初乳和乳外泌体中差异的候选免疫系统microrna鉴定。

Poonam Verma, Niharika Mohanty, Babita Pruseth, Sonali Sahoo, Amit Katiyar, Harpreet Singh, Saubhagya Kumar Jena, Rashmi Ranjan Das, Tapas Kumar Som, Sanjeeb Kumar Sahoo, Pranati Nanda, Amit Ghosh
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引用次数: 1

摘要

背景:胎儿在无菌子宫环境中生长。出生后,新生儿的免疫系统有两个直接的障碍需要清除。首先,立即抑制子宫相容免疫系统,并启动新生儿的免疫系统,以对抗抗原世界。乳微rna (miRNAs)免疫波动的潜在机制可能对危重新生儿或早产儿的治疗至关重要。方法:从哺乳母亲中各采集初乳和成熟乳各14份样品,各4份样品进行微阵列分析,另外10份样品采用实时PCR技术进行miRNA表达谱分析。结果:从微阵列中鉴定出154个差异表达的mirna,而根据文献研究发现49个mirna是免疫相关的mirna。在49个mirna中,33个已经被证明是强验证的免疫相关mirna(通过qPCR、Western Blot和荧光素酶测定验证),可以考虑进一步分析。实时PCR分析22个miRNA表达,因为它们的Ct值在相当大的范围内。与初乳相比,成熟乳中有12个mirna显著下调,这些mirna再次进行了生物信息学分析,以预测差异表达mirna背后的生物学机制。结论:本研究揭示了哺乳期间母乳外泌体miRNA的表达动态及其在新生儿免疫系统逐渐偏斜中的可能作用。这些信息对于新生儿重症监护病房早产儿脓毒症的发展和发病至关重要。
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Identification of Candidate Immune System MicroRNAs Differentially Found in Colostrum and Milk Exosomes.

Background: The fetus grows in a sterile womb environment. After birth, the newborn immune system has two immediate hurdles to clear. First immediate suppression of the womb compatible immune system and turn on the immune system of the newborn that can counter the antigenic world. The underlying mechanism of immune fluctuation by milk microRNAs (miRNAs) can be crucial for the treatment of critical or premature newborn.

Methods: We collected fourteen samples of each colostrum and mature milk from lactating mothers, four samples of each were used for microarray analysis, and the other ten were used for miRNA expression profiling by real-time PCR.

Results: From the microarray, 154 differentially expressed miRNAs were identified, whereas 49 miRNAs were revealed as immune-related miRNAs based on a literature study. Among the 49 miRNAs, 33 were already shown as strongly validated immune-related miRNAs (validated by qPCR, Western Blot, and Luciferase assay) and were considered for further analysis. Twenty-two miRNA expressions were analysed by real-time PCR as their Ct values were within considerable limits. Twelve numbers of miRNAs were significantly downregulated in mature milk compared to colostrum, which were again subjected to bioinformatics analysis to predict the biological mechanisms behind the differentially expressed miRNAs.

Conclusion: This study shed light on the human milk exosome miRNA expression dynamics during lactation and their possible role in the gradual skewing of the newborns' immune system. The information is crucial for the development and onset of sepsis in premature newborns in the NICU.

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