端粒酶逆转录酶促进缺氧缺血性脑损伤后新生大鼠的血管生成。
Telomerase reverse transcriptase promotes angiogenesis in neonatal rats after hypoxic-ischemic brain damage.
机构信息
Department of Pediatrics, West China Second University Hospital, Sichuan University, Chengdu, China.
Department of Cardiovascular Surgery, West China Hospital, Sichuan University, Chengdu, China.
出版信息
PeerJ. 2022 Oct 21;10:e14220. doi: 10.7717/peerj.14220. eCollection 2022.
BACKGROUND
Angiogenesis is an endogenous repair mechanism following hypoxic-ischemic brain damage (HIBD). Interestingly, recent studies have shown that angiogenesis can be regulated by telomerase reverse transcriptase (TERT), a critical component of telomerase. As telomerase reverse transcriptase can promote angiogenesis after stroke, we hypothesized that it could also promote angiogenesis after HIBD. To test this hypothesis, we developed and HIBD models in neonatal rats.
METHODS
TERT was overexpressed by lentivirus and adenovirus infection, and levels were measured using quantitative real-time polymerase chain reaction. We used a cell counting kit to quantify the proliferation rate of brain microvascular endothelial cells (BMECs), and immunofluorescence staining to measure CD34 expression levels. A microvessel formation assay was used to evaluate angiogenesis. Blood-brain barrier (BBB) integrity was assessed using immunohistochemical staining for ZO-1 and Evans Blue staining. Lastly, the expression level of Notch-1 was measured by western blotting.
RESULTS
Overexpression of promoted the proliferation of BMECs after hypoxic-ischemic damage . overexpression increased the formation of microvessels in the neonatal brain after HIBD both and . Overexpression of improved BBB integrity in the brains of neonatal rats after HIBD. In addition, the expression level of Notch-1 was increased in BMECs following oxygen glucose deprivation, and overexpression of TERT further increased Notch-1 expression levels in BMECs following oxygen glucose deprivation.
DISCUSSION
Our results reveal that telomerase reverse transcriptase promotes angiogenesis and maintains the integrity of the blood-brain barrier after neonatal hypoxic-ischemic brain damage. Furthermore, the Notch-1 signaling pathway appears to contribute to the angiogenic function of telomerase reverse transcriptase. This protective effect of telomerase reverse transcriptase opens new horizons for future investigations aimed at uncovering the full potential of telomerase reverse transcriptase as a promising new target for the treatment of hypoxic-ischemic encephalopathy.
背景
血管生成是缺氧缺血性脑损伤（HIBD）后内源性修复机制。有趣的是，最近的研究表明，血管生成可以通过端粒酶逆转录酶（TERT）调节，TERT 是端粒酶的关键组成部分。由于端粒酶逆转录酶可以促进中风后的血管生成，我们假设它也可以促进 HIBD 后的血管生成。为了验证这一假设，我们在新生大鼠中建立了 HIBD 模型。
方法
通过慢病毒和腺病毒感染过表达 TERT，并用实时定量聚合酶链反应测量其水平。我们使用细胞计数试剂盒来量化脑微血管内皮细胞（BMEC）的增殖率，并通过免疫荧光染色来测量 CD34 的表达水平。使用微血管形成测定法来评估血管生成。通过 ZO-1 的免疫组织化学染色和 Evans Blue 染色评估血脑屏障（BBB）的完整性。最后，通过 Western blot 测量 Notch-1 的表达水平。
结果
缺氧缺血损伤后过表达 TERT 促进了 BMEC 的增殖。过表达 TERT 增加了 HIBD 后新生大脑中微血管的形成。过表达 TERT 改善了 HIBD 后新生大鼠大脑的 BBB 完整性。此外，缺氧葡萄糖剥夺后 BMEC 中的 Notch-1 表达水平增加，而过表达 TERT 进一步增加了缺氧葡萄糖剥夺后 BMEC 中的 Notch-1 表达水平。
讨论
我们的结果表明，端粒酶逆转录酶促进了新生缺氧缺血性脑损伤后的血管生成，并维持了血脑屏障的完整性。此外，Notch-1 信号通路似乎有助于端粒酶逆转录酶的血管生成功能。端粒酶逆转录酶的这种保护作用为未来的研究开辟了新的视野，旨在揭示端粒酶逆转录酶作为治疗缺氧缺血性脑病有前途的新靶点的全部潜力。
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