Activity Of Testicular Hyaluronidase Research Articles

Anti-Inflammatory and Antifibrotic Effects of a Combination of Spiperone and Immobilized Hyaluronidase on Partially Reversible and Irreversible Toxic Pneumofibrosis

The possibility of boosting antifibrotic activity of testicular hyaluronidase immobilized on polyethylene oxide with spiperone was studied on the bleomycin models of a single (partially reversible pneumofibrosis) and repeated (irreversible pneumofibrosis) injuries to the alveolar epithelium in C57Bl/6 mice. The antifibrotic effect was more pronounced after successive treatment with immobilized hyaluronidase and spiperone than after individual treatment with each of the compounds: no collagen deposition in the parenchyma of bleomycin-damaged lungs was found. The decrease in inflammatory cell (lymphocytes, macrophages, neutrophils, plasma cells) infiltration of the alveoli and alveolar tracts interstitium in mice treated by immobilized hyaluronidase and spiperone did not differ from the anti-inflammatory effect of spiperone monotherapy.

Antifibrotic Effects of Immobilized Hyaluronidase in Repeated Bleomycin-Induced Lesions of the Alveolar Epithelium

Antifibrotic activity of testicular hyaluronidase, immobilized on polyethylenoxide and obtained by electron beam synthesis, was studied on the model of bleomycin injuries to the alveolar epithelium (irreversible pneumofibrosis) in C57Bl/6 mice and compared to the effect of testicular hyaluronidase. Intranasal therapy with immobilized and testicular hyaluronidases prevented the deposition of fibrotic mass in the parenchyma of "bleomycin" lungs. The effect of immobilized hyaluronidase was more pronounced than that of testicular hyaluronidase. The studied compounds were virtually inessential for infiltration of the alveolar interstitium and alveolar tracts by lymphocytes, macrophages, neutrophils, and plasma cells. Unchanged histoarchitectonics of bleomycin-damaged lungs in immobilized hyaluronidase therapy was due to suppression of the progenitor fibroblast cells (CD45(-)).

Effects of Ascorbic Acid and Analogs on the Activity of Testicular Hyaluronidase and Hyaluronan Lyase on Hyaluronan

We have evaluated the inhibition of testicular hyaluronidase and hyaluronan lyase by L-ascorbic acid and chemical analogs. We observed that L-ascorbic acid, D-isoascorbic acid and dehydroascorbic acid inhibited both types of enzymes, but showed stronger effects towards hyaluronan lyase. But these compounds were observed to degrade the substrate, hyaluronan, by themselves. Of the other ascorbic acid analogs tested, saccharic acid inhibited hyaluronan lyase, while not affecting the enzymatic activity of testicular hyaluronidase, nor affecting the physic-chemical stability of hyaluronan. This is the first compound, to our knowledge, to be shown to possess such selective inhibition. Therefore, we propose that saccharic acid could serve as a lead compound for the development of potent and selective inhibitors of bacterial hyaluronan lyase or of polysaccharide lyase enzymes in general as we observed this compound to be capable of inhibiting chondroitinase ABC in addition to hyaluronan lyase.

Inhibition of in vitro fertilization of mouse gametes by sulfated sialic acid polymers.

The effect of sialic acid (N-acetyl neuraminic acid), sialic acid dimer, sialic acid polymers (colominic acid) and sulfated colominic acid on the activity of hyaluronidase, on the dispersion of cumulus cells by mouse sperm and on in vitro mouse fertilization (sperm penetration of zona pellucida) were evaluated. Bovine testicular hyaluronidase activity was significantly inhibited by colominic acid and sulfated colominic acid, but not by sialic acid and its dimer. The dispersion of cumulus cells from eggs by mouse sperm was also inhibited by colominic acid and sulfated colominic acid. In vitro fertilization of mouse gametes was inhibited by sulfated colominic acid. The IC50 value of sulfated colominic acid-induced inhibition of fertilization was 0.3 mg/ml (ca. 0.9 mM). The value changed from 0.9 mM for cumulus-surrounded egg to 1.5 mM for cumulus free-egg. On the other hand, colominic acid showed little or no inhibitory effect on mouse in vitro fertilization at 0.5 mg/ml (ca. 1.6 mM). This antifertility activity by sulfated colominic acid did not appear to be due to an effect on sperm motility or on the oocytes. These results suggest that (1) the cumulus cells surrounding the eggs were dispersed by sperm hyaluronidase, (2) hyaluronidase was inhibited by colominic acid and by sulfated colominic acid, (3) sulfated colominic acid inhibits sperm penetration of zona pellucida by the inhibition of hyaluronidase and/or some enzymes required for mouse gametes fertilization.

Inhalation of diesel engine exhaust affects spermatogenesis in growing male rats.

We conducted experiments to determine whether diesel engine exhaust affects reproductive endocrine function in growing rats. The rats were assigned to three groups: a group exposed to total diesel engine exhaust containing 5.63 mg/m3 particulate matter, 4.10 ppm nitrogen dioxide, and 8.10 ppm nitrogen oxide; a group exposed to filtered exhaust without particulate matter; and a group exposed to clean air. Dosing experiments were performed for 3 months beginning at birth (6 hr/day for 5 days/week). Serum levels of testosterone and estradiol were significantly higher in animals exposed to total diesel exhaust and filtered exhaust (p < 0.05 for each group) as compared to the controls. Follicle-stimulating hormone was significantly decreased in the two groups exposed to diesel exhaust as compared to the control group (p < 0.05). Luteinizing hormone was significantly decreased in the total exhaust-exposed group as compared to the control and filtered groups (p < 0.05). Although testis weight did not show any significant difference among the groups, sperm production and activity of testicular hyaluronidase were significantly reduced in both exhaust-exposed groups as compared to the control group. Histological examination showed decreased numbers of step 18 and 19 spermatids in stage VI, VII, and VIII tubules in the testes of both diesel exhaust-exposed groups. This study suggests that diesel exhaust stimulates hormonal secretion of the adrenal cortex, depresses gonadotropin-releasing-hormone, and inhibits spermatogenesis in rats. Because these effects were not inhibited by filtration, the gaseous phase of the exhaust appears to be more responsible than particulate matter for disrupting the endocrine system.

Inhalation of Diesel Engine Exhaust Affects Spermatogenesis in Growing Male Rats

We conducted experiments to determine whether diesel engine exhaust affects reproductive endocrine function in growing rats. The rats were assigned to three groups: a group exposed to total diesel engine exhaust containing 5.63 mg/m3 particulate matter, 4.10 ppm nitrogen dioxide, and 8.10 ppm nitrogen oxide; a group exposed to filtered exhaust without particulate matter; and a group exposed to clean air. Dosing experiments were performed for 3 months beginning at birth (6 hr/day for 5 days/week). Serum levels of testosterone and estradiol were significantly higher in animals exposed to total diesel exhaust and filtered exhaust (p &lt; 0.05 for each group) as compared to the controls. Follicle-stimulating hormone was significantly decreased in the two groups exposed to diesel exhaust as compared to the control group (p &lt; 0.05). Luteinizing hormone was significantly decreased in the total exhaust-exposed group as compared to the control and filtered groups (p &lt; 0.05). Although testis weight did not show any significant difference among the groups, sperm production and activity of testicular hyaluronidase were significantly reduced in both exhaust-exposed groups as compared to the control group. Histological examination showed decreased numbers of step 18 and 19 spermatids in stage VI, VII, and VIII tubules in the testes of both diesel exhaust-exposed groups. This study suggests that diesel exhaust stimulates hormonal secretion of the adrenal cortex, depresses gonadotropin-releasing-hormone, and inhibits spermatogenesis in rats. Because these effects were not inhibited by filtration, the gaseous phase of the exhaust appears to be more responsible than particulate matter for disrupting the endocrine system.

A new non-hormonal antifertility drug DL 111-IT: II. Effects on testicular hyaluronidase activity in male rats

Effects of DL-111, [3-(2-ethylphenyl)-5-(3-methoxyphenyl-1H-1,2,4-triazole] a non-hormonal antifertility agent, on testicular hyaluronidase activity, an accurate biochemical marker for testicular function, were evaluated in male rats. Treatment of 21-day-old rats with DL-111 sc for 7 or 15 days resulted in a significant fall in testicular weight and complete suppression of hyaluronidase activity. During the 30-day post-treatment the enzyme activity was restored to normal levels. Treatment of 40-day-old rats for 7 or 15 days also produced a significant decrease in testicular weight and hyaluronidase activity. Simultaneous administration of LH, PMSG or T with DL-111 to 21-day-old rats blocked the inhibitory activity of the drug as the enzyme activity was restored to untreated control levels. Administration of FSH along with DL-111 had no effect on suppressive action of the drug. These results suggest that in male rats DL-111 inhibits testicular activity by reducing LH levels, thereby reducing T levels as observed by reduced weights of testes and accessory glands of reproduction and hyaluronidase activity.

Heparin inhibits bovine testicular hyaluronidase activity in myocardium of dogs with coronary artery occlusion

Bovine testicular hyaluronidase (BTH) reduces experimental myocardial infarct size and ameliorates electrocardiographic signs of ischemia. This study was done to determine if heparin, an in vitro inhibitor of hyaluronidase activity, blocks the action of BTH in the myocardium of dogs after coronary artery occlusion. BTH was administered intravenously as 5,000 NF units/kg at 0.5 and 2.5 hours after coronary occlusion. Heparin was administered intravenously as a 150-unit/kg loading dose, followed by 10 units/kg per hour i.v., beginning 15 minutes before coronary occlusion. The area of myocardial ischemia at risk was assessed by a radiolabeled microsphere technique; the area that developed necrosis was assessed by a histochemical technique. In vivo activity of BTH was assessed by a colorimetric analysis of the BTH substrate, i.e., hyaluronic acid (HA), extracted from myocardial tissue. For biochemical analysis of HA, the heart was divided into anterior myocardium, which included ischemic tissue and posterior nonischemic myocardium. The myocardial HA content of dogs treated with BTH plus heparin (anterior, 3.44 ± 0.40 μg HA/mg protein; posterior, 3.69 ± 0.33 μg HA/mg protein) was not significantly different from control (anterior, 3.61 ± 0.29 μg HA/mg protein; posterior, 3.55 ± 0.23 μg HA/mg protein). In contrast, BTH lowered myocardial HA content (anterior, 2.16 ± 0.21 μg HA/mg protein; posterior, 2.08 ± 0.14 μg HA/mg protein) compared with either BTH plus heparin or control groups in both anterior myocardium (p = 0.006) and posterior myocardium (p = 0.001). Significant salvage of ischemic myocardium at risk of necrosis was observed in BTH-treated dogs compared with control dogs (area necrosis = 66.4 ± 11.7%area risk vs area necrosis = 97.5 ± 5.4% area risk, p = 0.037). Treatment with BTH did not produce significant salvage of ischemic myocardium in heparinized dogs (area necrosis = 82.4 ± 15.0% area risk). Thus, heparin inhibits in vivo activity of BTH in myocardium of dogs after coronary artery occlusion.

Biological characteristics of a factor suppressing follicle stimulating hormone (inhibin) from ovine testes

Inhibin (follicle stimulating hormone suppressing factor) isolated from ovine testes has been characterized for its biological activity using a variety of tests. The bioassay used — inhibition of the human chorionic gonadotropin induced increment in the mouse uterine weight-demonstrates that there is a significant increment in specific activity (approx. 300-fold) with the progress of purification. Eventhough the final product has not been obtained in a homogenous state it has been possible to show that(a) [125I]-labelled inhibin is preferentially taken up and retained by the pituitary, pretreatment of rats with testosterone facilitating this uptake;(b) it is able to suppress specifically the levels of follicle stimulating hormone in castrated as well as immature intact rats and (c) treatment of immature male rats with inhibin preparation for ten days results in impairment of testicular function as judged by3H-thymidine incorporation into testicular DNA and testicular hyaluronidase activity.

Cell-substratum attachment and cell surface hyaluronate of Rous sarcoma virus-transformed chondrocytes.

Hyaluronate is associated with the cell surface of cultured Rous sarcoma virus-transformed chondrocytes. Detachment of these cells from their substratum by a variety of reagents is accompanied by release of 75-100% of this hyaluronate into solution. Treatment of the cells with 200 U/ml protease-free Streptomyces hyaluronidase at 37 degrees C cause release of greater than 90% of the cell surface hyaluronate and complete cell detachment. Treatment with a lower concentration of Streptomyces hyaluronidase (30 U/ml) at 25 degrees C or a corresponding activity of testicular hyaluronidase gives similar results, but only in the presence of mM EGTA. Treatment with the lower activities of either hyaluronidase or with 1 mM EGTA alone release only approximately 45% of the cell surface hyaluronate and does not cause significant cell detachment. It is concluded that there are two populations of cell surface hyaluronate differing in their accessibility or their resistance to dissociation from other components of the cell surface. It is proposed that the less readily released fraction is located between the transformed chondrocyte surface and substratum and is necessary for their interaction.

Unsuitability of indoxyl acetate as a substrate for the assay of testicular hyaluronidase.

The most recently published method for the assay of testicular hyaluronidase preparations was based on the premise that the enzyme also exhibited carboxylesterase activity towards indoxyl acetate. Studies on the relative enzyme activities of various hyaluronidase preparations towards hyaluronate and indoxyl acetate, the relative stabilities towards pH, temperature and mechanical shaking and the behaviour towards a variety of inhibitors, showed that the activities towards the two substrates reflected the presence of at least two different enzyme systems in the preparations. Gel chromatography and polyacrylamide-gel-electrophoresis experiments confirmed these conclusions and the collective findings clearly establish that methods based on the use of indoxyl acetate cannot be employed to measure testicular hyaluronidase activity.

Hormonal control of mammalian spermatogenesis.

Studies of the hormonal control of the spermatogenic process are reviewed. Most of the investigations used the techniques of ablation and replacement therapy and were performed on the rat. Results from the studies indicate that changes in the metabolism of proteins lipids and carbohydrates take place in the testes under the control of hormones. However it may be that the observed biochemical changes which occur are a reflection of the changes in cellular composition of testicular tissue rather than the result of metabolic changes within a specific cell type in response to the effect of a hormone. A hypothesis concerned with the qualitative aspects of hormonal control of spermatogenesis in the rat has been formed. It is proposed that the multiplication and growth of gonocytes prenatally and in the first few days after birth and formation of type A spermatogonia are under the control of testosterone. The formation of type A and B spermatogonia and primary spermatocytes and progression of the meiotic prophase may be influenced by growth hormone. Reduction division is under control of testosterone. The early steps of spermatid formation may be under no hormonal control or may require testosterone while the late stages of spermatid maturation require the presence of FSH.

Hormonal Regulation of Hyaluronidase during Spermatogenesis in the Rat

Abstract Hyaluronidase activity has been studied during hormone-dependent cell differentiation of the germinal epithelium in the rat testis. Hyaluronidase in rat testis is particulate and sediments with the lysosomal fraction, but can be released by treatment with cetyltrimethylammonium bromide. Rat testicular hyaluronidase activity is undetectable until 33 to 34 days of age, at which time it appears and subsequently increases 400-fold. This rapid increase is correlated with acrosome formation in the newly formed spermatids. Hypophysectomy completely prevents the formation of spermatids and of hyaluronidase, although prolonged treatment of the hypophysectomized rats with luteinizing hormone and follicle-stimulating hormone or with testosterone permits spermatid formation and restores the specific activity of hyaluronidase to normal. Hyaluronidase activity thus appears to represent an accurate biochemical marker for the biological development of testis cells at a specific, hormone-dependent transition in spermatogenesis.

Factors influencing the activity of testicular hyaluronidase

1. 1. The influence of dilution and of addition of polyelectrolytes, mainly of macromolecular nature, on the activity of testicular hyaluronidase was studied. 2. 2. Two samples of hyaluronidase, i.e., a commercial preparation and a product purified fivefold by zone electrophoresis, served for this study. Enzyme activity was measured by two independent procedures, namely by the viscosity-reducing and by the turbidity-reducing methods. The latter technique has been modified and adapted for use at high enzyme dilutions. 3. 3. Upon dilution, hyaluronidase is transformed into an enzymically inactive form, recognizable by the progressive loss of specific activity. This phenomenon is due to a reversible dissociation of the active enzyme into inactive components. 4. 4. Dissociation of hyaluronidase can be reversed by the addition of polycations, in particular by poly- l-lysine, protamine, chitin, and 1,10-diaminodecane, in the order of decreasing potency. These substances are, therefore, activators of dilute hyaluronidase. 5. 5. Macromolecular polyanions, such as polystyrene sulfonate, heparin, sulfated pectic acid, polymethacrylate, amylopectin sulfate, and deoxyribonucleate are inhibitors of hyaluronidase. They act by sequestrating the activator and, hence, they promote dissociation of the enzyme into inactive components. The inhibition is of the “activator-competitive” type, since it can be attenuated and even completely overcome by increasing the activator concentration.

Flavone-配糖體Rutinに關する研究

Rutin has a cardiotonic action on the isolated frog heart and contracts blood-vessels of toad's hind limbs and of rabbit's ears, but it has no significant effect on the blood pressure of rabbit. Rutin has also a suppressive action on the movement of the isolated rabbit's intestine as well as a slight diuretic action. Administered intravenously rutin is excreted in the urine at the rate of 22.61-44.16 % in 2.5 hours. Rutin has some influences on the blood-picture of rabbits : there is initial increase in erythrocytes and leucocytes count which is later decreased. No apparent changes are observed on the clotting time of the blood. Moreover, rutin has specific effects on adrenaline, e. g. the inhibition of its oxidation, the potentiation of its action and the sensitization to adrenaline. On the other hand rutin has an inhibitory effect on the Schwartzman phenomenon, testicular hyaluronidase activity, lung bleeding under low pressure and on the diffusion.with the Menkin factor. Furthermore it was proved that rutin was a stronger capillary stabilizer than hesperidin. Quercetin manifests similar but more stronger effect than rutin.

Inhibition of Testicular Hyaluronidase Activity by Rutin.

From the data it can be seen that rutin inhibits the spread of intradermally injected india ink to which hyaluronidase or saline is added. The effect is statistically highly significant. From the analysis of variance it is evident that the results are due to rutin treatment and not variations within the individual animals. Propylene glycol, which was used as the rutin solvent, inhibits the spreading power of hyaluronidase and this inhibition is increased by a highly significant factor when rutin is also present. Propylene glycol produces slight shock and the resultant liberation of adrenalin probably is responsible for inhibition of hyaluronidase activity. Favilli and Homburger have shown that adrenalin has this property.