S100 和 YKL40 在早产儿脑室出血中的作用以及 miR-138-siRNAs-HIF-1a 和 miR-21-siRNAs-HVCN1 在神经损伤新生鼠中的神经保护作用。

Role of S100 and YKL40 on Intraventricular Cerebral Hemorrhages in the Preterm Infant and the Neuroprotective Role of miR-138- siRNAs-HIF-1a and miR-21-siRNAs-HVCN1 in Neonatal Mice with Nerve Injury.

机构信息

Faculty of Reproductive Health, Golestan University of Medical Sciences, Gorgan, Iran.

Department of Cellular and Molecular Medicine, Faculty of Medicine, Royal's Institute of Mental Health Research, University of Ottawa, Ottawa, Canada.

出版信息

Curr Med Chem. 2024;31(34):5638-5656. doi: 10.2174/0929867331666230915103147.

DOI:10.2174/0929867331666230915103147

PMID:37724672

Abstract

BACKGROUND

Epilepsy and intraventricular-cerebral hemorrhage is a common complication irreversible in preterm infants. Inflammation leads to an increase in intracellular calcium, acidosis, and oxygen usage, and finally, may damage brain cells. Increases in HIF-1a and HVCN1 can reduce the complications of oxygen consumption and acidosis in infants with intraventricular hemorrhage (IVH). On the other hand, decreases in S100B can shield nerve cells from apoptosis and epilepsy by reducing brain damage.

OBJECTIVE

In this research, we investigated how miR-138-siRNAs-HIF-1a and miR-21- siRNAs-HVCN1 affect apoptosis in hypoxic mice.

METHODS

On the first and third days after delivery, the YKL40, HIF-1a, HVCN1, and S100b genes were compared between two groups of preterm infants with and without maternal inflammation. Afterward, the miRNAs were transfected into cell lines to monitor variations in YKL40, HIF-1a, HVCN1, and S100b gene expression and nerve cell apoptosis. We changed the expression of S100b, HVCN1, and HIF-1a genes by using specific siRNAs injected into mice. Using real-time PCR, Western blotting, flow cytometry (FCM), and immunofluorescence, and changes in gene expression were evaluated (IHC).

RESULTS

HVCN1 gene expression showed a strong negative correlation with epilepsy in both groups of infants (Pβ0.001). Significant correlations between epilepsy and the expression levels of the S100b, YKL40, and HIF-1a genes were found (Pβ0.001). According to FCM, after transfecting miRNA-431 and miRNA-34a into cell lines, the apoptosis index (A.I.) were 41.6 3.3 and 34.5 5.2%, respectively, while the A.I. were 9.6 2.7 and 7.1 4.2% after transfecting miRNA-21 and miRNA-138. MiR-138-siRNAs-HIF-1a and miR-21-siRNAs-HVCN1 were simultaneously injected into hypoxic mice, and IHC double-labeling revealed that this reduced apoptosis and seizures compared to the hypoxic group.

CONCLUSION

Our findings demonstrate that miR-138-siRNAs-HIF-1a and miR-21-siRNAs- HVCN1 injections prevent cerebral ischemia-induced brain damage in hypoxia mice by increasing HVCN1 and HIF-1a and decreasing S100b, which in turn lessens apoptosis and epilepsy in hypoxic mice.

摘要

背景

癫痫和脑室内出血是早产儿常见的不可逆并发症。炎症导致细胞内钙增加、酸中毒和耗氧量增加，最终可能导致脑细胞损伤。HIF-1a 和 HVCN1 的增加可以减少脑室内出血（IVH）婴儿的耗氧和酸中毒并发症。另一方面，S100B 的减少可以通过减少脑损伤来屏蔽神经细胞的凋亡和癫痫。

目的

本研究旨在探讨 miR-138-siRNAs-HIF-1a 和 miR-21-siRNAs-HVCN1 如何影响缺氧小鼠的细胞凋亡。

方法

在分娩后的第一天和第三天，比较两组早产儿中是否存在母体炎症对 YKL40、HIF-1a、HVCN1 和 S100b 基因的影响。然后，将 miRNA 转染到细胞系中，以监测 YKL40、HIF-1a、HVCN1 和 S100b 基因表达和神经细胞凋亡的变化。我们通过向小鼠注射特异性 siRNA 来改变 S100b、HVCN1 和 HIF-1a 基因的表达。使用实时 PCR、Western blot、流式细胞术（FCM）和免疫荧光法（IHC）评估基因表达的变化。

结果

HVCN1 基因表达与两组婴儿的癫痫均呈强负相关（Pβ0.001）。还发现癫痫与 S100b、YKL40 和 HIF-1a 基因表达水平之间存在显著相关性（Pβ0.001）。根据 FCM，将 miRNA-431 和 miRNA-34a 转染到细胞系后，凋亡指数（A.I.）分别为 41.6 3.3%和 34.5 5.2%，而转染 miRNA-21 和 miRNA-138 后 A.I.分别为 9.6 2.7%和 7.1 4.2%。将 miR-138-siRNAs-HIF-1a 和 miR-21-siRNAs-HVCN1 同时注射到缺氧小鼠体内，免疫组化双标记显示，与缺氧组相比，这两种方法均能减少细胞凋亡和癫痫发作。