Hypoxic Stress Induces Complement-Mediated Lysis of Mesenchymal Stem Cells by Downregulating Factor H and CD59.

IF 4.4 4区 医学 Q2 CELL & TISSUE ENGINEERING
Ramada R Khaswaneh, Ejlal Abu-El-Rub, Ayman Alzu'bi, Fatimah A Almahasneh, Rawan A Almazari, Heba F Ai-Jariri, Raed M Al-Zoubi
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Abstract

Background: Factor H and membrane inhibitor of reactive lysis (CD59) are key regulators of complement activation. Mesenchymal stem cells (MSCs) secrete Factor H and express CD59 to protect themselves from complement-mediated damage. Severe hypoxia found to decrease the survival chances of MSCs after transplantation; however, little is known about the impact of severe hypoxia on modulating the complement system activity and its effect on MSCs survival. Our study seeks to explore the effect of severe hypoxia on modulating the complement cascade in MSCs.

Methods: Human adipose tissue-derived MSCs (hAD-MSCs) were cultured under severe hypoxia using 400 μM Cobalt Chloride (CoCl2) for 48 h. The protein expressions of survival marker; Phosphoinositide 3-kinases (PI3K), and pro-apoptotic marker; Caspase-3 were assessed using western blotting. The level of complement system related factors; Factor H, CD59, C3b, iC3b, C5b, C9, and the complement membrane attack complex (MAC) were analyzed using Elisa assays, western blotting, and immunocytochemistry.

Results: Our results showed for the first time that severe hypoxia can significantly impair Factor H secretion and CD59 expression in MSCs. This has been associated with upregulation of MAC complex and increased level of cell lysis and apoptosis marked by downregulation of PI3K and upregulation of Annexin v and Caspase-3.

Conclusion: The loss of Factor H and CD59 in hypoxic MSCs can initiate their lysis and apoptosis mediated by activating MAC complex. Preserving the level of Factor H and CD59 in MSCs has significant clinical implication to increase their retention rate in hypoxic conditions and prolong their survival.

缺氧应激通过下调因子 H 和 CD59 诱导补体介导的间充质干细胞裂解
背景:因子H和反应性裂解膜抑制剂(CD59)是补体激活的关键调节因子。间充质干细胞(MSCs)分泌因子H并表达CD59,以保护自身免受补体介导的损伤。研究发现,严重缺氧会降低间充质干细胞移植后的存活机会;然而,人们对严重缺氧对调节补体系统活性的影响及其对间充质干细胞存活的影响知之甚少。方法:使用 400 μM 氯化钴(CoCl2)在严重缺氧条件下培养人脂肪组织来源间充质干细胞(hAD-MSCs)48 小时。使用 Elisa 检测法、Western 印迹法和免疫细胞化学法分析了补体系统相关因子 H、CD59、C3b、iC3b、C5b、C9 和补体膜攻击复合体(MAC)的水平:结果:我们的研究结果首次表明,严重缺氧会显著影响间充质干细胞中因子 H 的分泌和 CD59 的表达。这与 MAC 复合物的上调以及细胞裂解和凋亡水平的增加有关,细胞裂解和凋亡以 PI3K 的下调和 Annexin v 及 Caspase-3 的上调为标志:结论:缺氧间充质干细胞中因子 H 和 CD59 的缺失可通过激活 MAC 复合物介导细胞裂解和凋亡。保持间充质干细胞中因子 H 和 CD59 的水平对提高间充质干细胞在缺氧条件下的存活率和延长其存活时间具有重要的临床意义。
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来源期刊
Tissue engineering and regenerative medicine
Tissue engineering and regenerative medicine CELL & TISSUE ENGINEERING-ENGINEERING, BIOMEDICAL
CiteScore
6.80
自引率
5.60%
发文量
83
审稿时长
6-12 weeks
期刊介绍: Tissue Engineering and Regenerative Medicine (Tissue Eng Regen Med, TERM), the official journal of the Korean Tissue Engineering and Regenerative Medicine Society, is a publication dedicated to providing research- based solutions to issues related to human diseases. This journal publishes articles that report substantial information and original findings on tissue engineering, medical biomaterials, cells therapy, stem cell biology and regenerative medicine.
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