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目的研究邻苯二甲酸二(2-乙基己)酯[di-(2-ethylexyl)phthalate,DEHP]对人原代培养早孕绒毛滋养细胞浸润能力及基质金属蛋白酶(MMP)-2、MMP-9、基质金属蛋白酶抑制因子(TIMP)-1和TIMP-2表达的影响,探讨DEHP对妊娠的影响及其毒性机制。方法2006年12月于北京军区总医院采用不同浓度DEHP预处理人原代培养早孕绒毛滋养细胞,采用Transwell体外浸润实验检测滋养细胞浸润能力的改变。RT-PCR法检测滋养层细胞侵袭性相关基因MMP-2、MMP-9、TIMP-1和TIMP-2的mRNA表达,Western blot法检测MMP-2、MMP-9、TIMP-1和TIMP-2蛋白表达水平。结果DEHP作用后,滋养细胞浸润能力明显下降。当DEEP为50、100μmol/L时,MMP-2、MMP-9、TIMP-1的mRNA表达量分别为0.35±0.07、0.26±0.03、0.97±0.18和0.30±0.10、0.20±0.05、1.02±0.20,与DEEP为0时MMP-2、MMP-9、TIMP-1的mRNA的表达量(0.77±0.15、0.45±0.04、0.80±0.20)比较P<0.05;当DEEP为50、100μmol/L时,MMP-2、MMP-9、TIMP-1的蛋白表达量分别为0.35±0.03、0.23±0.05、0.69±0.08和0.24±0.02、0.17±0.02、0.86±0.10,与DEEP为0时MMP-2、MMP-9、TIMP-1的mR-NA的表达量(0.69±0.04、0.57±0.03、0.24±0.12)比较P<0.05。可见DEHP剂量≥50μmol/L时,滋养细胞MMP-2和MMP-9表达下降、TIMP-1表达增加,且呈剂量依赖性。结论DEHP可通过抑制MMP-2、MMP-9的表达并促进TIMP-1表达,影响滋养细胞浸润能力。
Objective To investigate the effect of di- (2-ethylexyl) phthalate (DEHP) on the trophoblast cell infiltration and matrix metalloproteinase (MMP) -2 and MMP- 9, the expression of matrix metalloproteinase inhibitor (TIMP) -1 and TIMP-2, to investigate the effect of DEHP on pregnancy and its toxic mechanism. Methods Preincubation of human trophoblasts in early pregnancy with different concentrations of DEHP was performed in Beijing Military Region General Hospital in December 2006. Transwell invasion assay was used to detect the changes of trophoblast invasion. The expression of MMP-2, MMP-9, TIMP-1 and TIMP-2 in trophoblastic cells was detected by RT-PCR and the expressions of MMP-2, MMP-9, TIMP-1 and TIMP- Protein expression levels. Results After DEHP treatment, the ability of trophoblast invasion decreased significantly. The mRNA expression levels of MMP-2, MMP-9 and TIMP-1 at the concentrations of 50,100μmol / L DEEP were 0.35 ± 0.07,0.26 ± 0.03,0.97 ± 0.18 and 0.30 ± 0.10,0.20 ± 0.05,1.02 ± 0.20 , P <0.05 compared with the expression of MMP-2, MMP-9 and TIMP-1 mRNA when DEEP was 0 (0.77 ± 0.15,0.45 ± 0.04,0.80 ± 0.20); when DEEP was 50 and 100μmol / L, The protein expression of MMP-2, MMP-9 and TIMP-1 were 0.35 ± 0.03, 0.23 ± 0.05, 0.69 ± 0.08 and 0.24 ± 0.02, 0.17 ± 0.02 and 0.86 ± 0.10, respectively. The expression levels of mR-NA of MMP-9 and TIMP-1 (0.69 ± 0.04, 0.57 ± 0.03 and 0.24 ± 0.12) were compared with P <0.05. It can be seen that when the dosage of DEHP≥50μmol / L, the expression of MMP-2 and MMP-9 in trophoblast cells decreased, and the expression of TIMP-1 increased in a dose-dependent manner. Conclusion DEHP can reduce the expression of MMP-2 and MMP-9, promote the expression of TIMP-1 and affect the ability of trophoblast invasion.