A Primary Human Trophoblast Model to Study the Effect of Inflammation Associated with Maternal Obesity on Regulation of Autophagy in the Placenta.
A Primary Human Trophoblast Model to Study the Effect of Inflammation Associated with Maternal Obesity on Regulation of Autophagy in the Placenta.
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DOI：
10.3791/56484
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发表时间：
2017-09
期刊：
影响因子：
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通讯作者：
Bailey Simon;Matthew Bucher;A. Maloyan
中科院分区：
文献类型：
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作者：
Bailey Simon;Matthew Bucher;A. Maloyan
母亲肥胖与围产期不良结局的风险增加有关，这可能是由于胎盘功能受损，部分原因是自噬功能失调。肥胖妊娠胎盘中自噬调节因子表达的异常变化可能受到与肥胖和妊娠相关的炎症过程的调节。这里描述了一种从人类胎盘中提取绒毛组织和分离绒毛细胞滋养层细胞以进行原代细胞培养的方法。然后是一种模拟肥胖宫内环境中炎症环境的方法，方法是用肿瘤坏死因子α(肿瘤坏死因子α)处理瘦身妊娠的初级滋养细胞，肿瘤坏死因子α是一种促炎细胞因子，在肥胖和怀孕期间会升高。通过实施本文描述的方案，研究发现，暴露于外源性肿瘤坏死因子α调节自噬的负性调节因子Rubcon在有女性胎儿的瘦身妊娠的滋养层中的表达。虽然在肥胖的宫内环境中，各种生物因素保持着调节滋养层细胞关键途径的潜力，但这个体外系统特别有助于确定在体内观察到的母体肥胖胎盘的表达模式是否是肿瘤坏死因子α信号的直接结果。最终，这种方法提供了机会来解析与母体肥胖相关的炎症对自噬和滋养层细胞中其他可能影响胎盘功能的关键细胞通路的调节和分子影响。
Maternal obesity is associated with an increased risk of adverse perinatal outcomes that are likely mediated by compromised placental function that can be attributed to, in part, the dysregulation of autophagy. Aberrant changes in the expression of autophagy regulators in the placentas from obese pregnancies may be regulated by inflammatory processes associated with both obesity and pregnancy. Described here is a protocol for sampling of villous tissue and isolation of villous cytotrophoblasts from the term human placenta for primary cell culture. This is followed by a method for simulating the inflammatory milieu in the obese intrauterine environment by treating primary trophoblasts from lean pregnancies with tumor necrosis factor alpha (TNFα), a proinflammatory cytokine that is elevated in obesity and in pregnancy. Through the implementation of the protocol described here, it is found that exposure to exogenous TNFα regulates the expression of Rubicon, a negative regulator of autophagy, in trophoblasts from lean pregnancies with female fetuses. While a variety of biological factors in the obese intrauterine environment maintain the potential to modulate critical pathways in trophoblasts, this ex vivo system is especially useful for determining if expression patterns observed in vivo in human placentas with maternal obesity are a direct result of TNFα signaling. Ultimately, this approach affords the opportunity to parse out the regulatory and molecular implications of inflammation associated with maternal obesity on autophagy and other critical cellular pathways in trophoblasts that have the potential to impact placental function.