Analysis The Related Factors Between Mothers And Babies Who Were Complicated By PIH With The Luminex System And Conventional Detection Methods

Objective and meaning:If intrauterine growth restriction (IUGR) don't get correct diagnosis and management, the morbidity and mortality of preterm delivery, asphyxia, low birth weight may be increased significantly. The important reason for IUGR is the insufficient uteroplacental and fetoplacental blood flow. There are also other reasons, such as the intrauterine infection of Cytomegalic virus and Rubella virus, as well as the Down's and Edward syndrome.Gestation hypertension is the most common factor for IUGR. The morbidity of pregnancy induced hypertension (PIH) complicated with IUGR is 20%-30% and the perinatal mortality of it is 6-to 9- fold than that of the normal birth weight newborns. The perinatal mortality of the IUGR babies delivered by mother with PIH is 3 times than by mother without PIH. Especially, the severe PIH can induce higher risk for IUGR, increase the perinatal mortality, and impact directly the prognosis of the fetus and newborns. Also, PIH is a risk factor for low birth weight; on the other hand, IUGR is a determinant factor for the outcomes of PIH. As follows: perinatal morbidity and mortality of the fetus, the management and treatment efficacy of the mother.PIH whose etiologic factor and mechanism still need further evaluation is the idiopathic syndrome at＞20 weeks' gestation and one of the important reasons for perinatal morbidity and mortality. Further, there are no effective prevention and treatment for it. Studies to date showed that the etiopathology of PIH and it's reasons of inducing IUGR may have relationship with the following factors:1. The imbalance of Th1/Th2 Romagnani confirmed that the Human CD⁴⁺ Th cells (assist T-lymphocytes) can be divided into Thl and Th2 subtypes in 1991. The Thl cells mostly secreted IL-2(interleukin-2), IFN-r(interferon-r), TNF-a(tumor necrosis factor-a), and mediate the cell immunity; The Th2 cells mostly secreted IL-4, IL-5, IL-6, IL-10 and i1-13, and mediate the humoral immunity. The Th1 and Th2 cells shared a common precursor cell-Th0 cell. The important inducer of the differentiation fromTh0 to Thl and Th2 is IL-12 and IL-4, respective. Lots of studies have confirmed that Thl and Th2 cell are one couple regulators. At the same time, they are the inhibitors to each other. IFN-r secreted by Th1 cells can prevent the differentiation and function of the Th2, and IL-10 and TGF-b secreted by Th2 can prevent the differentiation and function of the Th1.Todate, the studies showed that the ratio of Th1/Th2 which belonged to the patients with PIH was likely to increase. The imbalance of Th1 and Th2 was a important etiologic factor for PIH.Saito has reported that there were more IL-2, TNF-a and IFN-r and significant less IL-4 in the blood plasma of the mothers complicated PIH than of the normal mothers, also that the ratios of IL-2/IL-4, IFN-r/IL-4 were both higher in comparison with the normal pregnancy. Further, the blood pressure was significantly relative to the level of cell factors of the Thl cells. The proportion of the Thl cells significantly decreased in the late trimester of normal pregnancy and the ratio of Th1/Th2 also obviously fell in comparison with women without pregnancy. However, the proportion of Thl cells and the ratio of Th1/Th2 were higher in the women complicated with preeclampsia than the normal pregnant women. The reductive proportion of the Th2 cells, also the changes of the proportion of Thl, Th2 cells and changes of the ratio of Th1/Th2 had a relationship with the level of the cell factors. All aboved showed that the Thlimmunity had a advantage in the patients with preeclampsia. Another factor TNF-a which secreted by the Thl cells can induce the activation of vascular endothelial cells, stimulate the product of platelet-derived growth factor and mitotic activity, result in the break of renal endothelial cells, as well as prevent the product of vasodilator NO and reduce the sensitivity of any cell to the toxicity of oxygen free radicals.Placenta is the organ for substantial exchange between mothers and fetus, also a complex partial immune organs. Furture, placenta can secrete many kinds of cell factors. Some scholars believed that the damage of the immune protection which was from mother to trophoblast is the cause to PIH. IL-2 which secreted by Thl cells can active the NK cells of peripheral blood and the decidua, and the actived NK cells can attack the trophoblast.2. Vascular Endothelial Growth Factor(VEGF) express anormaly VEGF is specific mitogen coming endotheliocyte, which have facilitative intensively function to angiogenesis and can increase microvascular permeability, and play an important role in the formation of physiology new vessels net. Placenta VEGF distributed mainly over cytotroblast, syncytiotrophoblast, vassular endotheliocyte, villose mesenchymocyte. VEGF can effect particulaly endotheliocyte, promote blood vessel formation, and improve blood provision. It's main functions are follows: promoting endotheliocyte differentiation, accrementition, immigration, infiltrate;regulation vasculogenesis and maintaining the function of vascular endothelial cell. Establishment caduca capillary network and embryo's growth and development are based on angiogenesis and vascularize. Normal deciduate placenta histological sectiones are riched of VEGF, they express obviously at syneytiotrophoblast and caduca infiltrate trophocyte. However, the blood concentration of VEGF in patients suffered from PIH are very low, so did fetal umbilical vein.3.Placental Renin Angiotensin System(RAS) Some researches indicated that the patients suffering from PIH have more renin in villous trophoblast tissue. Takimoto make female rats which have been transferred angiotensinogen gene mate with male rats which have been transferred rennin gene, they discovered that the pregnant rats suffered from PIH, the renin coming from placenta enter maternal blood, higher renin's concentration in maternal plasma, and the placentas edema and organisms necrosis in histological sectiones. Above the all indicate that PIH is relate to RAS.Angâ…¡can increase vasopermeability, stimulate vascellum neogenesis, and play an important role in adjustment placental blood flow, too. Uterus-placenta vascular resistance and adjustment of blood flow is very sophisticated during pregnancy. Ang-â…¡can keep blood pressure stabilization and sustain uterus-placenta blood supply by stimulating vasodilator synthesis that prostaglandin-E,â… . So, low dose Angâ…¡can increase uterine blood flow by stimulating vasodilator synthesis,high dose Angâ…¡produce inverse role.The study use multifunction liquid-phase analytical system(Luminex 200), traditional ELISA, immunohistochemistry and fluorescent technique, detected PIH correlation factors (IL-2,4,6,10,12,TNF-a, IFN-r, ANGâ…¡,VEGF) expressing in maternal blood, cord blood, matemal surface, fetal surface, respectively. To analysis how they effect to pregnant mother and fetal, to explore etiopathogenisis of PIH and mechanism of IUGR, in order to decrease morbidity of IUGR and improve prognosises of IUGR and PTH.Methods:1.The selection and handling of samples:â‘ There are 30 peoples all of them are primipara both in pregnancy-induced hypertension syndrome group and in normal pregnant women group. Their aetas, gestational weeks and weight were not significant deviation;â‘¡peripheral vein blood of mothers and cord blood of newborn infants were selected 4ml, not anticoagulation, 2000r/min centrifulged, then supernate fluid was obtained and stored in refrigerator-70℃;â‘¢Two tissues weighting about lg both from maternal surface and child surface were taken down,residual blood was removed. Both the one fixed in paraformaldehyde, and the other weighed 0.5g and breakdown milled, then added lml normal sodium, 3000r/min centrifulged, and then supernate fluid was obtained and stored in refrigerator -70℃.2.To detect the contents of ANG II and VEGF in motermal blood, cord blood, maternal surface and child surface by classic ELISA.3. The placenta fixed by paraformaldehyde were maken into paraffin section to carry out immunohistochemistry stain and fluorescein stain.4. The contents of IL-2,IL-4,IL-6,IL-10,IL-12,IFN-r,TNF-a in motermal blood, cord blood, maternal surface and child surface by multipurpose liquid phase array analytical system (Luminex 200).5. Statistical analysis: experimental results were applied two independent-sample T test and Pearson correlation analysis with Excel 2003 software.Results:1.ANGâ…¡:â‘ It's strong positive at placental fetal surface of PIH by immunohistochemistry;â‘¡It's so by fluorescein stain.â‘¢By ELISA, Maternal surface, PIH: 8.51±4.01pg/ml, normal pregnancy: 7.76±3.47pg/ml, no statistical significance (P＞0.05); placental fetal surface, PIH: 11.82±3.92pg/ml, normal pregnancy: 9.64±2.63pg/ml, the difference has statistical significance (P＜0.005); peripheral blood,PIH: 46.44±8.48pg/ml, normal pregnancy: 32.43±5.87pg/ml, the difference hasstatistical significance(P＜0.005); cord blood, PIH: 68.83±8.68pg/ml, normalpregnancy: 72.47±8.51pg/ml, no statistical significance(P＞0.05). The coefficient ofcorrelation between cord blood and peripheral blood of PIH is 0.7379, P＜0.005.2.VEGF:â‘ It's strong positive at placental fetal surface of normal pregnancy byimmunohistochemistry;â‘¡It's so by fluorescein stain.â‘¢By ELISA, Maternal surface,PIH: 262.88±123.00pg/ml, normal pregnancy: 280.03±130.98pg/ml, no statisticalsignificance(P＞0.05); placental fetal surface, PIH: 200.11±110.32pg/ml, normalpregnancy: 261.52±82.84pg/ml, no statistical significance(P＞0.05); peripheral blood,PIH: 27.82±11.14pg/ml, normal pregnancy: 23.54±12.05pg/ml, no statisticalsignificance(P＞0.05); cord blood, PIH: 291.62±126.36pg/ml, normal pregnancy:209.27±141.21pg/ml, the difference has statistical significance(P＜0.005).3.TNF-a: maternal surface, the patients suffered from PIH: 17.40±8.08pg/ml,normal pregnancy: 10.70±5.55pg/ml, the difference has statistical significance(P＜0.005); placental fetal surface, PIH: 10.64±2.92pg/ml, normal pregnancy:5.94±3.42pg/ml, the difference has statistical significance (P＜0.005); peripheral blood,PIH: 13.34±6.50pg/ml, normal pregnancy: 7.78±4.73pg/ml, the difference hasstatistical significance (P＜0.005); cord blood, PIH: 4.50±2.12pg/ml, normalpregnancy: 5.36±2.13pg/ml, no statistical significance(P＞0.05).4.IFN-r: maternal surface, the patients suffered from PIH: 39.88±18.15pg/ml,normal pregnancy: 21.38±9.39pg/ml, the difference has statistical significance(P＜0.005); placental fetal surface, PIH: 20.99±6.60pg/ml, normal pregnancy:22.99±7.85pg/ml, no statistical significance(P＞0.05); peripheral blood, PIH:19.93±6.47pg/ml, normal pregnancy: 15.65±5.80pg/ml, the difference has statisticalsignificance (P＜0.01); cord blood, PIH: 15.32±6.31pg/ml, normal pregnancy: 10.70±4.34pg/ml, the difference has statistical significance (P＜0.01).5.IL-2: maternal surface, the patients suffered from PIH: 8.27±1.55pg/ml, normal pregnancy: 5.84±2.21pg/ml, the difference has statistical significance (P＜0.001); placental fetal surface, PIH: 5.24±2.45pg/ml, normal pregnancy: 5.49±2.35pg/ml, no statistical significance(P＞0.05); peripheral blood, PIH: 9.10±5.55pg/ml, normal pregnancy: 3.22±2.41pg/ml, the difference has statistical significance (P＜0.001); cord blood, PIH: 4.18±2.51pg/ml, normal pregnancy: 3.98±1.71pg/ml, no statistical significance(P＞0.05).6.IL-4: maternal surface, the patients suffered from PIH: 3.03±2.06pg/ml, normal pregnancy: 2.87±1.69pg/ml, they have no statistical significance(P＞0.05); placental fetal surface, PIH: 2.11±0.77pg/ml, normal pregnancy: 2.26±1.29pg/ml, no statistical significance(P＞0.05); peripheral blood, PIH: 2.37±1.06pg/ml, normal pregnancy: 5.60±4.81pg/ml, the difference has statistical significance (P＜0.005); cord blood, PIH: 2.15±1.33pg/ml, normal pregnancy: 1.66±0.59pg/ml, no statistical significance(P＞0.05).7.IL-6: maternal surface, the patients suffered from PIH: 33.02±12.57pg/ml, normal pregnancy: 47.51±25.95pg/ml, the difference has statistical significance (P＜0.01); placental fetal surface, PIH: 45.33±17.96pg/ml, normal pregnancy: 42.31±15.57pg/ml, no statistical significance(P＞0.05); peripheral blood, PIH: 5.41±0.79pg/ml, normal pregnancy: 2.77±1.11pg/ml, the difference has statistical significance (P＜0.001); cord blood, PIH: 2.65±0.53pg/ml, normal pregnancy: 1.52±0.92pg/ml, the difference has statistical significance (P＜0.001).8.IL-10: maternal surface, the patients suffered from PIH: 2.09±1.46pg/ml, normal pregnancy: 3.08±1.05pg/ml, the difference has statistical significance (P＜0.005); placental fetal surface, PIH: 3.07±0.99pg/ml, normal pregnancy: 3.35±1.24pg/ml, no statistical significance(P＞0.05); peripheral blood, PIH: 3.65±1.09pg/, normal pregnancy: 2.52±0.90pg/ml, the difference has statistical significance (P＜0.001); cord blood, PIH: 2.66±0.58pg/ml, normal pregnancy: 2.71+1.31pg/ml, no statistical significance(P＞0.05).9.IL-12: maternal surface, the patients suffered from PIH: 18.56±7.71pg/ml, normal pregnancy: 16.09±8.19pg/ml, they have no statistical significance(P＞0.05); placental fetal surface, PIH: 10.14±3.69pg/ml, normal pregnancy: 10.40±3.21pg/ml, no statistical significance(P＞0.05); peripheral blood, PIH: 6.57±2.35pg/ml, normal pregnancy: 4.81±0.80pg/ml, the difference has statistical significance (P＜0.05); cord blood, PIH: 4.25±1.72pg/ml, normal pregnancy: 3.73±1.46pg/ml, no statistical significance(P＞0.05).Conclusion:1.ANGâ…¡level of placenta closed with infants in PIH is increased while VEGF level decline. And which lead to chorionic vasoconstriction,Angiogenesis and vascular endothelial cell damaged, blood flow of placental closed with infants reduced.The impacts of fetal blood supply lead to Chronic hypoxia, which is the result of the important causes of IUGR. The increased ANGâ…¡levels of PIH serum can add to PIH. Meanwhile, ANGâ…¡levels of the cord blood are associated with them. The increased VEGF levels of cord blood in PIH are results of fetal hypoxia which stimulate the body to increase production.2.TNF-a,IFN-r,IL-2,IL-12,IL-6,IL-10 are increased and IL-4 is decreased in maternal blood of PIH. TNF-a,IFN-r,IL-2 increased and 1L-10,IL-6 decreasd in placenta closed with pregnant of PIH. IL-6,IFN-r rise in cord blood of PIH.The conclusions suggest that Th1/Th2 imbalance is the main reason of PIH and IUGR.