热休克蛋白 20 通过激活低氧条件下的 PI3K/Akt 信号通路促进内皮祖细胞血管生成。
Hsp20 Promotes Endothelial Progenitor Cell Angiogenesis via Activation of PI3K/Akt Signaling Pathway under Hypoxia.
机构信息
Guangxi Medical University, Nanning, Guangxi, 530021, People's Republic of China.
Department of Oral and Maxillofacial Surgery, College of Stomatology, Hospital of Stomatology, Guangxi Medical University, Nanning, Guangxi, 530021, People's Republic of China.
出版信息
Tissue Eng Regen Med. 2022 Dec;19(6):1251-1266. doi: 10.1007/s13770-022-00481-1. Epub 2022 Aug 30.
BACKGROUND
Mandibular distraction osteogenesis (MDO) is a kind of endogenous tissue engineering technology that lengthens the jaw and opens airway so that a patient can breathe safely and comfortably on his or her own. Endothelial progenitor cells (EPCs) are crucial for MDO-related angiogenesis. Moreover, emerging evidence suggests that heat shock protein 20 (Hsp20) modulates angiogenesis under hypoxic conditions. However, the specific role of Hsp20 in EPCs, in the context of MDO, is not yet known. The aim of this study was to explore the expression of Hsp20 during MDO and the effects of Hsp20 on EPCs under hypoxia.
METHODS
Mandibular distraction osteogenesis and mandibular bone defect (MBD) canine model were established. The expression of CD34, CD133, HIF-1α, and Hsp20 in callus was detected by immunofluorescence on day 14 after surgery. Canine bone marrow EPCs were cultured, with or without optimal cobalt chloride (CoCl) concentration. Hypoxic effects, caused by CoCl, were evaluated by means of the cell cycle, cell apoptosis, transwell cell migration, and tube formation assays. The Hsp20/KDR/PI3K/Akt expression levels were evaluated via immunofluorescence, RT-qPCR, and western blot. Next, EPCs were incorporated with either Hsp20-overexpression or Hsp20-siRNA lentivirus. The resulting effects were evaluated as described above.
RESULTS
CD34, CD133, HIF-1α, and Hsp20 were displayed more positive in the callus of MDO compared with MBD. In addition, hypoxic conditions, generated by 0.1 mM CoCl, in canine EPCs, accelerated cell proliferation, migration, tube formation, and Hsp20 expression. Hsp20 overexpression in EPCs significantly stimulated cell proliferation, migration, and tube formation, whereas Hsp20 inhibition produced the opposite effect. Additionally, the molecular mechanism was partly dependent on the KDR/PI3K/Akt pathway.
CONCLUSION
In summary, herein, we present a novel mechanism of Hsp20-mediated regulation of canine EPCs via Akt activation in a hypoxic microenvironment.
背景
下颌骨牵引成骨术（MDO）是一种内源性组织工程技术，可延长下颌骨并打开气道，使患者能够自主安全舒适地呼吸。内皮祖细胞（EPCs）对于与 MDO 相关的血管生成至关重要。此外，新出现的证据表明，热休克蛋白 20（Hsp20）在缺氧条件下调节血管生成。然而，Hsp20 在 MDO 中的具体作用在 EPC 中尚不清楚。本研究旨在探讨 Hsp20 在 MDO 过程中的表达以及 Hsp20 在缺氧条件下对 EPCs 的影响。
方法
建立下颌骨牵引成骨和下颌骨骨缺损（MBD）犬模型。术后第 14 天，通过免疫荧光法检测骨痂中 CD34、CD133、HIF-1α 和 Hsp20 的表达。培养犬骨髓 EPCs，并用或不用最佳浓度的钴氯化物（CoCl）。通过细胞周期、细胞凋亡、Transwell 细胞迁移和管形成实验评估 CoCl 引起的缺氧效应。通过免疫荧光、RT-qPCR 和 Western blot 评估 Hsp20/KDR/PI3K/Akt 的表达水平。接下来，将 EPCs 与 Hsp20 过表达或 Hsp20-siRNA 慢病毒结合。如上所述评估所得效果。
结果
与 MBD 相比，MDO 骨痂中 CD34、CD133、HIF-1α 和 Hsp20 的表达更为阳性。此外，0.1 mM CoCl 在犬 EPCs 中产生的缺氧条件加速了细胞增殖、迁移、管形成和 Hsp20 表达。EPCs 中 Hsp20 的过表达显著刺激细胞增殖、迁移和管形成，而 Hsp20 的抑制则产生相反的效果。此外，该分子机制部分依赖于 KDR/PI3K/Akt 通路。
结论
总之，本文提出了一种新的机制，即在缺氧微环境下，Hsp20 通过激活 Akt 调节犬 EPCs。
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