Journal
ACTA PHARMACOLOGICA SINICA
Volume 37, Issue 3, Pages 368-381Publisher
ACTA PHARMACOLOGICA SINICA
DOI: 10.1038/aps.2015.146
Keywords
ophiopogonin D; cardiomyocytes; angiotensin II; CYP2J3; epoxyeicosatrienoic acid; ER stress; Ca2+ homeostasis; BAPTA
Funding
- National Basic Research Program of China (973 Program) [2011CB505304, 2012CB518402]
- Scientific and Technological Major Special Project Major Creation of New Drugs [2009ZX09501-304]
Ask authors/readers for more resources
Aim: CYP2J3 in myocardium metabolizes arachidonic acid to 4 regioisomeric epoxyeicosatrienoic acids (EETs), which have diverse biological activities in rat heart. In this study we examined whether CYP2J3 was involved in cardioprotective effects of ophiopogonin D (OPD), a steroidal glycoside isolated from Chinese herb Radix ophiopogonis. Methods: Rat cardiomyoblast cell line (H9c2 cells) was tested. Intracellular Ca2+ concentrations ([Ca2+](i)) were measured using Fluo-4/AM. The expression of calcium-regulating molecules and ER stress signaling molecules was measured with qRT-PCR and Western blot analyses. Cell apoptosis was quantified with Hoechst 33258 staining and TUNEL assay. The level of 14,15-DHET, a stable metabolite of 14,15-EET, was assessed with ELISA. Results: Angiotensin II (10(-6) mol/L) significantly decreased the expression of calcium-regulating molecules (SERCA2a, PLB, RyR2 and FKBP12.6), and elevated [Ca2+](i) in H9c2 cells. Furthermore, angiotensin II markedly increased the expression of ER stress signaling molecules (GRP78, CHOP, p-JNK and cleaved caspase-12) and ER stress-mediated apoptosis. OPD (100, 250 and 500 nmol/L) dose-dependently increased CYP2J3 expression and 14,15-DHET levels in normal H9c2 cells. Pretreatment of H9c2 cells with OPD suppressed angiotensin II-induced abnormalities in Ca2+ homeostasis, ER stress responses and apoptosis. Overexpression of CYP2J3 or addition of exogenous 14,15-EET also prevented angiotensin II-induced abnormalities in Ca2+ homeostasis, whereas transfection with CYP2J3 siRNA diminished the effects of OPD on Ca2+ homeostasis. Furthermore, the intracellular Ca2+ chelator BAPTA suppressed angiotensin II-induced ER stress responses and apoptosis in H9c2 cells. Conclusion: OPD is a novel CYP2J3 inducer that may offer a therapeutic benefit in treatment of cardiovascular diseases related to disturbance of Ca2+ homeostasis and ER stress.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available