摘要
目的：检测核转录因子-κB亚基p65亲和肽(Nuclear Transcription Factor-KBp65, NF-κBp65)及白细胞介素-18(interleukin-18, IL-18)在未足月胎膜早破(preterm premature rupture the membrane, pPROM)、足月胎膜早破(premature rupture the membrane of term, tPROM)、足月胎膜未破孕妇(对照组)静脉血中浓度及分析二者在亚临床绒毛膜羊膜炎、非绒毛膜羊膜炎孕妇静脉血中水平,探讨NF-κBp65及IL-18在胎膜早破、亚临床绒毛膜羊膜炎发生、发展机制中的作用,及二者在胎膜早破、亚临床绒毛膜羊膜炎中的相关性,评价其预测亚临床绒毛膜羊膜炎的意义。
方法：采用酶联免疫吸附法(即ELISA法)分别检测未足月胎膜早破(pPROM)、足月胎膜早破(tPROM)及足月胎膜未破(对照组)各25例孕妇外周血中NF-κBp65和IL-18的水平；同时采用苏木精伊红(HE)染色的方法对所有胎膜早破产妇的胎膜进行亚临床绒毛膜羊膜炎的诊断,将其分为亚临床绒毛膜羊膜炎及非绒毛膜羊膜炎组,分析两组中的NF-κBp65和IL-18水平变化。用SPSS16.0软件统计包整理与分析数据。
结果：1、未足月胎膜早破(pPROM)、足月胎膜早破(tPROM)、对照组孕妇静脉血NF-κBp65浓度分别为(2127.5±380.84)ng/ml、(1732.54±441.81)ng/ml、(1432.96±371.40)ng/ml,与对照组比较,NF-κBp65水平在pPROM组与tPROM组明显增加(P<0.05,P<0.05),且pPROM组NF-κBp65水平较tPROM组明显增加(P<0.05)。
2、未足月胎膜早破(pPROM)、足月胎膜早破(tPROM)、对照组孕妇静脉血IL-18浓度分别为(51.16±6.52)ng/ml、(47.92±5.67)ng/ml、(42.20±6.42)ng/ml,与对照组比较,IL-18水平在pPROM组与tPROM组明显增加(P<0.05,P<0.05),且pPROM组IL-18水平较tPROM组明显增加(P<0.05)。
3、亚临床绒毛膜羊膜炎孕妇静脉血NF-Bp65浓度(2176.78±428.86)ng/ml明显高于非绒毛膜羊膜炎组中NF-κBp65的浓度(1664.72±311.14)ng/ml,经统计学分析,两组间有统计学意义(P<0.05)。
4、亚临床绒毛膜羊膜炎母血IL-18浓度(55.32±5.63)ng/ml明显高于非绒毛膜羊膜炎组中IL-18的浓度(46.29±5.10)ng/ml,两组间有统计学意义(P<0.05)。
5、NF-κBp65与IL-18在胎膜早破孕妇静脉血中的水平呈正相关关系(r=0.840,p<0.05),在正常对照组中NF-κBp65与IL-18无相关关系(r=0.382,P=0.059>0.05);母血NF-κBp65与IL-18在亚临床绒毛膜羊膜炎与非绒毛膜羊膜炎组呈正相关关系(r=0.806,P<0.05；r=0.776,P<0.05)。
结论：1、胎膜早破母血中NF-κBp65、IL-18的水平升高,推测二者可能参与了胎膜早破的发生与发展。
2、合并亚临床绒毛膜羊膜炎的未足月胎膜早破母血中NF-κBp65、IL-18的水平更高,推测二者水平升高除与胎膜早破发生有关还与感染的发生有关。
3、NF-κBp65、IL-18水平在有亚临床绒毛膜羊膜炎的胎膜早破孕妇静脉血中较无绒毛膜羊膜炎组升高,推测二者参与了亚临床绒毛膜羊膜的发生与发展。
4、母血NF-κBp65、IL-18水平升高在胎膜早破、亚临床绒毛膜羊膜炎中呈正相关关系,推测二者相互作用共同诱导胎膜早破及亚临床绒毛膜羊膜炎的发生,可作为预测亚临床绒毛膜羊膜炎的新指标。
Objectives:To investigate the pathogenesis of premature rupture of membrane (PROM)and of subclinical chorioamnionitis by detecting the level of Nuclear Transcription Factor-κBp65 (NF-κBp65) and interleukin-18 (IL-18) in maternal blood with premature rupture of membrane and subclinical chorioamnionitis.To discuss the relevance between the two factors, then to estimate their clinieal value in predieting subelinical chorioamnionitis.
Methods:pregnant women of preterm premature rupture the membrane、premature rupture the membrane of term and normal pregnant women in term with intact membranes, every 25 cases were studied.The concentration of NF-κBp65 and IL-18 in maternal blood were determined by solid phase sandwi-ch enzyme linked immuno-sorbent assay(ELISA).And subclinical chorioamnionitis or not subclinical chorioamnionitis were diagnosed by HE among fifty puerperants. They were divided into two groups. The level of NF-κBp65 and IL-18 were analysed between the two groups. These experimental data were analized by the software SPSS 16.0.
Results:(1) The level of NF-κBp65 in maternal blood pPROM group (2127.5±380.84)ng/ml and tPROM group (1732.54±441.81)ng/ml are significantly higher than the nomal group(1432.96±371.40)ng/ml,each group has statistically deviation(P＜0.05,P＜0.05),and the level of pPROM group is significantly higher than tPROM group(P＜0.05).
(2) The level of IL-18 in maternal blood of pPROM group (51.16±6.52)ng/ml and tPROM group(47.92±5.67)ng/ml are significantly higher than the nomal group(42.20±6.42)ng/ml, each group has statistically deviation(P＜0.05,P＜0.05), and the level of pPROM group is significantly higher than tPROM group(P＜0.05).
(3)The level of NF-κBp65 in maternal blood of subclinical chorioamnionitis group is (2176.78±428.86)ng/ml, The level of NF-κBp65 in maternal blood of Non-chorioamnionitis group is (1664.72±311.14)ng/ml, the level of subclinical chorioamnionitis group is significantly higher than Non-chorioamnionitis group, the two groups have statistically deviation (p＜0.05).
(4)The level of IL-18in maternal blood of subclinical chorioamnionitis group is (55.32±5.63)ng/ml, The level of IL-18 in maternal blood of Non-chorioamnionitis group is (46.29±5.10)ng/ml, the level of subclinical chorioamnionitis group is significantly higher than Non-chorioamnionitis group, the two groups have statistically deviation (p＜0.05).
(5)The level of NF-κBp65 and IL-18 inmaternal blood in premature rupture of membranes group is positive correlation (r=0.840,p＜0.05), but in the normal group is on correlation (r=0.382,P=0.059＞0.05); between subclinical chorioamnionitis group and Non-chorioamnionitis group is positive correlation (r=0.95,P＜0.05;r= 0.97, P＜0.05).
Conclution:1、The higher level of NF-κBp65 and IL-18 in maternal blood of premature rupture the membrane may lead to premature rupture of membrane.
2、The highest level of NF-κBp65 and IL-18 in maternal blood of pPROM group with subclinical chorioamnionitis, the highest level of these are correlated with infection expect PROM.
3、The higher level of NF-κBp65 and IL-18 in maternal blood of subclinical chorioamnionitis group may lead to subclinical chorioamnionitis.
4、The higher level of NF-κBp65 and IL-18 in maternal blood of PROM and subclinical chorioamnionitis,NF-κBp65 and IL-18 can interact with each other leading to PROM and subclinical chorioamnionitis,and identify the chorioamnionitis as valuable clinical index.
引文
[1]Murtha AP,Greig PC,Jimmerson CE,et al.Maternal serum interleukin-6 concentration in patients with preterm pemature rupture of membranes and evidence of infection [J]. Am J ObstetGynecol,1996;175(4):966-969.
[2]乐杰主编.妇产科学.第七版.北京：人民卫生出版社,2006,137.
[3]Fortunato SJ,Menon R,Lombardi SJ. Role of tumor necr osis factor-alpha in the p remature rupture of membranes and preterm labor pathways[J]. Am J Obstet Gynecol,2002;187(5): 1159-1162.
[4]Shobokshi A, Shaarawy M. Maternal serum and amniotic fluid cytokines in patients with preterm premature rupture of membranes with and without intrauterine infection[J]. Int J Gynaecol Obstet,2002;79 (3):209-215.
[5]Xu D,Trajkovic V,Hunter D,et al. IL-18 induces the differentiation of Thl or Th2 cells depending upon cytokine milieu and genetic background [J].Eur J Immunol,2000;30(11):3147-3156.
[6]PeniikainenV,SuomalainenL,Erkkila K,MartelinE,et al.Nuelear factor-kappa B activation in human testicular apoptosis[J].Am J Pathol,2002;160(1):205-218.
[7]Wenstrom KD, Andrews WW, Hauth JC, et al. Elevated second trimester amniotic fluid interleukin-6 levels predict preterm delievery [J].Am J Obstet Gynecol,1998,178(3):546-550
[8]Saji F,Samejima Y,Kamiura S,et al. Cytokine production in chorioamnionitis [J].JReprod Immunol,2000;47 (2):185-196.
[9]Aoraisham AS,Singhal N,McMillan DDet al.A multicenter study on the clinical outcome of chorioamnionitis in preterm infants. [J] Am J Obstet Gynecol 2009;200(4):1-6.
[10]Dammanno,O LevitonA.Matemal intrauter ineinfection, cytokines, and brain damage in the preterm newborn [J].Pediatr Res1997;42(1):1-8.
[11]MurphyDJ,SellersS,MackenzielZ,Yudkin P,et al..Case-control study of antenatal and intrapartum risk factors for cerebral palsy in very preterm singleton babies[J].Lancet 1995;346(8988):1449-1454.
[12]Osmers RG,Blaser J,KuhnW,et al.Interleukin-8 synthesis and the onset of labor[J].ObsetGynecol,1995;86(2):233-239.
[13]赵欣.CPR、IL-12在胎膜早破孕妇中的表达及意义[J].中华妇产科杂志,2010,(3)：320-322.
[14]McGregor JA,Schoon Maker J,Lunt BD, et al. Antibiotic inhibition of bacterially induced fetal membrane weakening [J]. Obstet Gynecol,1990;76(1):124-128.
[15]尹香花,朴贞姬,宋艳春.早产病人血c-反应蛋白总补体活性补体成分与亚临床绒毛膜羊膜炎的关系[J].中国实用妇科与产科杂志,2001,17(3)：152.
[16]郭智勇,郭新宇,李文玲等.胎膜早破孕c反应蛋白测定[J].南方医科大学学报,2006,26(12)：1762-1764
[17]Sen R,BaltimoreD et al. Tiple nuclear factors interactwith the immunoglobulin enhancer sequence. Cell,1986;46(3):705-708.
[18]Shanmugam N,Reddy MA,Guha M,et al.High glucose-induced expression of proinflammatroy cytokine and chemokine genes in monocytic cells[J].Diabetes,2003;52(5):1256-1264.
[19]Hilliard BA,Mason N,Xu L,et al.Critical roles of C-Rel in autoimmune inflammation and helper T cell differentiation[J].J Clin Invest,2002;110(6):843-850.
[20]Liou HC,Hsia CY et al.Distinctions between c-Rel and other NF-kappa B proteins in immunity and disease[J].Bioessays,2003;25(8):767-780.
[21]Artis D,Shapira S,Mason N,et al.Differential requirement for NF-kappa B family members in control of helminth infection and intestinal inflammation[J].J Immunol,2002;169(8):4481-4487.
[22]Aradhya S,Nelson DL.NF-kappaB signaling and human disease[J].Curr Opin Genet Dev,2001;11(3):300-306.
[23]Nakamura H,Kimura T,Ogita K,et al.NF-kappaB activation at implantation window of the mouse utems[J].Am J Reprod Immuno,2004;51(1):16-21.
[24]谢青贞,辛志敏.核因子-κB与生殖调节[J].国外医学妇产科学分册2002,29(4)：223-225.
[25]谢青贞,辛志敏,曹路敏等.白细胞介素-1β对人早孕细胞滋养层细胞核因子-κB的激活机制[J].生殖与避孕,2005,25(10)：605-612.
[26]Soloff MS,Cook DL Jr.,Jeng YJ,Anderson GD.In situ analysis of interleukin-1-induced transcription of cox-2 and il-8 in cultured human myometrial cells.Endocrinology.2004, 145:1248-1254.
[27]Astle S,Slater DM,Thornton S et al.The involvememt of progesterone in the onset of human
labour [J].Eur J Obstet Gynecol Reprod Biol,2003;108(2):177-181.
[28]Lappas M,Permezel M,Georgiou HM,et al.Regulation of phospholipase isozymes by nuclear factor-kappaB in human gestational tissues in vitro [J].J Clin Endocfinol Metab.2004;89(5):2365-2372.
[29]Chase AJ,Bond M,Crook MF,Newby AC.Role of nuclear factor-kappa B activation in metalloproteinase-l,-3,and-9 secretion by human macrophages in vitro and rabbit foam cells produced in vivo[J]. Arteriosclerosis Thrombosis and Vascular Biology.2002,22:765-771.
[30]Mccracken SA,Hadfield K,Rahimi Z,et al.NF-kappaB-regulated suppression of T-bet in T cells represses Thl immune responses in pregnancy[J].Eur J Immunol.2007;37(5):1386-1396.
[31]Mccracken SA,Gallery E,Morris JM.Pregnancy-specific down-regulation of NF-kappaB expression in T cells in humans is essential for the maintenance of the cytokine profile required forpregnancy success[J].J Immunol,2004,172(7):4583-4591.
[32]李丹,周昌菊,姚穗.早产孕妇血中核因子-κ BP65与亚临床绒毛膜羊膜炎的关系[J].中国妇幼保健,2008,23(26)：3725-3728.
[33]张艳.IL-18、iNOS和NF-B在妊娠和流产大鼠中的表达及意义.博士论文,2008
[34]Sakai M,Shiozaki A,Sasaki Y,et al.The ratio of interleukin(IL)-18 to IL-12 secreted by peripheralblood mononuclear cells is increased in normal pregnant subjects and decreased in preeclamptic patients[J].J Reprod Immunol,2004;61(2):133-143.
[35]Yoshimoto T,Takeda K,Tanaka T,et al.IL-12 up-regulates IL-18 receptor expression on T cells,Thl cells,and B cells:synergism with IL-18 for IFN-gamma production[J].J Immunol,1998;161(7):3400-3407.
[36]Ida A,Tsuji Y,Muranaka J,et al.IL-18 in pregnancy;the elevation of IL-18 in maternal peripheral blood during labour and complicated pregnancies[J].J Reprod Immunol,2000;47(1):65-74.
[37]张茗.母血清和胎膜白细胞介素18含量与胎膜早破的相关性研究[J].中国妇幼保健,2003,5(23)68.
[38]Shazlys E L,Makhseed M,Azizieh F,et al.Increased expression of pro-inflammatory cytokines in placentas of woman undergoing spontaneous preterm delivery or premature repture of membranes [J].Am J Reprod Immunol,2004;52 (1):45-52.
[39]Menon R,Lombardi SJ,Fortunato SJ.IL-18, a product of choriodecidual cells,incre ases during premature rupture of membranes but fails to turn on the Fas-FasL-mediated apoptosis pathway[J].Assist Reprod Genet,2001;18(5):276-284.
[40]王春,尚涛.白介素-18在预测早产中的价值[J].中华实用医学,2003,5(1)：52-66.
[41]王楠.IL-6、IL-18在胎膜早破孕妇母血、脐血、胎膜中的表达.中国妇幼保健,2009,4(12)：54-57.
[42]洪丽玲,陈惠池,潘莉莉等.胎膜早破NF-κB在胎膜的表达和母血中IL-15的含量[J]。剖科学进展,2005,11(3)：216-218.
[43]Gomez B,Romero R,Edwin SS,et al.Pathogenesis of preterm labor and preterm premature of membranes assoeiated with in traamniotie infection [J].Infect Dis Clin North Am,1997;11(1):135-176.
[44]JacobssonB,Holst RM,Mattsby-BaltZer et al.Interleukin-18 in cervieal mucus and amniotic fluid:relationship to microbial invasion of the arnniotic fluld, intra-amniotic inflamlnation and pretem delivery [J].BJOG,2003;110(6):598-603.
[45]Alla SPlichalova,Yoshihiro Muneta,et al.Effect of bacterial virulence on IL-18 expression in the amnion infeeted with Escherichia coli [J]. AMJ Reprod Immunol,2005,53(5):255-260.
[46]Abraham E.NF-kappa B activatin[J].Crit Care Med,2000;28(4):100-104.
[47]Vandenbroeck K,Alloza I,Gadina M,et al.Inhibiting cytokines of the interleukin-12 family:recent advances and novel challenges[J].J Pharmacol,2004,56(2):145-160.
[48]Grandjean-Laquerriere A, Antonicelli F, et al. UVB-induced IL-18 production in human keratinocytecellline NCTC 2544 throughNF-kappaBactivation[J].Cytokine.2007;37(1):76-83.
[49]陈玉熹,卢中秋,邱俏檬.创伤弧菌脓毒症大鼠肺组织NF-κB和IL-18的基因表达以及药物的干预效应[J].中华医院感染学杂志,2010,20(3)：320-323.
[50]曹静,徐丽瑾,刘桂芬.白介素-18水平及核因子-κB活性变化与早产儿脑室周围白质软化相关性研究[J].中国妇幼保健,2009,24(35)：5066-5068.
[1]Xu D,Trajkovic V,Hunter D,et al. IL-18 induces the differentiation of ThlorTh2 cells depending upon cytokine milieu and genetic background[J].Eur J Immunol,2000,30(11):3147-3156.
[2]Liu B,Mori I,Hossain MJ,et al.Interleukin-18 improves the early defence system against influenza virus infection by augmenting natural killer cell-mediated cytotoxicity [J].J Gen Virol,2004,85(2):423-428. [3] Huang HY,Chan SH,Yu HT,et al.Interleukin-18 system messenger RNA and protein expression in human endometrium during the menstrual cycle[J].Fertil Steril,2006,86(4):905-913.
[4]Murakami Y,Otsuki M,Kusumoto K,et al. Estrogen inhibits interleukin-18 mRNA expression in the mouse uterus[J].J Reprod Dev,2005,51(5):639-647.
[5]张艳李文献陈树林.白细胞介素-18在大鼠胚泡植入前后子宫的分布及表达[J]解剖学报,2008,39(1)：98-101
[6]Chaouat G, Zourbas S, Ostojic S, et al. A brief review of recent data on some cytokine expressions at the materno foetal interface which might challenge the classical Thl/Th2 dichotomy[J].J Reprod Immunol,2002,53(1-2):241-256.
[7]Guilbert LJ.There is a bias against type 1 (inflammatory)cytokine expression and function in perngnaey[J].J Reprod Immunol,1996,32(2):105-110
[8]Okamura H,Tsutsui H,Kashiwamura S,et al.Interleukin-18:a novel cytokine that augments both innate and acquired immunity[J] Adv Immunol.1998,70:281-312.
[9]Okamura H,Kashiwamura S,Tsutsui H,etal. Regulation of interferon-gamma production by IL-12 and IL-18[J]. Curr Opin Immunol,1998,10(3):259-264.
[10]Yoshimoto T,Takeda K,Tanaka T,et al.IL-12 up-regulates IL-18 receptor expression on T cells,Thl cells,and B cells:synergism with IL-18 for IFN-gamma production[J].J Immunol,1998,161(7):3400-3407.
[11]Xu D,Chan WL,Leung BP,et al.Selective expression and functions of interleukin 18 receptor on T helper(Th)type 1 but not Th2 cells[J].J Exp Med,1998,188(8):1485-1492.
[12]Kohno K,Kataoka J,Ohtsuki T,etal.IFN-gamma-inducing factor(IGIF)is a costimulatory factor on the activation of Th1 but not Th2 cells and exerts its effect independently of IL-12[J].J Immunol,1997,158(4):1541-1550.
[13]Ida A, Tsuji Y, Muranaka J, et al.IL-18 in pregnancy;the elevation of IL-18 in maternal peripheral blood during labour and complicated pregnancies[J].J Reprod Immunol,2000,47(1):65-74.
[14]Menon R,.Lombardi SJ.Fortunato SJ,et al.IL-18,a Product of Choriodecidual cells, increases during premature rupture of membranes but fails to turn to the Fas-FasL-mediated apoptosis pathway[J].J Assist Reprod Genet,2001,18(5):276-284.
[15]Jacobsson B, Hoist RM, Mattsby-Baltzer I, et al.Interleukin-18 in cervical mucus and amniotic fluid:relationship to microbial invasion of the amniotic fluid, intra-amniotic inflammation and preterm deliver[J].BGOG,2003,110(6):598-603
[16]Ekelund CK,Vogel I,Skogstrand K,et al. Interleukin-18 and interleukin-12 in maternal serum and spontaneous preterm delivery[J]. J Reprod Immunol,2008,77(2):179-185
[17]Wang X,Hagberg H,Mallard C,et al.Disruption of interleukin-18, but not interleukin-1, increases vulnerability to preterm delivery and fetal mortality after intrauterine iflammation[J]. Am J Pathol,2006,169(6):967-976
[18]Sakai M,hiozaki A,Sasaki Y,et al.The ratio of interleukin(IL)-18 to IL-12 secreted by peripheral blood mononuclear cells is increased in normal pregnant subjects and decreased in pre-eclamptic patients[J].J Reprod Immunol,2004,61(2):133-43.
[19]Huang X,Huang H,Dong M, et al.Serum and placental interleukin-18 are elevated in preeclampsia[J].J Reprod Immunol,2005,65(1):77-87.
[20]Ishihara N,Matsuo H, et al. Increased apoptosis in the syncytiotrophoblast in human term placentas complicated by either preeclampsia or intrauterine growth retardation[J].Am J Obstet Gynecol,2002,186 (1):158-166.
[21]Wilson R,Moor J,Jenkins C,et al.Abnormal first trimester serum interleukin 18 levels are associated with a poor outcome in women with a history of recurrent miscarriage[J]. Am J Reprod Immunol,2004,51(2):156-159.
[22]Ledee-Bataille N,Olivennes F,Kadoch J,et al. Detectable levels of interleukin-18 in uterine luminal secretions at oocyte retrieval predict failure of the embryo transfer[J].Hum Reprod,2004,19(9):1968-1973.
[23]Ostojic S,Volk M,Medica I, et al.Polymorphisms in the interleukin-12/18 genes and recurrent spontaneous abortion[J].Am J Reprod Immunol,2007,58(5):403-408.