Biomedical subjects

Y Shibasaki

Publications and source records attributed to Y Shibasaki.

At least 19 recordsLinked to original sources

PubMed2003

Measurement of mechanical strain on mandibular surface with mastication robot: influence of muscle loading direction and magnitude.

OBJECTIVES: To investigate the mechanical effects of mastication on the mandible, we developed computational controlled mastication robot system with human dry skull and analyzed the strain distribution on the mandibular bone surface. DESIGN: In the mastication robot, the mandible was suspended by eight wires, which simulated masticatory muscles. A non-linear spring damper generated viscoelastic properties, and tension sensors for simulation of jaw reflection to avoid unusual biting force were applied as a biological feedback mechanism. By using this robot system, various patterns of muscle loading (change of wire direction and magnitude) were performed. RESULTS: From the results, significant differences in the amount of principal strain and its distribution were demonstrated in each condition (ANOVA, post hoc test, and p < 0.05). The value of maximum principal strain ranged from 79.66 x 10(-6) [at anterior border of ramus (Buccal side), 128 N] to -1.42 x 10(-6) [at foramen mentale (Buccal side), 32 N]. CONCLUSION: These results suggested that the muscle loading generated the mechanical strain on the mandibular bone surface and it was affected by the changes in loading direction and magnitude.

Analysis of Variance↗

PubMed2003

Micro X-ray computed tomography analysis for the evaluation of asymmetrical condylar growth in the rat.

OBJECTIVES: To investigate the influence of forced lateral bite on mandibular growth, micro X-ray computed tomography (CT) was used for the purpose evaluating condylar cartilage and cancellous bone formation in 10 male Wister rats (3 weeks of age). SETTINGS AND SAMPLE POPULATION: The rats were divided into two groups--experimental and control. In experimental group, an inclined crown was cemented onto the maxillary incisors to produce 2.5 mm shift toward the left side during mastication. Right-left differences in whole mandibular length, mandibular height, condylar size, trabecular structure of the condylar head and three-dimensional (3-D) finite element analysis were assessed using 3-D images reconstructed from micro X-ray CT scans when the mice had reached 21 weeks. MEASUREMENTS AND RESULTS: Asymmetrical growth was found in the experimental group, in which the left condylar head became thicker and shorter than the right condylar head during development. When comparing the left and right condyles of the experimental animals, histomorphometric analysis from micro X-ray CT showed that the bone volume (BV) of the cancellous bone, the surface area of the cancellous bone (BS), the BS/BV ratio, the BV fraction (BV/TV), and the trabecular thickness and trabecular number were less for the right condyle than for the left condyle. CONCLUSIONS: These findings suggested that artificial changes in the mastication do influence the growth of condylar head, condylar bone trabecular structure, and mineralization.

Animals↗

PubMed2003

Hemichrome formation observed in human haemoglobin A under various buffer conditions.

AIM: To observe hemichrome formation in human haemoglobin A under various buffer conditions. METHOD: Hemichrome formation of human oxyhaemoglobin A (HbO2) was studied spectrophotometrically in 0.1 m buffer at various temperatures and pH values. RESULTS: Following autoxidation in ferrous HbO2, it was evident that formation of hemichrome, which tends to precipitate, occurred at various stages during the course of the autoxidation reaction namely at initial, intermediate or final stages, depending on temperature and pH of the solution. By varying temperature of the solution from 35 to 55 degrees C and pH from 4.5 to 10.5, it is shown here that HbO2 exhibits high susceptibility for hemichrome formation and its occurrence is a function of pH, temperature and progress of autoxidation of HbO2. Unlike HbO2 and its separated haemoglobin chains, monomeric bovine heart myoglobin (MbO2) did not easily form hemichrome. CONCLUSION: These findings provide a clue on the crucial role of haemoglobin molecule for senescent cell recognition or homeostasis in the blood circulation.

Animals↗

PubMed2003

Renal blood flow measurement with contrast-enhanced harmonic ultrasonography: evaluation of dopamine-induced changes in renal cortical perfusion in humans.

BACKGROUND: An accessible non-invasive method for evaluating renal regional blood flow in real time is highly desirable in the clinical setting. Recent progress in ultrasonography with microbubble contrast has allowed quantification of regional blood flow in animal models. AIMS: Goal ofthis study was to establish a convenient contrast--enhanced harmonic ultrasonography (CEHU) method for evaluating renal cortical blood flow in humans. METHODS: We carried out intermittent second harmonic imaging in 9 healthy volunteers. Pulse interval was progressively decreased from 4 s - 0.2 s during continuous venous infusion of the microbubble contrast agent. RESULTS: Pulse interval versus CEHU-derived acoustic intensity plots provided microbubble velocity (MV) and fractional vascular volume (FVV) during renal cortical perfusion in humans. Low-dose dopamine infusion (2 microg/min/kg) resulted in a significant increase in MV which correlated well with the increase in total renal blood flow (RBF) determined by a conventional study of p-aminohippurate clearance (C(PAH)) (r = 0.956, p < 0.0001). Although FVV was not significantly increased, alterations in CEHU-derived renal cortical blood flow calculated by the products of MV and FVV were also correlated with alterations in total RBF (r = 0.969, p < 0.0001). Thus, low-dose dopamine infusion increases renal cortical blood flow observed in CEHU, mainly by increasing MV. CONCLUSIONS: The present study shows that renal cortical blood flow in humans can be measured non-invasively by CEHU and that CEHU can be used for quantitatively evaluating changes induced by a therapeutic agent such as dopamine in flow velocity and in FVV.

Adult↗

PubMed2001

Immunolocalization of vacuolar-type H+-ATPase, cathepsin K, matrix metalloproteinase-9, and receptor activator of NFkappaB ligand in odontoclasts during physiological root resorption of human deciduous teeth.

To investigate the cellular mechanisms of physiological root resorption in human deciduous teeth, the authors examined the immunocytochemical localization of vacuolar-type H+-ATPase, a lysosomal cysteine proteinase, cathepsin K, matrix metalloproteinase-9 (MMP-9), and receptor activator of NFKB ligand (RANKL) in odontoclasts. H+-ATPase, cathepsin K, and MMP-9 are the most important enzymes for decalcification of apatite crystals and degradation of type-I collagen. In addition, RANKL is one of the key regulatory molecules in osteoclast formation and functions. Odontoclasts developed extensive ruffled borders and clear zones apposed to the resorbing root dentine surfaces. On immunoelectron microscopy, the expression of vacuolar-type H+-ATPase was detected along the limiting membranes of pale vacuoles and the ruffled border membranes of odontoclasts. Cathepsin K in odontoclasts was localized within pale vacuoles, lysosomes, the extracellular canals of ruffled borders, and the underlying resorbing dentine surfaces. MMP-9 localization in odontoclasts was similar to those of cathepsin K. RANKL was detected in both mononuclear stromal cells and odontoclasts located on resorbing dentine surfaces. These results suggest that (1) odontoclasts are directly involved in decalcification of apatite crystals by active extrusion of proton ions mediated by H+-ATPase and (2) extracellular degradation of dentine type-I collagen by both cathepsin K and MMP-9, and (3) odontoclast differentiation and activity are regulated, at least in part, by RANKL, possibly produced by mononuclear stromal cells and odontoclasts themselves in the resorbing tissues. Thus, the cellular mechanisms of physiological root resorption appear to be quite similar to those of osteoclastic bone resorption.

Carrier Proteins↗

PubMed2001

Effect of angiotensin II type 2 receptor on tyrosine kinase Pyk2 and c-Jun NH2-terminal kinase via SHP-1 tyrosine phosphatase activity: evidence from vascular-targeted transgenic mice of AT2 receptor.

Angiotensin II (Ang II) has two major receptor isoforms, AT1 and AT2. AT1 transphosphorylates Ca(2+)-sensitive tyrosine kinase Pyk2 to activate c-Jun NH2-terminal kinase (JNK). Although AT2 inactivates extracellular signal-regulated kinase (ERK) via tyrosine phosphatases (PTP), the action of AT2 on Pyk2 and JNK remains undefined. Using AT2-overexpressing vascular smooth muscle cells (AT2-VSMC) from AT2-transgenic mice, we studied these undefined actions of AT2. AT1-mediated JNK activity was increased 2.2-fold by AT2 inhibition, which was abolished by orthovanadate. AT2 did not affect AT1-mediated Pyk2 phosphorylation, but attenuated c-Jun mRNA accumulation by 32%. The activity of src-homology 2 domain-containing PTP (SHP-1) was significantly upregulated 1 min after AT2 stimulation. Stable overexpression of SHP-1 dominant negative mutant in AT2-VSMC completely abolished AT2-mediated inhibition of JNK activation and c-Jun expression. These findings suggest that AT2 inhibits JNK activity by affecting the downstream signal of Pyk2 in a SHP-1-dependent manner, leading to a decrease in c-Jun expression.

Angiotensin Receptor Antagonists↗

PubMed2001

Phosphatidylinositol 4,5-bisphosphate induces actin stress-fiber formation and inhibits membrane ruffling in CV1 cells.

Phosphatidylinositol 4,5 bisphosphate (PIP(2)) is widely implicated in cytoskeleton regulation, but the mechanisms by which PIP(2) effect cytoskeletal changes are not defined. We used recombinant adenovirus to infect CV1 cells with the mouse type I phosphatidylinositol phosphate 5-kinase alpha (PIP5KI), and identified the players that modulate the cytoskeleton in response to PIP(2) signaling. PIP5KI overexpression increased PIP(2) and reduced phosphatidylinositol 4 phosphate (PI4P) levels. It promoted robust stress-fiber formation in CV1 cells and blocked PDGF-induced membrane ruffling and nucleated actin assembly. Y-27632, a Rho-dependent serine/threonine protein kinase (ROCK) inhibitor, blocked stress-fiber formation and inhibited PIP(2) and PI4P synthesis in cells. However, Y-27632 had no effect on PIP(2) synthesis in lysates, although it inhibited PI4P synthesis. Thus, ROCK may regulate PIP(2) synthesis by controlling PI4P availability. PIP5KI overexpression decreased gelsolin, profilin, and capping protein binding to actin and increased that of ezrin. These changes can potentially account for the increased stress fiber and nonruffling phenotype. Our results establish the physiological role of PIP(2) in cytoskeletal regulation, clarify the relation between Rho, ROCK, and PIP(2) in the activation of stress-fiber formation, and identify the key players that modulate the actin cytoskeleton in response to PIP(2).

ADP Ribose Transferases↗

PubMed2001

A three-dimensional, quantitative computed tomographic study of changes in distribution of bone mineralization in the developing human mandible.

The three-dimensional distribution of highly mineralized cortical bone on the buccal surface of the mandible was evaluated using quantitative computed tomography. The study was completed in 34 living persons (21 females and 13 males) between the ages of 9 and 32 yr of age with symmetrical mandibles. The geometrical distribution of the highly mineralized areas (>1250 mg/cm(3)) was determined by three-dimensional reconstruction of images. The pattern of distribution was evaluated by superimposing a 9 by 9-mesh grid over the lateral view of the mandible and adapting it to the same anatomical landmarks for different aged participants. Individuals demonstrated four different patterns of distribution of highly mineralized buccal cortical bone. The youngest had highly mineralized bone in the corpus of the mandible and along the anterior ramal border. In older individuals, the mineralization changed to include the mid- and posterior ramus. Highly mineralized bone was not found within the condylar, coronoid, or gonial angle regions of any participant. A comparison of the mineralization pattern between the left and right lateral mandibular views showed bilateral symmetry with similar patterns of distribution. Comparison of the number of grids occupied by highly mineralized cortical bone with the age of the participant, mandibular length, gonial angle, and cross-sectional area of the masseter muscle produced the highest correlation coefficient with the size of the masseter muscle.

Adolescent↗

PubMed2001

Convenient synthesis of aliphatic polyesters by distannoxane-catalyzed polycondensation.

A convenient synthesis of aliphatic polyesters by the distannoxane-catalyzed polycondensation of aliphatic dicarboxylic acids and aliphatic diols in solvents under azeotropic conditions was developed. To determine the optimum conditions for polycondensation of succinic acid with 1,4-butandiol, the effect of various types of catalyst and their concentration was investigated. By choosing the appropriate solvent, the polymerization was made to proceed in a two-phase system of solvent and molten polymer, and in the presence of 0.001 mol % of 1-chloro-3-hydroxy-1,1,3,3-tetrabutyldistannoxane (CHTD), poly(butylene succinate) (PBS) with a weight-average molecular weight of 277,000 was obtained. This method was then successfully applied to the synthesis of various aliphatic polyesters, such as poly(ethylene succinate), poly(1,4-cyclohexanedimethylene succinate), and poly(ethylene sebacate).

Biodegradation, Environmental↗

PubMed2001

Angiotensin II signaling and HB-EGF shedding via metalloproteinase in glomerular mesangial cells.

BACKGROUND: Angiotensin II (Ang II) has been implicated in the development of glomerulosclerosis by stimulating fibronectin (FN) synthesis. The processing and release of heparin binding-endothelin growth factor (HB-EGF) are activated by protein kinase C (PKC) and Ca2+ signaling. We studied the roles of HB-EGF and endothelial growth factor (EGF) receptor (EGFR) in Ang II-induced FN expression using mesangial cells. METHODS: Mesangial cells were prepared from mouse kidneys by the explant method and cells were used at passages 4 and 5. RESULTS: Ang II stimulated FN mRNA levels dose-dependently with a maximal increase (3.4-fold) after 12 hours of incubation. This action was completely inhibited by PKC inhibitors and slightly blocked by Ca2+ chelating agents. FN mRNA accumulation by Ang II was abolished by tyrosine kinase inhibitors, a specific inhibitor for EGFR (AG1478) and extracellular signal-regulated kinase (ERK) inactivation. Addition of neutralizing anti-HB-EGF antibody, as well as pretreatment with heparin or the metalloproteinase inhibitor batimastat abolished induction of FN expression by Ang II. In mesangial cells stably transfected with a chimeric construct containing HB-EGF and alkaline phosphatase (ALP) genes, ALP activity in incubation medium was rapidly increased by Ang II (1.7-fold at 0.5 min) and reached a 4.1-fold increase at two minutes. Ang II phosphorylated EGFR (maximal at 2 min) and ERK (maximal at 8 min) in a PKC- and metalloproteinase-dependent manner. Ang II stimulated the expression and release of transforming growth factor-beta (TGF-beta) via EGFR-mediated signaling, and the released TGF-beta also contributed to Ang II-mediated FN expression via EGFR transactivation. CONCLUSIONS: Ang II-mediated FN expression was regulated by autocrine effects of HB-EGF and TGF-beta, suggesting a novel paradigm for cross-talk between Ang II and growth factor receptor signaling pathways.

Angiotensin II↗

PubMed2001

A successfully treated case of disseminated tuberculosis-associated hemophagocytic syndrome and multiple organ dysfunction syndrome.

We report the case of a patient who presented with disseminated tuberculosis-associated hemophagocytic syndrome (HPS). A 40-year-old man was admitted because of fatigue, fever, and renal dysfunction. Chest radiograph and computed tomography scan showed diffuse reticulonodular shadow, and Mycobacterium tuberculosis was identified. Peripheral blood counts decreased rapidly, and bone marrow aspiration revealed hemophagocytosis by macrophages. Despite antituberculous and steroid pulse therapy, multiple organ dysfunction syndrome developed. After plasma exchange and continuous hemodiafiltration were started, hypercytokinemia and vital signs improved dramatically. Although disseminated tuberculosis-associated HPS carries a poor prognosis, acute blood purification may be an effective means of treating HPS involving multiple organ dysfunction syndrome.

Adult↗

PubMed2001

A clinical and histological evaluation of titanium mini-implants as anchors for orthodontic intrusion in the beagle dog.

The aim of this study was to determine the anchorage potential of the titanium mini-implant for orthodontic intrusion of the mandibular posterior teeth. Six mini-implants were surgically placed around the mandibular third premolars on each side in 3 adult male beagle dogs. On the buccal site, three mini-implants were placed distal to the apex of the distal root of the third premolar, at the interradicular septa of the third premolar, and mesial to the apex of the mesial root of the third premolar, as linearly as possible. The same procedure was performed at the lingual site on both sides of the mandibular third premolars in each dog. Bilateral interradicular mini-implants on both the buccal and the lingual sites were used as the anchorage for the intrusion of the third premolars (loaded implants) and the other mini-implants were used as control (unloaded) implants. In 6 weeks, an intrusive force (150 g) was applied between the interradicular implants on the buccal and the lingual sites by closed coil springs run across the crowns of the third premolars. After 12 to 18 weeks of orthodontic intrusion, the animals were killed and their mandibles were dissected and prepared for histologic and fluorescent observation. The results indicated that the mandibular third premolars intruded 4.5 mm, on average, after 12 to 18 weeks of orthodontic force application, with mild root resorption at the furcation area as well as the root apex. All the mini-implants remained stable during orthodontic tooth movement without any mobility or displacement. The morphometrical findings indicated that the calcification of the peri-implant bone on the loaded implants was equal to or slightly greater than those of the controls. In addition, 6 of the 36 mini-implants were removed after tooth movement, and all of them were easily removed with a screwdriver. These findings suggest that mini-implants are effective tools for the anchorage of orthodontic intrusion in beagle dogs.

Alveolar Process↗

PubMed2001

Growth hormone receptor gene variant and mandibular height in the normal Japanese population.

This study was aimed at quantitatively evaluating the relationship between craniofacial morphology and the Pro561Thr (P56IT) variant in the growth hormone receptor gene (GHR), which is considered to be an important factor in craniofacial and skeletal growth. The subjects were unrelated individuals in a normal Japanese population and consisted of 50 men and 50 women. With the use of genomic DNA extracted from whole blood, the GHR gene P56IT variant was detected by the polymerase chain reaction-restriction fragment length polymorphism method (with the restriction enzyme StuI). The relationships of the genotypes to body height and 5 linear measurements from lateral cephalograms were examined for evaluation of craniofacial morphology. The normal Japanese population without P56IT had a significantly greater mandibular ramus length (condylion-gonion) than did those with P56IT. This suggests that the GHR gene P56IT variant may be associated with mandibular height growth and can be a genetic marker for it. Further studies about such genetic markers may expand our understanding of the genetic control in craniofacial morphological determinants and help in the prediction of craniofacial growth.

Adolescent↗

PubMed2001

Cortical bone mineral density in asymmetrical mandibles: a three-dimensional quantitative computed tomography study.

The three-dimensional distribution (3D) of the highest mineralized cortical bone was evaluated in 32 subjects between the ages of 8 and 30 years with asymmetrical mandibles using quantitative computed tomography (QCT). The geometrical distribution of the highest mineralized areas (> 1250 mg/cm3) representative of mandibular cortical bone was determined by 3D reconstruction of the images. The length of the mandible on each side was determined by defining a new linear measurement from the centre of the 3D reconstructed condyle to the midline of the symphysis as identified from a submental view. The cross-sectional areas of the masseter and medial pterygoid muscles were assessed from bilateral axial views through the middle of the muscles parallel to the Frankfort-Horizontal plane. Comparison between the lengths of the two mandibular sides (right-left = mm) indicated a range of asymmetries with an equal number of subjects with the left and right mandible longer. Comparison of the area of highest mineralized cortical bone between the right and left sides (R/L) to the ratio of the mandibular length (R/L) showed a high correlation coefficient (r = 0.629) suggesting that the shortest mandibular side had more highly mineralized bone. A comparison of the area of highest mineralized cortical bone between the right and left sides (R/L) to the ratio of cross-sectional areas of the muscles showed the highest correlation coefficient (r = 0.724) with the ipsilateral masseter muscle. These findings indicate that asymmetrical mandibles are associated with asymmetrical distributions of the highest mineralized cortical bone and that this is age dependent.

Adolescent↗

PubMed2001

Evaluation of PCR primers to screen for Streptococcus pneumoniae isolates and beta-lactam resistance, and to detect common macrolide resistance determinants.

Pneumococcal isolates (n = 148) from various countries (mostly from the USA) were tested by a primer set for PCR. Thirty-eight (86.4%) of the 44 penicillin G-susceptible isolates (MIC < or = 0.06 mg/L) had unaltered pbps, while six isolates (13.6%) had either one or two alterations in pbps. Of 47 penicillin G-resistant strains (MIC > or = 2 mg/L), 41 isolates (87.2%) had all three pbps altered, six isolates (12.8%) had altered pbp1a + 2x. Various combinations of altered pbp were seen in penicillin G-intermediate isolates. Prevalence of macrolide resistance genes mef(A) and erm(B) in isolates was clearly reflected by their MICs. All isolates were positive for lytA. The primers were useful for screening for Streptococcus pneumoniae and beta-lactam resistance, and for detection of common macrolide resistance determinants.

Anti-Bacterial Agents↗

PubMed2001

Localization of receptor activator of NFkappaB ligand, RANKL, in periodontal tissues during experimental movement of rat molars.

The receptor activator of NFkappaB ligand, RANKL, is one of the key regulatory molecules in osteoclast formation and function. We examined RANKL localization in the periodontal tissues during experimental movement of rat molars. To produce orthodontic force, an elastic band was inserted between the upper first and second molars for 4 days, and the dissected maxillae were subjected to light and electron microscopic immunocytochemical examination for RANKL. Expression of RANKL protein was detected in osteoblasts, osteocytes, fibroblasts, and osteoclasts mostly located in resorption lacunae. In osteoblasts, osteocytes, and fibroblasts, RANKL localization was mainly observed in the cytoplasm, the cisternae of rough endoplasmic reticulum and along plasma membranes. In osteoclasts, RANKL was expressed along the ruffled, border membranes and in the cytoplasm, including the clear zone. These results suggest that during tooth movement, osteoclast differentiation and activation are regulated, at least in part, by RANKL, possibly produced by osteoblasts/stromal cells and osteoclasts themselves in the periodontal tissues.

Animals↗

PubMed2001

Association of amino acid substitutions in penicillin-binding protein 3 with beta-lactam resistance in beta-lactamase-negative ampicillin-resistant Haemophilus influenzae.

The affinity of [(3)H]benzylpenicillin for penicillin-binding protein (PBP) 3A was reduced in 25 clinical isolates of beta-lactamase-negative ampicillin (AMP)-resistant (BLNAR) Haemophilus influenzae for which the AMP MIC was > or =1.0 microg/ml. The affinities of PBP 3B and PBP 4 were also reduced in some strains. The sequences of the ftsI gene encoding the transpeptidase domain of PBP 3A and/or PBP 3B and of the dacB gene encoding PBP 4 were determined for these strains and compared to those of AMP-susceptible Rd strains. The BLNAR strains were classified into three groups on the basis of deduced amino acid substitutions in the ftsI gene, which is thought to be involved in septal peptidoglycan synthesis. His-517, near the conserved Lys-Thr-Gly (KTG) motif, was substituted for Arg-517 in group I strains (n = 9), and Lys-526 was substituted for Asn-526 in group II strains (n = 12). In group III strains (n = 4), three residues (Met-377, Ser-385, and Leu-389), positioned near the conserved Ser-Ser-Asn (SSN) motif, were replaced with Ile, Thr, and Phe, respectively, in addition to the replacement with Lys-526. The MICs of cephem antibiotics with relatively high affinities for PBP 3A and PBP 3B were higher than those of AMP and meropenem for group III strains. The MICs of beta-lactams for H. influenzae transformants into which the ftsI gene from BLNAR strains was introduced were as high as those for the donors, and PBP 3A and PBP 3B showed decreased affinities for beta-lactams. There was no clear relationship between 7-bp deletions in the dacB gene and AMP susceptibility. Even though mutations in another gene(s) may be involved in beta-lactam resistance, these data indicate that mutations in the ftsI gene are the most important for development of resistance to beta-lactams in BLNAR strains.

Ampicillin↗

PubMed2001

Angiotensin II type 2 receptor inhibits epidermal growth factor receptor transactivation by increasing association of SHP-1 tyrosine phosphatase.

Angiotensin (Ang) II has 2 major receptor isoforms, Ang type 1 (AT(1)) and Ang type (AT(2)). AT(1) transphosphorylates epidermal growth factor receptor (EGFR) to activate extracellular signal-regulated kinase (ERK). Although AT(2) was shown to inactivate ERK, the action of AT(2) on EGFR activation remains undefined. Using AT(2)-overexpressing vascular smooth muscle cells from AT(2) transgenic mice, we studied these undefined actions of AT(2). Maximal ERK activity induced by Ang II was increased 1.9- and 2.2-fold by AT(2) inhibition, which was abolished by orthovanadate but not okadaic acid or pertussis toxin. AT(2) inhibited AT(1)-mediated EGFR tyrosine phosphorylation by 63%. The activity of SHP-1 tyrosine phosphatase was significantly upregulated 1 minute after AT(2) stimulation and association of SHP-1 with EGFR was increased, whereas AT(2) failed to tyrosine phosphorylate SHP-1. Stable overexpression of SHP-1-dominant negative mutant completely abolished AT(2)-mediated inhibition of EGFR and ERK activation. AT(1)-mediated c-fos mRNA accumulation was attenuated by 48% by AT(2) stimulation. Induction of fibronectin gene containing an AP-1 responsive element in its 5'-flanking region was decreased by 37% after AT(2) stimulation, corresponding to the results of gel mobility assay with the AP-1 sequence of fibronectin as a probe. These findings suggested that AT(2) inhibits ERK activity by inducing SHP-1 activity, leading to decreases in AP-1 activity and AP-1-regulated gene expression, in which EGFR dephosphorylation plays an important role via association of SHP-1.

Angiotensin II↗