Vascular Endothelium Has a Local Anti-Adenovirus Vector System and Glucocorticoid Optimizes Its Gene Transduction

Murata, Takahisa; Hori, Masaru; Lee, Sheng; Nakamura, Akio; Kohama, Kazuhiro; Karaki, Hideaki; Ozaki, Hiroshi

doi:10.1161/01.atv.0000174130.75958.b7

Cited by 12 publications

(9 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We used the appropriate siRNA. For specific silencing of GRK2 or β-arrestin 2 expression, we used a well-established method for the organ culture (15,16). After transfection with a given siRNA, aortas were incubated for 48 h, as described by Chen et al (17).…”

Section: Methodsmentioning

confidence: 99%

G Protein–Coupled Receptor Kinase 2, With β-Arrestin 2, Impairs Insulin-Induced Akt/Endothelial Nitric Oxide Synthase Signaling in ob/ob Mouse Aorta

et al. 2012

View full text Add to dashboard Cite

In type 2 diabetes, impaired insulin-induced Akt/endothelial nitric oxide synthase (eNOS) signaling may decrease the vascular relaxation response. Previously, we reported that this response was negatively regulated by G protein–coupled receptor kinase 2 (GRK2). In this study, we investigated whether/how in aortas from ob/ob mice (a model of type 2 diabetes) GRK2 and β-arrestin 2 might regulate insulin-induced signaling. Endothelium-dependent relaxation was measured in aortic strips. GRK2, β-arrestin 2, and Akt/eNOS signaling pathway proteins and activities were mainly assayed by Western blotting. In ob/ob (vs. control [Lean]) aortas: 1) insulin-induced relaxation was reduced, and this deficit was prevented by GRK2 inhibitor, anti-GRK2 antibody, and an siRNA specifically targeting GRK2. The Lean aorta relaxation response was reduced to the ob/ob level by pretreatment with an siRNA targeting β-arrestin 2. 2) Insulin-stimulated Akt and eNOS phosphorylations were decreased. 3) GRK2 expression in membranes was elevated, and, upon insulin stimulation, this expression was further increased, but β-arrestin 2 was decreased. In ob/ob aortic membranes under insulin stimulation, the phosphorylations of Akt and eNOS were augmented by GRK2 inhibitor. In mouse aorta, GRK2 may be, upon translocation, a key negative regulator of insulin responsiveness and an important regulator of the β-arrestin 2/Akt/eNOS signaling, which is implicated in diabetic endothelial dysfunction.

show abstract

Section: Methodsmentioning

confidence: 99%

G Protein–Coupled Receptor Kinase 2, With β-Arrestin 2, Impairs Insulin-Induced Akt/Endothelial Nitric Oxide Synthase Signaling in ob/ob Mouse Aorta

et al. 2012

View full text Add to dashboard Cite

show abstract

“…We used a well-established method for the organ culture (17,24), and the appropriate siRNA was transfected into aortas as described by Chen et al (25). Briefly, aortic rings (2 mm in length) were transfected with GRK2 siRNA (300 nM) or with control siRNA (siRNA scrambled control) with the aid of siRNA transfection reagent (6 l/ml transfection volume; Santa Cruz Biotechnology).…”

Section: Studies Using Small Interfering Rna (Sirna) In Organ Culturementioning

confidence: 99%

Inhibitor of G Protein-Coupled Receptor Kinase 2 Normalizes Vascular Endothelial Function in Type 2 Diabetic Mice by Improving β-Arrestin 2 Translocation and Ameliorating Akt/eNOS Signal Dysfunction

et al. 2012

View full text Add to dashboard Cite

In type 2 diabetes, although Akt/endothelial NO synthase (eNOS) activation is known to be negatively regulated by G protein-coupled receptor kinase 2 (GRK2), it is unclear whether the GRK2 inhibitor would have therapeutic effects. Here we examined the hypotensive effect of the GRK2 inhibitor and its efficacy agonist both vascular (aortic) endothelial dysfunction (focusing especially on the Akt/eNOS pathway) and glucose intolerance in two type 2 diabetic models (ob/ob mice and nicotinamide+streptozotocin-induced diabetic mice). Mice were treated with a single injection of the GRK2 inhibitor or vehicle, and the therapeutic effects were compared by examining vascular function and by Western blotting. The GRK2 inhibitor lowered blood pressure in both diabetic models but not in their age-matched controls. The GRK2 inhibitor significantly improved clonidine-induced relaxation only in diabetic (ob/ob and DM) mice, with accompanying attenuations of GRK2 activity and translocation to the plasma membrane. These protective effects of the GRK2 inhibitor may be attributable to the augmented Akt/eNOS pathway activation (as evidenced by increases in Akt phosphorylation at Ser(473) and at Thr(308), and eNOS phosphorylation at Ser(1177)) and to the prevention of the GRK2 translocation and promotion of β-arrestin 2 translocation to the membrane under clonidine stimulation. Moreover, the GRK2 inhibitor significantly improved the glucose intolerance seen in the ob/ob mice. Our work provides the first evidence that in diabetes, the GRK2 inhibitor ameliorates vascular endothelial dysfunction via the Akt/eNOS pathway by inhibiting GRK2 activity and enhancing β-arrestin 2 translocation under clonidine stimulation, thereby contributing to a blood pressure-lowering effect. We propose that the GRK2 inhibitor may be a promising therapeutic agent for cardiovascular complications in type 2 diabetes.

show abstract

“…Some of these cytokines are interferons (IFN) that have been linked to pregnancy recognition (IFN-α in pigs, IFN-α and IFN-β in humans, ω48 and IFN-γ in rabbits or IFN-τ in ruminants) (Cross & Roberts, 1989; Aboagye-Mathiesen et al ., 1995; Muscettola et al ., 2003; Spencer et al ., 2004). Moreover, IFNs are involved in the angiogenesis process and the activation of natural killer cells (IFN-α in mouse or IFN-γ in rabbits) (Krusche et al ., 2002; Murata et al ., 2005; Godornes et al ., 2007).…”

Section: Introductionmentioning

confidence: 99%

Oviductal and endometrial mRNA expression of implantation candidate biomarkers during early pregnancy in rabbit

Saeed¹,

Saenz-de-Juano

Marco‐Jiménez

et al. 2013

Zygote

View full text Add to dashboard Cite

SummaryPrenatal losses are a complex problem. Pregnancy requires orchestrated communication between the embryo and the uterus that includes secretions from the embryo to signal pregnancy recognition and secretion and remodelling from the uterine epithelium. Most of these losses are characterized by asynchronization between embryo and uterus. To better understand possible causes, an analysis was conducted of gene expression of a set of transcripts related to maternal recognition and establishment of rabbit pregnancy (uteroglobin, SCGB1A1; integrin α1, ITGA1; interferon-γ, IFNG; vascular endothelial growth factor, VEGF) in oviduct and uterine tissue at 16, 72 or 144 h post-ovulation and insemination. In the oviduct tissue, a significant decrease in the level of SCGB1A1 mRNA expression was observed from 144 h post-ovulation. In the case of ITGA1, the transcript abundance was initially lower, but mRNA expression increased significantly at 72 and 144 h post-ovulation. For IFNG, a huge decrease was observed from 16 to 72 h post-ovulation. Finally, no significant differences were observed in the VEGF transcript. For the endometrium, the results showed a significant decline in the level of SCGB1A1 mRNA expression from 16 to 144 h post-ovulation induction. The highest levels of ITGA1 transcript were detected at 144 h, followed by the 16 h group and lower at 72 h post-ovulation. For IFNG there were no significant differences among post-ovulation induction times. Finally, it was possible to observe that VEGF mRNA abundance was present at low levels at 16 h post-ovulation and remained low at 72 h, but increased at 144 h. The functional significance of these observations may provide new insights into the maternal role in prenatal losses.

show abstract

Vascular Endothelium Has a Local Anti-Adenovirus Vector System and Glucocorticoid Optimizes Its Gene Transduction

Cited by 12 publications

References 31 publications

G Protein–Coupled Receptor Kinase 2, With β-Arrestin 2, Impairs Insulin-Induced Akt/Endothelial Nitric Oxide Synthase Signaling in ob/ob Mouse Aorta

G Protein–Coupled Receptor Kinase 2, With β-Arrestin 2, Impairs Insulin-Induced Akt/Endothelial Nitric Oxide Synthase Signaling in ob/ob Mouse Aorta

Inhibitor of G Protein-Coupled Receptor Kinase 2 Normalizes Vascular Endothelial Function in Type 2 Diabetic Mice by Improving β-Arrestin 2 Translocation and Ameliorating Akt/eNOS Signal Dysfunction

Oviductal and endometrial mRNA expression of implantation candidate biomarkers during early pregnancy in rabbit

Contact Info

Product

Resources

About