Sae-Kwang Ku

Effects of ß-glucan fromAureobasidium pullulans on acute inflammation in mice

The effects of ß-glucan isolated fromAureobasidium pullulans were observed on acute xylene-induced inflammation, ß-glucan at a dose of 62.5, 125 or 250 mg/kg were administered once orally to xylene-treated mice (0.03 mL of xylene was applied on the anterior surface of the right ear to induce inflammation), and the body weight change, ear weight, histological profiles and histomorphometrical analyses of ear were conducted upon sacrifice. The xylene was topically applied 30min after dosing with ß-glucan. The results were compared to those of diclofenac, indomethacin and dexamethasone (15 mg/kg injected once intraperitoneally). All animals were sacrificed 2 h after xylene application. Xylene application resulted in marked increases in induced ear weights compared to that of intact control ear; hence, the differences between intact and induced ear were also significantly increased. The histological characteristics of acute inflammation, such as severe vasodilation, edematous changes of skin and infiltration of inflammatory cells, were detected in xylene-treated control ears with marked increase in the thickness of the ear tissues. However, these xylene-induced acute inflammatory changes were significantly and dose-dependently decreased by ß-glucan treatment. We conclude that ß-glu-can fromA. pullulans has a somewhat favorable effect in the reduction of the acute inflammatory responses induced by xylene application in mice.

Effect of Picrorrhiza Rhizoma Extracts on Early Diabetic Nephropathy in Streptozotocin-Induced Diabetic Rats

The hypoglycemic effect of an aqueous extract of Picrorrhiza Rhizoma (PR) was investigated on diabetic nephropathy. The body weight of each of eight rats per group was measured 7 days after streptozotocin (STZ) treatment. The blood glucose levels 3 days after STZ treatment and the changes in body weight, kidney weight, blood glucose levels, and serum blood urea nitrogen (BUN) were also measured, and the histopathology of the kidney was examined. Body weight decreases and increases of kidney weight, blood glucose levels, and serum BUN and creatinine levels were detected in the diabetic control with histopathological changes related to diabetic nephropathy in the kidney. However, the level of hyperglycemia was significantly lowered in all groups given the PR extract. In addition, the changes related to diabetic nephropathy with the body weight were also significantly lower in the captopril and all PR extract-dosing groups than in the diabetic control. In conclusion, an aqueous extract of PR has relatively good inhibitory effects on STZ-induced diabetes with early diabetic nephropathy with a similar efficacy to an equal dose of captopril.

Micronucleus Test of DHU001, a Polyherbal Formula, in Bone Marrow Cells of Male ICR Mice

The genotoxic effects of DHU001, a polyherbal formula were evaluated using the mouse micronucleus test. DHU001 was administered once a day for 2 continuous days by oral gavage to male ICR mice at doses of 2000, 1000 and 500 mg/kg. Cyclophosphamide was used as a known geno-toxic agent in a positive control. The appearance of a micronucleus is used as an index for genotoxic potential. In addition, the changes on the total white blood cells and differential counts on the lymphocytes, neutrophils, eosinophils, basophils and monocytes in the prepared blood smears were also conducted to observe the possible immunosuppression. The results indicats that DHU001 showed no genotoxicity effects up to 2000 mg/kg dosing levels and did not influenced on the total white blood cells and differential counts. In addition, it is also considered that there were no problems from cytotoxicity of DHU001 tested in this study because the polychromatic erythrocyte ratio was detected as > 0.41 in all tested groups.

Micronucleus Test of Picrorrhiza Rhizoma Aqueous Extract in Bone Marrow Cells of Male ICR Mice.

In this research, the genotoxic effect of Picrorrhiza Rhizoma (PR) aqueous extract was evaluated using the mouse micronucleus test. PR extract was administered once a day for 2 continuous days by oral gavage to male ICR mice at doses of 2000, 1000 and 500 mg/kg. Cyclophosphamide was used as a known genotoxic agent in a positive control. The appearance of a micronucleus (MN) in polychromatic erythrocyte (PCE) is used as an index for genotoxic potential, and PCE ratio is used as an index of cytotoxicity. Although significant (p < 0.01) increase of the number of PCE with one or more nuclei (MNPCE) was detected in cyclophosphamide treated groups, no significant increases of MNPCE numbers were observed in all three different dosages of PR extracts treated mice with over 0.39 of the individual polychromatic erythrocyte ratio in all mice used in this study. The results obtained indicated that PR extract shows no genotoxicity effects up to 2000 mg/kg dosing levels.

Immunohistochemistry of endocrine cells in the alimentary tract of the tree frog, hyla arborea japonica

The regional distribution and relative frequencies of endocrine cells were studied immunohistochemically (PAP methods) in the alimentary tract of the tree frog, Hyla arborea japonica, using specific antisera against serotonin, somatostatin, bovine Sp‐1/chromogranin (BCG), cholecystokinin (CCK)‐8, vasoactive intestinal polypeptide (VIP), gastrin, bombesin, secret in and pancreatic polypeptide (PP). Six kinds of endocrine cells were identified in this study. These immunoreactive cells were located in the gastric glands of stomach regions and in the basal portion of the epithelium of the intestinal tract or esophagus with variable frequencies. They were spherical or spindle‐shaped. Serotonin‐immunoreactive cells were observed in the whole alimentary tract including the esophagus. Somatostatin‐immunoreactive cells were also detected throughout the alimentary tract except the rectum. CCK‐8‐immunoreactive cells were observed from the pylorus to ileum. VIP‐immunoreactive cells were restricted to the rectum. Bombesin‐immunoreactive cells were restricted to the fundic gastric regions and gastrin‐immunoreactive cells were restricted to the pylorus. However, no BCG‐, secretin and PP‐immunoreactive cells were demonstrated in this study. In conclusion, the regional distribution and relative frequency of the endocrine cells in the alimentary tract of the tree frog were similar to other anuran species, but some differences which may be caused by feeding habits and species specification were also observed.

An immunohistochemical study on the neuropeptide-producing endocrine cells in the alimentary tract of wrinkled frog, Rana rugosa (Ranidae).

The regional distribution and relative frequency of neurohormonal peptides-producing endocrine cells were demonstrated in the alimentary tract of wrinkled frog, Rana rugosa, using eight types of specific antisera raised against mammalian regulatory peptides. The alimentary tract of frog was divided into six portions from esophagus to rectum. Most of the cells in the epithelial lining portion, between epithelial cells, were generally spherical or spindle shaped having long cytoplasmic process that was reached to the lumen (open-typed cell) while cells showing round shape (close-typed cell) were also found in the gastric, esophageal or intestinal glands occasionally. All of eight immunoreactive (IR) cells against serotonin, somatostatin, bovine Sp-1/chromogranin (BCG), gastrin, cholecystokinin (CCK)-8, bombesin, glucagon, and human pancreatic polypeptide (HPP) were observed in this study. Serotonin-IR cells were demonstrated throughout whole alimentary tract including esophagus and showed most predominant frequency in antrum. Somatostatin-IR cells were also detected throughout whole alimentary tract including esophagus and showed most predominant in pylorus and antrum. BCG-IR cells were restricted to stomach regions with relatively low frequencies. CCK-8-IR cells were observed from antrum to ileum and showed highest frequency in antrum. Gastrin-IR cells were restricted to antrum with low frequency and bombesin-IR cells were demonstrated from esophagus to antrum with various frequencies. Glucagon-IR cells were located throughout whole alimentary tract except for rectum and showed most predominant frequency at antrum. HPP-IR cells were detected from antrum to ileum with highest frequency in antrum. In conclusion, the regional distribution and relative frequency of these IR cells correspond well to the previous report in anuran species but somewhat peculiar patterns are also detected.

Mouse Single Oral Dose Toxicity Study of DHU001, a Polyherbal Formula.

This study was conducted to obtain acute information of the oral dose toxicity of DHU001, a polyherbal formula in male and female mice. In order to calculated 50% lethal dose (LD50) and approximate lethal dose (LD) , test material was once orally administered to male and female ICR mice at dose levels of 2000, 1000, 500, 250 and 0 (vehicle control) ml/kg (body weight) . The mortality and changes on body weight, clinical signs, gross observation, organ weight and histopathology of principle organs were monitored 14 days after treatment with DHU001. We could not find any mortalities, DHU001 treatment-related clinical signs, changes on the body and organ weights, gross and histopathological findings. The results obtained in this study suggest that LD50 and approximate LD in mice after single oral dose of DHU001 were considered over 2000 mg/kg in both female and male mice.

Synergistic Effect of Fermented Rice Extracts on the Probiotic and Laxative Properties of Yoghurt in Rats with Loperamide-Induced Constipation

Aim. The objective was to evaluate the synergistic effects of fermented rice extracts (FRe) on the laxative and probiotic properties of yoghurt in rats with loperamide-induced constipation. Methods. After constipation induction, yoghurt containing FRe (BFRe; 0.05%, 0.1%, or 1%) was administered orally once per day for 6 days. Results. Loperamide treatment caused marked decreases in fecal pellet numbers and water content discharged, as well as in the surface mucosal thickness of the colonic lumen, intestinal charcoal transit ratio, thickness, and number of mucous-producing goblet cells in the colonic mucosa, whereas it increased the remnant fecal pellet number and the mean diameter of the colonic lumen. However, this loperamide-induced constipation was ameliorated by treatment with FRe, yoghurt single formula, or 0.05%, 0.1%, or 1% BFRe (10u2009mL/kg). Additionally, the viable numbers of Lactobacillus in the cecal contents and feces were markedly higher than those in constipated rats. Moreover, greater probiotic and laxative effects were detected in BFRe-treated rats than in rats treated with equivalent doses of yoghurt or FRe single formula. Conclusion. The results suggest that addition of FRe to liquid yoghurt will enhance the probiotic and beneficial laxative effects of yoghurt in the digestive tract, without causing side effects.

Nephroprotective Effect of POLYCAN on Acute Renal Failure Induced by Cisplatin in Rats

We performed to evaluate the effect of POLYCAN (β-glucan) on cisplatin-(CDDP-)induced acute renal failure (ARF) in rats. POLYCAN was administered orally once a day for 32 days. Each of 8 rats per group was selected based on the body weight (BW) after acclimatization and they were sacrificed at 5 days after CDDP injection. There was significant (P < 0.05) increase of BW after CDDP dosing in all POLYCAN groups than vehicle control and significant (P < 0.01 or P < 0.05) decrease of absolute and relative kidney weight were detected in all POLYCAN groups compared with vehicle control. In addition, serum BUN and creatinine level in all POLYCAN groups were significantly (P < 0.01 or P < 0.05) lower than vehicle control and the percentage of degenerative regions significantly (P < 0.01) decreased in all POLYCAN groups. As the results of CDDP-induced ARF process, dramatic decrease of the BW, increase of the kidney weight, serum BUN, and creatinine level were detected in vehicle control group compared with sham control group. The changes by CDDP-induced ARF process in POLYCAN groups were significantly and dose-dependently improved compared with vehicle control group. Therefore, POLYCAN has enough potential to develop as a new agent of prevention or treatment for ARF.

Single Oral Dose Toxicity Study of Pinelliae Rhizoma Aqueous Extract in ICR Mice

This study was conducted to obtain acute information of the oral dose toxicity of lyophilized water extract of Pinelliae Rhizoma, a dried tuber of Pinellia ternata (PR) in male and female mice. In order to calculated 50% lethal dose (LD50) and approximate lethal dose (ALD), test material was once orally administered to male and female ICR mice at dose levels of 2000, 1000, 500, 250, 125 and 0 (vehicle control) ml/kg (body weight). The mortality and changes in body weight, clinical signs, gross observation, organ weight and histopathology of principle organs were monitored 14 days after treatment with PR extract. We could not find any mortalities, clinical signs, changes in the body and organ weights, gross and histopathological findings except for dose-dependent increases in the hepatic fatty change frequencies detected in PR extract 2000 and 1000mg/kg treated in both male and female mice. The results obtained in this study suggest that LD50 and approximate LD in mice after single oral dose of PR extracts were considered over 2000 mg/kg in both and female male mice, but more than 1000mg/kg of PR extracts treatment could induce slight hepatotoxicity the fatty changes in mice.