Transforming growth factor-β1 antisense oligodeoxynucleotides block interstitial fibrosis in unilateral ureteral obstruction

Transforming growth factor-β1 antisense oligodeoxynucleotides block interstitial fibrosis in unilateral ureteral obstruction.BackgroundInterstitial expression of transforming growth factor-β1 (TGF-β1) is important in tubulointerstitial fibrosis, a common process in most progressive renal diseases. However, no effective therapy for progressive interstitial fibrosis is known. Recently, we developed an artificial viral envelope (AVE)-type hemagglutinating virus of Japan (HVJ) liposome-mediated retrograde ureteral gene transfer method, which allowed us to introduce the genetic material selectively into renal interstitial fibroblasts.MethodWe introduced antisense or scrambled oligodeoxynucleotides (ODNs) for TGF-β1 into interstitial fibroblasts in rats with unilateral ureteral obstruction, a model of interstitial fibrosis, to block interstitial fibrosis by retrograde ureteral injection of AVE-type HVJ liposomes.ResultsTGF-β1 and type I collagen mRNA increased markedly in the interstitium of untreated obstructed kidneys, and those were not affected by scrambled ODN transfection. Northern analysis and in situ hybridization revealed that the levels of TGF-β1 and type I collagen mRNA were dramatically decreased in antisense ODN-transfected obstructed kidneys. Consequently, the interstitial fibrotic area of the obstructed kidneys treated with antisense ODN was significantly less than that of the obstructed kidneys untreated or treated with scrambled ODN.ConclusionThe introduction of TGF-β1 antisense ODN into interstitial fibroblasts may be a potential therapeutic maneuver for interstitial fibrosis

Increased release of nitric oxide in ischemic hearts after exercise in patients with effort angina

AbstractObjectives. The aim of this study was to determine whether the release of nitric oxide (NO) from the ischemic heart increases during exercise in patients with effort angina.Background. Myocardial ischemia increases NO production in the canine heart, but no such increase has been demonstrated in the ischemic human heart.Methods. Fifteen patients with effort angina underwent supine ergometer exercise tests. All patients had severe proximal stenosis (>90%) in the left anterior descending coronary artery. The control group consisted of 17 subjects without coronary artery disease or systemic hemodynamic abnormalities.Results. Neither the lactate extraction ratio (LER) nor the difference in NO concentration between coronary venous and arterial blood (ΔVA[NO]) was affected by exercise in the control subjects. In patients with effort angina, neither variable differed from that in the control group at rest; however, exercise markedly decreased LER and significantly increased ΔVA(NO) (from 4.7 ± 0.3 to 16.5 ± 1.6 μmol/liter, p < 0.001) in the patient group. The extent of decrease in LER was significantly correlated with the extent of increase in ΔVA(NO) in the patients with effort angina (r2= −0.837, p < 0.001).Conclusions. Provocation of myocardial ischemia by exercise stress increases NO production in the hearts of patients with effort angina

Role of intron 1 in smooth muscle α-actin transcriptional regulation in activated mesangial cells in vivo

Role of intron 1 in smooth muscle α-actin transcriptional regulation in activated mesangial cells in vivo.BackgroundThe activation of glomerular mesangial cells is one of the early, important features of progressive glomerular disease. Smooth muscle α-actin (SMαA) is an excellent marker of activated mesangial cells. However, the mechanisms of SMαA regulation are only available from in vitro investigation.MethodsWe examined in vivo promoter analysis of the SMαA gene-utilizing transgenic mice harboring different promoter regions of the SMαA gene fused to chloramphenicol acetyl transferase (CAT). CAT activities were tested in primary cultured mesangial cells and in glomerular legions of Habu venom glomerulonephritis.ResultsThe DNA sequence -891 to +3828, which contains exon 1, intron 1, and the first 14bp of exon 2 in addition to the 5′-flanking sequence of the SMαA gene, induced high levels of transcription in activated mesangial cells in in vivo habu venom glomerulonephritis and in cultured mesangial cells derived from transgenic mice. The DNA region -891 to -124 was a positive element in mesangial cells derived from transgenic mice. Deletions (3316 or 137bp) in intron 1 reduced transcription to undetectable levels. The 137bp sequence is highly conserved among several species, containing one CArG box element, which is one of the key motifs for transcriptional activation of contractile-related proteins. In vitro transfection analysis failed to demonstrate these positive effects of intron 1 and region -891 to -124.ConclusionsIn vivo promoter analysis of the SMαA gene provided new information about the transcriptional regulation of SMαA in activated mesangial cells. The DNA region -891 to -124 has a positive effect on SMαA transcription in cultured mesangial cells. The intron 1 region (+1088 to +1224) plays a pivotal role in SMαA transcription in activated mesangial cells in vivo. Further analysis of this conserved region in intron 1, including the CArG motif, will be of great value in understanding the molecular mechanisms of mesangial activation

Cyclooxygenase Regulates Angiogenesis Induced by Colon Cancer Cells

AbstractTo explore the role of cyclooxygenase (COX) in endothelial cell migration and angiogenesis, we have used two in vitro model systems involving coculture of endothelial cells with colon carcinoma cells. COX-2-overexpressing cells produce prostaglandins, proangiogenic factors, and stimulate both endothelial migration and tube formation, while control cells have little activity. The effect is inhibited by antibodies to combinations of angiogenic factors, by NS-398 (a selective COX-2 inhibitor), and by aspirin. NS-398 does not inhibit production of angiogenic factors or angiogenesis induced by COX-2-negative cells. Treatment of endothelial cells with aspirin or a COX-1 antisense oligonucleotide inhibits COX-1 activity/expression and suppresses tube formation. Cyclooxygenase regulates colon carcinoma-induced angiogenesis by two mechanisms: COX-2 can modulate production of angiogenic factors by colon cancer cells, while COX-1 regulates angiogenesis in endothelial cells

Effect of intracoronary thrombectomy on 30-day mortality in non-diabetic patients with acute hyperglycemia after acute myocardial infarction

SummaryBackgroundThere is limited evidence about useful therapeutic interventions for patients with acute hyperglycemia (AH) after acute myocardial infarction (AMI).MethodsWe studied 2433 consecutive non-diabetic AMI patients who underwent percutaneous coronary intervention (PCI) within 24h after the onset. Patients were divided into two groups according to the presence or absence of AH (admission serum glucose level ≥11.1mmol/l). We assessed the association between intracoronary thrombectomy and the clinical outcome in AMI patients with AH.ResultsPatients with AH had more risk factors than those without AH. The 30-day mortality rate of patients with AH was significantly higher than that of those without (11.7% vs 1.7%, p<0.001). Among patients with AH, the 30-day mortality rate was significantly lower for those with intracoronary thrombectomy than those without it (4.9% vs 17.2%, p=0.004). Among patients without AH, however, the 30-day mortality rate was similar between those with and without intracoronary thrombectomy (1.5% vs 1.9%, p=NS). Multivariate analysis showed that intracoronary thrombectomy was associated with an improved 30-day mortality rate for patients with AH (hazard ratio: HR 0.184, 95% CI 0.057–0.598, p=0.005).ConclusionsIn AMI patients with AH, intracoronary thrombectomy prior to PCI might improve the 30-day mortality rate