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甘氨酸通过 miR-19a-3p/AMPK/GSK-3β/HO-1 通路改善缺血性脑卒中。

Glycine Improves Ischemic Stroke Through miR-19a-3p/AMPK/GSK-3β/HO-1 Pathway.

机构信息

Neurological Intervention Department, Dalian Municipal Central Hospital, Dalian, Liaoning Province, People's Republic of China.

出版信息

Drug Des Devel Ther. 2020 May 25;14:2021-2031. doi: 10.2147/DDDT.S248104. eCollection 2020.

DOI:10.2147/DDDT.S248104
PMID:32546967
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7260540/
Abstract

PURPOSE

To explore the molecular mechanism of glycine in improving ischemic stroke.

PATIENTS AND METHODS

The serum samples of patients with ischemic stroke and healthy people were compared. The ischemic stroke model of PC12 cells was established by oxygen-glucose deprivation (OGD). qPCR quantified miR-19a-3p and AMPK mRNA, and protein expression was detected by Western blot. MTT was used to detect cell activity. Flow cytometry was used to detect cells. Glucose metabolism kit was used to detect glucose intake and formation amount of lactic acid.

RESULTS

Compared with the control group, OGD group (OGDG) showed lower cell activity and increased cell apoptosis. TNF-α, IL-1βI, L-6, Caspase 3, Caspase 9 and Bax were up-regulated, and Glut1, HK2, LDHA, PDK1, PKM2 and Bcl2 were down-regulated. At the same time, glucose intake, formation amount of lactic acid and cell apoptosis rate were reduced, and AMPK/GSK-3β/HO-1 pathway activity was down-regulated. Glycine could counteract the above phenomena in OGDG. miR-19a-3p and AMPK decreased and increased, respectively, during glycine therapy. AMPK was the target gene of miR-19a-3p. Rescue experiments demonstrated that glycine improved cell apoptosis, inflammatory response and glucose metabolism disorder of ischemic stroke through miR-19a-3p/AMPK/GSK-3β/HO-1 pathway.

CONCLUSION

Glycine improves ischemic stroke through miR-19a-3p/AMPK/GSK-3β/HO-1 pathway.

摘要

目的

探讨甘氨酸改善缺血性脑卒中的分子机制。

方法

比较缺血性脑卒中患者和健康人的血清样本。通过氧葡萄糖剥夺（OGD）建立 PC12 细胞缺血性脑卒中模型。qPCR 定量检测 miR-19a-3p 和 AMPK mRNA，Western blot 检测蛋白表达。MTT 检测细胞活性。流式细胞术检测细胞凋亡。葡萄糖代谢试剂盒检测葡萄糖摄取和乳酸生成量。

结果

与对照组相比，OGD 组（OGDG）细胞活性降低，细胞凋亡增加。TNF-α、IL-1βI、L-6、Caspase 3、Caspase 9 和 Bax 上调，Glut1、HK2、LDHA、PDK1、PKM2 和 Bcl2 下调。同时，葡萄糖摄取、乳酸生成量和细胞凋亡率降低，AMPK/GSK-3β/HO-1 通路活性下调。甘氨酸可拮抗 OGDG 中的上述现象。甘氨酸治疗过程中，miR-19a-3p 和 AMPK 分别降低和增加。AMPK 是 miR-19a-3p 的靶基因。挽救实验表明，甘氨酸通过 miR-19a-3p/AMPK/GSK-3β/HO-1 通路改善缺血性脑卒中细胞凋亡、炎症反应和葡萄糖代谢紊乱。

结论

甘氨酸通过 miR-19a-3p/AMPK/GSK-3β/HO-1 通路改善缺血性脑卒中。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6550/7260540/2ea04c0a2020/DDDT-14-2021-g0006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6550/7260540/c79a5d94f45e/DDDT-14-2021-g0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6550/7260540/2354a41bd8c7/DDDT-14-2021-g0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6550/7260540/1cc3471ec582/DDDT-14-2021-g0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6550/7260540/3a09e656864b/DDDT-14-2021-g0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6550/7260540/a7695beaa8c3/DDDT-14-2021-g0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6550/7260540/2ea04c0a2020/DDDT-14-2021-g0006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6550/7260540/c79a5d94f45e/DDDT-14-2021-g0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6550/7260540/2354a41bd8c7/DDDT-14-2021-g0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6550/7260540/1cc3471ec582/DDDT-14-2021-g0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6550/7260540/3a09e656864b/DDDT-14-2021-g0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6550/7260540/a7695beaa8c3/DDDT-14-2021-g0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6550/7260540/2ea04c0a2020/DDDT-14-2021-g0006.jpg

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