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目的研究子痫前期患者胎盘滋养细胞基质金属蛋白酶(MMP)9、2及其组织抑制物(TIMP)1、2,肿瘤转移抑制基因KiSS-1 mRNA、蛋白的表达变化及其与子痫前期发病的关系。方法采用RT-PCR、免疫印迹(western blot)法对30例正常足月妊娠妇女(正常妊娠组)和10例妊娠期高血压患者(高血压组)及27例子痫前期(子痫前期组)患者胎盘滋养细胞中MMP-9、MMP-2、KiSS-1、TIMP-1和TIMP-2基因mRNA及蛋白表达水平[均以相对吸光度(A)表示]进行检测;应用明胶酶谱分析法,检测3组妇女胎盘孵育液MMP-9、MMP-2活性。结果(1)胎盘滋养细胞浸润相关基因mRNA表达水平:子痫前期组MMP-9、MMP-2 mRNA表达水平分别为0．39±0．05和0．71±0．16,均明显低于正常妊娠组的0．78±0．11和1．63±0．31,两组分别比较,差异有统计学意义(P均<0．05)。高血压组MMP-9 mRNA的表达水平明显高于重度子痫前期患者,差异有统计学意义(P<0．05)。子痫前期组胎盘滋养细胞KiSS-1 mRNA和TIMP-1 mRNA的表达水平分别为1．97±0．21和1．11±0．18,均明显高于正常妊娠组的0．69±0．27和0．65±0．19,差异均有统计学意义(P<0．05);高血压组胎盘滋养细胞KiSS-1 mRNA表达水平低于子痫前期组,但与正常妊娠组比较,差异无统计学意义(P>0．05)。重度子痫前期患者胎盘滋养细胞TIMP-2 mRNA表达水平明显高于正常妊娠组,差异均有统计学意义(P<0．05)。(2)胎盘滋养细胞浸润相关基因的蛋白表达水平:子痫前期组MMP-9、MMP-2基因的蛋白表达水平分别为1．07±0．35和0．74±0．23,均明显低于正常妊娠组的2．43±0．92和1．48±0．78,差异均有统计学意义(P<0．05)。子痫前期组胎盘KISS-1和TIMP-1蛋白表达水平分别为2．46±0．39和1．51±0．40,均明显高于正常妊娠组的0．91±0．35和0．93±0．56,差异均有统计学意义(P<0．05)。子痫前期组胎盘滋养细胞TIMP-2蛋白表达水平与正常妊娠组及高血压组比较,差异均无统计学意义(P>0．05)。(3)胎盘MMP-9和MMP-2酶比活性:子痫前期组分别为(2．67±0．53)和(1．13±0．28)灰度·g-1·L-1,均明显低于正常妊娠组的(8．44±3．70)和(3．87±1．43)灰度·g-1·L-1,差异均有统计学意义(P<0．05)。结论子痫前期患者胎盘滋养细胞促进浸润基因。MMP-9、MMP-2表达降低和抑制浸润基因KiSS-1和TIMP-1表达升高,可能在子痫前期胎盘缺血缺氧中起重要作用。
Objective To investigate the expression of matrix metalloproteinase (MMP) 9,2 and its tissue inhibitor of metalloproteinase (TIMP) 1,2 and KiSS-1 mRNA and protein in placental trophoblast cells of preeclampsia and its relationship with preeclampsia Relationship. Methods Thirty normal full-term pregnant women (normal pregnancy group) and 10 pregnant women with hypertensive disorder (hypertension group) and 27 preeclampsia (preeclampsia group) were treated with RT-PCR and western blot. The mRNA and protein expression of MMP-9, MMP-2, KiSS-1, TIMP-1 and TIMP-2 in placental trophoblast cells were detected by the relative absorbance (A) The activity of MMP-9 and MMP-2 in the placenta incubation solution of 3 women were detected. Results (1) The mRNA expression levels of placental trophoblast invasion related genes: The levels of MMP-9 and MMP-2 mRNA in preeclampsia were 0.39 ± 0.05 and 0.71 ± 0.16, respectively, The normal pregnancy group 0.78 ± 0.11 and 1.63 ± 0.31, respectively, the difference between the two groups was statistically significant (P all <0.05). The expression of MMP-9 mRNA in hypertension group was significantly higher than that in severe preeclampsia, the difference was statistically significant (P <0.05). The expression of KiSS-1 mRNA and TIMP-1 mRNA in placental trophoblast cells of preeclampsia were 1.97 ± 0.21 and 1.11 ± 0.18, respectively, which were significantly higher than those in normal pregnancy group (0.69 ± 0 .27 and 0.65 ± 0.19, respectively (all P <0.05). The expression of KiSS-1 mRNA in placental trophoblast cells in hypertension group was lower than that in preeclampsia group, but compared with normal pregnancy group , The difference was not statistically significant (P> 0.05). The expression of TIMP-2 mRNA in placental trophoblast cells in patients with severe preeclampsia was significantly higher than that in normal pregnancy (P <0.05). (2) The protein expression levels of placental trophoblast invasion related genes: The protein expression levels of MMP-9 and MMP-2 in preeclampsia were 1.07 ± 0.35 and 0.74 ± 0.23, respectively Which was lower than that of normal pregnancy group (2.43 ± 0.92 and 1.48 ± 0.78, P <0.05). The expression levels of KISS-1 and TIMP-1 in preeclampsia placenta were 2.46 ± 0.39 and 1.51 ± 0.40, respectively, which were significantly higher than those in normal pregnancy group (0.91 ± 0.35 and 0 .93 ± 0.56, the difference was statistically significant (P <0.05). The expression of TIMP-2 protein in placental trophoblast cells in preeclampsia group was not significantly different from that in normal pregnancy group and hypertension group (P> 0.05). (3) The specific activity of MMP-9 and MMP-2 in the placenta: (2.67 ± 0.53) and (1.13 ± 0.28) grayscale · g-1 · L-1 (8.44 ± 3.70) and (3.87 ± 1.43) grayscale · g-1 · L-1 in normal pregnancy group, respectively, with statistical significance (P <0. 05). Conclusion Placental trophoblasts in patients with preeclampsia promote invasion of genes. Decreased expression of MMP-9 and MMP-2 and inhibition of the expression of infiltrating genes KiSS-1 and TIMP-1 may play an important role in placental ischemia and hypoxia.