促性腺激素释放激素激动剂对子宫腺肌病在位内膜细胞凋亡的影响

目的 研究促性腺激素释放激素激动剂(GnRHa)对子宫腺肌病(AM)在位内膜细胞凋亡的影响.方法 取本院妇科住院行全子宫切除术的32例AM患者在位子宫内膜细胞为研究组;同期因卵巢上皮性良性肿瘤行卵巢囊肿剔除术的20例患者正常子宫内膜细胞为埘照组.两组细胞进行体外培养,用GnRHa 10-7、10-5mol/L分别作用24、48、72 h后,用末端转移酶介导的缺口末端标记法(TUNEL)及流式细胞仪检测各组细胞的凋亡率.结果 (1)镜下可见两组部分细胞固缩、漂浮、核浓缩、碎裂,呈典型的凋亡小体;(2)不含GnRHa培养的AM在位内膜细胞凋亡率在各时间点均低于对照组,且两组都随时间的延长凋亡率增加(P<0.01);(3)GnRHa作用后,两组的凋亡率高于作用前,且均随浓度的增加而增加,且研究组符时间点、各浓度AM在位内膜细胞凋亡率均显著高于同时间、同浓度的对照组(P<0.01).结论 在位内膜细胞凋亡率异常可能是AM发病机制之一.GnRHa可能以自分泌或旁分泌机制直接作用于子宫在位内膜细胞,提高了AM在位内膜细胞的凋亡率.     

VEGF基因多态性与子宫内膜异位症和子宫腺肌病遗传易感性研究

目的 探讨血管内皮生长因子(vascular endothelial growth factor,VEGF)基因启动子区-460C/T和-1154G/A单核苷酸多态性与子宫内膜异位症和子宫腺肌病发病风险的关系.方法 采用聚合酶链反应-限制性片段长度多态方法检测344例子宫内膜异位症患者(内异症组)和360名对照妇女(对照组)、174例子宫腺肌病患者(腺肌病组)和199名对照妇女(对照组)的VEGF基因2个多态性位点的基因型频率分布情况.结果 VEGF-460C/T多态的基因型和等位基因频率分布在两病例组与其对照组间差异均无统计学意义(P>0.05).在内异症组和对照组中,VEGF-1154G/A多态的AA、GA、GG 3种基因型频率分别是1.7%、28.8%、69.5%和5.8%、32.8%、61.4%,两组比较差异有统计学意义(P=0.006);G、A等位基因频率分别是83.9%、16.1%和77.8%、22.2%,两组比较差异有统计学意义(P=0.004);与GA+AA基因型相比,携带GG基因型明显增加内异症的发病风险(OR=1.43,95%CI:1.05～1.96).在腺肌病组和对照组中,VEGF-1154G/A多态的AA、GA、GG 3种基因型频率分别是2.9%、23.6%、73.6%和7.0%、34.2%、58.8%.两组比较差异有统计学意义(P=0.007);G、A等位基因频率分别是85.3%、14.7%和75.9%、24.1%,两组比较差异有统计学意义(P=0.001);与GA+AA基因型相比,携带GG基因型明显增加腺肌病的发病风险(OR=1.95,95%CI:1.26～3.03).结论 VEGF基因启动子区-1154G/A多态与子宫内膜异位症和子宫腺肌病的发病风险明显相关,携带GG基因型显著增加子宫内膜异位症和子宫腺肌病的发病风险.     

经阴道三维超声测定子宫体积对GnRH-a结合LNG-IUS治疗子宫腺肌病的评估

子宫腺肌病（AM）发病逐渐年轻化,临床主要通过左炔诺孕酮宫内缓释系统（LNG-IUS）结合促性腺激素释放激素激动剂（GnRH-a）治疗。本研究选择在我院接受治疗的子宫腺肌病患者109例作为研究对象,对照组采用LNG-IUS治疗,观察组采用GnRH-a结合LNG-IUS治疗。分析经阴道三维超声测定子宫体积对GnRH-a结...     

GnRHa主动免疫对幼鼠子宫发育的作用

目的探讨促性腺激素释放激素激动剂(GnRHa)主动免疫对幼鼠子宫发育的作用。方法 60只昆明雌鼠随机均分为四组,分别颈部皮下注射不同剂量阿拉瑞林抗原,各组均连续注射7 d。在0 d、7 d、14 d和21 d测定体重,于21 d处理小鼠,显微镜观察子宫的组织结构变化,并用Motic imagles软件测定分析图像数据。结果阿拉瑞林能明显抑制子宫的发育,且剂量越大作用越明显。EG-Ⅲ的UWT明显缩小(P<0.05);实验组EET均小于对照组(P<0.05)。EG-Ⅰ子宫腔轻度缩小;EG-Ⅱ子宫腔和腺体管腔缩小,子宫管壁明显变薄;内膜皱襞减少,上皮变薄;EG-Ⅲ子宫壁变薄,子宫腺减少,内膜细胞胞核变小,上皮变薄,胞质明显减少。结论阿拉瑞林主动免疫能显著抑制幼鼠的子宫发育,且连续重复免疫对小鼠具有毒性作用,剂量越大,作用越明显。     

GnRHa联合孕三烯酮对子宫内膜异位症患者PRL、sHLA-G及对妊娠结局的影响

目的 分析促性腺激素释放激素激动剂（GnRHa）联合孕三烯酮对子宫内膜异位症患者泌乳素（PRL）、可溶性人类白细胞抗原-G(sHLA-G)及对妊娠结局的影响。方法 选取在我院2018年1月至2020年10月期间治疗子宫内膜异位症的患者110例,随机分为对照组和观察组,每组各55例。对照组治疗使用孕三烯酮胶囊进行治疗,观察组在服用孕三烯酮胶囊的同时进行GnRHa治疗。对比两组患者治疗前后黄体生成素（LH）、泌乳素（PRL）、雌二醇（E2）水平,纤溶酶原激活物抑制因子-1(PAI-1)、可溶性人类白细胞抗原-G(sHLA-G)、巨噬细胞移动抑制因子（MMIF）水平、白细胞介素（IL-1β）、肿瘤坏死因子（TNF-α）,妊娠结局,临床疗效等。结果 治疗后两组LH、PRL、E2水平降低,且观察组LH、PRL、E2水平低于对照组,具有统计学差异（P<0.05）,治疗后两组PAI-1、sHLA-G、MMIF水平降低,且观察组PAI-1、sHLA-G、MMIF水平低于对照组,具有统计学差异（P<0.05）,治疗后两组IL-1β、TNF-α水平降低,且观察组IL-1β、TNF-α水平低于对...     

促性腺激素释放激素激动剂改善子宫内膜异位症患者生育力的作用机制及应用

促性腺激素释放激素激动剂（GnRHa）为治疗子宫内膜异位症（EM）的常用药物之一。EM可多方面对女性生育力产生不利影响,而GnRHa可通过改善EM患者腹腔及卵泡微环境、提高子宫内膜容受性及改善输卵管功能等作用提高EM相关不孕患者生育力,并在联合腹腔镜手术及辅助生殖技术（ART）治疗改善EM不孕患者妊娠结局中具有一定价值。     

血清泌乳素(PRL)在子宫腺肌病发病中的作用

甾体性腺激素的异常分泌是导致子宫腺肌病（ＡＭ）的重要原因 ,其中泌乳素在ＡＭ的发生中有重要作用。本文探讨了子宫腺肌病患者的泌乳素分泌状态和可能的临床意义。对象和方法选择１９９７年７月～１９９９年７月两年间在本院住院手术治疗的子宫腺肌病患者４０例 ,平均年龄为４５±４．７岁 ,随机取同期住院并接受手术治疗的子宫肌瘤患者 ,平均年龄为４４．１±４．３岁 ,作为对照。两组病例均经病理证实且均除外内分泌疾病。应用放射免疫双抗体法测定ＰＲＬ,试剂盒由天津德普（ＤＰＣ）公司提供 ,样本采集于术前２～３天上午１０时安静平…     

促性腺激素释放激素激动剂治疗子宫内膜异位症的进展

促性腺激素释放激素激动剂（gonadotropin releasing hormone agonists,GnRH-a）有效的治疗子宫内膜异位症已得到普遍的认可及应用。随着对GnRH-a的作用机制及临床应用方面的不断认识,GnRH-a治疗子宫内膜异位症有了新的进展,随之产生的反加疗法及时减轻了GnRH-a治疗子宫内膜异位症的不良反应,即血管运动综合症及骨质疏松等。     

MMP9、TIMP1及VEGF在子宫内膜异位症患者在位内膜的表达

研究基质金属蛋白酶 9(matrixmetalloproteinase 9,MMP9)及其抑制剂 (tissueinhibitorofmetalloproteinase 1,TIMP1)和血管内皮生长因子 (vascularendothelialgrowthfactor,VEGF)在子宫内膜异位症 (endometriosis,EM)患者的在位内膜中的表达。采用免疫组化SP法分别检测 4 5例EM(研究组 )在位内膜及 32例子宫肌瘤 (对照组 )的在位内膜中MMP9、TIMP1及VEGF的表达。MMP9、TIMP1、MMP9 TIMP1和VEGF在研究组和对照组的表达分别为 0 336 ,0 2 76 ;0 2 5 3,0 2 6 7和 1 2 93,1 0 34;0 372 ,0 2 88。其中MMP9、MMP9 TIMP1和VEGF在研究组中的表达显著高于对照组内膜 (P <0 0 1)。EM患者的在位内膜中MMP9、VEGF高表达可能是EM发生发展的重要原因。诊刮筛选出高MMP9、VEGF表达的在位内膜可望成为预测和诊断EM的方法之一。     

Altered apoptosis and proliferation in endometrial stromal cells of women with adenomyosis

BACKGROUND: The eutopic endometrium in a woman suffering from adenomyosis is known to be biologically different from that of healthy women. The aim of this study was to examine the apoptosis and proliferation of eutopic endometrium from women with adenomyosis. METHODS: We enrolled 23 women with adenomyosis (study group) and 21 without (control group). Eutopic endometrium was obtained and separated into single endometrial stromal cells (ESCs). ESCs were treated in vitro with hydrogen peroxide (H(2)O(2)) to examine their apoptosis using a fluorescence-activated cell sorter. Cells were also treated with estradiol (E(2)), medroxyprogesterone acetate, interleukin (IL)-6, lipopolysaccharide and interferon-gamma (IFN-gamma) to test their proliferation using a non-radioactive cell proliferation assay. RESULTS: The percentage of annexin V ( + )/7-amino-actinomycin D ( + ) ESCs was much lower in women with adenomyosis after 24 h culture with and without H(2)O(2) treatment when compared with the control group. ESCs of adenomyosis proliferated more rapidly than those of the control group, whether they were cultured alone or were treated with E(2), MPA, IL-6 or IFN-gamma. The immunocytochemical Ki-67 labelling index was much more prominent in adenomyotic ESCs than that of the control group (7.7% versus 1.1%, P < 0.001). CONCLUSIONS: Altered apoptosis and proliferation of eutopic endometrium possibly elucidate some aspects of the pathophysiology of adenomyosis. A high Ki-67 labelling index in immunocytochemistry might be a potential indicator in predicting the occurrence of adenomyosis.     

Effect of GnRH analogues on apoptosis and release of interleukin-1beta and vascular endothelial growth factor in endometrial cell cultures from patients with endometriosis

BACKGROUND: The aim of the present study was to evaluate the effect of GnRH analogues on the in-vitro eutopic endometrial cell apoptosis and release of interleukin-1beta (IL-1beta) and vascular endothelial growth factor (VEGF). METHODS: Biopsy specimens of eutopic endometrium obtained from 16 women with untreated endometriosis and 14 controls were studied. Apoptosis, IL-1beta and VEGF release were evaluated in epithelial endometrial cell cultures after incubation with leuprolide acetate (LA) as GnRH agonist, antide as GnRH antagonist, and a combination of both. The percentage of apoptotic cells was evaluated by the acridine orange-ethidium bromide technique, and IL-1beta and VEGF concentrations were assessed by using commercial enzyme-linked immunosorbent assay (ELISA) kits. RESULTS: We found that LA (100 ng/ml) enhanced apoptosis in endometrial cell cultures from endometriosis patients and controls and this effect was reversed by antide at 10(-7) mol/l. IL-1beta and VEGF release was downregulated by LA in cultures from controls and endometriosis patients. The addition of antide 10(-7) mol/l reversed this inhibition. Endometrial cultures treated with antide at 10(-7) mol/l did not show any significant effects compared with basal conditions. CONCLUSIONS: GnRH agonists appear to have a direct effect in endometrial cells cultures, by enhancing the percentage of apoptotic cells and decreasing the release of pro-mitogenic cytokines such as IL-1beta and VEGF.     

Effect of GnRH-II on the ESC proliferation,apoptosis and VEGF secretion in patients with endometriosis in vitro

Objective: To study the effect of GnRH-II on the cell proliferation, apoptosis and secreting vascular endothelial growth factor (VEGF) of ectopic, eutopic and normal endometrial stromal cells (ESC) from patients with or without endometriosis (EMs) in vitro. Methods: The ectopic, eutopic and normal ESC were isolated, cultured and identified, then added 0 M, 10^-10 M, 10^-8 M, 10^-6 M GnRH-II. The growth and proliferation of three ESC were measured by MTT assay; the cell apoptosis were detected with the method of Hoechst staining and Flow Cytometry test; ELISA was used to measure the VEGF concentration change by three ESC secretion. Results: GnRH-II inhibited the proliferation of ectopic, eutopic ESC from patients with endometriosis and normal ESC from control patients, in a dose- and time-dependent manner (P<0.05); GnRH-II increased the apoptotic rate of three ESC in a dose-dependent manner (P<0.05); The concentration of VEGF in three ESC was significantly decreased after the treatment of GnRH-II, in a dose-dependent manner (P<0.01); And these above effects were the strongest on the ectopic than on the eutopic or normal, there were statistical significance (P<0.05); and three was no significantly difference between the eutopic and normal (P>0.05). Conclusions: GnRH-II significantly inhibited the cell proliferation, induced cell apoptosis and decreased the VEGF secreting of ectopic, eutopic and normal ESC in EMs in vitro, and these effects were the strongest on ectopic ESC, which suggested that GnRH-II may become a new effective treatment for endometriosis.     

外源性拟载脂蛋白E肽对脑血管痉挛后血管内皮生长因子介导的脑动脉内皮细胞凋亡的影响

目的:探讨外源性拟载脂蛋白E(apoE)肽对脑动脉痉挛后内皮细胞凋亡的调节作用及对血管内皮生长因子(VEGF)表达的影响.方法:采用非开颅血管内穿线法制备小鼠蛛网膜下腔出血模型并给予apoE-1410肽,术后行神经功能评分并检测大脑中动脉直径变化,应用原位杂交法、免疫印迹法检测各组右侧大脑动脉VEGF表达,TUNEL法检测大脑中动脉内皮细胞凋亡.结果:出血组小鼠神经功能评分均降低,大脑中动脉直径减小,VEGF mRNA及VEGF蛋白显著升高,TUNEL显色阳性.治疗组小鼠神经功能评分及大脑中动脉管径增加,VEGF表达降低,内皮细胞凋亡数目减少.VEGF mRNA表达与TUNEL呈正相关.结论:蛛网膜下腔出血后痉挛脑动脉内皮细胞凋亡与VEGF高表达有关;拟apoE-1410肽对脑血管痉挛具有一定的治疗作用,其机制之一可能是通过抑制VEGF活化,减少大脑中动脉内皮细胞凋亡.     

Lymphangiogenesis of normal endometrium and endometrial adenocarcinoma

BACKGROUND: Information about lymphatics and lymphangiogenesis in the human endometrium is limited. We investigated the distribution of endometrial lymphatic vessels during the normal menstrual cycle and in association with endometrial adenocarcinoma and investigated the expression of lymphangiogenic growth factors, vascular endothelial growth factor (VEGF)-C, VEGF-D and VEGF receptor-3 (VEGF-R3). METHODS AND RESULTS: Full thickness uterine samples (n = 23 proliferative; n = 23 secretory) and endometrial adenocarcinoma samples (n = 7 grade I; n = 10 grade III) were collected for the study and analysed by immunohistochemistry and western blotting. Lymphatic vessels of the functionalis were significantly reduced compared with basalis (P = 0.001) across the menstrual cycle with lymphatics of the basalis sometimes intimately associated with spiral arterioles. Lymphatic vessels of endometrial adenocarcinomas were located intra-tumoural and peri-tumoural with significant increases in the peri-tumoural lymphatic vessels compared with normal basalis (P = 0.02). Interestingly, high-grade adenocarcinoma vessels containing tumour emboli demonstrated a mixed blood/lymphatic endothelial cell phenotype. VEGF-C and VEGF-D were immunolocalized in glandular epithelium and some stromal cells with the staining intensity of this localization increasing in endometrial adenocarcinoma. Protein analysis identified VEGF-C (58, 41, 31 and 21 kD) and VEGF-D (56, 41, 31 and 21 kD) and VEGF-R3 (148 and 65 kD) peptides in normal endometrium, with significant increases in several of these peptides for VEGF-C and VEGF-D and no changes in protein expression for VEGF-R3 in endometrial adenocarcinoma. CONCLUSION: Endometrial lymphatics are significantly reduced in the functionalis, and increases in endometrial adenocarcinoma peri-tumoural lymphatics are associated with increases in VEGF-C and VEGF-D peptides.     

In vitro apoptosis effects of GnRHII on endometrial stromal cells from patients with endometriosis

Purpose: To study the effect of gonadotropin-releasing hormone II (GnRHII) on the cell apoptosis of ectopic, eutopic and normal endometrial stromal cells cultured in vitro from endometriosis patients, and to provide theoretical basis for exploring new treatments for endometriosis (EMs). Methods: Ectopic, eutopic and normal endometrial stromal cells were isolated, cultured and identified in vitro, then treated with different concentrations of GnRHII (0, 10^-10 M, 10^-8 M and 10^-6 M). Cell apoptosis was detected by Hoechst staining and flow cytometry. Results: GnRHII increased apoptosis in ectopic, eutopic and normal stromal cells in a dosage-dependent manner (P<0.05), and apoptosis of ectopic stroma cells was significantly higher than that of eutopic and normal cells (P<0.05); apoptosis in eutopic and normal cells had no different (P>0.05). Conclusion: GnRHII can significantly induce apoptosis in ectopic, eutopic and normal endometrial stromal cells from patients with endometriosis, especially to the ectopic.     

Effect of VEGF on Mouse Thymocyte Proliferation and Apoptosis <Emphasis Type="Italic">In Vitro</Emphasis>

Vascular endothelial growth factor is not only angiogenic, but also immunoregulatory factor. For evaluation of the possibility of its direct interaction with mouse thymocytes we studied the effect of vascular endothelial growth factor on proliferation and apoptosis of thymocytes and expression of genes for the corresponding receptor on these cells. Vascular endothelial growth factor modulated mitogen-induced proliferation of thymocytes and stimulated spontaneous apoptosis in intact thymus cells. Thymocytes express mRNA of type 2, but not type 1 vascular endothelial growth factor receptors.__________Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 139, No. 5, pp. 533–537, May, 2005     

阻断VEGFR-3表达对肿瘤细胞诱导的淋巴管内皮细胞增殖的影响

目的观察阻断VEGFR-3(F lt 4)表达对肿瘤细胞(前列腺癌细胞株PC3)诱导的淋巴管内皮细胞增殖的影响。方法实验分4组,第1组为对照组。每组有6孔,每孔具有相同细胞数2×105/m l,实验组每孔各加入试剂100 m l/L。第1组(对照组)加入淋巴管内皮细胞完全条件培养液(LEC)1 m l,第2组加入LEC 1 m l+兔血清100μl,第3组加入LEC 1 m l+PC3细胞上清100μl。第4组加入LEC+抗F lt 4抗体100μl。并于24、48、72、96 h观察各组淋巴管内皮细胞生长情况。各时间段计数第1~3组细胞数,第4组于72 h计数后,去除抗体,重新加入LEC+PC3细胞上清继续培养至120 h;除96 h外,其余各时间段均计数细胞数。比较各组细胞增殖情况。结果第1、2组淋巴管内皮细胞在加试剂后各时间段两组细胞数及形态无明显差别,第3组加入PC3细胞上清后,细胞数明显多于第1、2组。第4组加试剂后24、48、72 h细胞计数均少于前24 h。清除抗体后加入PC3细胞上清48 h计数细胞,细胞数仅略见增加。结论VEGF-C高表达的PC3细胞上清能显著刺激淋巴管内皮细胞增殖,阻断F lt4表达,可在一定程度上阻断PC3细胞上清促淋巴管内皮细胞增殖作用。     

光动力疗法对子宫内膜癌Ishikawa细胞系细胞周期和凋亡的影响

目的 探讨光动力疗法(PDT)对子宫内膜癌Ishikawa细胞系细胞周期及凋亡的影响.方法 以四甲基吡啶卟啉(TMPyP)为光敏剂的PDT处理Ishikawa细胞,显微镜观察细胞形态;细胞计数试剂盒CCK-8检测细胞的存活率;流式细胞术检测细胞周期;使用Annexin V-FITC/PI凋亡检测试剂盒及流式细胞术检测细胞的凋亡;蛋白免疫印迹法检测Bcl-2、NF-κB P65及磷酸化NF-κB P65的表达.结果 PDT可以改变Ishikawa细胞的形态,降低细胞存活率(P＜0.05),并与光敏剂TMPyP的浓度和激光能量密度有关;PDT引起细胞S期阻滞(P＜0.05),诱导Ishikawa细胞发生凋亡(P＜0.05),降低Bcl-2蛋白的表达和NF-κB P65的磷酸化水平(P＜0.05).结论 PDT可能通过下调Bcl-2、NF-κB的活性引起Ishikawa细胞S期阻滞和细胞凋亡.