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目的探讨胎盘滋养细胞中KiSS-1基因及基质金属蛋白酶(MMP)9在子痫前期发病中的作用及其与新生儿预后的相关性。方法采用RT-PCR和western blot(蛋白印迹法)检测40例子痫前期患者(其中轻度15例、重度25例,子痫前期组)和20例正常足月正常妊娠妇女(正常妊娠组)胎盘滋养细胞中KiSS-1mRNA和MMP-9mRNA及其蛋白的表达,并与其临床症状及其新生儿围产结局进行相关性分析。结果(1)子痫前期组滋养细胞中KiSS-1mRNA及其蛋白表达水平分别为1.73±0.24和(78.4±8.0)μg/100μg总蛋白,其中轻度患者为1.50±0.15和(72.4±6.9)μg/100μg总蛋白,重度患者为1.87±0.20和(83.5±3.6)μg/100μg总蛋白;均显著高于正常妊娠组的1.24±0.25和(63.4±2.7)μg/100μg总蛋白(P<0.01)。(2)子痫前期组MMP-9mRNA及其蛋白表达水平分别为0.09±0.06和(9.6±4.3)μg/100μg总蛋白,其中轻度患者为0.11±0.08和(10.0±3.2)μg/100μg总蛋白,重度患者为0.07±0.05和(7.8±2.0)μg/100μg总蛋白;均显著低于正常妊娠组的0.17±0.10和(17.9±7.3)μg/100μg总蛋白(P<0.01)。(3)子痫前期组KiSS-1mRNA及其蛋白表达水平与中心动脉压(MAP)和24h尿蛋白定量呈正相关,r分别为0.610(P=0.023)、0.713(P=0.011)和0.397(P=0.003)、0.638(P=0.002);与有、无眼底动脉痉挛呈显著正相关,r分别为0.499(P=0.000)和0.511(P=0.000)。子痫前期组MMP-9mRNA及其蛋白表达水平则与MAP、24h尿蛋白定量及有、无眼底动脉痉挛呈显著负相关,r分别为0.561(P=0.042)、0.571(P=0.022)、0.275(P=0.039)、0.375(P=0.048)、0.346(P=0.001)、0.543(P=0.000)。(4)子痫前期组MMP-9mRNA及其蛋白表达水平与新生儿体重呈显著正相关,r分别为0.651(P=0.000)和0.544(P=0.004);KiSS-1mRNA及其蛋白表达水平与其呈显著负相关,r分别为0.759(P=0.000)和0.865(P=0.000)。(5)随着新生儿窒息程度加重,MMP-9mRNA及其蛋白表达水平呈降低趋势,KiSS-1mRNA及其蛋白表达水平呈升高趋势(P均<0.05)。结论MMP-9和KiSS-1表达失衡在子痫前期发病中起重要作用,且与新生儿预后密切相关。
Objective To investigate the role of KiSS-1 gene and matrix metalloproteinase (MMP-9) in the pathogenesis of preeclampsia in placental trophoblast and its relationship with the prognosis of neonates. Methods Forty pregnant women with preeclampsia (mild 15, severe 25, preeclampsia) and 20 normal pregnant women (normal pregnancy group) were tested by RT-PCR and western blot (Western blot) The expression of KiSS-1 mRNA and MMP-9 mRNA and their proteins in trophoblast cells were analyzed with correlation with clinical symptoms and perinatal outcome of neonates. Results (1) KiSS-1mRNA and protein expression level of trophoblast cells in preeclampsia group were 1.73 ± 0.24 and (78.4 ± 8.0) μg / 100μg total protein respectively, with mild cases being 1.50 ± 0.15 and (72.4 ± 6.9) μg / 100μg total protein in severe patients were 1.87 ± 0.20 and (83.5 ± 3.6) μg / 100μg total protein, respectively, which were significantly higher than that of normal pregnancy group (1.24 ± 0.25 and 63.4 ± 2.7μg / 100μg total protein, P <0.01 ). (2) The levels of MMP-9 mRNA and protein in preeclampsia group were 0.09 ± 0.06 and (9.6 ± 4.3) μg / 100μg total protein, respectively, and the mild patients were 0.11 ± 0.08 and (10.0 ± 3.2) μg / 100μg total Protein, severe patients were 0.07 ± 0.05 and (7.8 ± 2.0) μg / 100μg total protein, respectively, which were significantly lower than that of the normal pregnancy group (0.17 ± 0.10 and 17.9 ± 7.3μg / 100μg total protein, P <0.01). (3) KiSS-1mRNA and protein expression in preeclampsia group were positively correlated with MAP and 24h urinary protein, with r values of 0.610 (P = 0.023), 0.713 (P = 0.011) and 0.397 = 0.003), and 0.638 (P = 0.002). There was a significant positive correlation with and without retinal artery spasm, with r values of 0.499 (P = 0.000) and 0.511 (P = 0.000), respectively. The levels of MMP-9mRNA and protein in preeclampsia group were significantly negatively correlated with the quantitative, and without retinal arterial spasm of MAP, 24h, r = 0.561 (P = 0.042), 0.571 (P = 0.039), 0.375 (P = 0.048), 0.346 (P = 0.001), 0.543 (P = 0.000). (4) There was a significant positive correlation between MMP-9 mRNA and protein expression in preeclampsia and neonatal body weight (r = 0.651, P = 0.000) and 0.544 (P = 0.004) There was a significant negative correlation with r = 0.759 (P = 0.000) and 0.865 (P = 0.000). (5) As neonatal asphyxia increased, the expression of MMP-9mRNA and its protein decreased, while the expression of KiSS-1mRNA and its protein increased (all P <0.05). Conclusion The imbalance of MMP-9 and KiSS-1 expression plays an important role in the pathogenesis of preeclampsia and is closely related to the prognosis of neonates.