尿皮素对体外培养的人脐静脉内皮细胞一氧化氮及一氧化氮合酶的影响

目的:通过检测尿皮素(UCN)对人脐静脉内皮细胞(HUVEC)一氧化氮(NO)生成及一氧化氮合酶(NOS)表达的影响,探讨UCN调节胎儿-胎盘血管张力的分子机制。方法:在离体培养的HUVEC中加入不同浓度的UCN(0、0.1、1、10nmol/L)、10nmol/L促肾上腺皮质激素释放激素(CRH)及向以上各组UCN/CRH同时加α-helical-CRH进行体外培养,用硝酸还原酶法检测细胞上清液中NO的水平,蛋白印迹法检测内皮型一氧化氮合酶(eNOS)和诱导型一氧化氮合酶(iNOS)蛋白表达水平的变化。结果:在与UCN共同培养4~8h后,HUVEC细胞上清液中NO水平呈时间和剂量依赖性变化,随着培养时间的延长,UCN浓度增加,NO水平逐渐升高;UCN组HUVEC iNOS表达水平比对照组明显升高(P<0.05),且呈浓度依赖性升高。而各组eNOS表达水平与对照组比较无明显变化(P>0.05)。UCN/CRH α-helical-CRH组HUVEC上清液中NO水平和HU-VEC iNOS表达水平与对照组比较无明显变化,差异无统计学意义(P>0.05),与相同浓度UCN/CRH组比较NO水平和iNOS蛋白表达水平明显降低,差异有统计学意义(P<0.05)。结论:UCN可促HUVEC合成及释放NO,NO生成增多与UCN通过促进肾上腺皮质激素释放激素2β受体(CRH-R2β)正相调节iNOS蛋白表达有关,这可能是UCN调节胎儿-胎盘血管张力的分子机制之一。     

一氧化氮及一氧化氮合酶与卵巢肿瘤凋亡的关系

目的探讨卵巢良、恶性肿瘤患者血清、肿瘤组织、腹水中一氧化氮(nitricoxide,NO)及其合酶(nitricoxidesynthase,NOS)含量与肿瘤凋亡的关系。方法用分光光度计测定NO、NOS、iNOS活性,流式细胞仪检测细胞凋亡。结果恶性卵巢肿瘤患者血清、肿瘤组织中NO、NOS、iNOS高于良性肿瘤患者(P<0.05);腹水或腹腔冲洗液中iNOS亦高于良性(P<0.05),但腹水中NO、NOS与良性无差异(P>0.05)。恶性肿瘤细胞凋亡率高于良性(P<0.05)。结论NO、NOS、iNOS与卵巢肿瘤的恶性行为及凋亡有关,检测患者血清、组织、腹水中的NO、NOS、iNOS对鉴别良、恶性肿瘤有一定参考价值,可作为观测卵巢癌疗效及预后的指标。     

Effect of gonadotropins on human endometrial stromal cell proliferation in vitro

It has long been known that endometrial regeneration and proliferation is regulated by sex steroids, cytokines and various growth factors. The mechanisms responsible for such organized growth are still under investigation. Human chorionic gonadotropin/luteinizing hormone (hCG/LH) receptors have been found to be localized in human endometrium by immunocytochemistry. Gonadotropins have been widely used for hyperstimulation during in vitro fertilization (IVF) procedures; however, the direct effect of gonadotropin on the endometrium has not been adequately investigated yet. This study attempted to define the effect of gonadotropins on the proliferation of human endometrial stromal cells in vitro. Human endometrial stromal cells were obtained from hysterectomy specimens and cultured in serum-containing media for up to 72 h. The effects of adding 7.5, 15, 30, 150 mIU/ml of human menopausal gonadotropin (hMG) and follicle stimulating hormone (FSH), and 10, 100, 1,000, 10,000 mIU/ml of hCG on cumulative [³H]-thymidine incorporation in endometrial stromal cells were assessed. This study demonstrated that FSH and hMG induced significant inhibition in [³H]-thymidine uptake at all concentrations, respectively (P<0.05). In contrast to the above two hormones, hCG exerted inhibitory effect at concentrations of 1,000 and 10,000 mIU/ml (P<0.05). There was no evidence of dose-response correlations in all three gonadotropin experiments. These data imply that gonadotropins at the concentrations studied inhibit the proliferation of human endometrial stromal cells, at least, in short-term culture in vitro. Accordingly, we cannot negate the possibility that administered gonadotropin during ovarian hyperstimulation may directly influence the proliferation of human endometrial cells. Received: 24 September 2001 / Accepted: 18 December 2001     

可溶性endoglin对人脐静脉内皮细胞产生一氧化氮及其合成酶磷酸化水平的影响

目的 观察可溶性endoglin(soluble endoglin,sEng)对体外培养的人脐静脉内皮细胞产生一氧化氮(nitric oxide,NO)、内皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)及其1177位点丝氨酸磷酸化程度的影响. 方法 将3代以内的人脐静脉内皮细胞接种于96孔培养板,分别加入完全细胞培养液(对照组)和不同浓度的sEng(1、10、100 μg/L),作用6、12和24 h后收取细胞培养液及细胞,硝酸酶还原法测定细胞培养液中NO代谢产物亚硝酸盐浓度反映NO含量,Western印记法检测各组细胞eNOS的相对表达量及其1177位点丝氨酸磷酸化情况,实时荧光聚合酶链反应技术检测各组细胞eNOS mRNA的相对表达量.采用方差分析、LSD法及Pearson相关分析法对各组进行比较. 结果 (1)1、10、100μg/L sEng作用6h,细胞培养液中NO浓度分别为(59.25±1.63)、(41.08±2.71)和(30.38±1.63)μmol/L;作用12 h,细胞培养液中NO浓度分别为(54.98±3.34)、(35.00±8.60)和(19.82±3.75)μmol/L;作用24 h,细胞培养液中NO浓度分别为(46.14±4.93)、(30.24±2.08)和(12.78±5.01)μmol/L,而对照组NO浓度随着时间的推移无明显改变(F=2.30,P=0.14).与sEng共培养后,细胞培养液中NO浓度明显降低,并与sEng作用时间(r=0.98,P＜0.05)及浓度(r=-0.88,P＜0.05)呈明显的负相关.(2)1、10、100 μg/L sEng作用6 h eNOS相对表达量分别为0.71±0.00、0.47±0.00和0.32±0.00;作用12 h,eNOS相对表达量分别为0.58±0.00、0.42±0.00和0.25±0.00;作用24 h,eNOS相对表达量分别为0.49±0.00、0.33±0.00和0.18±0.00.而对照组eNOS相对表达量及1177位点丝氨酸磷酸化eNOS吸光度/总eNOS吸光度随时间推移无明显改变(F分别为3.59和0.37,P分别为0.09和0.80).与sEng共培养后,细胞中eNOS蛋白表达量较对照组明显下降,其1177位点丝氨酸磷酸化程度明显减弱,与sEng作用时间(r分别为-0.98和-0.96,P均＜0.05)及浓度(r分别为-0.76和-0.79,P均＜0.05)呈明显负相关.(3)与sEng共培养后,细胞培养液中eNOS mRNA的表达量明显下降,亦与sEng作用时间(r=-0.51,P＜0.05)及浓度(r=-0.82,P＜0.05)呈明显的负相关. 结论 sEng 可能通过抑制eNOS 1177位点丝氨酸磷酸化,导致eNOS激活被抑制,NO生成减少,引起血管舒缩失调.     

一氧化氮与妊娠早期人巨细胞病毒宫内活动性感染的关系

目的　探讨一氧化氮与妊娠早期人巨细胞病毒 (HCMV)宫内活动性感染的关系。方法　选取既往有自然流产、死胎、胎儿畸形、胎儿生长受限等异常妊娠史并行人工流产术的早孕妇女 81例 ,应用逆转录 聚合酶链反应技术检测其胎盘绒毛组织HCMV mRNA的表达 ;酶联免疫吸附试验法测定其血清HCMV IgM ;原位杂交法检测其蜕膜巨噬细胞诱生型一氧化氮合酶 (iNOS) mRNA的表达并行定量分析。结果　 (1) 81例孕妇中有 6例胎盘绒毛组织HCMV mRNA阳性 ,14例血清HCMV IgM阳性 ,阳性率为 17 3 % (14/ 81)。依据HCMV mRNA与HCMV IgM检测结果 ,81例孕妇分为 3组 :Ⅰ组 :HCMV mRNA与HCMV IgM均阳性 5例 ,其宫内传播率为 3 5 7% (5 / 14) ;Ⅱ组 :HCMV mRNA阴性而HCMV IgM阳性 9例 ;Ⅲ组 :HCMV mRNA与HCMV IgM均阴性 66例。另有 1例HCMV IgM阴性者其胎盘绒毛组织HCMV mRNA阳性。 (2 )Ⅰ组患者蜕膜iNOS mRNA呈强表达 ,其吸光度值为 0 412±0 0 19,明显高于Ⅱ组的 0 172± 0 0 3 3与Ⅲ组的 0 167± 0 0 3 3 ,Ⅰ组与Ⅱ组及Ⅲ组比较 ,差异有显著性(P <0 0 5 ) ;Ⅱ组与Ⅲ组比较 ,差异无显著性 (P >0 0 5 )。 1例HCMV IgM阴性而胎盘绒毛组织HCMV mRNA阳性者 ,蜕膜iNOS mRNA也呈强表达 ,其吸光度值为 0 40 3。结论　一氧化氮可     

卵巢颗粒膜细胞一氧化氮的分泌及激素调节作用

目的探讨排卵前人卵巢颗粒膜细胞一氧化氮（NO）的分泌与白细胞介素1β（IL-1β）的关系,及其调节雌、孕激素的作用。方法收集体外受精-胚胎移植时穿刺卵巢抽吸的卵泡液,取出卵细胞后,分离出颗粒膜细胞进行培养,分别进行向其内添加IL-1β及NO供体的实验,测定各自培养液中的NO、雌二醇(E2)、孕酮(P)量及芳香化酶的活性。结果添加IL-1β组颗粒膜细胞的NO分泌量为（537±23）μmol/L,不添加IL-1β组为（411±15）μmol/L（P<0.01）;添加NO供体S-硝基乙酰基青霉胺（SNAP）组颗粒膜细胞的E2分泌量为（1234±136）μg/L,不添加NO供体SNAP组为（1792±103）μg/L（P<0.01）;添加NO供体SNAP组颗粒膜细胞的P分泌量为（988±103）μg/L,不添加NO供体SNAP组为（2486±123）μg/L（P<0.001）;添加NO供体SNAP组颗粒膜细胞的芳香化酶活性为（0.422±0.052）Ci/ml,不添加NO供体SNAP组为（0.833±0.012）Ci/ml（P<0.01）。结论在排卵前,卵巢颗粒膜细胞可分泌NO并可能受IL-1β的调节,NO与IL-1β共同参与类固醇激素合成的调节。     

一氧化氮与HPV感染的关系及其对子宫颈癌细胞增殖与凋亡的影响

目的 探讨一氧化氮(NO)与HPV感染的关系及其对宫颈癌细胞增殖与凋亡的影响.方法 选择2009年12月1日至2010年8月5日在复旦大学附属妇产科医院宫颈疾病诊疗中心行阴道镜检查的患者115例,应用HPV分型检测试剂盒(可检测21种HPV亚型)进行HPV分型,同时应用Griss法间接检测官颈局部NO含量.在体外培养的宫颈腺癌细胞株HeLa细胞和宫颈鳞癌细胞株Caski细胞中,加入不同浓度(终浓度分别为0.125、0.25、0.5 、1.0及2.0 mmol/L)NO供体--硝普钠(SNP)后培养24 h,采用四甲基偶氮唑蓝(MTT)比色法检测细胞生长情况,流式细胞仪检测细胞凋亡情况,并应用荧光定量逆转录PCR技术及蛋白印迹法检测SNP处理后细胞中HPVE6、E7 mRNA的表达及p53蛋白的表达.结果 (1)115例患者中,宫颈HPV感染93例,其中高危型50例,低危型43例;无HPV感染22例.115例患者的宫颈局部均检测到NO,其中高危型HPV感染患者的宫颈局部NO含量为(47.6±1.4)μmol/L,低危型HPV感染者为(24.1±1.2)μmol/L,无HPV感染者为(22.8±0.3)μmol/L,高危型HPV感染者高于低危型HPV感染者和无HPV感染者,差异有统计学意义(P＜0.05);而低危型HPV感染者与无HPV感染者比较,差异无统计学意义(P＞0.05).(2)不同浓度(0.125、0.25、0.5 1.0及2.0 mmol/L)SNP处理HeLa、Caski细胞24 h后,能抑制细胞生长、促进细胞凋亡,当SNP浓度≥1.0 mmol/L时,与SNP处理前比较,差异均有统计学意义(P＜0.05).1.0 mmol/L的SNP处理24h后,HeLa细胞中HPV18 F6、E7 mRNA的表达水平(分别为19.181±0.360、17.571±0.010)明显低于SNP处理前(分别为27.362±0.191、22.962±0.053;P＜0.05),p53蛋白的表达水平(1.17±0.03)明显高于SNP处理前(0.23±0.05;P＜0.05);但Caski细胞中HPV16E6、E7mRNA和P53蛋白的表达在SNP处理前、后无明显变化(P＞0.05).结论 宫颈局部微环境中NO含量增加与感染高危型HPV有一定的相关性;SNP能抑制宫颈癌细胞的生长,促进其凋亡,降低HPV18 E6、E7 mRNA的表达及活化p53蛋白,其机制可能是宫颈腺癌细胞株HeLa细胞对SNP更敏感.NO释放增加可能是官颈局部微环境清除HPV的免疫机制之一.

Abstract：

Objective To study the relationship between nitric oxide within cervical microenvironment and different HPV types as well as the effect of sodium nitroprusside( SNP), a nitric oxide donor, on the proliferation and apoptosis of cervical cancer cell lines. Methods HPV typing test was assessed from 115 women by using high-risk HPV (HR-HPV) 21 typing test and the release of cervical nitric oxide(NO) was assessed as nitrate, nitrite in cervical fluid. Cervical NO was then compared between women showing different HPV types. Proliferation of Caski and HeLa cervical cells was determined by methyl thiazolyl tetrazolium (MTT) assay, cell apoptosis was detected by flow cytometry after 24 hours treated by different final concentration of SNP (0. 125,0. 25,0. 5,1.0 and 2. 0 mmol/L, respectively). The expressions of HPV E6,E7 gene mRNA and p53 protein were detected by SYBR Green Ⅰ quantitative real-time PCR and western blot. Results ( 1 ) The cervical NO release of women with HR-HPV was higher compared to that in HPV negative women [ (47. 6 ± 1.4) μmol/L vs ( 22. 8 ± 0. 3 ) μmol/L; P ＜ 0. 05 ]; but there was no statistical difference between low-risk HPV (LR-HPV) group [ (24. 1 ± 1.2 ) μmol/L] and control group (P ＞0. 05 ). (2)After 24 hours treated by different final concentration of SNP, the results shown that SNP could inhibited the proliferation and increased apoptosis rate in Caski and HeLa cells, in which the concentration of SNP ≥ 1.0 mmol/L , there were significantly different ( P ＜ 0. 05 ), while when SNP ≥2. 0mmol/L, the proliferation of cells inhibited seriously. Treated by SNP ( 1.0 mmol/L ) 24 hours, the expressions of HPV18 E6, E7 mRNA in HeLa cells were reduced from 27. 362 ±0. 191,22. 962 ±0. 053 to19. 181 ±0. 360, 17. 571 ±0. 010 and the protein expression of p53 increased from 1. 17 ±0. 03 to 0. 23 ±0. 05, there were statistically significant differences between adding SNP group and the control group ( P ＜0. 05); but there were no statistically significant differences in HPV16 E6, E7 mRNA and that of p53 in Caski cells( P ＞ 0. 05 ). Conclusions The presence of HR-HPV is associated with an increased release of NO in the human uterine cervix; NO could inhibit the growth and proliferation and enhance the apoptosis of cervical cancer cells, inhibit the expression of HPV18 E6, E7 mRNA in HeLa cells and activate the expression of p53 protein, the mechanism may be due to higher sensitivity of HeLa cells (cervical adenocarcinoma cell) to SNP. The increasing release of NO may play a role in regulating the elimination of HPV in cervical microenvironment, which is a part of mucous membrane immunity.     

雌激素替代疗法对绝经后妇女血浆一氧化氮的影响

目的探讨雌激素替代疗法（ＥＲＴ）对绝经后妇女血浆一氧化氮（ＮＯ）含量的影响。方法测定１７例健康绝经后妇女（健康组）,２１例患高血脂症绝经后妇女（高血脂组）应用ＥＲＴ前及应用ＥＲＴ４周后血浆ＮＯ及血清雌二醇（Ｅ２）含量,并与１０例健康绝经后妇女应用安慰剂（对照组）进行对照。结果对照组应用安慰剂前后,ＮＯ含量无变化;健康组及高血脂组应用ＥＲＴ后,ＮＯ含量明显升高,健康组升高尤其明显;健康组应用ＥＲＴ４周后Ｅ２水平与ＮＯ升高有明显相关性。结论绝经后妇女补充雌激素,可通过升高ＮＯ含量发挥其对心血管的保护作用。     

Dual effect of anandamide on rat placenta nitric oxide synthesis

Anandamide (AEA) has been reported to have pleiotropic effects on reproduction, but the mechanism by which it exerts these effects is unclear. The aim of this study is to characterize rat placental endocannabinoid system and to analyze the possible functional role of AEA in the regulation of NO levels in rat placenta during pregnancy. We found that cannabinoids receptors (CB1 and CB2), FAAH and TRPV1 were expressed in chorio-allantoic placenta. NOS activity peaked at day 13 and decreased with progression of pregnancy. Both exogenous and endogenous AEA significantly decreased NOS activity. Although pre-incubation with AM251 (CB1 antagonist) or AM630 (CB2 antagonist) had no effect, co-incubation with both antagonists induced NOS activity. Furthermore, pre-incubation with exogenous AEA and both antagonists resulted in the induction of placental NOS activity and this effect was reverted with capsazepine (selective TRPV1 antagonist). Additionally, the enhanced NO synthesis caused by capsaicin was abrogated by co-treatment with capsazepine, illustrating that NOS activity could be modulated by TRPV1. Finally, the inhibition of TRPV1 receptor by capsazepine caused a significant fall in NOS activity. These data support the concept that AEA modulates NO levels by two independent pathways: (1) diminishing the NOS activity via CBs; and (2) stimulating NO synthesis via TRPV1. We hypothesized that AEA have an important implication in the normal function of placental tissues.     

胎盘生长因子与子痫前期发病及一氧化氮关系的研究

目的:探讨胎盘生长因子(PLGF)在子痫前期发病中的作用及其与一氧化氮的关系。方法:选择妊娠期高血压疾病患者45例,其中妊娠期高血压10例,轻度子痫前期12例,重度23例;选择同期正常妊娠妇女20例作为对照组。采用免疫组织化学染色法和逆转录-聚合酶链式反应(RT-PCR)检测两组患者胎盘PLGF蛋白及mRNA的表达。采用硝酸盐还原酶法测定两组胎盘组织NO浓度的变化。结果:(1)免疫组化结果显示,轻度和重度子痫前期的胎盘绒毛合体滋养细胞、绒毛间质PLGF表达均显著低于正常妊娠组(P<0.05),妊娠期高血压组与正常组无差别;PLGF在妊娠期高血压、子痫前期组及正常妊娠组分布范围基本一致,主要分布在绒毛合体滋养细胞和间质细胞胞浆,部分血管合体膜上也有表达;(2)轻、重度子痫前期胎盘组织PLGF mRNA平均灰度分别为3.33±0.39、1.97±0.29,显著低于正常妊娠组的平均灰度4.87±0.60(P<0.01);(3)轻、重度子痫前期胎盘组织中NO浓度分别为8.20±5.56μmol/g、6.46±2.25μmol/g,显著低于对照组18.10±7.12μmol/g(P<0.05);妊娠期高血压组胎盘组织NO浓度与对照组差异无显著性;(4)胎盘组织中胎盘生长因子表达水平与胎盘组织NO浓度呈显著正相关(r=0.54,P<0.05)。结论:子痫前期胎盘组织中胎盘生长因子水平降低,NO浓度下降,可能在子痫前期的发病中起一定作用。     

不同海拔地区健康儿童气体信号因子一氧化氮及硫化氢的变化

目的探讨气体信号因子一氧化氮(NO)及硫化氢(H2S)在不同海拔地区健康儿童中的含量差异。 方法2005-06—2006-02，将随机选取的北京市和青海省海北州体检时的健康儿童青少年，按照不同海拔分组：北京地区(平均海拔43.5m)17名， 男11名，女6名，年龄(7.28±4.79)岁；青海省海北州地区(平均海拔3200m以上)25名，男12名，女13名，年龄(10.42±0.49)岁。均空腹取肘静 脉血，采用Griess法测定血清NO浓度，敏感硫电极法测定血清H2S的浓度。 结果北京地区健康儿童血清中NO浓度(60.58±16.85)μmol/L，低于青海省海北州地区健康儿童血清中NO浓度(73.88±18.97)μmol/L；北京地 区健康儿童血清中H2S浓度(65.55±9.07)μmol/L，高于青海省海北州地区健康儿童血清中H2S浓度(46.92±6.46)μmol/L。 结论高海拔地区健康儿童血清中NO浓度明显高于低海拔地区，而H2S浓度则明显低于低海拔地区。     

Stimulated nitric oxide release and nitric oxide sensitivity in forearm arterial vasculature during normotensive and preeclamptic pregnancy

OBJECTIVE: We sought to determine whether the enhanced forearm vascular activity of nitric oxide during pregnancy and preeclampsia is associated with altered smooth muscle sensitivity to nitric oxide or with stimulated nitric oxide release. STUDY DESIGN: Forearm blood flow responses to brachial artery infusion of glyceryl trinitrate (a nitric oxide donor), serotonin (an endothelium-dependent nitric oxide-mediated agonist), and ritodrine (a beta-adrenergic receptor agonist) were studied in 10 nonpregnant women, 12 pregnant women, and 7 women with preeclampsia by means of strain-gauge plethysmography. Responses to each drug (maximum dilator response and the sum of the percentage of dilator responses to each drug) were compared by analysis of variance. RESULTS: Compared with nonpregnant women, pregnant subjects showed reduced responses to serotonin (summary response, 117 +/- 19 vs 221 +/- 30; P <.05). Responses to serotonin were reduced in the group with preeclampsia compared with those in the nonpregnant group (summary response, 71 +/- 28; P <.05) but did not differ from the responses in pregnant women. There were no differences between responses to glyceryl trinitrate and responses to ritodrine in any of the groups. CONCLUSION: Vascular smooth muscle sensitivity to nitric oxide is not altered in normal pregnancy or preeclampsia, but dilator responses to serotonin appear blunted. Alterations in serotonin receptor coupling to nitric oxide synthase, or a limitation of availability of the substrate for nitric oxide synthase (L-arginine) during pregnancy, could account for the reduction in stimulated nitric oxide release.     

抑制一氧化氮合成对孕鼠血管调节因子的影响

目的 研究抑制一氧化氮（ＮＯ）合成对孕鼠血浆内皮素（ＥＴ）、血栓烷素（ＴＸＡ２）、前列环素（ＰＧＩ２）及血管紧张素Ⅱ（ＡＮＧⅡ）水平变化的影响;并探讨这些因子在妊高征发病中的作用。方法 选择清洁级Ｗｉｓｔａｒ大鼠,随机分为两组,（１）对照组１２只,于妊娠第１４天开始皮下注射生理盐水直至分娩。（２）亚硝基左旋精氨酸甲酯（Ｌ－ＮＡＭＥ）组,１２只,于妊娠第１４天开始皮下注射一氧化氮合酶（ＮＯＳ）抑制剂     

细胞外信号调节蛋白激酶通路在胎盘生长因子1诱导脐静脉内皮细胞释放一氧化氮中的作用

目的 探讨细胞外信号调节蛋白激酶(ERK)通路在胎盘生长因子1(PLGF-1)诱导脐静脉内皮细胞(HUVEC)释放一氧化氮(NO)中的作用.方法 选择因头盆不称、胎儿窘迫或胎位异常行剖宫产分娩的新生儿50例,采用胰蛋白酶消化法进行脐带HUVEC原代培养.(1)培养成功后.用免疫组化法通过Ⅷ因子鉴定细胞形态;(2)用蛋白印迹法和RT-PCR技术检测HUVEC中酪氨酸激酶受体1(Fit-1)蛋白和mRNA表达;(3)用浓度为10 ng/ml的PLGF-1培养细胞(观察A组),并在培养前及培养后2.5、5、10、20 min收集细胞,提取蛋白,用蛋白印迹法检测ERK蛋白的表达;(4)用浓度为10 ng/ml的PLGF-1培养细胞,收集培养后20、40、160、360、480、720、1440 min的细胞培养液,用硝酸盐还原酶法检测培养液中NO的含量;(5)先用含ERK特异性抑制剂--PD98059(100μmo/L)的培养基培养细胞60 min后,换含PLGF-1的培养基继续培养(观察B组),收集培养后20、40、160、360、480、720、1440 min的细胞培养液,用硝酸盐还原酶法检测培养液中NO的含量.以无血清培养基培养的细胞为对照组,细胞处理时间、培养条件和收集细胞液时间同观察组.实验重复3次.结果 (1)免疫组化鉴定培养的细胞为HUVEC.(2)HUVEC中有Fit-1 mRNA和蛋白的表达.(3)观察A组PLGF-l培养HUVEC后2.5 min即有ERK蛋白表达强度的升高,5 min时表达强度达到高峰,10 min时表达强度降低.(4)PLGF-l培养20 min后HUVEC培养液中NO含量开始升高,为(6.96±0.34)μmol/L,培养40 min时NO含量为(9.45 4-0.59)μmol/L,培养360 min时NO达(15.82±0.69)μmo/L,各时间点NO含量比较,差异均有统计学意义(P<0.05).(5)观察B组NO释放显著受到抑制,从培养160 min到1440 min,NO含量下降达50%以上.结论 ERK通路可能是PLGF诱导HUVEC释放NO的重要信号通路之一.     

妊高征孕妇胎盘NOS及血清中NO变化的研究

目的 探讨一氧化氮(nitric oxide, NO)在妊高征发病中的作用。方法 选取妊娠晚期或足月重度妊高征孕妇32例为研究对象(PIH组),正常足月妊娠孕妇30例为对照组;用生化法测定胎盘组织一氧化氮合成酶(NOS)活性、母血及脐血一氧化氮(NO)的终末代谢产物亚硝酸盐/硝酸盐(NO_2~-/NO_3~-)含量。结果 PIH组胎盘组织NOS活性明显低于对照组(P<0.01);PIH组母亲静脉血及脐静脉血NO_2~-/NO_3~-含量与对照组比较差异均无显著性(P>0.05),而两组内母亲静脉血NO_2~-/NO_3~-含量较脐静脉血均明显增高,差异有显著性(P<0.01)。结论 NO合成障碍可能在PIH发病中起一定的作用,母血和脐血中NO_2~-/NO_3~-含量无明显差异,尚需进一步探讨。     

米非司酮对早孕妇女血清一氧化氮和子宫局部血流的影响

探讨米非司酮对早孕妇女血清一氧化氮及子宫局部血流的影响。方法 利用镀铜镉还原法测定服用米非司酮前,后的早孕妇女及正常非孕妇女血清ＮＯ浓度,同时利用彩色多普勒超声显像监测子宫局部血流变化,并观察流产后绒毛和蜕膜组织的超微结构。结果早孕妇女血清ＮＯ浓度明显高于正常非孕妇女,服米非司酮后血清ＮＯ浓度明显低于服药前;服米非司酮后子宫动脉和滋养层动脉血流的指数,收缩期血流峰值与舒张期血流最小值的比值,较服药