摘要
目的探讨地塞米松(DEX)对原代培养的早产胎盘绒毛滋养细胞11β-羟基类固醇脱氢酶2(11β-HSD2)表达的影响。方法采用酶消化法和组织块培养法进行早产胎盘原代绒毛滋养细胞培养、纯化和鉴定;用DEX(100nmol/L)模拟两疗程糖皮质激素给药干预原代培养的绒毛滋养细胞,荧光定量PCR和Western blot免疫印迹法检测滋养细胞11β-HSD2mRNA及蛋白表达情况;此外在DEX干预的同时加入RU486(1μmol/L),检测滋养细胞11β-HSD2mRNA和蛋白表达情况。结果DEX干预原代培养滋养细胞,培养1d后11β-HSD2mRNA表达开始上升,第2d和第3d继续升高(P<0.05),第4d改为无药物培养液后表达明显下降,第5d^7d重复给药后再次升高(P<0.05);在DEX干预的同时加入RU486(1μmol/L)共同作用,可见每天滋养细胞11β-HSD2蛋白和mRNA表达下降不显著(P>0.05)。结论重复给予DEX具有刺激原代培养的早产胎盘绒毛滋养细胞增加11β-HSD2蛋白和mRNA表达的作用;故早产胎盘滋养细胞可以通过调节11β-HSD2表达起到保护胎儿的屏障作用。
Objective To study the influence of dexamethasone (DEX) on the expression of 11β-hydroxysteroid dehydrogenase type 2 (11β-HSD2) in primary cultured cytotrophoblasts from human preterm placenta. Methods Placenta villous eytotrophobiasts from preterm birth were cultured with the protocol of enzymatic dissociation and tissue incubation method. After isolation and identification, eytotrophoblasts were treated with the repeated administration of DEX (100nmol/L), or DEX+RU486 (1μmol/L) for 7 days, in which DEX was not added into the culture at 4th day. Cytotrophoblasts were collected everyday, and the expression levels of 11β-HSD2 mRNA and protein were determined by real-time fluorescence quantitative PCR and Western blot method. Results After the treatment of DEX (100 nmol/L), the expression of 11β-HSD2 mRNA and protein in cytotrophoblast increased in the first three days (P〈0.05). At 4th day, 11β-HSD2 mRNA and protein declined in the absence of DEX. In 5th-7th day, the increase of 11β-HSD2 expression were resumed when eytotrophoblasts received DEX again (P〈0. 05). With the treatment of DEX and RU486 (1μmol/L), both mRNA and protein level of 11β-HSD2 in eytotrophoblasts were lower than those with DEX alone, but there was not significantly different (P〉0.05). Conclusion Repeated administration of DEX can upregulate the expression level of 11β-HSD2 in primary cultured cytotrophoblasts from preterm placenta. Cytotrophoblasts from preterm birth may have the ability to protect the infants by the mechanism of 11β-HSD2 regulation.
出处
《四川大学学报(医学版)》
CAS
CSCD
北大核心
2009年第1期116-120,共5页
Journal of Sichuan University(Medical Sciences)