• Title/Summary/Keyword: Sprague Dawley rat

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Effect of Dietary Cadmium Level on Clinical Toxicity, Sperm Capacity and Histopathological Changes in Rats (카드뮴 투여수준이 흰쥐의 임상적인 중독증과 정자능력 및 장기조직에 미치는 영향)

  • 한찬규;노정해;이남형;채찬희;김광식;이복희
    • Korean Journal of Animal Reproduction
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    • v.23 no.1
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    • pp.1-12
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    • 1999
  • The present study was conducted to find the effects of different cadmium(Cd) levels in diets on clinical toxicity, sperm capacity and histopathological changes in rats. Thirty male rats of Sprague-Dawley weighing 125.3$\pm$15.2g were randomly blocked into five groups according to body weights. Five levels of Cd in AIN-76 purified diet(0, 25, 50, 100 and 250 ppm) had been fed for 8 weeks. Cadmium was supplemented with a form of CdCl$_2$. 1. After 8 weeks of Cd intake had resulted in apparent cadmium intoxication; reduced growth rate, enlarged kidney and testis, decreased hematocrit value and hemoglobin content in response to supplemented Cd levels in the diets. 2. Cadmium accumulation in liver and kidney showed a tendency to increase in cadmium-exposed groups. The levels of metallothionein were also significantly elevated in the tissues of liver in response to the levels of Cd supplemented(P<0.05). 3. Although sperm motility was not significantly different among treatments, rats fed Cd tended to have reduced sperm motility but sperm concentration of Cd supplemented groups were significantly lower than that of control(p<0.05). 4. Based on the findings from gross lesion, rats fed 250ppm of Cd were externally emaciated, had exposed penis and observed atrophies of kidney and testis. Histopathological observation seemed that the liver of groups feeding Cd supplemented diets showed cellular degeneration and accumulation of eosinophilic materials in the capillaries. In kidney, rats fed Cd diets had shown tubular epithelium degeneration and lesions of basophilic materials, while testes were weakened in numbers of spermatid and sporadically enlarged of giant cells.

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Effects of Deer Antler on the Regeneration of Peripheral Nerves; About Sprout Formation of Experimentally Transected Sciatic Nerves in Rat (말초신경의 재생에 대한 녹용의 효과; 랫드에서 실험적 절단 좌골신경의 Sprout 형성에 관해)

  • Chang, Byung-Joon;Cho, Ik-Hyun;Choi, Hye-Young;Won, Hui-Young;Park, Chang-Hyun;Bae, Chun-Sik;Choe, Nong-Hoon
    • Applied Microscopy
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    • v.32 no.1
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    • pp.67-80
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    • 2002
  • This study was carried out to investigate the effects of deer antler extract on the regeneration of peripheral nerves. Sprague-Dawley male rats weighing about 300 gm were fed deer antler extract for 1, 2, and 3 weeks per oral (1.5 ml/100 gm B.W.), respectively, once a day and transected both sides of sciatic nerve of each leg. After keeping for 6 hours, sciatic nerves taken from proximal part of transected region were treated with conventional transmission electron microscopical method and then observed with electron microscope. The results obtained were summarized as follows; 1. Sciatic nerves of normal control group were not showing any sprouts and electron dense axolemmal projections were frequently observed. 2. Sciatic nerves of saline treated groups were showing axonal sprouts at the nodes of Ranvier. The length of them was usually short, and numerous vesicles, vacuoles and organelles including neurofilament were contained. The number of nodes of Ranvier containing sprouts from 100 longitudinal sectioned nerve fibers was 29 (29%) in 1 week treated group, 32 (32%) in 2 weeks treated group, and 30 (30%) in 3 weeks treated group, respectively. 3. Sciatic nerves of deer antler treated groups were showing axonal sprouts at the node of Ranvier as well. Although most of the sprouts were short, some sprouts of 2 weeks and 3 weeks treated groups were quite long. Sprouts usually contained numerous vesicles, vacuoles and cell organelles such as neurofilaments and mitochondria. The number of nodes of Ranvier containing sprouts from 100 longitudinal sectioned nerve fibers was 38 (38%) in 1 week treated group, 46 (46%) in 2 weeks treated group, and 48 (48%) in 3 weeks treated group respectively. The results described above explain pretreatment of deer antler extract improves the sprout formation of transected sciatic nerves, and then it suggests deer antler may be effective for the regeneration of peripheral nerves.

Evaluation of Macrophage Activity and Repeated Oral Dose Toxicity in Sprague-Dawley Rats on Multivitamin (종합비타민의 랫드에서 반복투여독성 시험과 대식세포 기능 활성 평가)

  • Kim, Hye-Ri;Jang, Hye-Yeon;Lee, Hae-Nim;Park, Young-Seok;Park, Byung-Kwon;Kim, Byeong-Soo;Kim, Sang-Ki;Cho, Sung-Dae;Nam, Jeong-Seok;Choi, Chang-Sun;Chang, Soon-Hyuk;Jung, Ji-Youn
    • Journal of Food Hygiene and Safety
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    • v.28 no.4
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    • pp.360-366
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    • 2013
  • The objective of this study is to investigate the effect of multivitamin on macrophage activity in Raw 264.7 cell and repeated oral dose toxicity in Sprague-Dawely rat of multivitamin. Raw 264.7 cells were treated with 50 and $100{\mu}g/mL$ multivitamin for 24 h. To measure the activity of macrophages, NO and TNF-${\alpha}$ assays were performed in Raw 264.7 cells. Treatment with 50 and $100{\mu}g/mL$ multivitamin for 24 h significantly increased production of NO and TNF-${\alpha}$ compared with control groups, indicating activation of macrophages. The female rats were treated with multivitamin of control group, low group (0.24 g/kg), medium group (1 g/kg) and high group (2 g/kg) intragastrically for 4 weeks, respectively. We examined the body weight, the feed intake, the clinical signs and serum biochemical analysis. We also observed the histopathological changes of liver, ovary, brain, adrenal gland, spleen, kidney, heart and lung in rats. No significant differences in body weights, feed intake, biochemical analysis and histopathological observations between control and multivitamin treatment group were found. In conclusion, multivitamin is physiologically safe and improve macrophage activity.

THE EXPRESSION OF NITRIC OXIDE SYNTHETASE IN THE EXPERIMENTAL TOOTH MOVEMENT IN RATS (백서의 실험적 치아이동시 Nitric Oxide Synthetase의 발현 양상)

  • Park, Dong-Kwon;Kim, Sang-Cheol
    • The korean journal of orthodontics
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    • v.31 no.1 s.84
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    • pp.107-120
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    • 2001
  • Nitric oxide(NO) has been reported to be one of the mediators relating to bone remodelling. Nitric oxide is synthesized from L-arguinine by nitric oxide synthetase(NOS), which is largely divided Into two groups. One group which is composed of $NOS_1\;and\;NOS_3$, is dependent of calcium or calmodulin. The other consisted of $NOS_2$, which is independent of calcium or calmodulin. NOS is thought to be a possible intermediate affecting in the course of tooth movement. This study was designed to evaluate the expression of nitrous oxide synthetase(NOS) in periodontal tissue during the experimental movement of rat incisors, by LSAB(labelled streptavidine biotin) immunohistochemical staining for $NOS_2\;and\;NOS_3$. Twenty seven Sprague-Dawley rats were divided into a control group(3 rats), and 6 experimental groups(24 rats), to which 75g of force was applied, with helical springs across the maxillary incisors. Rats of experimental groups were sacrificed at 12 hours, 1, 4, 7, 14 and 28 days after force application, respectively. After that, the tissues of the control group and experimental groups were studied immunohistochemically. The results were as follows: 1. In control group, the expression of $NOS_3$ was rare in gingiva, dentin, periodontal ligament and alveolar bone, and was mild in the capillaries of pulp and intermaxillary suture. And the expression of $NOS_2$ showed similar pattern to that of $NOS_3$. 2. There were no differences in the expression of $NOS_2\;or\;NOS_3$ in dentin, gingiva, cementum, cementoblast and odontoblast, between control and experimental groups, regardless of the duration of the force application. 3. The expression of $NOS_3$ began to increase at 4 days and showed to the highest degree at 7 days after force application, in the apical region of pressure side of periodontal ligament in experimental groups. 4. The expression of $NOS_3$ in alveolar bone was rare until 7 days, after which it increased to mild degree at 14 days through 28 days in experimental group. But there was no difference between pressure and tension side of periodontal ligament. 5. The expression of $NOS_2$ in periodontal ligament was mild from 7 days after force application, regardless of the side of periodontium, which was generally more evident than that of $NOS_3$. 6. The expression of $NOS_2$ in alveolar bone increased to mild degree at 14 days after force application, and it was evident in osteoblasts, osteoclasts and osteocytes. And the expression of $NOS_2$ was little more stronger in the tension side than that of pressure side of alveolar bone.

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The Effect of Heat Shock Response on the Tumor Necrosis Factor-$\alpha$-induced Acute Lung Injury in Rats (Tumor Necrosis Factor-$\alpha$로 유도되는 백서의 급성 폐손상에 열충격반응이 미치는 효과)

  • Koh, Youn-Suck;Lim, Chae-Man;Kim, Mi-Jung;Cho, Won-Kyung;Jeoung, Byung-O;Song, Kyu-Young;Shim, Tae-Sun;Lee, Sang-Do;Kim, Woo-Sung;Kim, Dong-Soon;Kim, Won-Dong
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.6
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    • pp.1343-1352
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    • 1997
  • Background : Heat-treated cells are known to be protected from lysis by TNF, which is considered to play a central role in the pathogenesis of sepsis-induced acute lung injury. The objective of the study was to investigate the effect of heat shock response by heat-pretreatment on the acute lung injury of the rats induced by intratracheally administered TNF-$\alpha$, Methods : We intratracheally instilled either saline or TNF (R&D, 500ng) with and without heat pretreatment in Sprague-Dawley rats weighing 250~350 g. The heated rats were raised their rectal temperature to $41^{\circ}C$ and was maintained thereafter for 13 minutes at 18 h before intratracheal administration of saline or TNF. After 5 h of intratracheal treatment, lung leak, lung myeloperoxidase activity (MPO) and heat shock proteins were measured in rats. Lung leak index was defined as counts per minute of $I^{25}$ in the right lung divided by counts per minutes of $I^{25}$ in 1.0 ml of blood. All data are expressed as means ${\pm}$SE. Results : There is no difference in acute lung leak index ($0.099{\pm}0.024$ vs $0.123{\pm}0.005$) among the rats given saline intratracheally with and without heat pretreatment, but MPO activity showed a decreased tendency in heat-pretreated rats ($4.58{\pm}0.79\;U/g$) compared with heat-unpretreated rats ($7.32{\pm}0.97\;U/g$) (P=0.064). Rats administered TNF intratracheally with heat-pretreatment had decreased lung leak index ($0.137{\pm}0.012$) and lung MPO activity ($5.51{\pm}1.04\;U/g$) compared with those of heat-unpretreated and TNF-administered rats ($0.186{\pm}0.016$, $14.34{\pm}1.22\;U/g$) (P<0.05 in each). There were no significant difference of lung leak index and MPO activity between TNF-treated rats with heat-pretreatment and saline-treated rats with and without heat-pretreatment. Conclusion : The heat shock response attenuated neutrophil recruitment and acute lung leak induced by intratracheal instillation of TNF-in rats.

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Effects of FK224, a $NK_1$ and $NK_2$ Receptor Antagonist, on Plasma Extravasation of Neurogenic Inflammation in Rat Airways (미주 신경의 전기적 자극으로 유발된 백서의 기도내 혈장 유출에 대한 FK224의 효과)

  • Shim, Jae-Jeong;Lee, Sang-Yeub;Lee, Sang-Hwa;Park, Sang-Myun;Seo, Jeong-Kyung;Cho, Jae-Yun;In, Kwang-Ho;Yoo, Se-Hwa;Kang, Kyung-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.42 no.5
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    • pp.744-751
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    • 1995
  • Background: Asthma is an inflammatory disease because there are many inflammatory changes in the asthmatic airways. Axon reflex mechanisms may be involved in the pathogenesis of asthma. Sensory neuropeptides are involved in this inflammation, which is defined as neurogenic inflammation. Substance p, neurokinin A, and neurokinin B may be main neuropeptides of neurogenic inflammation in airways. These tachykinins act on neurokinin receptors. Three types of neurokinin receptors, such as $NK_1$, $NK_2$, and $NK_3$, are currently recognized, at which substance p, neurokinin A, and neurokinin B may be the most relevant natural agonist of neurogenic inflammation in airways. The receptor subtypes present in several tissues have been characterized on the basis of differential sensitivity to substance p, neurokinin A, and neurokinin B. Plasma extravasation and vasodilation are induced by substance p more potently than by neurokinin A, indicating NK1 receptors on endothelial cells mediate the response. But airway contraction is induced by neurokinin A more potently than by substance P, indicating the $NK_2$ receptors in airway smooth muscles. These receptors are used to evaluate the pathogenesis of brochial asthma. FK224 was identified from the fermentation products of Streptomyces violaceoniger. FK224 is a dual antagonist of both $NK_1$ and $NK_2$ receptors. Purpose: For a study of pathogenesis of bronchial asthma, the effect of FK224 on plasma extravasation induced by vagal NANC electrical stimulation was evaluated in rat airway. Method: Male Sprague-Dawley rats weighing 180~450gm were anesthetized by i.p. injection of urethane. Plasma extravasation was induced by electrical stimulation of cervical vagus NANC nerves with 5Hz, 1mA, and 5V for 2 minutes(NANC2 group) and for sham operation without nerve stimulation(control group). To evaluate the effect of FK224 on plasma extravasation in neurogenic inflammation, FK224(1mg/kg, Fujisawa Pharmaceutical Co., dissolved in dimethylsulphoxide; DMSO, Sigma Co.) was injected 1 min before nerve stimulation(FK224 group). To assess plasma exudation, Evans blue dye(20mg/kg, dissolved in saline) was used as a plasma marker and was injected before nerve stimulation. After removal of intravascular dye, the evans blue dye in the tissue was extracted in formamide($37^{\circ}C$, 24h) and quantified spectrophotometrically by measuring dye absorbance at 629nm wavelength. Tissue dye content was expressed as ng of dye per mg of wet weight tissue. The amount of plasma extravasation was measured on the part of airways in each groups. Results: 1) Vagus nerve(NANC) stimulation significantly increased plasma leakage in trachea, main bronchus, and peripheral bronchus compared with control group, $14.1{\pm}1.6$ to $49.7{\pm}2.5$, $17.5{\pm}2.0$ to $38.7{\pm}2.8$, and $12.7{\pm}2.2$ to $19.1{\pm}1.6ng$ of dye per mg of tissue(mean ${\pm}$ SE), respectively(p<0.05). But there was not significantly changed in lung parenchyma(p>0.05) 2) FK224 had significant inhibitory effect upon vagal nerve stimulation-induced airway plasma leakage in any airway tissues of rat,such as trachea, main bronchus, and peripheral bronchus compared with vagus nerve stimulation group, 49%, 58%, and 70%, respectively(p<0.05). Inhibitory effect of FK224 on airway plasma leakage in neurogenic inflammation was revealed the more significant in peripheral bronchus, but no significant in lung parenchyma. Conclusion: These results suggest that FK224 is a selective NK receptor antagonist which effectively inhibits airway plasma leakage induced by the endogenous neurotransmitters relased by neurogenic inflammation in rat airway. Tachykinin receptor antagonists may be useful in the treatment of brochial asthma.

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Study on the changes of sulfamethnzine residues in serum and practical organs of rats orally administrated with sulfamethnzine sodium (Rat에 sulfamethazine sodium 경구투여 후 혈청 및 실질장기내 sulfamethazine의 잔류량 추이에 관한 연구)

  • 도재철;이영미;조민희;신상희;박희주;송희종;정종식
    • Korean Journal of Veterinary Service
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    • v.23 no.4
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    • pp.321-333
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    • 2000
  • In order to know the depletive changes of sulfamethazine residues in senlm and practical organs of rats orally administered with sulfamethazine sodium(SMS), the concentration of sulfamethazine was measured in serum and tissue(kidney, liver, spleen, testis, and skeletal muscle) of rats with using high performance liquid chromatography(HPLC). SMS was orally administrated to sprague-dawley male rats(body weight, 200~300g) with using sonde at the rate of 20mg/100g body weight(recommended therapeutic dose) on once a day for 3 days. There were investigated the depletive changes of the sulfamethazine in serum, kidney, liver, spleen, testis and skeletal muscle of rat at the time 8 hours, 1st, 2nd, 3rd, 4th, 5th and 6th day after administration SMS, respectively. The results obtained were summarized as follows; 1. After oral administration of the SMS, the mean concentrations of sulfamethazine in serum according to the time lapsed were showed 215.53$\pm$42.99ppm at the 8 hours after withdrawal of medicated sulfamethazine. And gradually according to the time lapsed, the concentrations of sulfamethazine residues in serum were significantly (p<.05) decreased 25.87$\pm$5.18ppm at 1st day, 2.30$\pm$0.61ppm at 3rd day and 0.11$\pm$0.02ppm at 6th day respectively. 2. The mean concentrations of sulfamethazine in kidney, liver, spleen, muscle and testis according to the time lapsed after administration SMS were showed 83.82$\pm$12.16, 81.77$\pm$12.88, 36.96$\pm$5.35, 35.96$\pm$TEX>$\pm$1.39 and 27.89$\pm$1.92 ppm at the 8 hours, respectively. And gradually according to the time lapsed, the concentrations of sulfamethazine residues in the each of samples were significantly(p<.05) decreased such as 7.15$\pm$0.26, 5.62$\pm$0.72, 2.43$\pm$0.29, 1.99$\pm$0.14 and 3.11$\pm$0.48 ppm at 1st day, 0.52$\pm$0.04, 1.32$\pm$0.22, 0.13$\pm$0.03, 0.15$\pm$0.06 and 0.26$\pm$0.11ppm at 3rd day, and 0.03$\pm$0.01, 0.11$\pm$0.03, 0.02$\pm$0.01, 0.009$\pm$0.001 and 0.02$\pm$0.01 ppm at 6th day, respectively. 3. After oral administration of the SMS to rats, the residual concentrations of sulfamethazine in skeletal muscle were significantly (p<.05) decreased 35.96$\pm$1.39 to 0.009$\pm$$\pm$0.001 ppm between 8 hours and 6th day, respectively From the 4th day, the residual concentrations of sulfamethazine were showed 0.10$\pm$0.04 ppm below 0.1 ppm at the permitted limit concentration of muscle in Korea. In conclusion, this study could be suggested the relationship between administrated period, doses of sulfonamides and residual aspects of serum and practical organs, and the importance of observing ceasing period of antibiotic drugs before forwarding livestocks to slaughter.

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Neuroprotective effects of geneticin (G418) via apoptosis in perinatal hypoxic-ischemic brain injury (주산기 저산소성 허혈성 뇌손상에서 항고사를 통한 geneticin (G418)의 신경보호 효과)

  • Ju, Mi;Lee, Hyun Ju;Lee, Sun Ju;Seo, Eo Su;Park, Hye Jin;Lee, Kye Yang;Lee, Gyeong Hoon;Choi, Eun Jin;Kim, Jin Kyung;Lee, Jong Won;Chung, Hai Lee;Kim, Woo Taek
    • Clinical and Experimental Pediatrics
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    • v.51 no.2
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    • pp.170-180
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    • 2008
  • Purpose : Some antibiotics were known to exert neuroprotective effects in the animal model of hypoxic-ischemic (H-I) brain injury, but the mechanism is still unclear. A recent study reported that geneticin (G418), an aminoglycoside antibiotic, increased survival of human breast cancer cells by suppressing apoptosis. We investigated the neuroprotective effects of systemically administrated geneticin via anti-apoptosis following the H-I brain injury Methods : Seven-day-old Sprague-Dawley rat pups were subjected to unilateral (left) common carotid artery occlusion followed by 2.5 hours of hypoxic exposure and the cortical cell culture of rat brain was done under a hypoxic incubator. Apoptosis was measured in the injured hemispheres 7 days after H-I insult and in the injured cells from hypoxic chamber using morphologic analysis by Terminal dUTP Nick-end Labeling(TUNEL) assay and immunohistochemistry for caspase-3, and cytologic analysis by western blot and real time PCR for bax, bcl-2, and caspase-3. Results : The gross appearance and hematoxylin and eosin stain revealed increased brain volume in the geneticin-treated animal model of perinatal H-I brain injury. The TUNEL assay revealed decreased apoptotic cells after administration of geneticin in the cell culture model of anoxia. Immunohistochemistry showed decreased caspase-3 expression in geneticin-treated cortical cell culture. Western blot and real-time PCR showed decreased caspase-3 expression and decreased ratio of Bax/Bcl-2 expression in geneticin-treated animal model. Conclusion : Geneticin appears to exert a neuroprotective effect against perinatal H-I brain injury at least via anti-apoptosis. However, more experiments are needed in order to demonstrate the usefulness of geneticin as a preventive and rescue treatment for H-I brain injuries of neonatal brain.

The Efficacy Evaluation of Tourmaline-Ionized Water in Animal Study (투어마린이온활성수의 효능 평가)

  • Yoon, Yang-Suk;Kim, Dong-Heui;Qi, Xu-Feng;Song, Soon-Bong;Jung, Jong-Ho;Joo, Kyung-Bok;Teng, Yung-Chien;Lee, Kyu-Jae
    • Applied Microscopy
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    • v.39 no.4
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    • pp.311-317
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    • 2009
  • This study was performed using animals to confirm the effect of tourmaline-ionized water (TIW) the properties of which were changed by tourmaline energy and electric discharge. In the ICR mice fed high-fat diet, body weight increasing rate of the TIW-treated group (Exp) was generally decreased and moreover exhibited significance at 11th week (P<0.05) compared with the control (Con) group fed distilled water, although water intake of the Exp group was lower than that of the Con group. In the ICR mice with $CCl_4$-induced hepatotoxicity, AST and ALT activities of the Exp group were not significant but showed some decreasing trend, and histological damage of liver was less compared with thatof the Con group. On the study of ethanol-induced hangovers in Sprague-Dawley rat, blood alcohol concentration was significantly decreased (P<0.01), activity of GST, antioxidant enzyme related to the alcohol metabolism, was increased in liver tissue (P<0.05), and AST and ALT show a tendency to be decreasedin the Exp group. These results suggest that drinking TIWhas not only some obesity preventing effect but also an alcohol detoxification effect and liver protecting effect in vivo. It is supposed due to a structural change of water cluster and a property which maintains the changed structure through tourmaline energy and electric discharge. Therefore, TIW has a potentiality to be developed as functional water with several beneficial effects as well as for daily drinking, but further study on the mechanism related with efficacy will be necessary.

Some Observations on the Intercellular Junctions between the Hepatocytes in Fasting States as Revealed by Freeze Fracture Replica (기아가 간세포막결합장치에 미치는 영향에 관한 Freeze Fracture Replica법에 의한 관찰)

  • Ahn, Tae-Soon;Shin, Young-Chul
    • Applied Microscopy
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    • v.25 no.2
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    • pp.53-64
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    • 1995
  • This study was designed to investigate the morphological alterations of zonula occludens, macula adherences and gap junctions between the hepatocytes in the fasting conditions. Animals (Sprague Dawley, $250{\sim}280g$) were divided into two groups: normal and fasting. The latter were fasted for eight days prior to sampling. Liver tissues were sectioned and replicated after freeze fracturing for the transmission electron microscopy. In the normal rat liver, the interhepatocellular space at the area of some zonula occludens appeared to be widened in thin sections. On the freeze fracture replicas., the zonula occludens appeared as an anastomosing network of $2{\sim}4$ strands or grooves on P or E faces. Free ends and fragments of the strands were observed. In the rat fasted for eight days, the hepatocytes were diminished in size and the organelles were decreased in number and size. The intercellular space was wide at many areas of zonula occludens in thin section. On the freeze fracture replicas, the zonula occludens showed diminution or disappearence of anastomosing network of strands or grooves. Free ends and small fragments of the strands or grooves were frequently encountered. The macula adherens was markedly increased in number in thin sections, although they could not be found on the freeze fracture replicas. The gap junctions were increased in number in thin sections. Small aggregations of the intramembranous particles appeared with larger ones on the freeze fracture replica. The evidences may suggest the followings: (1) The disassembly of zonula occludens in the fasting states is led from the diminished mechanical stress on the luminal surface of bile canaliculus with the impaired secretion of bile components from the hepatocytes. (2) The increase of macula adherens is necessary to maintain the liver parenchyma integrity in the fasting state which leads the hepatocyte to be diminished and finally the intercellular space to be separated. (3) The rise in both number and size of gap junctions is owing to the need of increasing intercellular communication between the hepatocytes during the fasting. (4) The alteration of zonula occludens is easily led by the physiological condition of hepatocytes even in the normal ones.

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