子痫前期患者滋养细胞浸润相关基因mRNA及蛋白表达的研究

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。

Expression of trophoblast invasion related genes mRNA and protein in human placenta in preeclampsia

OBJECTIVE: To study the mRNA and protein expression of matrix metalloproteinase (MMP) 9, 2, metastasis suppressor gene KiSS-1, tissue inhibitor of metalloproteinase-1 (TIMP-1) and TIMP-2 in placenta of preeclampsia patients and their relation to the pathogenesis of preeclampsia. METHODS: Expression of MMP-9, MMP-2, KiSS-1, TIMP-1, TIMP-2 mRNAs and proteins in placenta from 27 cases of preeclampsia, 10 cases of gestational hypertension and 30 cases of normal term pregnant women was detected by semi-quantitative RT-PCR, western blotting, and enzyme activity of MMP-9 and MMP-2 was measured by gelatin zymography. RESULTS: (1) Expression of MMP-9 and MMP-2 mRNA in placenta of preeclampsia (0.39 +/- 0.05 and 0.71 +/- 0.16) was significantly lower than that in normal term pregnancy (0.78 +/- 0.11 and 1.63 +/- 0.31, P < 0.05). Expression of KiSS-1 and TIMP-1 mRNAs in placenta of preeclampsia (1.97 +/- 0.21 and 1.11 +/- 0.18) was significantly higher than that in normal term pregnancy (0.69 +/- 0.27 and 0.65 +/- 0.19) (P < 0.05.). There was no significant difference in TIMP-2 mRNA level between preeclampsia and normal pregnancy (P > 0.05). (2) Expression of MMP-9 and MMP-2 proteins in preeclampsia (1.07 +/- 0.35 and 0.74 +/- 0.23) was significantly lower than that in normal term pregnancy (2.43 +/- 0.92 and 1.48 +/- 0.78) (P < 0.05). Expression of KiSS-1 and TIMP-1 proteins in placenta of preeclampsia (2.46 +/- 0.39 and 1.51 +/- 0.40) was significantly higher than that in normal pregnancy (0.91 +/- 0.35 and 0.93 +/- 0.56) (P < 0.05). There was no significant difference in TIMP-2 protein level between preeclampsia and normal pregnancy (P > 0.05). (3) Enzyme activity of MMP-9 and MMP-2 in placenta of preeclampsia [(2.67 +/- 0.53) gray level.g(-1).L(-1) and (1.13 +/- 0.28) gray level.g(-1).L(-1))] was significantly lower than that in placenta of normal pregnancy [(8.44 +/- 3.70) gray level.g(-1).L(-1) and (3.87 +/- 1.43) gray level.g(-1).L(-1)] (P < 0.05). CONCLUSION: Abnormal expression of MMP-9, MMP-2, KiSS-1 and TIMP-1 can cause insufficiency invasion of trophoblast in preeclampsia and superficial placentation, which may play an important role in the pathogenesis and development of preeclampsia.