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确定 miR-451a 在恶性疟原虫红内期、氧化应激和蛋白质组学特征中的作用。

Determination of the role of miR-451a on Plasmodium falciparum red blood cell stages, oxidative stress, and proteomic profiling.

机构信息

Department of Biochemistry, School of Sciences, Gujarat University, Ahmedabad, Gujarat, 380009, India.

Department of Zoology, BMTC, Human Genetics and WLC, School of Sciences, Gujarat University, Ahmedabad, Gujarat, 380009, India.

出版信息

Mol Biol Rep. 2024 Oct 7;51(1):1041. doi: 10.1007/s11033-024-09938-z.

Abstract

BACKGROUND

This study examines the feasibility and effects of introducing microRNA mimic into red blood cells (RBCs) at the initial phases of Plasmodium falciparum 3D7 (Pf3D7) infection. The aim is to determine the correlation between increased expression of miR-451a and parasitaemia.

METHODS

In this study miR-mimic-451a labelled with Cy3 and transfected into control and infected RBCs using lipofectamine and analysed using the fluorescence microscopy and flow cytometry. The study demonstrated the efficacy of miR-451a by treating pre-and post-transfected control RBCs and Pf3D7-infected RBCs with miR-mimic-451a. We also examined its impact on % growth inhibition of Pf3D7, oxidative stress markers (Luminometry, LPO, SOD, CAT, GSH and GPx). Additionally, determination of pH, haemoglobin (Hb), and proteomic profile performed using SDS-PAGE.

RESULTS

Modified expression level of mir-451a has the potential to change the progression of the infection and yielded a 50% decrease in parasitaemia within 48 h. Moreover, transfected samples were shown to be efficacious in counteracting the oxidative stress-induced alterations during Pf3D7 infection and enable to return the cells towards the normalcy. Modified proteomic profile of transfected iRBCs demonstrates the correlation between overexpression of miRNA and protein expression. where, the major changes were observed in the heavy molecular weight proteins more than 57 kDa.

CONCLUSION

The study reveals promising effects of miR-mimic-451a enrichment during RBC stages of Pf3D7, offering insights into potential malaria therapeutic strategies and potential biomedical research implications.

摘要

背景

本研究旨在探讨在恶性疟原虫 3D7(Pf3D7)感染初期将 microRNA 模拟物导入红细胞(RBC)的可行性及其影响。目的是确定 miR-451a 表达增加与寄生虫血症之间的相关性。

方法

本研究使用脂质体将 Cy3 标记的 miR-mimic-451a 转染到对照和感染的 RBC 中,并使用荧光显微镜和流式细胞术进行分析。通过用 miR-mimic-451a 处理转染前后的对照 RBC 和 Pf3D7 感染的 RBC,研究证明了 miR-451a 的功效。我们还研究了其对 Pf3D7 的生长抑制百分比(%)、氧化应激标志物(发光测定法、LPO、SOD、CAT、GSH 和 GPx)的影响。此外,使用 SDS-PAGE 进行 pH 值、血红蛋白(Hb)和蛋白质组学图谱的测定。

结果

miR-451a 的修饰表达水平有可能改变感染的进展,并在 48 小时内使寄生虫血症降低 50%。此外,转染的样本在对抗 Pf3D7 感染引起的氧化应激改变方面显示出有效性,并使细胞恢复正常。转染 iRBC 的修饰蛋白质组学图谱表明 miRNA 过表达与蛋白质表达之间存在相关性。其中,在大于 57 kDa 的重分子量蛋白质中观察到主要变化。

结论

本研究揭示了在 Pf3D7 的 RBC 阶段富含 miR-mimic-451a 的有希望的效果,为潜在的疟疾治疗策略和潜在的生物医学研究意义提供了新的见解。

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