Oral administration of wild plant-derived minerals and red ginseng ameliorates insulin resistance in fish through different pathways.

Many kinds of fish are characterized by a limited efficiency to use carbohydrates. For this reason, raw fish and mixed feed containing a lot of fish meal have been used as feed for fish farming. However, continuing to use high-protein diets not only increases the cost of fish farming, but may also fuel animal protein shortages. Furthermore, carbohydrates are added to improve the texture of the feed and act as a binding agent and are usually contained at 20% in the feed. It makes sense, therefore, to find ways to make good use of carbohydrates rather than wasting them. The physiological mechanisms of glucose intolerance in fish are not yet well understood. Therefore, we investigated the glucose utilization of fish, omnivorous goldfish Carassius auratus and carnivorous rainbow trout Oncorhynchus mykiss. Furthermore, the effects of oral administration of wild plant-derived minerals and red ginseng on the glucose utilization in these fish muscle cells were investigated. As a result, we found the following. (1) An extremely high insulin resistance in fish muscle and the symptom was more pronounced in carnivorous rainbow trout. (2) Administration of wild plant-derived minerals promotes the translocation of the insulin-responsive glucose transporter GLUT4 to the cell surface of white muscle via activation of the PI3 kinase axis, whereas administration of red ginseng not only promotes GLUT4 transfer and translocation to the cell surface of white muscle via AMPK activation as well as promoting glucose uptake into muscle cells via a pathway separate from the insulin signaling system. (3) In fish, at least goldfish and rainbow trout, both PI3K/Akt and AMPK signaling cascades exist to promote glucose uptake into muscle cells, as in mammals.

Radioprotection of mice by lactoferrin against irradiation with sublethal X-rays.

The influence of a host defense protein, lactoferrin (LF), contained in exocrine secretions such as milk, on radiation disorder was investigated. A total of 25 C3H/He mice in each of two groups were maintained with 0.1% LF-added and LF-free diets, respectively, for one month. The mice were then treated with single whole-body X-ray irradiation at a sublethal dose (6.8 Gy), and the survival rate after irradiation was investigated. The survival rate at 30 d after irradiation was relatively higher in the LF group than in the control group (LF-free), (85 and 62%, respectively). The body weight 15 d after X-ray irradiation was also significantly greater in the LF group than in the control group. The hemoglobin level and hematocrit value were higher in the LF group at 5 d before X-ray irradiation. Another 52 mice underwent whole-body X-ray irradiation at the sublethal dose (6.8 Gy), and then LF was intraperitoneally injected once at 4 mg/animal to half of them. The survival rate in LF-treated mice 30 d after irradiation was 92%, significantly higher than in mice treated with saline (50%) (P = 0.0012). In addition, LF showed hydroxyl radical scavenger activity in vitro. These findings suggest that LF may inhibit radiation damage.

[Chemopreventive effects of 5-fluorouracil and lactoferrin on goldfish intestinal carcinogenesis induced by 1,2-dimethylhydrazine].

The present study was carried out to examine the chemopreventive effects of 5-fluorouracil (5-FU) and lactoferrin (LF) on goldfish intestinal carcinogenesis induced by 1,2-dimethylhydrazine (DMH). DMH was given to fish by intraperitoneal injection in a dosage of 15 mg/kg body weight once a week for 6 weeks. Eight weeks after the initial DMH injection, fish were randomly divided into 2 groups, control and LF-treated groups. Control fish fed a commercial diet. LF- treated fish fed a commercial diet with bovine lactoferrin (oral administration at 200 mg/kg body weight/day). Ten weeks after the initial DMH injection, each was divided into 2 groups, saline- and 5-FU- treated groups. Physiological saline for freshwater fish (0.75% NaCl solution) in the saline-treated fish and 5-FU dissolved in 0.75% NaCl solution in the 5-FU-treated (75 mg/kg body weight) fish were injected intramuscularly three times every other day, respectively. The mean number of precancer cell foci (PCF) per intestine was 2.7 in DMH treated fish. PCF showed broader distribution in the entire intestine derived from DMH-treated fish. LF-only-treatment has no effect on the number of PCF. Mean number of PCF in 5-FU-only-treated fish decreased in comparison with that of the saline-treated control group, though no statistically significant reduction in PCF was found. But if 5-FU treatment was added to LF pretreatment, a statistically significant reduction in the number of PCF was observed. Pretreatment with LF for 2 weeks also reduced the deleterious side effects of 5-FU.

[Lactoferrin reduces physiological dysfunctions of goldfish induced by chemotherapeutic agents].

We investigated whether the deleterious side effects of chemotherapeutic agents on the physiologic functions of fish could be modulated by lactoferrin (LF). Goldfish, weighing about 25 g, were treated intramuscularly with methotrexate (MTX: 2.5 mg/kg body weight) and fluorouracil (FU: 15 or 50 mg/kg body weight) three times every other day. In control fish fed a commercial diet, MTX induced severe immunosuppression, increased the number of total bacteria and Enterobacteriaceae in the intestinal tract, and caused intestinal damage such as lowered and thickened mucosa and thinned muscularis externa, with moderate renal dysfunction. A few fish treated with MTX died. In fish injected with FU or FU plus MTX, the side effects were slightly less in comparison with those in the MTX group. Pretreatment with LF (oral administration at 200 mg/kg body weight/day) for 3 weeks reduced the deleterious side effects of MTX and FU. One intraperitoneal injection of LF (200 mg/kg body weight) immediately after the first MTX injection also reduced the side effects. These results show that LF reduces the physiologic dysfunction of fish treated with chemotherapeutic agents.

Absence of tissue-bound semicarbazide-sensitive amine oxidase activity in carp tissues.

We have previously reported that carp (Cyprinus carpio) tissue mitochondria contain a novel form of monoamine oxidase (MAO), which belongs neither to MAO-A nor to MAO-B of the mammalian enzyme. This conclusion results from the findings that the carp MAO was equally sensitive to a selective MAO-A inhibitor clorgyline and to the MAO-B selective inhibitor l-deprenyl, when tyramine, a substrate for both forms, serotonin or beta-phenylethylamine, a substrate for either A or B-form of mammalian MAO, was used. In the present study, we tried to detect another amine oxidase, termed tissue-bound semicarbazide-sensitive amine oxidase (SSAO), activity in carp tissues. As definition of SSAO was used, such as insensitivity to inhibition of the kynuramine oxidizing activity by an MAO inhibitor pargyline and high sensitivity to the SSAO inhibitor semicarbazide. The results indicated that the oxidizing activity was selectively and almost completely inhibited by 0.1 mM pargyline alone or a combination of 0.1 mM pargyline plus 0.1 mM semicarbazide, but not by 0.1 mM semicarbazide alone. We also tried to detect any SSAO activity by changing experimental conditions, such as lower incubation temperature, higher enzyme protein concentration, a lower substrate concentration and different pH's in the reaction, as the enzyme source. However, still no SSAO activity could be detected in the tissues. These results conclusively indicate that carp tissues so far examined do not contain SSAO activity.

Effects of moderate and substantial hypoxia on erythropoietin levels in rainbow trout kidney and spleen.

Erythropoietin (EPO) is a glycoprotein hormone that regulates the proliferation and differentiation of erythroid progenitor cells in mammals. Although EPO has been identified in fish, the specific function and effects of hypoxia have not been investigated previously. In this study, we have demonstrated a relationship between increases in renal EPO levels and decreases in spleen EPO levels and spleen-somatic index (SSI), with increases in haemoglobin (Hb) concentration in the blood during hypoxia exposure in rainbow trout. Splenic contraction and the subsequent red blood cell release accounts for the initial increase in Hb concentration in the blood, whereas EPO action probably accounts for the later increases in hemoglobin concentration in the blood. Our data indicate that fish and mammalian erythropoietic systems are similar in response to hypoxia, in that erythropoiesis in fish is influenced by EPO.

Interrenal stress responsiveness of marbled sole, Pleuronectes yokohamae, inhabiting Tokyo Bay.

The interrenal stress responsiveness of marbled sole (Pleuronectes yokohamae) captured in the interior (Tokyo) and at the mouth (the Uraga Strait; Uraga) of Tokyo Bay, in the coastal region of Onjuku (the eastern coastal region of Chiba prefecture; Pacific) and in Ishinomaki Bay (Ishinomaki) in Japan was investigated. That is, the effects of air-exposure on plasma cortisol and glucose levels in the fish from the four different regions, and the effect of in vitro ACTH treatment on cortisol release from the head kidney tissue of the fish held under resting conditions were examined. In this study, it was found that (i) there were no significant differences in plasma cortisol and glucose levels among the fish from the four different regions under resting conditions, (ii) each air-exposed fish increased plasma cortisol and glucose levels, (iii) the cortisol response in marbled sole inhabiting Tokyo Bay, in particular the interior, was lower, delayed and prolonged, and (iv) cortisol release from the head kidney to in vitro ACTH treatment was lower in the fish from Tokyo compared with the fish from the reference (not so polluted) sites, Pacific and Ishinomaki. These results suggest that marbled sole inhabiting Tokyo Bay, in particular the interior, have been altered in their responsiveness of cortisol-producing interrenal cells to the stressor in the hypothalamus-pituitary-interrenal (HPI) axis.