Mol Pharmacol 2005 67: 1018-1025

誰も敗血症の遺伝子治療など考えていないときに発案した思い出深い研究です。
この第3弾，第4弾は，さらに驚くべき結果となっており，後世に残る研究となると考えております。

Nuclear Factor-kB Decoy Oligodeoxynucleotides Prevent Acute Lung Injury in Mice with Cecal Ligation and Puncture-Induced Sepsis
Naoyuki Matsuda, Yuichi Hattori and Satoshi Gando
Departments of Anesthesiology and Critical Care Medicine, Hokkaido University Graduate School of Medicine, Sapporo, Japan


Abstract

The transcription factor nuclear factor-kB (NF-kB) plays a key role in expression of many inflammatory genes responsible for the pathophysiology of sepsis-induced acute lung injury. We investigated whether the introduction of synthetic double-stranded oligodeoxynucleotides (ODNs) with consensus NF-kB sequence as transcription factor decoy can prevent acute lung injury with suppression of pulmonary expression of multiple genes involved in its pathological process in a cecal ligation and puncture septic mouse model. NF-kB decoy ODNs were introduced with the aid of the hemagglutinating virus of Japan-envelope vector method. Northern blot analysis indicated that transfection of NF-kB decoy ODN, but not of its scrambled form, resulted in a significant inhibition of sepsis-induced gene overexpression of inducible nitric-oxide synthase (iNOS), cyclooxygenase-2, histamine H1-receptor, platelet-activating factor receptor, and bradykinin B1 and B2 receptors in lung Histological damage in lungs tissues. (wall thickening, inflammatory infiltrate, and hemorrhage), increased pulmonary vascular permeability, and blood gas exchange impairment were clearly documented in mice after cecal ligation and puncture. These changes were strongly eliminated by the introduction of NF-kB decoy but not of scrambled ODN. The effects of the iNOS inhibitor FR260330 on these histological and functional derangements compared unfavorably with those of NF-kB decoy ODN transfection. Our results suggest that ODN decoy, acting as in vivo competitor for the transcription factor's ability to bind to cognate recognition sequence, may represent an effective strategy in the treatment of septic acute lung injury.

J Clin Invest. 2005 Feb;115(2):221-4. 血管平滑筋傷害

Defining smooth muscle cells and smooth muscle injury.

Mahoney WM, Schwartz SM.

Department of Pathology, University of Washington, Seattle, Washington 98195-7335, USA.

For 3 decades, terms such as synthetic phenotype and contractile phenotype have been used to imply the existence of a specific mechanism for smooth muscle cell (SMC) responses to injury. In this issue of the JCI, Hendrix et al. offer a far more precise approach to examining the mechanisms of SMC responses to injury, focused not on general changes in phenotype but on effects of injury on a single promoter element, the CArG [CC(A/T)6GG] box, in a single gene encoding smooth muscle (SM) alpha-actin. Since CArG box structures are present in some, but not all, SMC genes, these data suggest that we may be progressing toward establishing a systematic, molecular classification of both SMC subsets and the response of SMCs to different injuries.

敗血症病態の血管平滑筋傷害をこの半年，再び，検討し始めております。

Br J Pharmacol 144: 715-726, 2005 スタチンとエンドセリン

Effects of statins on vascular function of endothelin-1
Fatima Mraiche1, Jonathan Cena1, Debarsi Das1 and Bozena Vollrath1

1Department of Pharmacology, Faculty of Medicine, University of Alberta, Edmonton, Alberta, Canada T6G 2H7
Correspondence: Bozena Vollrath, E-mail: bozena.vollrath@ualberta.ca

Although statins have been reported to inhibit the prepro-endothelin-1 (ET-1) gene transcription in endothelial cells, their effects on the vascular function of ET-1 have not been explored. We, therefore, examined the effects of statins on contraction and DNA synthesis mediated by ET-1 in vascular smooth muscle. The effects of statins on contraction induced by ET-1 were compared to those mediated by noradrenaline (NA) and KCl.
Simvastatin (SV) induced a concentration-dependent relaxation of tonic contraction mediated by ET-1 (10 nM) (IC50 value of 1.3 M). The relaxation was also observed in rings precontracted with NA (0.1 M) and KCl (60 mM). In contrast, pravastatin did not have any effect on the contractions.
Endothelial denudation or pretreatment with L-NAME did not prevent the relaxation, but did reduce the relaxant activity of SV.
SV prevented Rho activation caused by ET-1 and KCl in aortic homogenates, as assessed by a Rho pulldown assay.
The Rho kinase inhibitor HA-1077 mimicked the effects of SV on tonic contractions induced by ET-1, NA and KCl.
Pretreatment with the Kv channels inhibitor, 4-aminopyridine, attenuated the ability of SV to relax contractions mediated by ET-1 and NA.
In quiescent VSM cells, SV significantly inhibited DNA synthesis and Rho translocation stimulated by ET-1, as assessed by [3H]thymidine incorporation and Western blot, respectively.
Inhibition of Rho geranylgeranylation by GGTI-297, or treatment with HA-1077, mimicked the effects of SV on DNA synthesis stimulated by ET-1.
The results show that the statin potently inhibits both ET-1-mediated contraction and DNA synthesis via multiple mechanisms. Clinical benefits of statins may result, in part, from their effects on vascular function of ET-1.

僕の敗血症病態におけるスタチンの研究はほぼ完結に近づいています。

敗血症の定義に関する重要文献集 Dｒ.Marshallの論文もあります

 敗血症の定義に関する重要文献集 

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J. Biol. Chem. 2005 280: 10040-10046

僕と異なるデータである。培養細胞系で検証しなければない。
しかし，血管内皮細胞にはCD14がない。LPSの反応は極端に弱いか，ないはずであるので，どういうメカニズムでiNOSが上昇するのか自体が疑問である。レフリーはどういう審査をしたのだろうか？

Resistance to Endotoxic Shock in Endothelial Nitric-oxide Synthase (eNOS) Knock-out Mice
A PRO-INFLAMMATORY ROLE FOR eNOS-DERIVED NO IN VIVO*
Linda Connelly, Melanie Madhani, and Adrian J. Hobbs
From the Wolfson Institute for Biomedical Research, University College London, Cruciform Building, Gower Street, London WC1E 6AE, United Kingdom

The expression of inducible nitric-oxide synthase (iNOS) and subsequent "high-output" nitric oxide (NO) production underlies the systemic hypotension, inadequate tissue perfusion, and organ failure associated with septic shock. Therefore, modulators of iNOS expression and activity, both endogenous and exogenous, are important in determining the magnitude and time course of this condition. We have shown previously that NO from the constitutive endothelial NOS (eNOS) is necessary to obtain maximal iNOS expression and activity following exposure of murine macrophages to lipopolysaccharide (LPS). Thus, eNOS represents an important regulator of iNOS expression in vitro. Herein, we validate this hypothesis in vivo using a murine model of sepsis. A temporal reduction in iNOS expression and activity was observed in LPS-treated eNOS knock-out (KO) mice as compared with wild-type animals; this was reflected in a more stable hemodynamic profile in eNOS KO mice during endotoxaemia. Furthermore, in human umbilical vein endothelial cells, LPS leads to the activation of eNOS through phosphoinositide 3-kinase- and Akt/protein kinase B-dependent enzyme phosphorylation. These data indicate that the pathogenesis of sepsis is characterized by an initial eNOS activation, with the resultant NO acting as a co-stimulus for the expression of iNOS, and therefore highlight a novel pro-inflammatory role for eNOS.

J Clin Invest. 2005 Feb;115(2):233-6

Knock your SOCS off!

Leroith D, Nissley P.

Diabetes Branch, National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK), National Cancer Institute, NIH, Bethesda, Maryland 20892-1758, USA. derek@helix.nih.gov

The growth hormone/IGF-1-signaling (GH/IGF-1-signaling) system is involved in numerous physiological processes during normal growth and development and also in the aging process. Understanding the regulation of this system is therefore of importance to the biologist. Studies conducted over the past decade have shown that the JAK/STAT pathways are involved in GH signaling to the nucleus. More recently, evidence has been presented that a member of the SOCS family, SOCS2, is a negative regulator of GH signaling. This story began several years ago with the dramatic demonstration of gigantism in the SOCS2-knockout mouse. A more specific definition of the role of SOCS2 in GH signaling is provided in this issue of the JCI by the demonstration that the overgrowth phenotype of the SOCS2-/- mouse is dependent upon the presence of endogenous GH and that administration of GH to mice lacking both endogenous GH and SOCS2 produced excessive growth.

Figure 1
Model of JAK/STAT signaling and negative feedback by SOCS proteins. Cytokine signaling involves ligand binding and activation of the cell surface cytokine receptor. Recruitment and activation of JAK in turn facilitates phosphorylation of a STAT tyrosine residue, and subsequent STAT activation induces dimerization. This activation is tightly controlled by multiple negative regulators of phosphorylation such as phosphatases, SOCS, and PIAS. SOCS proteins are also induced by cytokine signaling and form a closed-loop, negative-feedback mechanism.

Figure 2
GH activates the GHR, which leads to multiple tissue effects, including IGF-1 gene expression. In turn, IGF-1, via IGF-1R, induces cellular effects. SOCS2 may negatively regulate these events at multiple levels. A series of papers from the Walter and Eliza Hall Institute of Medical Research, including the paper by Greenhalgh et al. in this issue of the JCI (3), provide convincing evidence that SOCS2 is a negative regulator of GH receptor signaling. Negative regulation of IGF-1R signaling by SOCS2 is more speculative.

Cancer Res. 2003 Oct 1;63(19):6424-31

Interferon regulatory factor 5, a novel mediator of cell cycle arrest and cell death.

Barnes BJ, Kellum MJ, Pinder KE, Frisancho JA, Pitha PM.

Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, The Johns Hopkins University School of Medicine, 1650 Orleans Street, Baltimore, MD 21231, USA. barnebe@jhmi.edu

We have previously shown a critical role for IFN regulatory factor 5 (IRF-5) in the innate immune response to virus infection. For the first time, we now show that although IRF-5 is a direct target of p53, its cell cycle regulatory and proapoptotic effects are p53 independent. IRF-5 inhibits both in vitro and in vivo B-cell lymphoma tumor growth in the absence of wild-type p53. The molecular mechanism(s) of IRF-5-mediated growth inhibition is associated with a G(2)-M cell cycle arrest and modulation of growth regulatory and proapoptotic genes, including p21, Bak, DAP kinase 2, and Bax. Taken together, these data indicate that although IRF-5 is a downstream target of p53, its growth inhibitory and proapoptotic effects are independent of p53.