The Comparative Effect of Plasma Exosomes of Young and Old People on the Expression of BCL-2 and BAX Genes in Hematopoietic Stem Cells

Roya Moradi, Reza Afrisham, Zahra Kashanikhatib, Seyed Hadi Mousavi, Masoud Soleimani, Shaban Alizadeh
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Abstract

Apoptosis may disrupt differentiation of hematopoietic stem cells (HSCs), which can affect aging. Thus, the main goal of this study was to compare the effect of plasma exosomes from young and old people on the expression of Bcl-2-associated X (BAX) and B-cell lymphoma 2 (BCL-2) genes in the HSCs. Plasma samples were acquired from four elderly adults and four younger adults, referring to Blood Transfusion Organization of Tehran-Iran during August 2022– September 2022.Then, the exosomes of the samples were extracted and analyzed using DLS, TEM, and CD63 surface marker. HSCs were isolated from umbilical cord blood cells. The MTT test was used to assess the viability of exosomes-treated HSCs at doses of 5 and 10 μg/ml. The expression of BAX and BCL-2 genes in the cells was examined using real-time PCR. A one-way analysis of variance (ANOVA) was performed to examine the distinctions among five groups. The viability of HSCs was not affected by the exosomes from young and old people than the control group (P = 0.453). Exosomes from young people (doses 5 and 10 µg/ml) did not have any significant impacts on BAX (P = 0.746, and P = 0.345, respectively) and BCL-2 (P = 0.773, and P = 0.461, respectively) expression in the HSCs compared to the control group. The BAX gene was significantly upregulated and the BCL-2 gene was significantly downregulated after utilizing the exosomes derived from the plasma of elderly individuals (dose 10 µg/ml) compared to the control (P = 0.001, P = 0.002, respectively). The current research shows that aged people's exosomes can increase BAX/ BCL-2 ratio in umbilical cord blood-derived HSCs compared to control and young groups.

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年轻人和老年人血浆外泌体对造血干细胞中 BCL-2 和 BAX 基因表达的影响比较
细胞凋亡可能会破坏造血干细胞(HSCs)的分化,从而影响衰老。因此,本研究的主要目的是比较年轻人和老年人的血浆外泌体对造血干细胞中Bcl-2相关X(BAX)和B细胞淋巴瘤2(BCL-2)基因表达的影响。研究人员于2022年8月至2022年9月期间从伊朗德黑兰输血组织获取了4名老年人和4名年轻人的血浆样本,然后提取样本中的外泌体,并使用DLS、TEM和CD63表面标记物进行分析。从脐带血细胞中分离出造血干细胞。MTT试验用于评估外泌体处理的造血干细胞的活力,剂量分别为5和10微克/毫升。使用实时 PCR 检测细胞中 BAX 和 BCL-2 基因的表达。进行单因素方差分析(ANOVA)以检验五组间的差异。与对照组相比,来自年轻人和老年人的外泌体对造血干细胞的活力没有影响(P = 0.453)。与对照组相比,年轻人的外泌体(剂量分别为5和10 µg/ml)对造血干细胞中BAX(P = 0.746,P = 0.345)和BCL-2(P = 0.773,P = 0.461)的表达没有显著影响。与对照组相比,利用从老年人血浆中提取的外泌体(剂量为10微克/毫升)后,BAX基因明显上调,BCL-2基因明显下调(分别为P = 0.001和P = 0.002)。目前的研究表明,与对照组和年轻组相比,老年人的外泌体能提高脐带血造血干细胞中的BAX/ BCL-2比率。
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期刊介绍: Indian Journal of Hematology and Blood Transfusion is a medium for propagating and exchanging ideas within the medical community. It publishes peer-reviewed articles on a variety of aspects of clinical hematology, laboratory hematology and hemato-oncology. The journal exists to encourage scientific investigation in the study of blood in health and in disease; to promote and foster the exchange and diffusion of knowledge relating to blood and blood-forming tissues; and to provide a forum for discussion of hematological subjects on a national scale. The Journal is the official publication of The Indian Society of Hematology & Blood Transfusion.
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