% confidence interval90.4–95.5%) was identical in the two groups. There wasno significant difference in the number of women who passedthe fetus within 4 h of receiving the prostaglandin (64 versus74%), the days of bleeding (14.6 ± 1.1 versus 15.3 ±0.9) nor in the onset of the next period (39.7 ± 1.3versus 36.7 ± 1.3) respectively between the groups receiving200 or 600 mg mifepristone. However, the complete abortion ratewas significantly higher in women 49 days compared to women50–63 days amenorrhoea (97.5 versus 89.1% respectively;P < 0.02). There was no difference in any of the other parametersat different weeks of gestation. We conclude: (i) that the recommendeddose of mifepristone could be reduced from 600 to 200 mg withoutloss of clinical efficacy, (ii) that the combination of mifepristoneand 600 µg misoprostol is a highly effective alternativeto vacuum aspiration for inducing abortion in women < 50days amenorrhoea and (iii) at gestation >56 days, this combinationmay result in too many incomplete abortions to be clinicallyacceptable.     

西红花酸对H2O2诱发心肌细胞凋亡及相关调控蛋白caspase-3、Bcl-2表达改变的影响

目的：探讨西红花酸对过氧化氢（H₂O₂）诱导的培养心肌细胞凋亡及相关调控蛋白caspase-3、Bcl-2表达改变的作用。 方法： 通过光镜观察细胞形态、碘化丙啶（PI）染色法和流式细胞术相结合检测培养细胞凋亡率、免疫荧光染色法和流式细胞术相结合检测细胞中caspase-3、Bcl-2蛋白。 结果： 在本实验使用浓度范围内，各浓度H₂O₂组细胞形态明显改变、凋亡率明显高于正常对照组，1×10^-4 mol·L^-1 H₂O₂可使培养心肌细胞Bcl-2蛋白表达明显减少，而caspase-3表达明显增多；各剂量西红花酸组细胞形态学改变减少、凋亡率明显低于1×10^-4 mol·L^-1 H₂O₂组，细胞中Bcl-2蛋白减少幅度与caspase-3增加幅度均减小，且较高浓度（5×10^-5 mol·L^-1）西红花酸组比较低浓度（5×10^-7 mol·L^-1）西红花酸组作用也更明显（P<0.05）。 结论： 西红花酸能够减轻H₂O₂对培养心肌细胞的损伤性凋亡作用，可能与稳定细胞内凋亡相关调控蛋白caspase-3、Bcl-2的功能有关。