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Biomedical subjects

T Kaji

Publications and source records attributed to T Kaji.

At least 127 records · Page 7Linked to original sources
PubMed1992

Rhodamine B inhibits collagen synthesis by human lip fibroblasts in culture.

To investigate the effect of the cosmetic dye, rhodamine B, on the metabolism of collagen in fibroblasts, confluent KD cells, an established cell line of fibroblasts from human lip, were cultured for 6 h in a serum-free medium in the presence of the dye at 100 micrograms/ml and below. It was found that rhodamine B significantly decreased the content of procollagen type I C-terminal peptide antigen in both the cell layer and the medium with an only slight decrease in the cell number. Rhodamine B significantly decreased the incorporation of [3H]proline into either the collagen-digestible protein or the non-collagen protein in the cell layer. The incorporation of both [3H]thymidine and [14C]leucine into the acid-insoluble fraction of the cell layer was significantly decreased by rhodamine B; the activity of lactate dehydrogenase that leaked into the medium was not changed by the dye. From these results, it was suggested that rhodamine B has the capacity of decreasing the collagen content of the fibroblast cell layer of the human lip, which may result from a non-specific inhibition of protein synthesis without non-specific cell damage. Rhodamine B may impair the formation of extracellular matrix which is important for the maintenance of the lip tissue.

Cell Count↗
PubMed1992

Protective effect of copper against cadmium cytotoxicity on cultured vascular endothelial cells.

We investigated the effect of copper on cadmium-induced cytotoxicity on vascular endothelial cells from bovine aorta in a culture system. Cytotoxicity was evaluated by the [3H]adenine release assay and the histological observation. After a 24-h incubation, cadmium exhibited a significant cytotoxicity on confluent cultures of endothelial cells in a dose-dependent manner, while copper only slightly did after a 24-h incubation. It was found that copper (5 microM) significantly decreased cadmium (1 and 2 microM) cytotoxicity; histologically, formation of de-endothelialized areas in the cell layer caused by cadmium was reduced by copper. The accumulation of cadmium in the cell layer was significantly decreased by copper; however, that of copper was unaffected by cadmium. It was therefore suggested that copper significantly protects cadmium-induced cytotoxicity on cultured endothelial cells primarily through decreasing the cellular cadmium accumulation.

Animals↗
PubMed1992

Endothelin modulation of tissue plasminogen activator release from human vascular endothelial cells in culture.

To clarify a possible involvement of the vasoconstrictive peptide endothelin in the regulation of endothelial cell-mediated fibrinolytic system, confluent cultures of vascular endothelial cells from human umbilical vein were incubated in serum-free medium in the presence of endothelin-1 at 100 nM and below, and tissue plasminogen activator antigen (t-PA:Ag) in the medium was determined by enzyme immunoassay. Endothelin-1 at 1 nM and above significantly decreased the release of t-PA:Ag from the endothelial cells after a 24 h incubation. The t-PA:Ag release was also decreased by either endothelin-2 or endothelin-3 at 10 nM. The activity of lactate dehydrogenase in the medium was not changed by endothelin-1 at 100 nM and below, suggesting that the peptide did not cause nonspecific cell damage. The decrease in the t-PA:Ag release induced by endothelin-1 occurred in the presence or absence of 8-bromo cyclic AMP, which is an active congener of cyclic AMP; 3-isobutyl-1-methylxanthine, which is an inhibitor of phosphodiesterase; and forskolin, which is a stimulator of adenylate cyclase. These results strongly indicated that cyclic AMP which is known to down-regulate t-PA:Ag release was not involved in the endothelin-1 effect. However, endothelin-1 failed to decrease the t-PA:Ag release in the presence of either calcium ionophore A23187 or EGTA; the ionophore itself markedly decreased the release. The cytosolic calcium accumulation was significantly increased by endothelin-1. These results suggest that endothelin-1 decreases the release of t-PA:Ag from human endothelial cells through an excess accumulation of intracellular, especially cytosolic which would be mediated by an extracellular, calcium-dependent mechanism.

1-Methyl-3-isobutylxanthine↗
PubMed1992

Stimulants from gardeniae fructus for cultured endothelial cell proliferation.

Bioassay-guided fractionation of Gardeniae Fructus extract (GFE), which stimulates the proliferation of cultured endothelial cells, led to the isolation of glycerol and D-mannitol. Both compounds significantly increased the incorporation of [3H]thymidine and [14C]leucine into the acid-insoluble fraction of bovine aortic endothelial cell layers in culture. This clearly indicated that glycerol and D-mannitol are active components of GFE on endothelial cell proliferation. On the other hand, they did not change the number of cultured vascular smooth muscle cells from bovine aorta. Glycerol and D-mannitol may be beneficial drugs for vascular disorders.

Animals↗
PubMed1992

[Activities of antithrombin III and heparin cofactor II in patients with pathologic blood coagulation conditions].

To investigate the physio-pathological functions of HC-II, assays for HC-II and AT-III were performed simultaneously on the samples from patients with DIC, liver dysfunction or renal disease from the three view points of consumption, production and loss of AT-III and HC-II. For the AT-III activity, two kinds of assays were applied: the automatic chromogenic substrate method and a newly developed clotting method which receives no effects from HC-II activity. The activity of HC-II was significantly lower than that of AT-III in patients with either DIC or liver dysfunction. However, no significant difference between HC-II and AT-III activities in patients with either thrombosis or renal disease. There were high correlations between HC-II and AT-III activities were found in the patients with liver dysfunction, suggesting that low activity was due to decreased production of HC-II and AT-III in the liver. It will be necessary that elucidation of the significant functions of HC-II not only in coagulation and hemostasis but also in regulation of local inflammation and invasion of neoplasm is necessary.

Adult↗
PubMed1991

Inhibitory effect of heparin cofactor II on thrombin-stimulated prostacyclin production by cultured vascular smooth muscle cells.

We investigated the effect of heparin cofactor II (HCII) on thrombin-induced prostacyclin (PGI2) production by A10 cells, an established cell line of vascular smooth muscle cells from murine aorta. Confluent monolayers of A10 cells were incubated with 0.1 NIH U/ml of thrombin for 30 min in the presence of antithrombin III (ATIII) or HCII, and PGI2 production by the cells was measured by radioimmunoassay as 6-keto-prostaglandin F1a, the stable metabolite of PGI2. ATIII at 40 mInh.U/ml and more significantly inhibited thrombin-induced PGI2 production by A10 cells, but HCII at the same doses did only slightly. However, when A10 cells were preincubated with HCII for 30 min before exposure to thrombin, the PGI2 production was markedly inhibited. The medium conditioned by A10 cells for 30 min did not enhance the inhibitory effect of HCII on thrombin-induced PGI2 production by the cells. On the other hand, A10 cells synthesized both dermatan sulfate and heparan sulfate which are capable of activating HCII. From these results, it was suggested that HCII would be activated by glycosaminoglycans (GAGs) such as dermatan sulfate of the cell layers and could inhibit thrombin-induced PGI2 production. HCII may be a modulator of thrombin on the physiological functions of vascular smooth muscle cells, reacting to the cell surface GAGs.

Antithrombin III↗
PubMed1991

Antithrombin III modulates the effect of thrombin on the metabolism of glycosaminoglycans in cultured endothelial cells.

We previously reported that a treatment of cultures of endothelial cells from bovine aorta with thrombin resulted in a less accumulation of glycosaminoglycans (GAG) in the cell layer. In the present study, we found that thrombin-induced decrease in the accumulation of [35S]sulfate-labeled GAG (35S-GAG) such as heparan sulfate was prevented by antithrombin III (AT III) but not by heparin cofactor II (HC II). However, AT III did not show a significant effect on the 35S-GAG accumulation individually. Pretreatment of the cell layer with neither AT III nor HC II showed any preventive effect. When GAG in the cell layer was labeled with both [35S]sulfate and [3H]glucosamine, neither thrombin nor a combination of thrombin with AT III changed the ratio of the radioactivity of 35S to that of 3H. Although thrombin stimulated the release of 35S-GAG from the cell layer, AT III completely prevented the stimulatory effect. In conclusion, it was suggested that AT III may inhibit the thrombin action on GAG metabolism of endothelial cells to prevent thrombosis in vivo.

Animals↗
PubMed1991

Effect of thrombin on the production of glycosaminoglycans by cultured endothelial cells.

We investigated the effect of thrombin on the production process of glycosaminoglycans (GAG) in cultured bovine aortic endothelial cells. It was revealed that the trichloroacetic acid-insoluble [35S]sulfate-labeled GAG (35S-GAG) was decreased by thrombin. The thrombin-induced decrease in the accumulation of 35S-GAG in the cell layer occurred even in the presence of actinomycin D or cycloheximide. The incorporation of both [35S]sulfate and [3H]glucosamine into the GAG was decreased by thrombin, while the ratio of 35S to 3H was not changed. Also, the incorporation of both [3H]glucosamine and [14C]UDP-xylose into the GAG was inhibited by thrombin. From these results, it was suggested that thrombin decreased GAG in the endothelial cell layer through an suppression of formation of polysaccharide chains rather than that of core protein.

Animals↗
PubMed1991

Thrombin decreases glycosaminoglycans content of endothelial cells in culture.

The effect of thrombin on the metabolism of glycosaminoglycans (GAG) in cultured bovine aortic endothelial cells was investigated. Incubation of confluent endothelial cell cultures with thrombin at 1.0 NIH U/ml for 8 h and above caused a significant decrease in the accumulation of [35S]sulfate-labeled GAG (35S-GAG) in the cell layer. After a 24 h incubation, thrombin at 0.5 U/ml and above decreased the accumulation of 35S-GAG in both cell layer and medium. The percentage of 35S-GAG released into the medium during the last 3 h of 24 h incubation was significantly increased by thrombin. In thrombin-treated cell layer, both heparan sulfate and the other GAG was significantly decreased at the same degree. The incorporation of both [3H]thymidine and [14C]leucine was significantly increased by thrombin. Gabexate mesilate, a serine protease inhibitor, suppressed the thrombin inhibition. From these results, it was suggested that thrombin decreased 35S-GAG in the endothelial cell layer without a nonspecific damage of the cells through mainly an inhibition of GAG production and additionally a stimulation of GAG release.

Animals↗
PubMed1991

Rhodamine B inhibition of glycosaminoglycan production by cultured human lip fibroblasts.

Rhodamine B (Rh B) is a dye which is used in cosmetics such as lipsticks. We investigated the effect of the dye on the metabolism of [3H]glucosamine-labeled glycosaminoglycans ([3H]-GAG) in confluent cultures of human lip fibroblast KD cells. It was found that Rh B at 10 micrograms/ml and above significantly decreased the accumulation of [3H]-GAG in both the cell layer and the medium after a 24-hr culture. This Rh B (50 micrograms/ml) effect was recognized in the cell layer and the medium after 8 and 24 hr, respectively, and longer. The Rh B at the 25 micrograms/ml-induced decrease in the [3H]-GAG accumulation was reversible in both the cell layer and the medium. On the other hand, the release of [3H]-GAG from the cell layer was unaffected by Rh B. A characterization of [3H]-GAG revealed that all components of the GAG including hyaluronic acid, heparan sulfate, chondroitin 4- and/or 6-sulfate, and dermatan sulfate were significantly decreased by 50 micrograms/ml Rh B in both the cell layer and the medium. Rh B significantly decreased the accumulation of [3H]-GAG in the presence of either 10 microM cycloheximide or 1 mM p-nitrophenyl-beta-D-xyloside, suggesting that Rh B inhibited the sugar chain formation rather than core protein synthesis. Although the number of confluent KD cells was significantly decreased by Rh B at 10 micrograms/ml and above, this Rh B effect was much weaker than that on the [3H]-GAG accumulation. The release of lactate dehydrogenase from the cell layer was significantly increased by 100 micrograms/ml Rh B but not by the dye at 75 micrograms/ml and below. From these results, it was suggested that Rh B decreases the GAG content of human lip fibroblasts through a functional suppression of polysaccharide chain formation in the process of the GAG production.

Cell Count↗
PubMed1991

Inhibitory effect of rhodamine B on the proliferation of human lip fibroblasts in culture.

The effect of the cosmetic dye rhodamine B on the proliferation of human lip fibroblasts (KD cells) was investigated in a culture system. Rhodamine B at 25 micrograms/ml and above significantly decreased the number of the cells after a 72 h culture. A time course study revealed that 50 micrograms/ml of rhodamine B-induced decrease in the cell number occurred after 48 h and longer, suggesting that the dye inhibited cell proliferation without a decrease in cell attachment. The detachment of [3H]thymidine-labeled cells from the monolayer was unaffected by rhodamine B at 100 micrograms/ml and below. The incorporation of [3H]thymidine and [14C]leucine into the acid-insoluble fraction of the cell layer was significantly inhibited by 50 micrograms/ml rhodamine B treatment. Histologically, the damage of KD cells was not marked, however, a degenerative change of nuclei and an irregular shape of the cells as well as a decrease in the cell number were caused by 50 micrograms/ml rhodamine B. Rhodamine 6G caused a severe damage of the cells, and rhodamine B significantly decreased the cell number; rhodamine 123 had no significant effect; rhodamine 116 significantly increased the cell number. Furthermore, rhodamine B decreased the number of both vascular endothelial cells from bovine aorta and vascular smooth muscle cells from murine aorta after a 72 h culture. It is concluded that rhodamine B inhibits the proliferation of human lip fibroblasts. This rhodamine B effect may be a warning sign for the dye toxicity.

Cell Division↗
PubMed1991

Effect of lead on the glycosaminoglycans metabolism of bovine aortic endothelial cells in culture.

We investigated the effect of lead nitrate (0.1, 1.0, 10 or 20 microM) on the metabolism of glycosaminoglycans (GAG) in confluent cultures of bovine aortic endothelial cells. It was found that lead at 10 and 20 microM significantly decreased the accumulation of [35S]sulfate-labeled GAG ([35S]GAG) both in the cell layer and the medium after a 24-h culture. A time course study showed that 10 microM lead decreased the accumulation of [3H]glucosamine-labeled GAG both in the cell layer and the medium after 24 h and longer. The release of [35S]GAG from the cell layer during the last 3 h of a 24 h culture was not changed by lead. The detachment of [3H]thymidine-labeled cells from the monolayer was unaffected by lead. It was shown that lead at 10 microM decreased both heparan sulfate and the other GAG in the cell layer; the former was more sensitive to lead treatment. Lead at 20 microM and below failed to increase the release of lactate dehydrogenase, suggesting that non-specific cell damage was not caused by lead. From these results, it was suggested that lead decreases endothelial cell heparan sulfate content through a decrease in the GAG production without a non-specific cell damage. Lead may be a risk factor of vascular disorders.

Animals↗
PubMed1991

Interaction between cadmium and copper on ossification of embryonic chick bone in tissue culture.

To investigate the interaction between cadmium and copper in ossification, femurs from 9-day-old chick embryos were cultured for 6 days in the presence of 2 microM cadmium and/or 1 microM copper. It was found that cadmium + copper treatment caused interactively severe damage to osteogenic mesenchymal cells in the periosteum and a severe degenerative change in osteoblasts around the trabecula, resulting in severe impairment of ossification in the diaphysis. Cadmium content was increased by copper; however, copper content was unaffected by cadmium in the diaphysis. It was therefore considered that the copper-induced increase in cadmium content was a primary factor in the interactive toxic effect of the cadmium + copper treatment in ossification.

Animals↗
PubMed1991

Stimulation of heparan sulphate release from cultured endothelial cells by plasmin.

We investigated the effect of plasmin on the release of glycosaminoglycans (GAG) from bovine aortic endothelial cells in culture. Plasmin significantly increased the release of [35S]sulphate-labelled GAG (35S-GAG) from the cell layer, although plasminogen did not show such a stimulatory effect. The percentage of heparan sulphate in the released 35S-GAG (about 80%) was not changed by plasmin. Plasmin enhanced the detachment of endothelial cells from the monolayer only slightly. From these results, it was suggested that the local anticoagulant activity on endothelium mediated by the anticoagulant heparan sulphate will be increased by plasmin; the conversion of plasminogen to plasmin may be mediated by plasminogen activators released from thrombin-stimulated endothelial cells.

Animals↗
PubMed1991

Gardenia fruit extract does not stimulate the proliferation of cultured vascular smooth muscle cells, A10.

We investigated the effect of a hot water extract from Gardenia fruit (Gardenia jasminoides Ellis) (GFE), which has a stimulatory effect on endothelial cell proliferation, on the proliferation of A10 cells, an established cell line of vascular smooth muscle cell from murine aorta in a culture system. GFE did not change the number of A10 cells after a 48 h culture. GFE significantly increased the incorporation of [3H]thymidine and [14C]leucine into the acid-soluble fraction of bovine aortic endothelial cell layers, but significantly decreased that of A10 cells. These results suggested that GFE stimulates the proliferation of endothelial cells but not of A10 cells. In the endothelial cell culture, GFE significantly increased the accumulation of basic fibroblast growth factor, which is an autocrine for endothelial cell proliferation in medium and low-affinity (glycosaminoglycans-binding) fractions, while A10 cells did not produce a significant amount of the factor. Since it is postulated that a selective stimulation of endothelial cell proliferation by increasing the production of basic fibroblast growth factor is appropriate for prevention of arteriosclerosis and thrombosis, GFE may contain a beneficial component as a useful drug.

Animals↗
PubMed1991

Low molecular weight heparin enhances prostacyclin production by cultured human endothelial cells.

Confluent cultures of vascular endothelial cells derived from the human umbilical vein were incubated in a serum-free medium in the presence of low molecular weight heparin (LMWH) with molecular weights of 4000-6000 dalton (Da), or of unfractionated heparin (UFH) with average molecular weight 12,000 Da, and prostacyclin production was determined by radioimmunoassay for 6-keto-prostaglandin F1 alpha, the stable metabolite of prostacyclin. LMWH at 1 U/ml as anti-factor Xa activity significantly increased prostacyclin production after 6h or longer; however, UFH at 1 USP U/ml did not induce such a significant change. The LMWH-induced increase in prostacyclin production occurred at 0.1 U/ml and above after 6 h of treatment. Since prostacyclin is both a potent inhibitor of platelet aggregation and a vasodilator, it was suggested that the increased endothelial cell prostacyclin production induced by LMWH may be a component of the anticoagulant activity of the drug.

Cells, Cultured↗
PubMed1991

Role of histamine in motion sickness in Suncus murinus.

The levels of histamine (HA) and tele-methylhistamine (t-MH) were determined in five brain regions of Suncus murinus (suncus) and the effects of motion stimulus or drugs influencing the turnover of these amines were studied to elucidate the role of histamine in motion sickness. Shaking the animals for 2 min increased HA contents in telencephalon and diencephalon without significantly changing the t-MH levels. alpha-Fluoromethylhistidine (alpha-FMH), which is presumed to deplete the neuronal HA, tended to raise the HA levels. alpha-FMH slightly alleviated the vomiting response to motion stimulus and suppressed the HA increase in diencephalon caused by shaking. Compound 48/80, which releases HA from mast cells, did not alter the control HA levels, but effectively prevented the motion sickness and completely suppressed the motion-induced rises in HA levels. These results provide further evidence that brain HA plays an important role in the development of motion sickness.

Animals↗