Optimization of trophoblast cell model in preeclampsia induced by hypoxia in vitro and metabonomics analysis

Yang Xiaotao; Han Dingli; Philip N.Baker; Qi Hongbo; Tong Chao; Yang Zhu

doi:10.13406/j.cnki.cyxb.001343

您当前的位置：

首页 >

文章列表页 >

Optimization of trophoblast cell model in preeclampsia induced by hypoxia in vitro and metabonomics analysis

更新时间：2024-01-17

- Optimization of trophoblast cell model in preeclampsia induced by hypoxia in vitro and metabonomics analysis
- Journal of Chongqing Medical University Vol. 42, Issue 8, Pages: 1045-1052(2017)
- 作者机构：
  
  重庆医科大学附属第二医院妇产科重庆医科大学"中国-加拿大-新西兰"联合母胎医学实验室
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.001343
  CLC： R714.244
- Published：2017，
扫描看全文
[1]杨晓涛,韩顶立,Philip N.Baker,漆洪波,童超,杨竹.体外缺氧诱导子痫前期滋养细胞模型优化及代谢组学鉴定[J].重庆医科大学学报,2017,42(08):1045-1052.

Yang Xiaotao, Han Dingli, Philip N.Baker, et al. Optimization of trophoblast cell model in preeclampsia induced by hypoxia in vitro and metabonomics analysis. [J]. Journal of Chongqing Medical University 42(8):1045-1052(2017)
[1]杨晓涛,韩顶立,Philip N.Baker,漆洪波,童超,杨竹.体外缺氧诱导子痫前期滋养细胞模型优化及代谢组学鉴定[J].重庆医科大学学报,2017,42(08):1045-1052. DOI： 10.13406/j.cnki.cyxb.001343.

Yang Xiaotao, Han Dingli, Philip N.Baker, et al. Optimization of trophoblast cell model in preeclampsia induced by hypoxia in vitro and metabonomics analysis. [J]. Journal of Chongqing Medical University 42(8):1045-1052(2017) DOI： 10.13406/j.cnki.cyxb.001343.

摘要

目的:探讨滋养细胞体外缺氧不同处理方法模拟子痫前期模型的优化研究。方法:HTR8/SVneo滋养细胞分别在1%O2（缺氧环境）和21%O2（常氧环境）培养箱中培养1 h（记为H1、N1）或2 h（记为H2、N2）

缺氧1 h后复氧1 h（记为H1R1）和缺氧2 h后复氧2 h（记为H2R2）

H1R1后再缺氧1 h（记为H1R1H1）

H2R2后再缺氧2 h（记为H2R2H2）

H1R12个循环（记为H1R1H1R1）和H2R22个循环（记为H2R2H2R2）

缺氧2 h后复氧6 h（记为H2R6）

持续缺氧24 h

以及持续常氧4 h（记为N4）、8 h（记为N8）和24 h（记为Normoxia）

然后提取蛋白以Western blot检测腺苷酸活化蛋白激酶（AMP-activated protein kinase

AMPK）磷酸化水平

以验证各组处理对于诱导滋养细胞能量应激的效果;HTR8/SVneo滋养细胞在常氧或者缺氧环境培养24 h后经气相质谱技术（gas chromatography mass spectrum

GS-MS）进行细胞代谢组学分析

以验证AMPK磷酸化水平上调是否与滋养细胞代谢组变化具有一致性。结果:缺氧组AMPK的活性（1.615±0.111）明显高于常氧组（1.000±0.107）和缺氧复氧组（1.277±0.113）;缺氧时显著增加的代谢物有4-甲基-2-戊酮酸（log22.597为1.377）、水合乙醛酸（log22.483为1.312）、组氨酸（log21.188为0.248）、苯丙氨酸（log21.262为0.335）、缬氨酸（log21.518为0.602）、正亮氨酸（log21.519为0.603）、乙酰丝氨酸（log21.691为0.758）、丝氨酸（log21.783为0.834）、半胱氨酸（log21.851为0.889）、蛋氨酸（log22.072为1.051）、鸟氨酸（log22.251为1.170）等;而棕榈反油酸（log20.127为-2.983）、11

17-廿碳三烯酸（log20.334为-1.583）、二十四单烯酸（log20.600为-0.738）、共轭亚油酸（log20.680为-0.557）、顺式十八碳烯酸（log20.711为-0.492）、9-十七碳烯酸（log20.782为-0.355）、二十二碳六烯酸（log20.829为-0.271）、二十碳五烯酸（log20.841为-0.250）、芥酸（log20.844为-0.244）、二十二碳五烯酸（log20.898为-0.156）、柠檬酸（log20.279为-1.842）、苹果酸（log20.208为-2.264）、琥珀酸（log20.254为-1.980）、顺乌头酸（log20.260为-1.946）、β-柠檬酸-左旋谷氨酸（log20.093为-3.430）、β-丙氨酸（log20.139为-2.851）、胱硫醚（log20.267为-1.904）、顺式-4-羟脯氨酸（log20.500为-1.000）等在缺氧时显著降低。代谢途径结果分析显示缺氧时滋养细胞中核苷酸代谢[log2（1.811、1.149）分别为0.857、0.201]、能量代谢[log2（1.510、1.173、1.149）分别为0.595、0.230、0.201]、维生素代谢[log2（1.045、1.052、1.125）分别为0.064、0.073、0.170]、氨基酸代谢[log2（1.245、1.020、1.027、1.123、1.127、1.076）分别为0.316、0.028、0.039、0.167、0.173、0.106]、信号转导（log21.046为0.065）、蛋白翻译（log21.026为0.037）等途径被激活

而碳水化合物[log2（0.857、0.857、0.799）分别为-0.222、-0.222、-0.323]、脂肪酸[log2（0.944、0.912、0.826）分别为-0.083、-0.133、-0.276]、内分泌代谢[log2（0.885、0.799、0.799）分别为-0.176、-0.323、-0.323]和其他第二代谢物生物合成[log2（0.947、0.871、0.743）分别为-0.079、-0.199、-0.428]等代谢途径显著受到抑制。结论:单纯缺氧模型更适合用于子痫前期病理生理机制的体外研究。

Abstract

Objective:To investigate the optimal in vitro hypoxia cell mode for preeclampsia study. Methods:Western blot was used to determine the phosphorylation of AMPK after HTR8/SVneo trophoblast cell being treated with 1% O2(hypoxia condition) or21% O2(normoxia condition)for various duration. GC-MS was used for assessing metabolome of HTR8/SVneo trophoblast cell after being incubated in normoxia or hypoxia environment

to validate the synchronous alteration in AMPK phosphorylation and metabolome. Results:The activity of AMPK was higher in hypoxia group(1.615 ±0.111) than in normoxia group(1.000 ±0.107) and hypoxia/reoxygenation group(1.277 ±0.113). The in-creased metabonomics of the HTR8/SVneo trophoblast cell induced by hypoxia were 4-methyl-2-oxopentanoic acid(log22.597 =1.377)

glyoxylic acid(log22.483=1.312)

histidine(log21.188 =0.248)

phenylalanine(log21.262 =0.335)

valine(log21.518 =0.602)

norleucine(log21.519=0.603)

O-aacetylserine(log21.691=0.758)

serine(log21.783=0.834)

cysteine(log21.851=0.889)

methionine(log22. 072 = 1. 051)

ornithine(log22. 251 = 1. 170). But palmitelaidic acid(log20. 127 =-2. 983)

17-eicosatrienoic acid(log20.334=-1.583)

nervonic acid(log20.600=-0.738)

conjugated linoleic acid(log20.680=-0.557)

cis-vaccenic acid(log20.711=-0.492)

9-heptadecenoic acid(log20.782=-0.355)

DHA(log20.829=-0.271)

EPA(log2 0.841=-0.250)

erucic acid(log20.844=-0.244)

DPA(log20.898=-0.156)

citric acid(log20.279=-1.842)

malic acid(log20.208=-2.264)

succinic acid(log20.254=-1.980)

cis-aconitic acid(log20.260=-1.946)

beta-citryl-L-glutamic acid(log20.093=-3.430)

beta-alanine(log20.139=-2.851)

cystathionine(log20.267 =-1.904)

cis-4-hydroxyproline(log20.500 =-1.000) were decreased in hypoxia group. The metabolic pathway analysis shown the nucleotide metabolism [log2(1.811、1.149)=0.857

0.201]

the energy metabolism[log2(1.510、1.173、1.149)=0.595

0.230

0.201]

the metabolism of cofactors and vitamins [log2(1.045、1.052、1.125)=0.064

0.073

0.170]

the amino acid metabolism[log2(1.245、1.020、1.027、1.123、1.127、1.076)=0.316

0.028

0.039

0.167

0.173

0.106]

the signal transduction(log21.046 =0.065)

the translation(log21.026=0.037)were activated in hypoxia group

but the carbohydrate metabolism[log2(0.857、0.857、0.799)=-0.222

-0.222

-0.323]

the lipid metabolism[log2(0.944、0.912、0.826)=-0.083

-0.133

-0.276]

the endocrine metabolism[log2(0.885、0.799、0.799)=-0.176

-0.323

-0.323] and the biosynthesis of other secondary metabolites[log2(0.947、0.871、0.743)=-0.079

-0.199

-0.428] were inhibited. Conclusion:Hypoxia alone treated HTR8/SVneo cell model is more suitable for studying the physiopathologic mechanism of preeclampsia in vitro.

关键词

子痫前期HTR8/SVneo滋养细胞缺氧腺苷酸活化蛋白激酶气相质谱技术

Keywords

preeclampsiaHTR8/SVneo trophoblast cellhypoxiaAMP-activated protein kinasegas chromatography mass spectrum

references

Views

260

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Expression of protein kinase SGK1 in preeclampsia placenta tissue and effects of hypoxia on the expression of SGK1 in placental trophoblast cells

Application value of placental growth factor，soluble fms-like tyrosine kinase-1，and glycosylated fibronectin in the prediction of preeclampsia

Study on the diagnostic value of inflammation score system in preeclampsia

Mechanisms of mitochondrial Drp1 inducing excessive opening of mPTP channel through LRRK2-HK2 after hypoxia

Effect of GPR30 protein on function of trophoblast cells and its relationship with the development of preeclampsia

Related Author

Zhan Yufeng

Dong Xiaojing

Zhang Xuerui

Ren Yan

Yang Lan

Xiao Jianping

Shi Hao

Su Jingna

Related Institution

Department of Obstetrics and Gynecology，The Second Affiliated Hospital，Chongqing Medical University

Department of Medical Genetics and Prenatal Diagnosis，Wuxi Maternal and Child Health Hospital

Department of Obstetrics and Gynecology，Third Affiliated Hospital of Zunyi Medical University， Zunyi First People’s Hospital

Department of Anesthesiology，The Second Affiliated Hospital of Chongqing Medical University

Department of Obstetrics and Gynecology,The First Affiliated Hospital of Chongqing Medical University

⁰