胎盘合体滋养细胞中硫化氢对激活素A影响的研究

Reguation of Hydrogen Sulfide on Activin A in Human Syncytiotrophoblast Cells

目的:探讨胎盘合体滋养层细胞中硫化氢(H2S)对激活素A(Activin A)的调控作用。方法:利用改良的Klim法原代培养正常胎盘合体滋养层细胞,并给予不同浓度的H2S供体NaHS、前体L-半胱氨酸,H2S合成酶[胱硫醚-β-合成酶(CBS)和胱硫醚-γ-裂解酶(CSE)]抑制剂(AOAA、PAG)和D-半胱氨酸处理细胞,同时利用RNA干扰技术,将CBS和CSE的表达干扰后,再给予L-半胱氨酸处理细胞。ELISA方法检测细胞上清中Activin A的浓度。结果:给予胎盘合体滋养细胞NaSH、L-半胱氨酸处理后,随NaSH、L-半胱氨酸浓度的增加Activin A浓度明显减少(P<0.01),且这种作用被H2S合成酶抑制剂AOAA所阻断(P<0.01)。转染干扰CBS表达的小干扰RNA片段-siCBS后,可逆转L-半胱氨酸对Activin A的下调作用(P<0.01);而加入D-半胱氨酸后,Activin A表达变化无明显差异(P>0.05)。结论:在人胎盘合体滋养细胞中,H2S可能主要在H2S合成酶之一CBS的作用下产生,并发挥对Activin A的下调作用,进而可能在预防妊娠期高血压疾病的发生中发挥作用。

Objective ：To explore the regulation of hydrogen sulfide on Activin A in human syncytiotropho- blast cells. Methods：Normal human syncytiotropho-blast cells were cultivated by improved Klim method, and were treated with different concentrations of NaSH, L-cysteine, CBS, CSE, D-cysteine. Meanwhile RNA inter- ference was used to interfere with the expressions of CBS and CSE, after that the cells were treated with L-cysteine,The expression of Activin A was determined through ELISA. Results：After the treatment of sodi- um hydrosulfide and L-cysteine,Activin A concentration significantly decreased by the increase of sodium hy- drosulfide and L-cysteine concentration （ P 〈 0.01 ）, and this effect was disappearing in the presence of AOAA （P 〈0.01 ）. After suppressing the expression of cystathionine 15-synthase （CBS）and cystathionine γ-lyase （CSE） by RNA interference, the effects of L-cysterine on inhibiting activin A expression was ineffectiveness （P〈0. 01 ）. Adding D-cysteine has no effects on the activin A expression regulation （P 〉0.05）. Conclu- sions.Endogenous H2S may be mainly produced by CBS and may regulate the expression of activin A in human syncytiotrophoblast cells,and consequently prevent the onset of disorders complicating pregnancy.