| PurposeSevere preeclampsia(SPE) is considered as a severe pregnancycomplication secondary to placental hypoxia. It is performed ashypertension, proteinuria and multiple tissue paralysis after20weeks ofpregnancy, which poses a grave threat to the life and health of gravidaand fetus. As an intermediate filament cytoskeleton protein, Nestin is amarker of neural stem cell, and participates in the development of nervecells. It also plays an important role in vasculogenesis and vascularmaintenance in hypoxia environment.In this study, hypoxia inducer was used to create an in vitro cellhypoxia environment, and the expression of Nestin in two differentembryonic stem cells: JEG-3cells and human umbilical vein endothelialcell (HUVEC) under different hypoxia times and degrees of hypoxia wasobserved to provide an evidence for the effect of placental hypoxia inSPE occurrence and development.Materials and MethodsCobalt dichloride (CoCl2) was used to mimic a cell hypoxia in vitroto induce JEG-3and HUVEC chemical hypoxia respectively. The mRNAof cells under different hypoxia conditions were extracted to test theexpression level of Nestin gene at the nucleic acid level. Meanwhile, cellculture medium of each group was collected, and the expression level ofNestin protein in cell culture media was tested using Nestin ELISA kitand statistical analysised. Results1． Expression of Nestin mRNA and protein in JEG-3cells underhypoxia environment.1).The Nestin mRNA expression of JEG-3cells cultured underchemical hypoxia with300umol/L CoCl2for12h,24h and48hrespectively were statistically significantly higher than the expressionwhen they were cultured for6h (P<0.01). Compared with under thenormal oxygen condition, the Nestin mRNA expression under24hhypoxia with300umol/L CoCl2was up-regulated significantly (P<0.05).The Nestin protein level in culture media gradually decreased with theextension of hypoxia duration, and the difference in Nestin protein levelswas statistically significant between cultured for6h and48h (P<0.05).Compared with the normal oxygen condition, the Nestin expression under24h hypoxia with300umol/L CoCl2was decreased without statisticallysignificant (P>0.05).2).Under24h CoCl2induced hypoxia, there was no significantdifferencein Nestin mRNA expression with the increasing of CoCl2concentration in JEG-3cells (P>0.05). There was no significantdifference in the Nestin protein level in culture media with the increase ofCoCl2(P>0.05).2． Expression of Nestin mRNA and protein in HUVEC underhypoxia environment.1).The Nestin mRNA expression of HUVEC cultured underchemical hypoxia with300umol/L CoCl2for12h,24h and48hrespectively were statistically significantly higher than the expressionwhen they were cultured for6h (P<0.01). Compared with under thenormal oxygen condition, the Nestin mRNA expression under24hhypoxia with300umol/L CoCl2was up-regulated significantly (P<0.05). there was no obvious change of Nestin protein concentration with theextension of hypoxia duration (P>0.05). Compared with the normaloxygen condition, the Nestin expression under24h hypoxia with300umol/L CoCl2was decreased without statistically significant (P>0.05).2). Under24h CoCl2-induced hypoxia, the Nestin mRNA expressionin HUVEC under24h hypoxia with400umol/L CoCl2was significantlyhigher than the expression with CoCl2concentrations of200umol/L and600umol/L (P<0.05; P<0.01), and the Nestin mRNA expression under600umol/L CoCl2was significantly lower than that under200umol/LCoCl2(P<0.01). and there was no obvious change of Nestin proteinconcentration (P>0.05).Conclusion1）. A short time chronic hypoxia can enhance the expression ofNestin mRNA in trophoblastic cells and placental vascular endothelialcells， and it may be related with the protective mechanism of theorganism in response to hypoxia in the progress of preeclampsia.2）. The extension of the chronic hypoxia time (within certain scope)is a negative factor influencing the expression of Nestin in trophoblasticcells and it may be related with shallow placental implantation anddysfunction of vascular remodeling in the preeclampsia patients.3）. The extension of the chronic hypoxia time (within certain scope)plays a supportive role in the long time expression of Nestin in placentalvascular endothelial cells and its cytoprotection function.4）. The extent of chronic hypoxia (within certain scope) has a weakinfluence on the expression of Nestin in trophoblastic cells. A slightaggravating of chronic hypoxia will enhance the expression of NestinmRNA in placental vascular endothelial cells. However, an obviousaggravating of hypoxia may be a negative factor limiting the expression of Nestin mRNA in placental vascular endothelial cells. |
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