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初乳和乳汁外泌体中差异发现的候选免疫系统微小RNA的鉴定

Identification of Candidate Immune System MicroRNAs Differentially Found in Colostrum and Milk Exosomes.

作者信息

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

机构信息

Department of Physiology, All India Institute of Medical Sciences, Bhubaneswar, Odisha 751019, India.

ICMR-AIIMS Computational Genomics Centre, Division of Biomedical Informatics, Indian Council of Medical Research, Ansari Nagar, New Delhi-110029, India.

出版信息

Microrna. 2022;11(3):216-226. doi: 10.2174/2211536611666220630102316.

Abstract

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.

摘要

背景

胎儿在无菌的子宫环境中生长。出生后,新生儿免疫系统面临两个亟待克服的障碍。首先是立即抑制与子宫相容的免疫系统,并开启能够应对抗原世界的新生儿免疫系统。母乳微小RNA(miRNA)引起免疫波动的潜在机制对于危重新生儿或早产儿的治疗可能至关重要。

方法

我们从哺乳期母亲那里收集了初乳和成熟乳各14份样本,其中4份用于微阵列分析,另外10份用于通过实时PCR进行miRNA表达谱分析。

结果

通过微阵列鉴定出154个差异表达的miRNA,而根据文献研究,有49个miRNA被鉴定为免疫相关miRNA。在这49个miRNA中,有33个已被充分验证为免疫相关miRNA(通过定量PCR、蛋白质免疫印迹和荧光素酶测定法验证)并被考虑进一步分析。对22个miRNA的表达进行了实时PCR分析,因为它们的Ct值在合理范围内。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人乳外泌体miRNA表达动态及其在新生儿免疫系统逐渐偏向中的可能作用。这些信息对于新生儿重症监护病房(NICU)中早产儿败血症的发生和发展至关重要。

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