Ma Liping, Cao Yanjing, Zhang Lin, Li Ketao, Yan Laixing, Pan Yizhan, Zhu Jianhua
Department of Cardiology, Shulan (Hangzhou) Hospital, Hangzhou, China.
Department of Neurology, Hangzhou Third People's Hospital, Hangzhou, China.
J Gene Med. 2020 Sep;22(9):e3201. doi: 10.1002/jgm.3201. Epub 2020 May 8.
Celastrol (Cel) has been corroborated as an anti-inflammatory and anti-apoptotic agent in multiple cell damage models. However, the protective effect of Cel in high glucose (HG)-induced cardiomyocyte injury is still unclear. The present study aimed to determine whether Cel can mitigate HG-stimulated cardiomyocyte injury via regulating the miR-345-5p/growth arrest-specific 6 (Gas6) signaling pathway.
Cardiomyocytes were exposed to normal glucose (NG; 5 mmol/l) or HG (30 mmol/l) and then administered with Cel. Cell counting kit-8 and flow cytometry assays were used to detect cell proliferative activity and apoptosis. mRNA and protein expression were analyzed using a quantitative reverse transcriptase-polymerase chain reaction and western blotting, respectively. A bioinformatics algorithm and a luciferase reporter gene assay were used to determine whether Gas6 is a direct target of miR-345-5p.
The present study confirmed the inhibitory effects of Cel in HG-induced inflammation in cardiomyocytes. Moreover, Cel exhibited the ability to antagonize HG-induced cardiomyocyte apoptosis and suppress the elevated Bax/Bcl-2 ratio elicited by HG stimulation. Intriguingly, Cel treatment revoked the HG-triggered repression of Gas6 protein expression, and Gas6 loss-of-function accelerated HG-induced cardiomyocyte apoptosis. HG-triggered up-regulation of miR-345-5p expression was depressed following Cel treatment. Importantly, we validated that Gas6 is a direct target of miR-345-5p. Transfection with miR-345-5p inhibitors restrained HG-induced release of pro-inflammatory cytokines and cell apoptosis.
The findings of the present study demonstrate that Cel administration antagonized HG-induced cardiomyocyte apoptosis and inflammation through up-regulating Gas6 expression by restraining miR-345-5p.
在多种细胞损伤模型中，雷公藤红素（Cel）已被确认为一种抗炎和抗凋亡剂。然而，Cel在高糖（HG）诱导的心肌细胞损伤中的保护作用仍不清楚。本研究旨在确定Cel是否能通过调节miR-345-5p/生长停滞特异性6（Gas6）信号通路减轻HG刺激的心肌细胞损伤。
将心肌细胞暴露于正常葡萄糖（NG；5 mmol/L）或HG（30 mmol/L）中，然后给予Cel。使用细胞计数试剂盒-8和流式细胞术检测细胞增殖活性和凋亡。分别使用定量逆转录聚合酶链反应和蛋白质免疫印迹分析mRNA和蛋白质表达。使用生物信息学算法和荧光素酶报告基因检测来确定Gas6是否是miR-345-5p的直接靶点。
本研究证实了Cel对HG诱导的心肌细胞炎症具有抑制作用。此外，Cel表现出拮抗HG诱导的心肌细胞凋亡的能力，并抑制HG刺激引起的Bax/Bcl-2比值升高。有趣的是，Cel处理可消除HG引发的Gas6蛋白表达抑制，而Gas6功能丧失会加速HG诱导的心肌细胞凋亡。Cel处理后，HG引发的miR-345-5p表达上调受到抑制。重要的是，我们验证了Gas6是miR-345-5p的直接靶点。转染miR-345-5p抑制剂可抑制HG诱导的促炎细胞因子释放和细胞凋亡。
本研究结果表明，给予Cel通过抑制miR-345-5p上调Gas6表达，从而拮抗HG诱导的心肌细胞凋亡和炎症。
