摘要
目的研究全氟辛烷磺酸(perfluorooctane sulfonate,PFOS)对人胎盘合体滋养细胞中糖皮质激素代谢酶11β-羟基类固醇脱氢酶Ⅱ型(11β-HSD2)和胎盘合体滋养细胞凋亡的影响,探讨PFOS对胎儿的影响。方法利用改良的Kliman法分离提纯原代人胎盘滋养层细胞,体外培养3 d,待滋养层细胞完全合体化为合体滋养细胞后,使用不同浓度的PFOS(0、0.001、0.01、0.1、1、10μmol/L)处理细胞,利用实时荧光定量聚合酶链反应(qRT-PCR)、免疫印迹等方法检测PFOS对胎盘合体滋养细胞11β-HSD2的mRNA和蛋白质表达水平,以及酶活性的影响,检测PFOS对天冬氨酸蛋白水解酶3(caspase3)蛋白质表达水平和酶活性的影响。结果体外培养第1、2、3天,胎盘滋养层细胞中11β-HSD2的mRNA和蛋白质相对表达量逐渐升高,与未处理时(即第0天)比较的差异具有统计学意义(P值均<0.01)。0.01μmol/L PFOS处理的人胎盘合体滋养细胞11β-HSD2的mRNA和蛋白质相对表达量较未处理时显著降低,且11β-HSD2的mRNA和蛋白质相对表达量随着PFOS剂量的增加(0.1、1、10μmol/L)显著降低(P值均<0.01)。0.01μmol/L PFOS处理的人胎盘合体滋养细胞11β-HSD2酶活性较未处理时显著降低,并随着PFOS剂量的增加,活性抑制程度显著增强(P值均<0.01)。0.01μmol/L PFOS处理的人胎盘合体滋养细胞pro-caspase3蛋白质相对表达量较未处理时显著降低,且蛋白质相对表达量随着PFOS剂量的增加(0.1、1、10μmol/L)显著降低(P值均<0.01)。同时,0.01μmol/L PFOS处理的人胎盘合体滋养细胞cleaved-caspase3的蛋白质相对表达量较未处理时显著升高,并随着PFOS剂量的增加显著升高(P值均<0.01)。进一步实验结果显示,0.01μmol/L PFOS处理的人胎盘合体滋养细胞caspase3的酶活性较未处理时显著增强,并随着PFOS剂量的增加活性显著增强(P值均<0.05)。结论PFOS能够呈浓度依赖性地抑制人胎盘合体滋养细胞中11β-HSD2的表达和酶活性,促进胎盘合体滋养细胞凋亡,这可能是PFOS导致病理妊娠,对胎儿产生不良影响的重要机制之一。
Objective To study the effect of perfluorooctane sulfonic acid(PFOS)on 11-beta-hydroxysteroid dehydrogenase type 2(11β-HSD2)which is responsible for human placental glucocorticoid metabolism in syncytiotrophoblasts and apoptosis of placental syncytiotrophoblasts,and to discuss the effects of PFOS on pregnancy and fetus.Methods Primary human placental cytotrophoblast cells were prepared using a modified Kliman method and cultured for 3 days in vitro.After syncytialization,the levels of 11β-HSD2 and apoptosis-related protein caspase3 were examined by quantitative real-time(qRT)-polymerase chain reaction(PCR)and western blotting after treatment with PFOS(0,0.001,0.01,0.1,1,and 10μmol/L).Results The mRNA and protein expression levels of 11β-HSD2 in placental trophoblast cells increased gradually on day 1,2 and 3 of in vitro culture,which were significantly different from those on day 0(P<0.01).The mRNA and protein expression levels of 11β-HSD2 in human placental syncytiotrophoblasts treated with 0.01μmol/L PFOS were significantly lower than those in untreated group,and the mRNA and protein expression levels of 11β-HSD2 were gradually decreased with the increase of PFOS dose(0.1,1 and 10μmol/L,all P<0.01).The activity of 11β-HSD2 in human placental syncytiotrophoblasts treated with 0.01μmol/L PFOS was significantly lower than that in the untreated group,and the inhibition degree was significantly increased with the increase of PFOS dose(all P<0.01).The expression level of pro-caspase3 protein in human placental syncytiotrophoblasts treated with 0.01μmol/L PFOS was significantly lower than that in untreated group,and the protein expression level was gradually decreased with the increase of PFOS dose(0.1,1 and 10μmol/L,all P<0.01).But the expression level of cleaved-caspase3 protein in human placental syncytiotrophoblasts treated with 0.01μmol/L PFOS was significantly higher than that in the untreated group,and it was increased gradually with the increase of PFOS dose(all P<0.01).Further experiments showed that the activity of caspase3 in 0.01μmol/L PFOS treated human placental syncytiotrophoblasts was significantly increased compared with that in untreated group,and the activity was significantly increased with the increase of PFOS dose(all P<0.05).Conclusion PFOS can inhibit the expression and enzyme activity of 11β-HSD2 and increase the apoptosis of human placental syncytiotrophoblasts in a concentration-dependent manner,which may be one of the important mechanisms that PFOS leads to pathological pregnancy and has adverse effects on the fetus.
作者
张楠
王育
程山山
ZHANG Nan;WANG Yu;CHENG Shanshan(Department of Obstetrics and Gynecology,Renji Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200120,China;不详)
出处
《上海医学》
CAS
2022年第6期380-386,共7页
Shanghai Medical Journal
基金
国家自然科学基金(82072866、82102856)。
