Abstract

Background

Ischemic stroke (IS) is a major public health issue causing mortality and disability and is more difficult to treat than other cerebral diseases. Previous study reported that miR-376a was upregulated in the serum of stroke patients, indicating that miR-376a played potential role in occurrence and development of stroke.

Methods

IS cell model was induced by oxygen-glucose deprivation (OGD) exposed HCMEC/D3 cells. The mRNA level of SNHG1, miR-376a and inflammatory cytokines were detected by q-PCR. Protein level of CBS, apoptotic proteins were examined by Western blot. Apoptosis was analyzed by flow cytometry, and H₂S level was measured by kit. Interaction among lncRNA, miRNA and target gene was validated by luciferase assay.

Results

Our research revealed that mRNA level of SNHG1 and CBS in HCMEC/D3 cells was downregulated while miR-376a was upregulated under OGD conditions. Further results demonstrated that miR-376a overexpression promoted apoptosis and inflammation while SNHG1 overexpressing alleviated such processes. Mechanistically, SNHG1 directly targeted miR-376a, and CBS was a target of miR-376a. Moreover, SNHG1 exert its function via inhibiting miR‐376a to regulate CBS expression.

Conclusion

LncRNA SNHG1 depressed apoptosis and inflammation of IS cell model via inhibiting miR-376a and upregulating CBS/H₂S signal. These results show light on underlying mechanisms of IS and provide potential targets for IS therapy.

中文翻译：

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LncRNA SNHG1 通过靶向 miR-376a 和调节 CBS/H2S 通路减轻缺血性卒中期间的细胞凋亡和炎症

摘要

背景

缺血性中风 (IS) 是导致死亡和残疾的主要公共卫生问题，并且比其他脑部疾病更难治疗。既往研究报道，中风患者血清中miR-376a表达上调，提示miR-376a在中风的发生发展中发挥潜在作用。

方法

IS 细胞模型由暴露于缺氧葡萄糖 (OGD) 的 HCMEC/D3 细胞诱导。通过q-PCR检测SNHG1、miR-376a和炎性细胞因子的mRNA水平。通过Western印迹检查CBS的蛋白水平、凋亡蛋白。流式细胞仪检测细胞凋亡，试剂盒检测H ₂ S水平。通过荧光素酶测定验证lncRNA、miRNA和靶基因之间的相互作用。

结果

我们的研究表明，在 OGD 条件下，HCMEC/D3 细胞中 SNHG1 和 CBS 的 mRNA 水平下调，而 miR-376a 上调。进一步的结果表明，miR-376a 过表达促进了细胞凋亡和炎症，而 SNHG1 过表达缓解了这些过程。机制上，SNHG1 直接靶向 miR-376a，而 CBS 是 miR-376a 的靶标。此外，SNHG1通过抑制 miR-376a 发挥其功能，从而调节 CBS 的表达。

结论

LncRNA SNHG1通过抑制miR-376a和上调CBS/H ₂ S信号抑制IS细胞模型的凋亡和炎症。这些结果揭示了 IS 的潜在机制，并为 IS 治疗提供了潜在的目标。