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母亲哮喘对胎盘和脐带血蛋白质谱的影响。

The effect of maternal asthma on placental and cord blood protein profiles.

作者信息

Murphy Vanessa E, Johnson Renee F, Wang Yung-Chih, Akinsanya Karen, Gibson Peter G, Smith Roger, Clifton Vicki L

机构信息

Mothers and Babies Research Centre, Hunter Medical Research Institute, Newcastle NSW, Australia.

出版信息

J Soc Gynecol Investig. 2005 Jul;12(5):349-55. doi: 10.1016/j.jsgi.2005.01.024.

Abstract

OBJECTIVE

We conducted a comparative proteomic analysis of placental and umbilical cord blood proteins using surface-enhanced laser desorption ionization-time of flight mass spectrometry (SELDI-TOF MS) to examine the associations among asthma, fetal gender, and protein profiles.

METHODS

Placental tissue and umbilical vein plasma were collected from 10 healthy and 20 asthmatic women. Placental proteins were extracted using phosphate-buffered saline containing protease inhibitors. Samples were applied to the surfaces of strong anion exchange (SAX2), weak cation exchange (WCX2) and immobilized metal affinity capture (IMAC-Cu(2+)) chips. Mass analysis was conducted using a Ciphergen Protein Biology System IIc (Freemont, CA), and differences in individual peak intensities between groups were determined.

RESULTS

Fourteen placental peaks were significantly different between asthmatic and non-asthmatic women (seven more highly expressed and seven less highly expressed). Ten umbilical cord blood peak differences were identified, with four peaks more highly expressed and six peaks less highly expressed in asthmatics. Four placental and three umbilical cord blood proteins differed significantly between male and female fetuses. Two placental and five umbilical cord blood peaks were specifically increased in a subgroup of samples collected from asthmatic women who did not use inhaled glucocorticoids and were pregnant with a female fetus, a group previously found to have altered placental function.

CONCLUSIONS

This study demonstrates the abilities of the SELDI technique as a tool for protein profiling in tissue or plasma. Further work to positively identify the candidate peptides found in this study may provide a greater understanding of the placental mechanisms leading to alterations in fetal growth in patients with bronchial asthma.

摘要

目的

我们使用表面增强激光解吸电离飞行时间质谱(SELDI-TOF MS)对胎盘和脐带血蛋白质进行了比较蛋白质组学分析,以研究哮喘、胎儿性别与蛋白质谱之间的关联。

方法

从10名健康女性和20名哮喘女性中收集胎盘组织和脐静脉血浆。使用含有蛋白酶抑制剂的磷酸盐缓冲盐水提取胎盘蛋白质。将样品应用于强阴离子交换(SAX2)、弱阳离子交换(WCX2)和固定化金属亲和捕获(IMAC-Cu(2+))芯片表面。使用Ciphergen Protein Biology System IIc(加利福尼亚州弗里蒙特)进行质谱分析,并确定组间个体峰强度的差异。

结果

哮喘女性和非哮喘女性之间有14个胎盘峰存在显著差异(7个高表达,7个低表达)。在脐带血中鉴定出10个峰差异,哮喘患者中有4个峰高表达,6个峰低表达。男性和女性胎儿之间有4种胎盘蛋白和3种脐带血蛋白存在显著差异。在未使用吸入性糖皮质激素且怀有女性胎儿的哮喘女性亚组中,有2个胎盘峰和五个脐带血峰特别升高,该亚组先前被发现胎盘功能发生改变。

结论

本研究证明了SELDI技术作为组织或血浆中蛋白质谱分析工具的能力。进一步对本研究中发现的候选肽进行阳性鉴定的工作可能会更深入地了解导致支气管哮喘患者胎儿生长改变的胎盘机制。

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