细胞凋亡与细胞增殖对重度妊娠高血压综合征的影响

细胞凋亡是自主性的细胞死亡过程，影响着正常妊娠孕卵的着床及胚胎的发育，病理状态下异常的细胞凋亡与多种疾病及妊娠并发症有关^[1]。本研究从细胞存活与凋亡的角度阐述导致重度妊娠高血压综合征(重度妊高征)发生发展的病理生理机制。     

胎盘细胞凋亡与病理妊娠

细胞凋亡是由基因编码决定的细胞主动死亡,参与机体生理过程及疾病发生过程。妊娠作为一个复杂的生理过程,胎盘的形成与发育至关重要,细胞凋亡参与了它的发生发展。现就正常妊娠与病理妊娠时胎盘中细胞的凋亡作一综述。     

细胞凋亡与妊高征的研究

细胞凋亡是一种在基因调控下的细胞主动死亡过程,以形态和生化上出现一定的改变为特征,与机体的生长发育、细胞分化和病理状态有着十分密切的关系,是细胞死亡的一种形式.生理状态下参与正常妊娠孕卵的着床及胚胎的发育,而在病理状态下异常的细胞凋亡又与自然流产、胎膜早破、输卵管妊娠、胎儿宫内发育迟缓等的发病有关.并在妊高征的发病过程中参予了胎盘、脐带血管内皮细胞的脱落、坏死,以及血管的粥样硬化、纤维样坏死和退形性变.     

妊高征胎盘细胞凋亡

细胞凋亡存在于正常妊娠胎盘,但胎儿宫内发育迟缓(IUGR)胎盘细胞凋亡增多。细胞凋亡的分子机制涉及某些细胞外配体及其受体如Fas和Fas配体、bcl-2基因家族等。bcl-2基因家族包括凋亡促进因子(Bax)和凋亡抑制因子(bcl-2和bcl-x)。bcl-2定位于人绒毛合体滋养叶细胞,随妊娠进展表达减弱,提示与正常胎盘老化有关。Fas配体在人滋养叶细胞表面持续表达,与Fas受体结合诱导细胞凋亡。     

妊娠高血压综合征胎盘细胞凋亡现象及凋亡相关基因表达的研究

妊娠高血压综合征 (简称妊高征 )的病因和发病机理至今未完全阐明。国外学者认为细胞凋亡在胎盘发生、发育、老化过程中起重要作用 ,并介导许多病理妊娠的发生 [1 ]。本实验旨在研究妊高征患者胎盘细胞凋亡发生率以及凋亡相关基因在胎盘部位的表达 ,从而探讨细胞凋亡在妊高征发     

滋养细胞凋亡与妊娠

滋养细胞凋亡可见于正常妊娠的各个时期和各种病理妊娠的胎盘组织.正常妊娠时滋养细胞凋亡发生率随孕龄增加,可能与胎盘老化和分娩发动过程中的生物学变化关联.病理妊娠中因炎症细胞因子释放、血液灌注不足、营养缺乏、激素水平低下等使滋养细胞凋亡大量发生,反映了疾病程度并可改变妊娠结局.滋养细胞可能通过自身凋亡和诱导母体免疫活性细胞凋亡而维护妊娠.     

细胞凋亡的基因表达和调节与妊娠相关性研究

细胞凋亡是细胞的主动死亡过程,方式与细胞坏死截然不同.细胞凋亡受自身基因控制,促细胞凋亡基因表达增多,细胞凋亡;抑制凋亡基因增多则细胞增殖.随研究深入,细胞凋亡基因表达及调控与产科的关系也逐渐被证明,正常妊娠随着孕周增加细胞凋亡比率增加,而各种病理妊娠细胞凋亡比率与正常对照组存在差异,其基因表达水平也相应改变,体现了基因调节与形态变化的一致.深入对其研究对病理妊娠的机制和临床治疗有重要意义.     

Bax/Bcl-2基因与细胞凋亡在母-胎界面的表达及与病理妊娠的关系

细胞凋亡是由基因编码决定的细胞主动死亡,其启动和进程受一定程序的控制,故也称程序性死亡。细胞凋亡在胎盘的发生、发展过程中起着举足轻重的作用。Bcl-2基因蛋白家族在细胞凋亡通路中起着重要的调节作用,Bax/Bcl-2基因表达异常可能参与流产、妊高症(PIH)、胎儿宫内发育迟缓(FGR)等病理妊娠的发生、发展。     

滋养细胞凋亡与妊娠

滋养细胞凋亡可见于正常妊娠的各个时期和各种病理妊娠的胎盘。正常 妊娠时滋养细胞凋亡发生率随孕龄增加，可能与胎盘老化和分娩发动过程中的生物学变化关联。病理妊娠中因炎症细胞因子释放、血液糖注不足、营养缺乏、激素水平低下等使滋养细胞凋亡大量发生，反映了疾病程度并可改变妊娠结局。滋养细胞可能通过自身凋亡和诱导母体免疫活性细胞凋亡而维护妊娠。     

细胞凋亡的基因表达和调节与妊娠相关性研究

细胞凋亡是细胞的主动死亡过程，方式与细胞坏死截然不同。细胞凋亡受自身基因控制，促细胞凋亡基因表达增多，细胞凋亡；抑制凋亡基因增多则细胞增殖。随研究深入，细胞凋亡基因表达及调控与产科的关系也逐渐被证明，正常妊娠随着孕周增加细胞凋亡比率增加，而各种病理妊娠细胞凋亡比率与正常对照组存在差异，其基因表达水平也相应改变，体现了基因调节与形态变化的一致。深入对其研究对病理妊娠的机制和临床治疗有重要意义。     

颗粒细胞凋亡率联合卵泡液孕激素预测IVF-ET结局

目的:探讨颗粒细胞凋亡率联合卵泡液中孕激素水平检测对体外受精-胚胎移植(IVF-ET)结局的预测价值。方法:选取2011年3月～2012年6月在我院行IVF-ET患者34例,分离收集颗粒细胞及卵泡液,33258荧光染料着色,记录颗粒细胞凋亡率,放射免疫法测定外周血及卵泡液中雌孕激素水平。比较妊娠组与非妊娠组的年龄、基础FSH、Gn用量及持续时间、卵泡数、取卵数、颗粒细胞凋亡率、雌孕激素水平、受精率。对颗粒细胞凋亡率及卵泡液雌孕激素水平进行相关性分析。以丘颗粒细胞凋亡率≤4‰,卵泡液孕激素浓度≤5.0ng/ml为阳性标准,进行联合检测。结果:妊娠组与非妊娠组的年龄、基础FSH、Gn用量及持续时间、卵泡数、取卵数、受精率及雌激素水平均无显著差异(P>0.05),颗粒细胞凋亡率、外周血及卵泡液中孕激素水平差异显著(P<0.05)。相关性分析发现,颗粒细胞凋亡率随孕激素含量升高而升高,但无显著差异(P>0.05)。联合检测阳性组妊娠率为75.8%。结论:颗粒细胞凋亡率、外周血及卵泡液孕激素水平与IVF-ET结局有关。联合检测能较好提高对妊娠结局的预测。     

在妊娠合并免疫性血小板减少症动物模型中骨髓间充质干细胞的改变#br#

Objective?To establish a method for isolation and culture of bone marrow mesenchymal stem cells (BM-MSCs) in mice with immune thrombocytopenia (ITP) during pregnancy. Methods?Mesenchymal stem cells were isolated and cultured from mouse bone marrow of normal pregnancy, ITP and ITP pregnancy group. Cell growth and proliferation were observed under optical microscope. The immunophenotype of mesenchymal stem cells was analyzed by flow cytometry. Oil red 0 and alizarin red staining were used to evaluate the ability of lipid osteogenesis differentiation. CCk-8 test was used to detect cell proliferation. The PI test measures the cell cycle.Annexin-v /PI staining was used to detect apoptosis. Results?①BM-MSCs derived from pregnant mice with ITP group grew slowly and were irregular in shape. BM-MSCs from different sub-groups were highly expressed in stem-cell related antigens CD44, CD29 and scal-1, partially expressing CD90, but not expressing leukocyte surface antigen CD45 and lymphocyte surface antigen CDllb. They can be induced to differentiate into osteoblasts and lipoblasts in vitro. ②Compared with the normal pregnancy group, the proliferation of BM-MSCs in the ITP group and the ITP pregnancy group was significantly inhibited (P<0.05), the cell proportion in G0/G1 phase was significantly increased (P<0.05), and the apoptosis rate was significantly increased (P<0.05). Compared with the ITP group, the growth rate of BM-MSCs in the ITP pregnancy group was inhibited and the apoptosis rate was increased (P<0.05). Conclusions?The BM-MSCs derived from mice in the ITP pregnancy group showed abnormal morphology, impaired proliferation ability and excessive apoptosis.     

先兆子痫患者母胎界面上细胞凋亡的研究

目的:研究先兆子痫患者母胎界面上细胞凋亡水平及凋亡相关分子的表达变化。方法:分别收集妊娠25、27、29、30和31周及足月分娩的正常妊娠者和先兆子痫患者的胎盘组织,采用脱氧核糖核酸末端转移酶介导的dUTP刻痕末端标记法(TUNEL)检测母胎界面上细胞凋亡情况, 并行凋亡细胞定位;RT-PCR检测正常妊娠和先兆子痫胎盘组织中P53,Bcl-2和Bax的mRNA表达水平的差异。结果:正常妊娠和先兆子痫患者胎盘中发生凋亡的细胞数目都随孕周延长而逐渐增高,但先兆子痫的胎盘中细胞凋亡率明显高于正常妊娠者;正常妊娠胎盘中凋亡细胞主要为合体滋养层细胞,足月胎盘绒毛内血管内皮细胞亦有少量细胞凋亡,而先兆子痫胎盘中细胞滋养层细胞、合体滋养层细胞和绒毛内血管内皮细胞均发生明显的凋亡;RT-PCR显示先兆子痫胎盘组织中P53和Bax的mRNA水平明显高于同期妊娠的正常胎盘,而Bcl-2水平显著降低。结论:正常妊娠过程中胎盘组织中只在有限部位出现少量细胞凋亡,而先兆子痫胎盘组织中细胞过度凋亡, 且凋亡细胞分布广泛,并伴有凋亡相关分子的表达上调。表明先兆子痫的发生与胎盘组织中滋养层细胞大量凋亡密切相关。     

miR-520 promotes DNA-damage-induced trophoblast cell apoptosis by targeting PARP1 in recurrent spontaneous abortion (RSA)

The establishment and maintenance of successful pregnancy mainly depends on trophoblast cells. Their dysfunction has been implicated in recurrent spontaneous abortion (RSA), a major complication of pregnancy. However, the underlying mechanisms of trophoblasts dysfunction remain unclear. DNA-damage-induced cell apoptosis has been reported to play a vital role in cell death. In this study, we identified a novel microRNA (miR-520) in RSA progression via regulating trophoblast cell apoptosis. Microarray analysis showed that miR-520 was highly expressed in villus of RSA patients. By using flow cytometry analysis, we observed miR-520 expression was correlated with human trophoblast cell apoptosis in vitro, along with decreased poly (ADP-ribose) polymerase-1 (PARP1) expression. With the analysis of clinic samples, we observed that miR-520 level was negatively correlated with PARP1 level in RSA villus. In addition, overexpression of PARP1 restored the miR-520-induced trophoblast cell apoptosis in vitro. The status of chromosome in trophoblast implied that miR-520-promoted DNA-damage-induced cell apoptosis to regulate RSA progression. These results indicated that the level of miR-520 might associate with RSA by prompting trophoblast cell apoptosis via PARP1 dependent DNA-damage pathway.     

米非司酮对人早孕蜕膜组织细胞凋亡的影响

细胞凋亡（ａｐｏｐｔｏｓｉｓ）同胚泡植入及胎盘发育密切相关,为了进一步探讨米非司酮终止早孕的机制,本文随机选择要求人工流产的妇女２５名,服药组１３名,对照组１２名。应用流式细胞仪对经碘化丙锭（ＰＩ）染色后的蜕膜组织细胞ＤＮＡ进行分析。结果两组细胞ＤＮＡ直方图上均出现细胞凋亡峰,其细胞所占比例（ｘ±ｓ）％服药组为６．８２±３．０９,对照组为３．４３±２．１３,服药组明显高于对照组（Ｐ＜０．０１）。二倍体峰细胞比例服药组为７６．００±７．０９,对照组为８４．４５±３．８４,服药组低于对照组（Ｐ＜０．０５）。四倍体峰细胞比例两组分别为３．７０±１．６４、３．２９±０．６７,无明显差别（Ｐ＞０．０５）。结果表明人早孕蜕膜组织中存在着细胞凋亡,米非司酮终止早孕的作用可能是通过拮抗孕激素作用促进蜕膜细胞凋亡而实现的。     

妊娠与细胞凋亡

凋亡是近年研究较多的一种细胞死亡方式 ,受 p53、bcl-2、ICE、Fas等基因的调控 ,并受一些细胞因子的影响。本文就发生在卵泡、黄体及胎盘中的细胞凋亡及可能的调控机理与妊娠的关系作一阐述     

Apoptosis in maternal peripheral blood during pregnancy

OBJECTIVE: To investigate the mononuclear cell apoptosis rate during pregnancy. MATERIALS AND METHODS: Apoptosis was quantitated by EtBr staining in whole peripheral blood samples of 135 women in different gestational weeks and 85 nonpregnant women used as controls. Apoptosis was also qualitated by TUNEL assay. RESULTS: The apoptosis rate increased during pregnancy according to gestational age. In chromosomally abnormal fetuses apoptosis was 2.5-fold higher than that found in pregnancies with normal embryos matched for gestational age. FISH in TUNEL-positive cells using X, Y and 21 chromosome probes verified the fetal origin of part of the apoptotic population. CONCLUSION: Apoptosis is stimulated in maternal peripheral blood during pregnancy, possibly accounting partly for the presence of free fetal DNA in maternal serum. The increased apoptosis rate in pregnancies with chromosomally abnormal fetuses may have additional clinical importance.     

Patterns of uterine cellular proliferation and apoptosis in the implantation site of the rat during pregnancy

During gestation, the balance between cell proliferation and death is crucial for successful embryo implantation and maintenance of pregnancy. The uterine endometrium responds to blastocyst implantation with extensive proliferation and differentiation of stromal cells into decidual cells, forming the antimesometrial and mesometrial decidua, which regress by apoptosis. In the latter region it is also observed the growth of metrial gland. To elucidate the events underlying this tissue remodelling we investigated the spatial and temporal pattern of expression of the proliferating cell nuclear antigen (PCNA) and localized the apoptotic cells, by the TUNEL assay and by the expression of active caspase-3. We found that PCNA is expressed at high levels during decidualization until day 12 of gestation declining thereafter abruptly. On the contrary, the appearance of apoptotic cells was detected, by the TUNEL and active caspase-3 expression, in the mesometrial decidua on day 12, increasing from days 14 to 16 in the decidua and metrial gland. In the antimesometrial decidua apoptosis was observed from early to day 12 of pregnancy. However, on day 13 only cell debris and neutrophils were observed, indicating also the presence of necrosis. These results suggest that decidual cells undergo, in distinct regions and at different stages of pregnancy, cell death by apoptosis and secondary necrosis.     

Apoptosis of extravillous trophoblast cells limits the trophoblast invasion in uterine but not in tubal pregnancy during first trimester

During the first trimester of pregnancy extravillous trophoblast cells (EVT) invade the maternal decidua. Invasion normally is reduced from the second trimester onwards and stops in the inner third of the myometrium. By contrast, in extrauterine tubal pregnancy, trophoblast invasion may even penetrate the tubal wall, which ultimately leads to the rupture of the fallopian tube. Induction of apoptosis of EVT cells, by maternal immune competent cells, may be an important mechanism to limit EVT invasion in uterine pregnancy. Tissue specimens from first and second trimester uterine pregnancy and first trimester tubal pregnancy were analyzed for apoptosis by TUNEL- and M30-staining. By immunohistochemical double labelling, maternal leukocyte subtypes were co-localized to apoptotic cells and in this context, the number of CD56(+)NK cells was analyzed. Our data show that apoptosis is confined to the decidua basalis. Most apoptotic cells are single cytokeratin-positive epithelial cells residing in the stromal compartment. Consequently these cells can only be EVT cells. Maternal leukocytes are not apoptotic. They are located in close contact to apoptotic cells. The number of apoptotic cells in the second trimester (1.8+/-0.7 per cent) is reduced compared to first trimester (5.6+/-0.7 per cent) of uterine pregnancy. In parallel, the number of NK cells declines from first (24.4+/-2.9) to second (12.4+/-1.8) trimester. Furthermore, apoptosis is significantly reduced in ectopic (0.9+/-0.3 per cent) compared to eutopic first trimester pregnancies. Consequently, we suggest that in first trimester uterine pregnancy, induction of EVT cell apoptosis by the maternal immune system is one mechanism to limit EVT invasion. During the second trimester, in parallel to declining numbers of NK cells, the mechanism changes. However, in tubal pregnancy due to differing immunological microenvironments at the ectopic implantation site, apoptosis induction fails, which deleteriously may result in uncontrolled invasion and penetration of the tubal wall.     

卵泡液中TNF-α和IFN-γ与卵丘颗粒细胞凋亡关系的探讨

目的:探讨卵泡液中肿瘤坏死因子α(TNF-α)和γ干扰素(IFN-γ)与颗粒细胞凋亡的关系。方法:应用TUNEL方法测定卵丘颗粒细胞凋亡率,应用酶联免疫方法(ELISA)测定成熟卵泡液中TNF-α和IFN-γ的浓度,探讨其相关性。结果:妊娠组颗粒细胞凋亡率为12.25±4.84%,明显低于未妊娠组的15.96±7.04%(P<0.05);TNF-α浓度与颗粒细胞凋亡率的相关系数r=0.56,P<0.05,两者存在显著的正相关关系;IFN-γ浓度与颗粒细胞凋亡率的相关系数r=-0.57,P>0.05,两者不存在相关关系。结论:卵泡液中TNF-α浓度与颗粒细胞凋亡呈正相关,IFN-γ浓度与颗粒细胞凋亡无相关关系,颗粒细胞凋亡率高的患者不易获得妊娠。