Susumu Tsurufuji

The effect of locally injected anti-inflammatory drugs on the synthesis of collagen and non-collagen protein of carrageenin granuloma in rats

Abstract The effect of steroid and non-steroid anti-inflammatory drugs was studied on protein synthesis of the granulation tissue induced by carrageenin on the dorsum of rats. The drugs were injected daily into the pouch of preformed granuloma for 4 days from day 5 up to day 8. Immediately after the last injection of the drugs the rats were injected s.c. with 3H-proline, and 24 hr later the granuloma was harvested on day 9. The total content, specific activity and total radioactivity of collagen and non-collagen protein of the granuloma were determined. Betamethasone disodium phosphate, a steroid drug, markedly inhibited the incorporation of 3H-proline into collagen hydroxyproline as well as into non-collagen protein. Among the non-steroids tested, indomethacin and phenylbutazone failed to show any significant changes in the synthesis of these proteins. Sodium salicylate significantly reduced the synthesis of collagen in the granulomatous tissue without affecting non-collagen protein synthesis.

The characteristic feature of glucocorticoids after local application with reference to leucocyte migration and protein exudation.

Abstract The carboxymethyl cellulose (CMC) pouch method which was developed to measure leucocyte migration and protein exudation simultaneously in acute experimental inflammation was used for the quantitative analysis of the characteristic feature of glucocorticoids after local application. The order of inhibitory potency of the steroidal anti-inflammatory agents on leucocyte migration was in good agreement with data obtained in other laboratories based on different biological responses. The glucocorticoids did not show any potent inhibitory effect on the exudation of protein even at higher dose levels in which 70–80% of leucocyte migration was inhibited. This may be because of the short treatment time. From the above results, it is suggested that the anti-inflammatory action of glucocorticoids after local application is much more potent in inhibiting the migration of leucocyte as compared with the inhibitory action on vascular permeability. Anti-inflammatory drugs Leucocyte migration CMC pouch method Protein exudation

Differences in the mode of exudative reaction between early phase and late phase of carrageenin-induced inflammation in rats

Differences in the mode of vascular permeability change induced by some stimulants such as chemical mediators between the early phase and the late phase of the carrageenin-induced granulomatous inflammation were investigated with the aid of 131I-human serum albumin as an indicator for the measurement of vascular permeability. In the early pregranulomatous phase of the inflammation, histamine injection into the inflammation locus markedly elevated local vascular permeability, while prostaglandins E1 and E2 showed no effect. On the other hand, in the late phase of the inflammation, where permanent granuloma tissue had been formed, prostaglandins E1 and E2 significantly enhanced the local vascular permeability, while histamine was inert.

Protocollagen proline hydroxylase in isolated rat liver cells.

Abstract 1. 1. Liver parenchymal cells from rats were dispersed by collagenase and hyaluronidase digestion, and by application of the bovine serum albumin flotation method. The viable cells, which were not stained with trypan blue, could be isolated with more than 95% viability. Contamination by non-parenchymal cells was less than 4% 2. 2. The endothelial cell fraction from rat liver was isolated by a combination of pronase digestion and bovine serum albumin flotation methods. Contamination by parenchymal cells was less than 0.4% and viability estimated with trypan blue was more than 99.8%. 3. 3. Protocollagen proline hydroxylase activity was assayed in both the parenchymal cell and the endothelial cell fractions. The parenchymal cells showed hydroxylase activity 100 times higher than the endothelial cells in terms of the number of cells. Trypan blue-stained parenchymal cells had about a quarter of the hydroxylase activity of viable parenchymal cells.

Biosynthesis of collagen in skin minces in relation to the mechanism of the formation of insoluble collagen

Abstract 1. 1. Incubation experiments for the synthesis of collagen were carried out with minced skin from growing rats using tritium-labeled proline as a tracer. 2. 2. Neutral salt-soluble, acid-soluble,and insoluble collagens were fractionated almost quantitatively. 3. 3. Neutral salt-soluble collagen was formed at an almost constant rate for 8 h in the normal incubation, whereas the rate of maturation of this fraction into insoluble collagen declined rapidly with the passage of time. 4. 4. Replacing oxygen in the gas phase by nitrogen, freezing and thawing of minced skin, or removal of radioactive proline from the medium in the course of normal incubation, all interrupted the maturation of neutral salt-soluble collagen into insoluble collagen. 5. 5. These results suggest that maturation of collagen depends on aerobic cellular activity which declines more rapidly than the capacity to synthesize tropocollagen. 6. 6. The idea of spontaneous maturation of soluble into insoluble collagen may be refuted by these findings.

Degradation and turnover of collagen in the mouse skin and the effect of whole body x-irradiation.

1. 1. Male mice were given L-[3H]proline intraperitoneally at 4 weeks of age. The total radioactivity of hydroxyproline in the whole body skin increased with time up to the 6th week after the injection, and then gradually decreased to 19.8% of the peak value in the following 50 weeks. The half-life of the labeled skin collagen was calculated as 80 days in the mice at 18–30 weeks of age, 130 days at 30–45 weeks of age and 200 days at 45–60 weeks of age, respectively. 2. 2. The degredation of skin collagen in adult mice was accelerated by whole body exposure to a total of 900 or 1200 R X-ray. Exposure to 300 R 4 times resulted in a 30% increase in the degradation of labeled collagen in the 30–60-week-old mice. 3. 3. The total amount of collagen in the whole body skin was also significantly decreased by a total of 900 or 1200 R irradiation. 4. 4. The turnover rate of 1 M NaCl-soluble collagen was higher than that of insoluble collagen and this tendency was amplified by X-irradiation. 5. 5. The decrease in the solubility of skin collagen with ageing was accelerated by X-irradiation.

Quantitative aspects of the formation and maturation of collagen in rat skin

Abstract 1. 1.|In studies on the uptake of uniformly labeled l -[3H]proline into rat skin collagen fraction the total radioactivity of hydroxyproline in the skin increased by 20% during the 2 weeks following intraperitoneal injection. This increase can be attributed to the reutilization of radioactive proline resulting from degradation of metabolically active proteins. 2. 2.|It is probable that the majority of neutral-salt-soluble collagen matures into acid-soluble and insoluble collagen; this process is so slow as to take several weeks. 3. 3.|Acid-soluble collagen does not appear to be an active precursor of insoluble collagen.