As far as we know, there were no studies of the effect of L-arginine on bone metabolism in post-menopausal women or ovariectomized rats. The primary objective of the current study was to determine whether arginine supplementation was associated with alterations in femoral and spinal bone mineral density (BMD) and bone markers in ovariectomized (Ovx) rats. Forty female Sprague-Dawley rats were divided into two groups, Ovx and sham groups, which were each randomly divided into two subgroups that were fed control and arginine supplemented diet. All rats were fed on experimental diet and deionized water ad libitum for 9 weeks. Bone formation was measured by serum osteocalcin and alkaline phosphatase (ALP) concentrations. Bone resorption was measured by deoxypyridinoline (DPD) crosslinks immunoassay and corrected for creatinine. Serum osteocalcin, growth hormone, insulin-like growth factor-1 (IGF-1), parathyroid hormone (PTH) and calcitonin were analyzed using radioimmunoassay kits. Bone mineral density (BMD) and bone mineral content (BMC) were measured using PIXImus (GE Lunar Co, Wisconsin, USA) in spine and femur. The serum and urine concentrations of Ca and P were determined. The plasma was analyzed for arginine. Diet did not affect weight gain, mean food intake, and plasma arginine concentration. Urinary Ca excretion was decreased by arginine supplementation in Ovx rats, but statistically not significant. The Ovx rats fed arginine-supplemented diet were not significantly different in ALP, osteocalcin, crosslinks value, PTH, calcitonin and IGF-1 compared to those fed control diet. The arginine-supplemented group had significantly higher serum Ca and growth hormone than control group. Spine and femur BMD were significantly increased by arginine supplementation on 5th and 9th weeks after feeding. Our findings indicate that dietary L-arginine supplementation decreased bone mineral density loss in Ovx rats. Therefore, dietary arginine supplementation may represent a potentially useful strategy for the management of osteoporosis.
Objectives: The aim of present study was to investigate recovery effects of Hirudo, which has been used clinically in diabetes therapy. Methods: We established three groups: normal, control, Hirudo, and assigned 6 rats to each group. The normal group was not treated by any process and fed by normal saline. The control & Hirudo groups were administered streptozotocin (STZ) to induce diabetes. Hirudo extract was orally administered to the Hirudo group for 10 days. After 8 weeks, the rats were sacrificed and their body weight, 24hrs urinary protein excretion, glucose, albumin, BUN, creatinine, total-cholesterol, LDL-cholesterol, triglyceride in blood, and level of glycation end-product (AGE) and transforming growth factor (TGF-${\beta}1$) in serum were measured. Morphological profiles and morphometric studies of the kidney cortex, renal transforming growth factor (TGF-${\beta}1$) expression, and renal receptor for advanced glycation end-products (RAGE) expression were studied. Results: The following results were obtained. The protein amount in urine per 24hrs of the Hirudo-treated group as compared to the control group was significantly reduced. The BUN and creatinine level in serum of the Hirudo-treated group as compared to the control group was significantly inhibited. The construction change in kidney of the Hirudo-treated group as compared to the control group was significantly inhibited. The factor of the Hirudo-treated group as compared to the control group was significantly inhibited, which induced the construction change in kidney. Conclusions: The above results suggest that Hirudo partially improved the function of the kidney.
To study the effect of menopause and dietary protein level on Ca metabolism, ovariectomy (OVAX) and sham operations were performed in 16-week old female rats. Each treatment group was fed for 16 weeks either 5%(L) or 50%(H) casein diets, forming SH, SL, OH, OL groups. High protein groups(SH, OH) showed higher Ca and hydroxyproline excretion in urine. Urinay hydroxyproline was also higher in OVAX, which tells the possibility of increased bone resorption by OVAX and by high dietary protein. At 16th wee, however, urinary Ca and hydroxyproline of SH caught up with OH group, whereas those of OL remained higher than SL. Therefore it seems that high dietary protein overrides the effect of OVAX with time. Urnary protein measured at 8th week was higher in high protein groups, especially in OH. GFR was not differ significantly among groups at 8th week. At 16th week, however, high protein groups showed twice the GFR value of low protein groups. Therefore the increase of urinary Ca and hydroxyproline in SH and OH groups can be explained partly by the increased GFR. The tendency of increased GFR and urinary excretion of protein, Ca, and hydroxyproline was most obvious in OH group. It seems that the effect of high protein diet is likely to accelerated by ovariectomy. The effect of Ovax and dietary protein on the composition of femur, scapular, and lumbar bones, was not pronounced. However, when only the high protein groups were compared, OVAX resulted in the reduction of bone weight, ash and Ca contents, especially in femur. The reason that was no significant effect on bone might be due to the short experimental period to induce that was no significant effect on bone might be due to the short experimental period to induce the changes on bone composition and dietary Ca content used in this experiment may have been high enough to prevent bone loss.
Metabolism of DWP401, recombinant juman epidermal growth factor, was examined in vivo and in vitro in rats. When $^{125}I$-labeled DWP401 was administered at a dose of 50 ${\mu}g$/kg by i.v. injection. $^{125}I$-labeled DWP401 was rapidly degraded within 30 minytes above 93%. Thin layer chromatography analysis of urine collected for 24 hr after i.v. administration of $^{125}I$-labeled DWP401 showed ohly one spot on a X-ray film which was considered as diiodo-tyrosine. This result suggests tha $^{125}I$-labeled DWP401 was completely digested into free amino acids without any specific intermediate polypeptides. About 42.1% of the administered iodine was recovered in 24 hr. For in vitro degradation study, $^{125}I$-labeled DWP401 was added to plama and tissue homogenates of rats and incubated at $37^{\circ}C$. Almost 98% of the added radioactivity recovered from the protein fraction of the liver, kidey, small intestine, stomach and spleen decreased rapidly. For examplem the recovery rates of $^{125}I$-labeled DWP401 were 58.6, 63.2, 39.9, 52.9 and 66.8% after 4hrs of incubation in respective organ homogenates.
Objectives : To evaluate the toxicity of KOB03, polyherbal medicine for the treatment of allergic rhinitis, we performed the dose-range finding test of KOB03 by oral supplementation for 4 weeks in Sprague-Dawley rats. Methods : The water extract of KOB03 consisting of five different herbs was supplied from GLP company. KOB03 was supplemented by gavage at 0, 500, 2,500 and 5,000 mg/kg/day for 4-week consecutive days. We recorded the clinical signs of toxicity, body weight, organ weights, hematology, gross and histological changes in target organs of rats, and clinical chemistry analysis. Results : KOB03 at all doses was shown no mortality or abnormal clinical signs in rats during at the observation period. Furthermore, there was no difference in body weight and food-take consumption, organ weight, gross pathological findings, and urine analysis among the groups of rats treated with different doses of KOB03. The hematological analysis and clinical blood chemistry data were revealed no toxic effects from KOB03-supplemented rats. Conclusions : The results suggest that KOB03 is a wide margin of safety on dose-range toxicity in rats. The no observable adverse effect level (NOAEL) of the test, KOB03 in rats is no less than 5,000 mg/kg/day.
This Study was conducted to assess the single dose toxicity of DA-125, a new anthracycline anti-cancer agent, in rats and mice. The Drug was administered once intravenously to both sexes of rats and mice. Then followed a 14-day period of observation. The $LD_{50}$ Values (95% confidence limit) were estimated to be 60.9 mg/kg (57.5~64.3 mg/kg) for male rats and 60.2 mg/kg (56.2~64.5 mg/kg) for female rats, and 85.8 mg/kg (81.0~90.9 mg/kg) for male mice and 84.5 mg/kg (78.2~91.9 mg/kg) for female mice. Both sexes of rats and mice given the drug revealed the clinical sign of decreased locomotor activity, emaciation, hair loss, red-dish brown urine, salivation, and watery diarrhea. In addition, body weight from the next day to the 7th day tended to be decreased slightly in rats and mice treated with DA-125. Death occurred from the next day after administration to the 12th day. Macroscopically, congestion of gastrointestinal organ, lung, and adrenal glands were found in both sexes on the dead rats and mice. Histopathological examination of dead rats manifested atrophy of spleen, hypoplasia of bone marrow, hypcplasia and necrosis of lymphocyte in thymus, atrophy of villi in small intestine (duodenum, jejunum, and ileum), hyperplasia of granular epithelium in small intestine, degeneration of germinal epithelium in testis, defer oration of tubular epithelium in kidney, and vacuolation and myolysis of myocardium in heart. Histopathological examination of dead mice revealed hypoplasia of spleen and mesenteric lymph node, local necrosis of liver, atrophy of villi in small intestine, hyperplasia of glandular epithelium in small and large intestine, degeneration of tubular in kidney, degeneration of germinal cells in testis, and slight vacuolar degeneration of myocardium in heart.
Aucubin, an irdoid g1ucoside, was investigated to determine whether it has a stimulating effect on $\alpha$-amanitin excretion in $\alpha$-amanitin intoxicated rats, and whether there is binding activity to calf thymus DNA. High-performance liquid chromatography (HPLC) analysis of $\alpha$-amanitin in rat urine allowed quantitative measurement of the $\alpha$-amanitin concentration with a detection limit of 50${mu}g/ml$. In this system, a group treated with both $\alpha$-amanitin and aucubin showed that o(-amanitin was excreted about 1.4 times faster than in the $\alpha$-amanitin only treated group. Our previous results showed that the toxicity of $\alpha$-amanitin is due to specific inhibition of RNA polymerase activity and the resultant blockage of the synthesis of certain RNA species in the nucleus. However, no significant activity change on RNA polymerase from Hep G2 cells was observed when aucubin was treated with $\alpha$-amanitin at any concentration tested. Nevertheless, aucubigenin inhibited both DNA polymerase (IC50, 80.5${mu}g/ml$) and RNA polymerase (IC50, 135.0${mu}g/ml$) from the Hep G2 cells. The potential of both $\alpha$-amanitin and aucubin to interact with DNA were examined by spectrophotometric analysis. $\alpha$-Amanitin showed no significant binding capacity to calf thymus DNA, but aucubin was found to interact with DNA, and the apparent binding constant ($K_{app}$) and apparent number of binding sites per D7A phosphate ($B_{app}$) were 0.45$0.45{\times}$$10^4$$M^{-1}$ and 1.25, respectively.
The therapeutic effect of AS2-006A, a derivative of asiaticoside, has been studied and is being developed as a new wound-healing agent. In the present study, the general pharmacological effects on 1) central nervous system, 2) autonomic nervous system, 3) respiratory system, 4) gastrointestinal system. 5) cardiovascular system. and 6) urinary system were assessed in experimental animals and in in vitro models. 1. In vivo animal study: External applications of the 1 % gel ointment of AS2-006A to rats at the doses of 200. 600 or 2000 mg/kg body weight showed no observable pharmacological effects. The effects on the central nervous system were assessed by observation of behavior, hexobarbital-induced sleeping time, pentetrazole-induced convulsion assay, body temperature measurements, and observations on spontaneous activity and catalepsy. The gel ointment exhibited no effects on the cardiovascular system (i.e. blood pressure and heart rate), renal physiology (i.e. urine volume and electrolytes excretion) and gas-trointestinal physiology (i.e. intestinal charcoal propulsion and gastric mucosal irritation). 2. In vitro experiments: The effects of AS2-006A on the physiology of smooth and cardiac muscles were assessed. Muscle contractions were isotonically and isometrically measured in organ chambers using a physiograph. Cumulative additions of AS2-006A (10$^{-9}$ -10$^{-5}$ M) induced no changes in the tension of isolated guinea pig ileum and tracheal muscles. AS2-006A only slightly increased contractility of rat atrial and papillary muscles at 10$^{-2}$ M, which was not statistically different from control. These data showed that the gel ointment of AS2-006A could be externally applied as a wound-healing agent with no potential side effects.
This study was carried out to examine a part of the mechanism for the etiology of diabetic complications. Thirty normal and forty streptozotocin(STZ)-induced diabetic rats were used as the animal models. The animals were sacrificed at the time points of 3 days, 1,2,4 and 6 weeks after STZ-injection and a time course changes in the concentrations of thiobarbituric acid-reactive substances(TBARS) in blood, urine, and tissues, along with the levels of conjugated dienes in tissues were measured as indices of in vivo lipid peroxidation. The activities of antioxidant enzymes, catalase, superoxide dismutase, glutathione peroxidase and the levels of blood retinol and alpha-tocopherol were also measured. The diabetic rats maintained a slightly higher plasma TBARS level throughout the experiment. The urinary TBARS level was significantly higher in diabetic group and gradually increased with time. Concentrations of TBARS in liver, heart, and kidney tissues from diabetic animals were higher than those from the normal group. An increase of conjugated dienes was also observed in the all tissues examined. The kidney tissue of diabetic animals revealed more significant lipid peroxidation state than any other organ tissues. The activities of hepatic antioxidant enzymes such as catalase, superoxide dismutase, glutathione peroxidase were higher in diabetic animals compared to the control ones and increased with the duration of diabetes mellitus. The plasma levels of vitamin A and E were loser in diabetic animals than in normal controls throughout the experimental period. The level of vitamin E in diabetic animals was significantly decreased with the duration of the disease. The results of this study suggest that an effective regimen to suppress the adverse changes in lipid peroxidation and antioxidant defense system is required from the early stage of the disease to prevent the development of diabetic complications. (Korean J Nutrition 34(3) : 253∼264, 2001)
This study was carried out to investigate the effect of addition of cellulose in the diet on the metabolism in rat fed high and low level of zinc. The experimental animals were consisted of 24 male weaning rats of Sprague-Dawley strain(mean weight 72.3g), and they were devided into 4 groups of 6 rats and fed experimental diets for four weeks. Dietary zinc levels used were 10 ppm, and 300ppm and cellulose levels were 2.5% and 10% of diet by weight. Throughout the experimental period, feed consumption and body weight gain were measured and feed efficiency ratio was calculated. The weight of live, kidney and spleen were measured, and the contents of zinc in feces, urine, liver, kidney, spleen and serum were determined. The results obtained are summarized as following ; 1. Body weight gain in high zinc-adequate cellulose group was significantly higher than the other groups. Feed consumptions were significantly higher in high zinc groups and no significant difference was found with dietary cellulose levels. 2. Fecal zinc excretions of four groups were not different at the first week, but at the end of fourth week, high zinc groups experince significantly more zinc excretion than low zinc groups, and also high cellulose groups had higher zinc contents in the feces than the adequate ones within the same zinc levels(p<0.05). There was no significant difference in the urinary zinc excretion. 3. The weights of liver, kidney and spleen were heavier in the high zinc groups than the lower ones, and higher in the high cellulose groups(p<0.05). The liver zinc contents were significantly lower in the low zinc and high cellulose groups. However zinc contents in the kidney and serum were not influenced by dietary zinc level but by cellulouse level. High cellulose diet lowered serum and kidney zinc concentrations(p<0.05).
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