This paper describes the cloning and sequence analysis of the cDNAs encoding the canine homologues of interleukin-3 (IL-3) and interleukin-6 (IL-6). The coding sequences for canine IL-3 and IL-6 were obtained by using the reverse transcription polymerase chain reaction (RT-PCR) with RNA harvested from canine peripheral blood mononuclear cells (PBMCs). Canine IL-3 cDNA includes a single open reading frame of 432 nucleotides, which encodes a 143 amino acid polypeptide and has 44.7, 42.4, 37 and 23.7% homology with the cow, sheep, human and rat IL-3 sequences, respectively. Canine IL-6 cDNA (GenBank accession number; AF275796) encodes a putative 20-amino acid signal peptide followed by a 187-amino acid mature protein. The predicted amino acid sequence of canine IL-6 shares 60.4, 77.2, 71.0, 55.8 and 42.0% sequence identity with those of human, feline, porcine, sheep and rat IL-6, respectively.
Amino Acid Sequence ; Animals ; Base Sequence ; Cloning, Molecular ; Concanavalin A/pharmacology ; DNA, Complementary/*chemistry ; Dogs/blood/genetics/*immunology ; Interleukin-3/chemistry/*genetics ; Interleukin-6/chemistry/*genetics ; Leukocytes, Mononuclear/chemistry/drug effects ; Molecular Sequence Data ; Open Reading Frames/genetics ; Protein Sorting Signals/genetics ; RNA/blood/genetics ; Reverse Transcriptase Polymerase Chain Reaction/veterinary ; Sequence Homology, Amino Acid ; Sequence Homology, Nucleic Acid

BACKGROUND: Interfering RNA (iRNA) represents a recent breakthrough in effective blocking of the target genes in mammalian cells. Angiotensin-converting enzyme (ACE) has been shown to play an important role in the pathogenesis of hypertension. The purposes of this study were to investigate the effects on blood pressure, myocardial hypertrophy and gene expressions of iRNA targeting ACE. METHODS: Twelve week old male Wistar-Kyoto rats were grouped as follows: control group (C group), spontaneously hypertensive rat (SHR) group (H group), and ACE-iRNA group (A group) in which SHR was treated with recombinant lentiviral vectors carrying small hairpin RNA targeting ACE. Reverse transcription-polymerase chain reaction and western blot analysis of ACE, endothelin (ET)-1, angiotensin (AT) II receptor type 1A, neutrophil cytosolic factor, caspase 3, Bax, and Bcl-2 were performed in the heart tissues. Serum AT, ACE, and high sensitive-C reactive protein were estimated. RESULTS: Systolic blood pressure was significantly decreased in the A group compared with the H group in weeks 3 and 5. Serum AT level was significantly lower on day 1, weeks 3 and 5 after ACE-iRNA treatment. ACE protein contents were significantly lower after ACE-iRNA treatment in week 5. ET-1 and Bcl-2 protein contents were significantly lower after ACE-iRNA treatment in weeks 3 and 5. Bax protein contents were significantly lower after ACE-iRNA treatment in week 3. CONCLUSIONS: Recombinant lentiviral vectors carrying shRNA targeting ACE prevented hypertension. Serum AT and gene expressions such as ACE, ET-1, Bax, and Bcl-2 were significantly decreased after ACE-iRNA treatment.
Angiotensins ; Animals ; bcl-2-Associated X Protein ; Blood Pressure ; Blotting, Western ; Caspase 3 ; Cytosol ; Endothelins ; Gene Expression ; Heart ; Humans ; Hypertension ; Hypertrophy ; Lentivirus ; Lifting ; Male ; Neutrophils ; Rats ; Rats, Inbred SHR ; RNA ; RNA Interference ; RNA, Small Interfering

BACKGROUND AND OBJECTIVES: The renin angiotensin system seems to play an important role in the development of cardiac and vascular hypertrophy in hypertension. The changes in pathology, and gene expressions of the angiotensin II receptor type 1A (ATIA) and angiotensin converting enzyme (ACE) were investigated in order to explore the effects of losartan in spontaneously hypertensive rat (SHR) models. MATERIALS AND METHODS: Twelve week-old male Wistar rats were grouped as follows: control (C) group, hypertension (H) group, and losartan (L) group in which SHR was treated with losartan (10 mg/kg/day). Western blot and reverse transcription-polymerase chain reaction analysis regarding seven genes such as endothelin-1, ACE, ATIA, neutrophil cytosolic factor, brain natriuretic peptide, troponin I, endothelial nitric oxide synthase were performed. RESULTS: Systolic blood pressure was significantly decreased in the L group compared with the H group in weeks 3 and 5. ACE and ATIA proteins in the L group were lower than H group in week 5. CONCLUSION: Losartan reduced blood pressure, cardiac hypertrophy and protein expressions of ACE and ATIA. Changes of protein expressions were more sensitive than changes in pathology. Further study is needed for the differing doses of losartan in SHR models.
Blood Pressure ; Blotting, Western ; Cardiomegaly ; Cytosol ; Endothelin-1 ; Gene Expression ; Humans ; Hypertension ; Hypertrophy ; Losartan ; Male ; Natriuretic Peptide, Brain ; Neutrophils ; Nitric Oxide Synthase Type III ; Peptidyl-Dipeptidase A ; Proteins ; Rats, Inbred SHR ; Rats, Wistar ; Receptors, Angiotensin ; Renin-Angiotensin System ; Troponin I