Emodin Inhibits Doxorubicin-Induced Cardiotoxicity by Regulating miR-29a-5p/VEGFB Mediated Apoptosis
- VernacularTitle:大黄素通过调控miR-29a-5p/VEGFB介导的细胞凋亡抑制多柔比星诱导的心脏毒性
- Author:
Zhaofeng ZHANG
1
;
Ruohan TANG
;
Liang ZHANG
Author Information
- Keywords: Emodin; Doxorubicin; H9C2 cells; Apoptosis; miR-29a-5p; VEGFB
- From: World Science and Technology-Modernization of Traditional Chinese Medicine 2023;25(6):2025-2035
- CountryChina
- Language:Chinese
- Abstract: Objective To investigate the effect and possible mechanism of Emodin on Doxorubicin(DOX)-induced cardiotoxicity.Methods H9C2 cells were preincubated with low,medium and high doses(5,15 and 25 μmol·L-1)of Emodin for 8 h,and then cardiomyocytes were treated with 2.5 μmol·L-1 DOX for 16 h to establish a cardiotoxic injury model.MTT assay was used to detect the cell survival rate.The levels of cardiac troponin(cTnT),lactate dehydrogenase(LDH),creatine kinase(CK),superoxide dismutase(SOD),reactive oxygen species(ROS),malondialdehyde(MDA),reduced glutathione(GSH),glutathione peroxidase(GSH-Px)and the activities of Caspase-3 and Caspase-9 in H9C2 cells were detected by kit;RT-qPCR was used to detect miR-29a-5p and vascular endothelial growth factor-B(VEGFB)expression levels.The expression levels of VEGFB protein and other apoptosis-related factors were detected by Western blot.After transfection of miR-29a-5p mimics and inhibitors,the targeting of VEGFB to miR-29a-5p was detected by RT-qPCR and Western blot.H9C2 cells were transfected with miR-29a-5p mimics,Under the dose of Emodin 15 μmol·L-1,the activity of H9C2 cells was detected by MTT method,and the expression levels of miR-29a-5p and VEGFB were detected by RT-qPCR and Western blot.Results Compared with the DOX group,Emodin low,medium and high dose groups significantly increased the activity of H9C2 cells(P<0.05,P<0.01),and decreased the levels of cTnT,LDH and CK(P<0.05,P<0.01);RT-qPCR indicated that the expression level of miR-29a-5p decreased significantly and the expression level of VEGFB increased significantly(P<0.05,P<0.01);The levels of ROS and MDA decreased significantly,and the levels of GSH and GSH-Px increased significantly(P<0.05,P<0.01);TUNEL positive cells decreased significantly,and the activities of Caspase-3 and Caspase-9 decreased significantly.Western blot showed that the expression level of VEGFB increased significantly,the expression level of apoptosis-related factor Bax decreased significantly,and the expression levels of Bcl-2 and Bcl-xl increased significantly(P<0.05,P<0.01).After transfection with miR-29a-5p mimics and inhibitors,the expression of VEGFB decreased and increased respectively(P<0.05,P<0.01).After transfection of miR-29a-5p mimics and treatment with Emodin 15 μmol·L-1 of H9C2 cells,the significant decrease in the activity of H9C2 cells induced by DOX was reversed,and the significant decrease in the expression level of VEGFB was also reversed(P<0.05,P<0.01).Conclusion Emodin significantly reduces DOX-induced cardiotoxicity by regulating miR-29a-5p/VEGFB-mediated cardiomyocyte apoptosis.
