Untangling Autophagy Measurements

Gottlieb, Roberta A.; Andres, Allen M.; Sin, Jon; Taylor, David P.J.

doi:10.1161/circresaha.116.303787

Cited by 134 publications

(112 citation statements)

References 101 publications

Supporting

Mentioning

108

Contrasting

Order By: Relevance

“…Another protein involved in autophagy regulation is AMPK. AMPK is phosphorylated, leading to its activation and autophagy induction [9]. As shown in Fig.…”

Section: Resultsmentioning

confidence: 99%

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

Cao

Chen

Dong

et al. 2017

Cell Physiol Biochem

View full text Add to dashboard Cite

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.

show abstract

“…Another protein involved in autophagy regulation is AMPK. AMPK is phosphorylated, leading to its activation and autophagy induction [9]. As shown in Fig.…”

Section: Resultsmentioning

confidence: 99%

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

Cao

Chen

Dong

et al. 2017

Cell Physiol Biochem

View full text Add to dashboard Cite

show abstract

“…If only markers of the early autophagic processes are measured, an investigator might be misled and before any firm conclusions about increased or decreased autophagy are reached, it is important to measure the overall activity of the entire process. 64 …”

Section: Autophagy and Cardiac Diseasementioning

confidence: 99%

Proteotoxicity and Cardiac Dysfunction

McLendon

Robbins

2015

Circulation Research

View full text Add to dashboard Cite

Baseline physiological function of the mammalian heart is under the constant threat of environmental or intrinsic pathological insults. Cardiomyocyte proteins are thus subject to unremitting pressure to function optimally and this depends upon them assuming and maintaining proper conformation. This review explores the multiple defenses a cell may employ for its proteins to assume and maintain correct protein folding and conformation. There are multiple quality control mechanisms to ensure that nascent polypeptides are properly folded and mature proteins maintain their functional conformation. When proteins do misfold, either in the face of normal or pathologic stimuli or because of intrinsic mutations or post-translational modifications, they must either be refolded correctly or recycled. In the absence of these corrective processes, they may become toxic to the cell. Herein, we explore some of the underlying mechanisms that lead to proteotoxicity. The continued presence and chronic accumulation of misfolded or unfolded proteins can be disastrous in cardiomyocytes as these misfolded proteins can lead to aggregation or the formation of soluble peptides that are proteotoxic. This in turn leads to compromised protein quality control and precipitating a downward spiral of the cell's ability to maintain protein homeostasis. Some underlying mechanisms are discussed and the therapeutic potential of interfering with proteotoxicity in the heart is explored.

show abstract

“…Duration/severity of diabetes is also unable to explain the variable findings [e.g., high fat-fed mouse 18 -20 wk duration: increased LC3BII (102) vs. decreased LC3BII (82)]. New work involving evaluation of autophagic flux will be important in gaining mechanistic understanding to resolve these variable findings, and we have recently highlighted the importance of flux measurement in relation to interpreting data from a dietary intervention model (21). Suppressed insulin signaling through the PI3K/Akt/mTOR pathway is a common observation in most diabetic models due to either lower insulin ligand stimulation of the receptor (T1D) or tissue insulin resistance (T2D).…”

Section: )mentioning

confidence: 99%