子宫平滑肌RhoA/Rho激酶表达与宫缩乏力性产后出血的关系

目的：探讨产妇子宫平滑肌组织RhoA/Rho激酶表达与宫缩乏力性产后出血的关系。方法：选择30例宫缩乏力性产后出血产妇为病例组,同期无产后出血产妇30例为对照组。采用肌条等长张力测定法检测子宫平滑肌自发收缩活动力、加入Rho激酶抑制剂Y-27632后子宫平滑肌收缩活动力;采用实时荧光定量反转录聚合酶链反应(RT-PCR)法及蛋白质印迹法检测子宫平滑肌组织RhoA,ROCKⅠ和ROCKⅡ mRNA及蛋白表达水平。结果：①病例组子宫平滑肌自发收缩活动力水平低于对照组,差异有统计学意义（P＜0.01）;病例组和对照组加入Y-27632后收缩活动力水平均低于加入前,差异有统计学意义（P＜0.01）。②病例组子宫平滑肌组织RhoA,ROCKⅠ和ROCKⅡmRNA表达水平均低于对照组,差异均有统计学意义（P＜0.05或P＜0.01）。③病例组子宫平滑肌组织RhoA,ROCKⅠ和ROCKⅡ蛋白表达水平均低于对照组,差异有统计学意义（P＜0.05或P＜0.01）。④2组子宫平滑肌组织RhoA mRNA及蛋白与ROCKⅠ和ROCKⅡ mRNA及蛋白表达水平均呈正相关。⑤2组子宫平滑肌组织RhoA,ROCKⅠ和ROCKⅡ mRNA及蛋白水平均与子宫平滑肌基础收缩活动力水平呈正相关。结论：宫缩乏力性产后出血产妇子宫平滑肌组织RhoA,ROCKⅠ和ROCKⅡ水平均降低,且均与子宫平滑肌收缩活动力呈正相关,提示子宫平滑肌组织RhoA/Rho激酶信号通路表达降低可能是发生宫缩乏力性产后出血的分子机制之一。     

子宫平滑肌大电导钙激活钾通道α、β 亚基表达与产后出血的关系

目的 探讨产妇子宫平滑肌大电导钙激活钾通道（BKCa）α、β亚基表达变化在宫缩乏力性产后出血发病中的作用。方法 2009年10月至2011年2月福建省妇幼保健院产科,采用肌条等长张力实验测定30例宫缩乏力性产后出血产妇（病例组）和30例宫缩正常无产后出血产妇（对照组）子宫平滑肌收缩频率、幅度及收缩活动力;应用蛋白印迹法（Western blot）测定两组产妇子宫平滑肌BKCa通道α、β亚基蛋白表达水平;应用实时荧光定量聚合酶链反应（real-time RT-PCR）测定两组产妇子宫平滑肌BKCa通道α、β亚基mRNA表达水平。结果 （1）病例组子宫平滑肌自发宫缩收缩频率、幅度、收缩活动力均低于对照组,分别比较差异均有统计学意义（P<0.05）。（2）病例组子宫平滑肌BKCa通道α、β亚基蛋白表达水平及其mRNA表达水平均高于对照组,分别比较,差异均有统计学意义（P<0.05）。（3）两组产妇子宫平滑肌收缩活动力与BKCa通道α、β亚基蛋白表达水平及BKCa通道α、β亚基mRNA表达水平均呈明显负相关（病例组：r分别为-0.401、－0.727及－0.723、－0.895;对照组：r分别为－0.541、－0.378及－0.703、－0.625;[WTBX]P[WTBZ]均＜0.05）。结论 宫缩乏力性产后出血产妇子宫平滑肌BKCa通道α、β亚基表达水平升高可能是发病的重要因素之一。     

缝隙连接蛋白43在子宫平滑肌细胞的表达调控

缝隙连接是细胞间的直接通讯方式。缝隙连接蛋白43(CX43)是构成细胞缝隙连接的主要蛋白,在子宫平滑肌呈特异性表达,可调控子宫平滑肌细胞内信号通路,影响子宫收缩的协调性和节律性。就缝隙连接与CX43的关系、CX43在子宫平滑肌细胞内的表达及通过对其磷酸化位点的磷酸化作用调控子宫平滑肌细胞内信号通路等综述。探讨CX43如何调控子宫收缩协调性、同步性和极性,为早产和宫缩乏力性产后出血的预防和治疗提供理论支持。     

缝隙连接蛋白43在子宫平滑肌细胞的表达调控

缝隙连接是细胞间的直接通讯方式。缝隙连接蛋白43（CX43）是构成细胞缝隙连接的主要蛋白,在子宫平滑肌呈特异性表达,可调控子宫平滑肌细胞内信号通路,影响子宫收缩的协调性和节律性。就缝隙连接与CX43的关系、CX43在子宫平滑肌细胞内的表达及通过对其磷酸化位点的磷酸化作用调控子宫平滑肌细胞内信号通路等综述。探讨CX43如何调控子宫收缩协调性、同步性和极性,为早产和宫缩乏力性产后出血的预防和治疗提供理论支持。     

子宫平滑肌组织Hsp27表达与宫缩乏力性产后出血的关系

目的:探讨宫缩乏力性产后出血产妇子宫平滑肌组织中热休克蛋白27(Hsp27)的表达及意义。方法:收集30例宫缩乏力性产后出血产妇为病例组,同期无产后出血产妇30例为对照组。采用肌条等长张力测定法检测两组子宫平滑肌基础收缩活动力;采用实时荧光定量RT-PCR法检测两组子宫平滑肌组织Hsp27mRNA表达水平;采用蛋白印迹法检测两组子宫平滑肌组织Hsp27、Ser15位点磷酸化的Hsp27(p-Hsp27)蛋白表达水平;采用酶联免疫吸附试验(ELISA)检测两组血清中Hsp27水平。结果:①病例组子宫平滑肌基础收缩活动力水平低于对照组,差异有高度统计学意义(P<0.01);②病例组子宫平滑肌组织Hsp27mRNA表达水平低于对照组,差异有高度统计学意义(P<0.01);③病例组子宫平滑肌组织Hsp27、p-Hsp27蛋白水平低于对照组,差异有统计学意义及高度统计学意义(P<0.05,P<0.01);④病例组和对照组血清中Hsp27表达水平比较,差异无统计学意义(P>0.05);⑤两组子宫平滑肌组织Hsp27mRNA、Hsp27、p-Hsp27蛋白表达水平均与子宫平滑肌基础收缩活动力水平呈正相关(P<0.05)。结论:子宫平滑肌组织Hsp27、p-Hsp27水平降低可能是宫缩乏力性产后出血发病的分子机制之一。     

蛋白激酶Cα、β及Rho激酶对宫缩乏力性产后出血产妇子宫平滑肌收缩功能影响研究

目的探讨蛋白激酶C(PKC)α和PKCβ特异性抑制剂Go6976、PKCβ特异性抑制剂LY333531和Rho激酶特异性抑制剂Y-27632对宫缩乏力性产后出血产妇子宫平滑肌收缩的抑制作用。方法 2010年11月至2011年12月于福建省妇幼保健院选择剖宫产分娩的初产妇60例,分宫缩乏力性产后出血组(病例组,n=30)和宫缩良好无产后出血组(对照组,n=30)。采用肌条等长张力测定方法检测两组产妇子宫下段平滑肌分别在Go6976、LY333531和Y-27632作用下自发性收缩功能变化及病例组在LY333531、Y-27632作用下梯度浓度缩宫素诱导的收缩潜能变化。结果 Go6976、LY333531和Y-27632均显著抑制两组产妇子宫平滑肌收缩幅度及活动力(均P0.05);对病例组抑制作用均小于对照组(均P0.05),而对两组的收缩频率无影响(P0.05)。LY333531和Y-27632均可显著降低病例组收缩潜能(P0.05)。结论 PKCα、PKCβ和Rho激酶影响子宫平滑肌收缩,可能参与宫缩乏力性产后出血的发生。     

分娩机制中细胞间隙连接蛋白CX43瞬时表达定位、定量分析及其意义(激光扫描共聚焦显微镜研究)

目的：探讨细胞间隙连接蛋白CX43在临产前后子宫平滑肌组织中的瞬时表达及其在分娩机制中的重要作用。方法：应用间接免疫荧光染色,结合激光扫描共聚焦显微镜技术,对19例足月妊娠未临产孕妇和23例临产孕妇,剖宫产子宫下段平滑脱组织中CX43蛋白表达的进行定位和定量分析。结果;未临产子宫平滑肌细胞胞质中CX43蛋白染色较弱,临产子宫平滑肌细胞CX43蛋白表达呈现强阳性信号。定量分析CX43在临产与未临产子宫中滑肌细胞中平均荧光强度分别为100像素线以上、100像素线以下;CX43蛋白表达阳性率分别为82．6％和36．8％,二者表达阳性率比较差并存在显著性（P＜0．05）。细胞间隙连接蛋白CX43在临产子宫平滑肌细胞中的时时高表达可能是宫缩过程中重要的事件,时阐明复杂的分娩机制提供了新的理论依据。     

子宫平滑肌大电导钙激活钾通道与小窝蛋白相互作用和宫缩乏力性产后出血的关系研究

目的:探讨产妇子宫平滑肌大电导钙激活钾通道(BKCa)与小窝蛋白相互作用在宫缩乏力性产后出血发病中的作用。方法:采用免疫荧光双染法及激光共聚焦显微镜观察30例宫缩乏力性产后出血产妇(病例组)和30例宫缩正常无产后出血产妇(对照组)子宫平滑肌BKCa(α亚基)与小窝蛋白-1、BKCa(α亚基)与小窝蛋白-2的共定位情况;采用免疫共沉淀法检测两组产妇子宫平滑肌BKCa(α亚基)与小窝蛋白-1、BKCa(α亚基)与小窝蛋白-2的相互作用关系。结果:(1)两组子宫平滑肌细胞均有BKCa(α亚基)、小窝蛋白-1和小窝蛋白-2表达,其中BKCa(α亚基)主要位于细胞膜表面,而小窝蛋白-1和小窝蛋白-2在细胞膜表面和细胞质均有表达;且BKCa(α亚基)与小窝蛋白-1、BKCa(α亚基)与小窝蛋白-2均有部分区域共表达于细胞膜。(2)病例组子宫平滑肌BK-Ca(α亚基)免疫沉淀物中小窝蛋白-1、小窝蛋白-2蛋白水平均高于对照组,差异均有统计学意义(P<0.05)。结论:宫缩乏力性产后出血产妇子宫平滑肌细胞中BKCa(α亚基)免疫沉淀物中小窝蛋白-1、小窝蛋白-2蛋白水平升高,提示小窝蛋白与BKCa(α亚基)相互作用增强可能是宫缩乏力性产后出血发病的重要原因之一。     

米索前列醇治疗顽固性产后出血13例报告

目的探讨米索前列醇对宫缩乏力性产后出血的治疗效果.方法对13例宫缩乏力性产后出血产妇,在采用缩宫素、麦角新碱治疗无效后,给予口服米索前列醇800 μg.结果13例产妇口服米索前列醇后,在3～12 min内子宫收缩加强,出血得到控制,无一例子宫切除.结论应用米索前列醇治疗顽固性产后出血,效果显著,可推广使用.     

产后出血原因及高危因素分析

产后出血是产科常见的严重并发症之一 ,为产妇死亡的首要原因。引起产后出血的原因主要有子宫收缩乏力、胎盘因素、软产道损伤和凝血功能障碍。其中以子宫收缩乏力性出血占首位 ,占产后出血总数的 70 %～ 80 %。产后出血的病因及高危因素如下。1　子宫收缩乏力任何影响产后子宫平滑肌收缩功能的因素均可能导致子宫收缩乏力 ,引起产后出血 ,诱发因素有 :1.1　全身因素1.1.1　产前精神过度紧张、恐惧分娩 ,或产时精神受到刺激 ,交感神经功能亢进 ,抑制子宫收缩 ;临产后过度疲劳 ,甚至衰竭 ,或进食进水不足。1.1.2　产程长、滞产、难产、急产。…     

血清PLT、Hb及D-D水平预测产后出血的临床价值

目的 分析血清血小板计数(PLT)、血红蛋白(Hb)及D-二聚体(D-D)水平在预测产后出血(PPH)中的临床价值.方法 100例临产孕妇依据产后是否出血(出血量≥500 mL)分为PPH组(n=25)、对照组(n=75),比较两组产前24h内血清PLT、Hb及D-D水平及其与出血程度关系,分析PLT、Hb及D-D对P...     

The mitogen-activated protein kinases (MAPK) p38 and JNK are markers of tumor progression in breast carcinoma

OBJECTIVE: To investigate the activation of mitogen-activated protein kinases (MAPK) in breast carcinoma effusions and to analyze its relationship to anatomic site and clinical parameters. METHODS: Activated MAPK (p-ERK, p-JNK, and p-p38) expression was studied in 42 effusions and 51 corresponding solid tumors (23 primary, 28 metastases) using immunohistochemistry (IHC). Hormone receptor and HER2 status, proliferation (Ki-67), and apoptosis (p85-PARP fragment) were assessed. MAPK levels, activity, and activation ratio (phospho/pan-MAPK ratio) were analyzed in 19 effusions using immunoblotting (IB). RESULTS: Nuclear expression of p-p38 and p-JNK was significantly higher in effusions compared with both primary tumors (P < 0.001 for p-JNK, P = 0.011 for p-p38) and lymph node metastases (P = 0.003 for p-JNK, P = 0.025 for p-p38) but was not accompanied by apoptosis. IB showed pan-ERK and p-ERK in 18/19 effusions, pan-JNK and p-JNK in 18/19 and 17/19 effusions, respectively, and pan-p38 and p-p38 in 19/19 and 17/19 specimens, respectively. In univariate survival analysis of all cases, advanced disease stage (P = 0.041), previous chemotherapy (P = 0.004), and radiation (P = 0.001) and higher Ki-67 scores (P = 0.01) correlated with worse overall survival (OS). In Cox multivariate analysis, stage (P = 0.018), chemotherapy (P = 0.024), radiation (P = 0.017), and ER status (P = 0.002) were independent prognosticators of OS. Quantitative analysis of IB data showed that higher p38 activation ratio correlates with shorter OS (P = 0.01). CONCLUSIONS: This study presents first evidence of in vivo activation of MAPK in breast carcinoma effusions. The elevated JNK and p38 activation in effusions may be a stress-related mechanism providing breast carcinoma cells with survival advantages rather than a drive towards apoptosis. p38 and Ki-67 may be new prognostic markers for patients with breast cancer effusions.     

MAPK and AP-1 proteins are increased in term pre-labour fetal membranes overlying the cervix: regulation of enzymes involved in the degradation of fetal membranes

Fetal membranes overlying the cervix (i.e. supracervical site, SCS) are characterised by increased extracellular matrix (ECM) degradation. In non-gestational tissues, the mitogen activated protein kinase (MAPK) and activator protein (AP)-1 family are involved in the regulation of the ECM degrading enzyme metalloproteinase (MMP)-9. The aims of this study were (i) to compare the expression of AP-1 proteins in fetal membranes from the SCS and a distal site (DS), and (ii) determine if the MAPK/AP-1 pathway is involved in the regulation of MMP-9. Fetal membranes overlying the cervix were identified in situ in women undergoing term elective Caesarean section. Immunohistochemistry (n = 6) was used to localise the expression of the MAPK proteins ERK (total and phosphorylated), JNK (total and phosphorylated) and p38 MAPK (total and phosphorylated), and the AP-1 proteins JunB, cJun (total and phosphorylated), JunD, cFos and FosD. There was no difference in JNK, p38 MAPK, FosB, cJun and JunD protein expression between SC and distal fetal membranes. However, when compared to DS, the intensity and/or extent of staining of ERK, p-ERK, p-JNK, p-p38 MAPK, cFos, JunB and p-cJun were greater in amnion and chorion obtained from the SCS. In order to elucidate a role for these proteins in ECM degradation, pharmacological inhibitors of MAPK protein activation were utilised in primary amnion cells. The ERK inhibitor U0126, JNK inhibitor SP600125 and p38 MAPK inhibitor SB202190 all significantly decreased IL-β-induced MMP-9 gene expression and pro MMP-9 in human primary amnion cells. In summary, at term, non laboured SC fetal membranes are characterised by increased expression of MAPK and AP-1 proteins. MMP-9 expression and production was significantly suppressed by inhibitors of three key enzymes in the signalling cascades leading to AP-1 formation, ERK 1/2, JNK and p38 MAPK. Thus, the MAPK/AP-1 pathway may play a role in the degradation of the ECM at the SCS making it more susceptible to membrane rupture.     

Effect of parathyroid hormone-related peptide on human and rat myometrial contractility in vitro

OBJECTIVES: The aims of this study were primarily to investigate the effects of parathyroid hormone-related peptide (human fragment 1-34) on human nonpregnant and pregnant (nonlabor and labor) myometrial contractility in vitro and secondarily to compare these effects with those of parathyroid hormone-related peptide on rat myometrial contractility. STUDY DESIGN: Isometric tension recording was performed under physiologic conditions in isolated myometrial strips obtained at hysterectomy and cesarean delivery and from Sprague-Dawley rats. The effect of cumulative additions of parathyroid hormone-related peptide (1, 10, and 100 nmol/L) on myometrial contractility was measured and the significance of results was assessed by 2-way analysis of variance. RESULTS: Parathyroid hormone-related peptide exerted a statistically significant net relaxant effect on myometrial contractility in human nonpregnant myometrium (34.71%; P<.01), in human pregnant myometrium obtained before (18.27%; P <.05) but not after (10.32%; P>.05) the onset of labor, and in rat tissue (31.60%; P<.01). CONCLUSIONS: Parathyroid hormone-related peptide exerts a relaxant effect on human and rat myometrial tissue. In human myometrium, sensitivity to parathyroid hormone-related peptide is reduced in pregnancy and abolished by the onset of labor.     

干扰CLDN6基因表达通过调控MAPK/ERK信号通路对多囊卵巢综合征大鼠卵巢颗粒细胞凋亡的影响

目的:探讨Claudin-6(CLDN6)在多囊卵巢综合征(PCOS)大鼠中的表达及其对卵巢颗粒细胞(GC)增殖与凋亡的影响和可能作用机制.方法:SD雌性大鼠皮下注射脱氢表雄酮(DHEA)诱导建立PCOS模型,采用实时荧光定量PCR(RT-qPCR)和West-ern blot法检测卵巢组织中CLDN6表达.分离培养原...