Niclosamide Inhibits Aortic Valve Interstitial Cell Calcification by Interfering with the GSK-3β/β-Catenin Signaling Pathway

The most common heart valve disorder is calcific aortic valve stenosis (CAVS), which is characterized by a narrowing of the aortic valve. Treatment with the drug molecule, in addition to surgical and transcatheter valve replacement, is the primary focus of researchers in this field. The purpose of this study is to determine whether niclosamide can reduce calcification in aortic valve interstitial cells (VICs). To induce calcification, cells were treated with a pro-calcifying medium (PCM). Different concentrations of niclosamide were added to the PCM-treated cells, and the level of calcification, mRNA, and protein expression of calcification markers was measured. Niclosamide inhibited aortic valve calcification as observed from reduced alizarin red s staining in niclosamide treated VICs and also decreased the mRNA and protein expressions of calcification-specific markers: runt-related transcription factor 2 and osteopontin. Niclosamide also reduced the formation of reactive oxygen species, NADPH oxidase activity and the expression of Nox2 and p22 phox . Furthermore, in calcified VICs, niclosamide inhibited the expression of β-catenin and phosphorylated glycogen synthase kinase (GSK-3β), as well as the phosphorylation of AKT and ERK. Taken together, our findings suggest that niclosamide may alleviate PCM-induced calcification, at least in part, by targeting oxidative stress mediated GSK-3β/β-catenin signaling pathway via inhibiting activation of AKT and ERK, and may be a potential treatment for CAVS.

Isolation and Activity Evaluation of Peptides with Anti-hypertensive Activity from Commercial Enzymatic Flounder Fish Hydrolysate

The potent antioxidant and anti-hypertension activities have evidenced gastric enzymatic hydrolysates from flounder fish and their derived peptides. However, peptide composition and functional effect in various enzymatic hydrolysates differ by enzyme types, hydrolyzed times temperatures, etc. Therefore, we determined potential anti-hypertensive effect of hydrolysates produced from flounder fish using commercial enzymes such as Protamex, Flavourzyme, and Kojizyme which are common food grade proteases and characterized on its derived peptides. In this study, Protamex-mediated hydrolysate showed a more potent anti-hypertension effect than other commercial enzymes. Protamex-mediated hydrolysate was fractionated into three ranges of molecular weight (above 10 kDa (FPH-I), 5-10 kDa (FPH-II), and below 5 kDa (FPH-III)). The FPH-III exhibited the strongest anti-hypertensive effect, and it was revealed that three active peptides, valine-phenylalanine-serine-glycinetryptophan-alanine-alanine (VFSGWAA), leucine-histidine-phenylalanine (LHF) and tryptophan-proline-tryptophan (WPW) were contained. The activities were confirmed via angiotensin-converting enzyme (ACE) inhibition and molecular docking simulation. Among the three peptides, LHF and WPW have a molecular structure stability against the gastrointestinal digestion. LHF showed a significant anti-hypertension effect at 9 h after oral administration in spontaneously hypertensive rats (SHRs). Therefore, we suggest that Protamex-mediated hydrolysate would be an excellent anti-hypertensive agent due to the existence of stabilized functional peptides, including LHF and WPW.

Surgically Metabolic Resection of Pericardial Fat to Ameliorate Myocardial Mitochondrial Dysfunction in Acute Myocardial Infarction Obese Rats

Background@#Pericardial fat (PF) is highly associated with cardiovascular disease but the effectiveness of surgical resection of PF is still unknown for myocardial mitochondrial structure and function in acute myocardial infarction (AMI) with obesity. The aim of this study was to demonstrate the difference in myocardial mitochondrial structure and function between obese AMI with additionally resected PF and those without resected PF. @*Methods@#Obese rats with 12-week high fat diet (45 kcal% fat, n = 21) were randomly assigned into 3 groups: obese control, obese AMI and obese AMI with additionally resected PF. One week after developing AMI and additional resection of PF, echocardiogram, myocardial mitochondrial histomorphology, oxidative phosphorylation system (OXPHOS), anti-oxidative enzyme and sarcoplasmic reticulum Ca 2+ ATPase 2 (SERCA2) in the non-infarcted area were assessed between these groups. @*Results@#There was significant improvement of systolic function in AMI with PF resection compared with the AMI group in the echocardiogram. Even though the electron microscopic morphology for the mitochondria seems to be similar between the AMI with PF resection and AMI groups, there was an improved expression of PGC-1α and responsive OXPHOS including NDUFB3, NDUFB5 and SDHB are associated with the ATP levels in the AMI with PF resection compared with those in the AMI group. In addition, the expression levels of antioxidant enzymes (MnSOD) and SERCA2 were improved in the AMI with PF resection compared with those in the AMI group. @*Conclusion@#Surgical resection of PF might ameliorate myocardial mitochondria dysfunction in obese AMI.