ABSTRACT
BACKGROUND: Abnormalities of the L-arginine-nitric oxide pathway induce hypertension. 5-Lipoxygenase (5-LO) is the key enzyme involved in synthesis of leukotrienes (LTs). However, whether nitricoxide synthase dysfunction induces hypertensive vascular remodeling by regulating 5-LO activity and its downstream inflammatory metabolites remains unknown. METHODS AND RESULTS: Six-week L-NAME treatment significantly induced hypertension and vascular remodeling in both wild-type (WT) and 5-LO-knockout (5-LO-KO) mice, and blood pressure in caudal and carotid arteries was lower in 5-LO-KO than WT mice with L-NAME exposure. On histology, L-NAME induced less media thickness, media-to-lumen ratio, and collagen deposition and fewer Ki-67-positive vascular smooth muscle cells (VSMCs) but more elastin expression in thoracic and mesenteric aortas of 5-LO-KO than L-NAME-treated WT mice. L-NAME significantly increased LT content, including LTB4 and cysteinyl LT (CysLTs), in plasma and neutrophil culture supernatants from WT mice. On immunohistochemistry, L-NAME promoted the colocalization of 5-LO and 5-LO-activating protein on the nuclear envelope of cultured neutrophils, which was accompanied by elevated LT content in culture supernatants. In addition, LTs significantly promoted BrdU incorporation, migration and phenotypic modulation in VSMCs. CONCLUSION: L-NAME may activate the 5-LO/LT pathway in immune cells, such as neutrophils, and promote the products of 5-LO metabolites, including LTB4 and CysLTs, which aggravate vascular remodeling in hypertension. 5-LO deficiency may protect against hypertension and vascular remodeling by reducing levels of 5-LO downstream inflammatory metabolites.
Subject(s)
Arachidonate 5-Lipoxygenase/genetics , Hypertension/prevention & control , Vascular Remodeling , Animals , Aorta/metabolism , Aorta/pathology , Arachidonate 5-Lipoxygenase/deficiency , Blood Pressure/drug effects , Cell Movement/drug effects , Cell Proliferation/drug effects , Hypertension/chemically induced , Hypertension/pathology , Leukotriene A4/blood , Leukotriene A4/pharmacology , Mice , Mice, Inbred C57BL , Mice, Knockout , Muscle, Smooth, Vascular/cytology , Muscle, Smooth, Vascular/metabolism , NG-Nitroarginine Methyl Ester/metabolism , NG-Nitroarginine Methyl Ester/toxicity , Neutrophils/immunology , Neutrophils/metabolism , Nitric Oxide Synthase Type II/metabolism , Rats , Rats, Sprague-Dawley , Vascular Remodeling/drug effectsABSTRACT
SIRT6, a member of the NAD(+)-dependent class III deacetylase sirtuin family, has been revealed to play important roles in promoting cellular resistance against oxidative stress. The formation of reactive oxygen species (ROS) and oxidative stress are the crucial mechanisms underlying cellular damage and dysfunction in cardiac ischemia/reperfusion (I/R) injury, but the role of SIRT6 in I/R-induced ROS and oxidative stress is poorly understood. In this study, by using heterozygous SIRT6 knockout (SIRT6(+/-)) mice and cultured neonatal cardiomyocyte models, we investigated how SIRT6 mediates oxidative stress and myocardial injury during I/R. Partial knockout (KO) of SIRT6 aggravated myocardial damage, ventricular remodeling, and oxidative stress in mice subjected to myocardial I/R, whereas restoration of SIRT6 expression by direct cardiac injection of adenoviral constructs encoding SIRT6 reversed these deleterious effects of SIRT6 KO in the ischemic heart. In addition, partial deletion of the SIRT6 gene decreased myocardial functional recovery following I/R in a Langendorff perfusion model. Similarly, the protective effects of SIRT6 were also observed in cultured cardiomyocytes following hypoxia/reoxygenation. Intriguingly, SIRT6 was noticed to up-regulate AMP/ATP and then activate the adenosine 5'-monophosphate-activated protein kinase (AMPK)-forkhead box O3α (FoxO3α) axis and further initiated the downstream antioxidant-encoding gene expression (manganese superoxide dismutase and catalase), thereby decreasing cellular levels of oxidative stress and mediating cardioprotection in the ischemic heart. These results suggest that SIRT6 protects the heart from I/R injury through FoxO3α activation in the ischemic heart in an AMP/ATP-induced AMPK-dependent way, thus upregulating antioxidants and suppressing oxidative stress.
Subject(s)
Forkhead Transcription Factors/metabolism , Myocardial Reperfusion Injury/metabolism , Sirtuins/metabolism , AMP-Activated Protein Kinases/metabolism , Adenosine Monophosphate/metabolism , Adenosine Triphosphate/metabolism , Animals , Animals, Newborn , Antioxidants/metabolism , Apoptosis , Catalase/metabolism , Cells, Cultured , Down-Regulation , Forkhead Box Protein O3 , In Vitro Techniques , Male , Mice , Myocytes, Cardiac/metabolism , Oxidative Stress , Rats, Sprague-Dawley , Reactive Oxygen Species/metabolism , Sirtuins/genetics , Superoxide Dismutase/metabolism , Ventricular RemodelingABSTRACT
OBJECTIVE: To investigate the function of Parkinson's disease (PD)-related protein Pink1 in autophagy. METHODS: Pink1, autophagy inhibitor Bcl-XL and autophagy induced-factor Beclin1 were amplified with RT-PCR and contructed into pcDNA3. 1 (+) vector. The stable HEK293 cell line of Pink1 expression was established through the lentivirus system. Pink1, Bcl-XL and Beclin1 were tansfected into the HEK293 Cell Line. Co-Immunoprecipitation and Western blot were performed to determine the interaction between Pink1 and Bcl-XL, the effect of Pink1 on the interaction between Bcl-XL and Beclin1, and the function of Pink1 in autophagy. RESULTS: There was a new interaction between Pink1 and antophagy inhibitor Bcl-XL. Overexpressed Pink1 promoted the dissociation of autophagy induced-factor Beclin1 from Bcl-XL. Overexpressed Pink1 increased the autophagic protein LC3 II/LC3 I. CONCLUSION: Pakinson's disease related protein Pink1 promotes the dissociation of autophagy induced-factor Beclin1 from antophagy inhibitor Bcl-XL and promotes autophagy.
