Stachydrine Protects Against Pressure Overload-Induced Cardiac Hypertrophy by Suppressing Autophagy.

Abstract

BACKGROUND:

Autophagy is required for the maintenance of cardiomyocyte homeostasis. However, excessive autophagy plays a maladaptive role in pressure overload-induced heart failure. To identify mechanisms by which Stachydrine inhibits pressure overload-induced cardiac hypertrophy, we determined inhibitory activities against activation of NADPH oxidase, reactive oxygen species(ROS) production and excessive activation of autophagy.

METHODS:

Stachydrine was administered intragastrically to Wistar rats after Transverse aortic constriction(TAC) and H9c2 cells were treated with Stachydrine after Angiotension II stimulation. The activation of NADPH oxidase2 required the membrane translocation of p47phox and p67phox. Cell membrane fraction was isolated by ultracentrifuge in sucrose. The expression of p67phox, p47phox, gp91phox subunit in the cell membrane were determined by western blot. The combination of p67phox and gp91 phox subunit was detected by immunofluorescence staining. The expression of phosphorylated p47phox subunit was determined by western blot. The intracellular ROS were measured with DCF-DA fluoresence. The autophagic flux was measured by recording the fluorescence emission of the fusion protein mRFP-GFP-LC3 by dynamic live-cell imaging. Reuslts: We report here that stachydrine, a major constituent of Leonurus heterophyllus Sweet, inhibited AngII-induced excessive autophagy within H9c2 cells. Stachydrine blocked the over phosphorylation of the p47phox subunit, decreased the translocation of p47phox and p67phox to the membrane, inhibited the activity of NOX2, and reduced the generation of ROS. We also demonstrated that stachydrine ameliorated TAC-induced cardiac hypertrophy, dysfunction and excessive autophagy in vivo.

CONCLUSIONS:

Our study highlights the importance of regulating NOX2 when autophagy is obviously activated. By inhibiting NOX2, Stachydrine inhibits ROS production, thus exerting a remarkable activity of inhibiting hypertrophy, which could have considerable effect on clinical practice.

• Authors: Tong-Tong Cao, Hui-Hua Chen, Zhiwei Dong, Yan-Wu Xu, Pei Zhao, Wei Guo, Hong-Chang Wei, Chen Zhang, Rong Lu
• Journal: Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology (Cell Physiol Biochem) Vol. 42 Issue 1 Pg. 103-114 ( 2017) ISSN: 1421-9778 [Electronic] Germany
• PMID: 28494450 (Publication Type: Journal Article)
• Copyright: © 2017 The Author(s). Published by S. Karger AG, Basel.

Chemical References

• LC3 protein, rat
• Membrane Glycoproteins
• Microtubule-Associated Proteins
• Protective Agents
• Reactive Oxygen Species
• Angiotensin II
• Proline
• NADH, NADPH Oxidoreductases
• NCF2 protein, rat
• Cybb protein, rat
• NADPH Oxidase 2
• NADPH Oxidases
• neutrophil cytosolic factor 1
• stachydrine

Topics

• Angiotensin II (pharmacology)
• Animals
• Autophagy (drug effects)
• Cardiomegaly (etiology, metabolism, prevention & control)
• Cell Line
• Heart (diagnostic imaging, drug effects)
• Male
• Membrane Glycoproteins (metabolism)
• Microtubule-Associated Proteins (metabolism)
• Myocardium (metabolism, pathology)
• NADH, NADPH Oxidoreductases (metabolism)
• NADPH Oxidase 2
• NADPH Oxidases (antagonists & inhibitors, metabolism)
• Phosphorylation (drug effects)
• Pressure
• Proline (analogs & derivatives, pharmacology, therapeutic use)
• Protective Agents (pharmacology, therapeutic use)
• Rats, Wistar
• Reactive Oxygen Species (metabolism)