• Title/Summary/Keyword: rat liver cell

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Effects of carbendazim on DNA, gene and chromosome (살균제 carbendazim이 DNA, 유전자 및 염색체에 미치는 영향)

  • Lee, Je-Bong;Sung, Pil-Nam;Jeong, Mi-Hye;Shin, Jin-Sup;Kang, Kyu-Young
    • The Korean Journal of Pesticide Science
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    • v.8 no.4
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    • pp.288-298
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    • 2004
  • Benzimidazole pesticide carbendazim that is effective against a wide range of fungal plant pathogens is a protective, eradicant, and systemic fungicide. For genetic toxicity evaluation of carbendazim on DNA, genes and chromosome, were investigated with chromosome aberration, bacterial reverse mutation, micronucleus test in mouse born marrow and DNA damage assay by single cell microgel electrophoresis. Substitution and frameshift mutation were not induce at variable concentration of carbendazim on Ames test with or without rat liver microsomal activation. For the result of chromosome aberration test, numerical changes of chromosome were detected at the concentrations higher than $4.0{\mu}g/m{\ell}$, but structural aberration was not induced. Positive control, Mitomycin-C and captafol made a structural aberration, but numerical change of chromosome did not appear. In the micronucleus test for mouse born marrow, carbendazim was negative, but was weak positive in DNA damage assay by single cell microgel electrophoresis because of increased DNA moving length of 20% to control.

Cytoprotective Effects of Polyamines Against Oxidative Stress (산화 스트레스에 대한 폴리아민의 세포보호 효과)

  • Ahn Seoni;Lee Ji Young;Chung Hae Young;Yoo Mi-Ae;Kim Jong-Min;Kim Byeong Gee
    • Journal of Life Science
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    • v.15 no.4 s.71
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    • pp.626-632
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    • 2005
  • The polyamines are essential components of all eukaryotic cells and absolutely necessary for cell growth. In the present study, the cytoprotective role of polyamine was characterized. When $Ac_2F$ rat liver cells were treated with 1M 2,2'-azobis (2-amidinopropane) dehydrochloride (AAPH), a water soluble free radical initiator, viability of the cells was noticeably decreased due to the increase of reactive oxygen species (ROS). The cytotoxic effect of AAPH as well as ROS generation were significantly inhibited by the treatment of polyamines. Among polyamines, especially spermine at $20{\mu}M$ concentration exerted over $45\%$ inhibition of AAPH-induced ROS generation. Western blotting was performed to determine whether superoxide dismutase(SOD) or catalase (CAT) expression was involved in oxidative stress. The AAPH treatment blocked both SOD and CAT protein expressions. Spermine could recover those protein expressions to the untreated control levels. According to the result of cycline E measurement, AAPH might block the entry of the cells into S phase of the cell cycle. The reduced expression of cyclin E protein could be fully recovered by the addition of spermine. The antioxidative effects of spermine was also further proved by the apopotitic morphological analysis using ethidium bromide and acridine orange.

Absorption, Excretion and Antioxidative Effect of Rebamipide on Reproductive Organ (Rebamipide의 생식기관 내 흡수, 배설 및 항산화제로서 불임치료효과)

  • Kim, Jong Il;Park, Hyun Jun;Park, Nam Cheol
    • Clinical and Experimental Reproductive Medicine
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    • v.32 no.4
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    • pp.301-314
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    • 2005
  • Objective: Rebamipide is a propionic acid derivative that has an action of the inhibition of superoxide production and removal of hydroxyl radical with the sperm incubation and cryopreservation. In the present study, to investigate whether rebamipide is useful to treat male infertility and sterility, the author observed the antioxidative effects in patient with male infertility and also examined its absorption and distribution in rat genital organ. Methods: To measure the distribution of rebamipide in reproductive organ in the rat, carbon indicated rebamipide, $^{14}C-OPC-12759$, was orally administered to 10 Spraque-Dawley rats and its organ concentration in serum, liver, kidney, stomach, duodenum, colon, urinary bladder, seminal vesicle, epididymis and testicle were measured each time after 0.5, 1, 2, 4, 8 and 24 hours by using HPLC fluorescent method. The concentrations in semen were measured by HPLC fluorescent method in a sample of 50 infertile males who took 900 mg of rebamipide daily for 3 months. To measure the antioxidative effect and fertility rate for 3 months, each month before and after the treatment, sperm motility, vitality, the oxygen free radical formation, level of peroxidation, fetilizing capacity of semen sample which were obtained from infertile male patients by masturbation after at least 48 hours abstinence were analyzed by computer assisted semen analyzer, eosin-nigrosin stain, chemiluminescence, thiobarbituric acid method and hypo-osmotic swelling test. Simultaneously in a sample that wanted baby, both pregnancy and delivery were researched. Results: The $^{14}C-OPC-12759$ concentration in the body of white rats was highest in gastrointestinal organ like stomach, smal intestine and duodenum and followed by genital organ like seminal vesicle, testis and epididymis. The rebamipide concentration in semen of infertile males was $220.77{\pm}327.84ng/mL$ (SD) which showed a large deviation but it was higher than serum which was $126{\pm}76ng/mL$ (SD). In the infertile males, after the treatment with rebamipide, the level of seminal reactive oxygen species (ROS) and lipid peroxidation have significantly decreased in duration of the treatment (p<0.05) and sperm vitality and fertilizing capacity except sperm motility significantly improved on post treatment of 2~3 months (p<0.05). Out of the 41 cases who hoped for pregnancy, 15 cases (36.6%) became pregnant and 12 cases had childbrith, 2 cases had miscarriage and one case is ongoing. The side effect was observed in 1 case (2%) which experienced diarrhea but it was lost spontaneously. Conclusions: We conclude from this study that rebamipide showed relatively high tendancy of absorption and excretion in the genital organ. In infertile males who had elevated ROS in semen, by specifically inhibiting the cell damage from the antioxidation, a way to preserve sperm motility, vitality and fertilizing capacity was confirmed.

Electron Microscopic Study on the Role of Actin Filaments during the Formation of Bile Canaliculi in Isolated Rat Hepatocyte Culture System (흰쥐에서 분리 배양한 간세포의 담세관 형성에 있어서 액틴미세섬유의 역할에 관한 전자현미경적 연구)

  • Park, Chang-Hyun;Chang, Byung-Joon;Uhm, Chang-Sub
    • Applied Microscopy
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    • v.29 no.4
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    • pp.437-450
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    • 1999
  • Bile canaliculi are the structure delivering bile secreted by hepatocytes into the bile passage. Bile secretion is mainly controlled by the cytoskeletal elements, mainly of actin in the microvilli, pericanalicular web. Most studies on the bile secretion have been done in viva situation, however, to control the various parameters in vitro culture system seem to be more useful. To set up an in vitro experimental system, the investigator isolated hepatocytes with an enzymatic method using a mixture of collagenase and hyaluronidase from normal Sprague-Dawley rat liver and cultured. Isolated hepatocytes were round and formed cords in culture. Microvilli covered the whole surface of hepatocytes. Bile canaliculi were formed between hepatocytes and were characterized by the presence of microvilli of various lengths and shapes mainly arising from small surface mounds. Actin filament core in the microvilli and pericanalicular actin web were incomplete. After cytochalasin D treatment, cultured hepatocytes were round but the surface were irregular with surfacen blebs, folds and grooves. Microvilli on the surface were scarce. Bile canaliculi were markedly dilated often with the detached junctional complexes. Bile canaliculi lacks microvilli almost completely and extended into the pericanalirular cytoplasm showing complex vacuolar and tubular structures by transmission electron mciroscopy. Pericanalicular actin web, intermediate filaments were hardly identified. Subsurface actin filaments were scattered scarcely under the cell membranes. These results suggest that hepatocytes isolated from rats can survive and form bile canaliculi in culture and the actin filaments are involved in the formation and/or maintenance of the bile canaliculi.

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Effect of Lonicerae Japonicae Flos on Bone Density in Ovariectomized Rat Model of Osteoporosis (난소 적출 흰쥐 골다공증 모델에서 금은화(金銀花)가 골밀도 증가에 미치는 효과)

  • Lee, SungYub;Kim, Minsun;Hong, SooYeon;Kim, Jae-Hyun;Kim, Hongsik;Lee, Chungho;Jung, Hyuk-Sang;Sohn, Youngjoo
    • The Korea Journal of Herbology
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    • v.36 no.5
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    • pp.81-91
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    • 2021
  • Objectives : Osteoporosis is a systemic skeletal disease that decreases bone density and increases the risk of fractures. Bisphosphonates and SERMs are mainly used to treat osteoporosis, but, long-term use increases the risk of side effects such as jaw bone necrosis and breast cancer. Therefore, it is necessary to develop a therapeutic agent for a natural product with few side effects. Water extract of Lonicerae Japonicae Flos (wLF) was mainly found to have anti-cancer and anti-inflammatory effects. However, the effect of wLF on osteoporosis has not been elucidated. Therefore, this experiment investigated the effect of wLF on osteoclasts, osteoblasts and osteoporosis models. Methods : In order to study the effect of wLF on osteoporosis, the OVX-induced rat model was used for in vivo study. After 8 weeks, we measured body weight, uterine weight, liver weight, femur weight, bone density, trabecular area and tibia ash weight. To determine the effect of wLF on osteoclast differentiation, we measured the number of TRAP-positive cells and TRAP activity. To examine the effect of wLF on the expression of osteoblast-related genes, we measured the mRNA expression of alkaline phosphatase (ALP, Alpl) and osteocalcin (OCN, Bglap2). Results : In vivo experiment, wLF inhibited the reduction of femur weight, trabecular area, bone density and tibia ash weight. In vitro experiment, wLF had no significant effect on osteoclast differentiation. However, wLF increased the mRNA expression of Alpl and Bglap2 in MC3T3-E1 cell. Conclusions : This result suggested that wLF may be used for the treatment and prevention of postmenopausal osteoporosis.

Protective Effects of Saururus chinensis Baill Extracts on Liver Cell (삼백초 건초 추출물의 간세포 보호 효과 연구)

  • Ha, Bae-Jin;Ha, Jong-Myung;Lee, Sang-Hyeon;Lee, Jin-Young;Park, Sung-Yi
    • Journal of Food Hygiene and Safety
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    • v.18 no.4
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    • pp.177-182
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    • 2003
  • Saururus chinensis Baill (Saururaceae) is a perennial plant that has been used for therapy of edema, jaundice and gonorrhea in Korean folk medicine. This study was carried out to investigate the inhibitory effects of Saururus chinensis Baill(SCB) on hepatoprotective effects in accutely Carbone Tetrachloride ($CCl_4$) treated rats. Saururus chinensis Baill (100 mg/kg) was administered into rats intraperitoneally for 2 weeks, after the injection of CCl4 (3.3 ml/kg). We dxamined the biochemical parameters by measuring the levels of Aspartate aminotransferase (AST), Alanine aminotransferase (ALT), Total cholesterol, Triglyceride (TG), HDL-cholesterol, and LDL-cholesterol in serum and Malonedialdehyde (MDA) in liver tissue of rats. cholesterol was siginificantly elevated in $CCl_4$-treated abnormal group (CTA). The higher level of HDL-cholesterol was found is Saururus chinensis Baill and CCl4 administered group (SCT), which showed the lower levels of Total-cholesterol and LDL-cholesterol. TG content in the SCT was decreased compared to CTA by 0.93 times. MDA content in the SCT was inhibited compared to CTA by 33.00%. These findings indicate that Saururus chinensis. Baill may have a protective effect against CCl4-treated hepatotoxicity in rats.

Effect of Dietary Fat Sources and L-arginine Supplementation on Endothelial function and Lipid Metabolism in Streptozotocin-Induced Diabetic Rats (식이지방의 종류와 L-arginine 보충이 당뇨쥐의 ENdothelium 기능 및 체내 지질대사에 미치는 영향)

  • 장문정;김연중;김명환
    • Journal of Nutrition and Health
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    • v.35 no.1
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    • pp.5-14
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    • 2002
  • Alteration in the syntesis or enhanced inactivation of nitric oxide(NO) can induce impairment of endothelial cell function. Insulin dependent diabetes mellitus(IDDM) is characterized by impaired endothelial function and vascular disease. NO is produced through L-arginine pathway To elucidate the hypothesis that the decreased production on NO in IDDM reflects vascular damage and the NO production can be manipulated by either dietary fat(7% of kg diet) or the oral supplementation with L-arginine(2g/kg bw), plasma markers for vascular endothelial damage and plasma lipid profiles were measured in streptozotocin(STZ)-induced diabetic rats. Diabetic or normal Sprague-Dawley rats were fed 6 different experimental diets for 4 weeks(SO : soybean oil, SOA: soybean oil + L-arginine supplementation, BT : beef tallow, BTA_ beef tallow + L-arginine supplementation, OV olive oil, OVA : olive oil + L-arginine supplementation). Plasma glucose, total cholesterel, HDL-cholesterol, LDL-cholesterol and triglyceride were measured. Endothelial markers, plasma von Willebrand factor(vWf), thromboxane B$_2$, and 6-keto PGF1$\alpha$ of aorta were measured by ELISA. Plasma NO production was evaluated through the measurement of nitrite by EIA. Feeding saturated fatty acid(SFA, BT) increased relative liver size(RLS) in diabetic rats compared to either polyunsatunted fatty acid(PUFA, SO) or monounsaturated fatty acid(MUFA, OV) The supplementation of L-arginine inhibited the liver and kidney enlargement in olive oil find diabetic rats. Plasma glucose was lower in diabetic animal find the olive oil compared to fed beef tallow and the supplementation L-arginine decreased it in diabetic rats find beef tallow significantly(p < 0.05). Plasma TXB$_2$ levels were increased due to diabetes and the value of beef tallow group showed highest value. Plasma vWf concentration of beef tallow group was higher value in normal rats and was elevated more in diabetes. In diabetic groups, the vWf concentration of olive oil group was lower than beef tallow or soybean oil group. The supplementation of L-arginine in diabetic rats decreased plasma TXB$_2$ and vWf levels significantly(p < 0.05). NO production was higher in normal olive oil fed rats and was tend to be decreased in diabetic rats and the supplementation of L-arginine recovered to normal value(p < 0.05), Olive oil supplemented with L-arginine tended to lower plasma total cholesterol and LDL-cholesterol after 4 week treatment. These results suggest that generalized vascular endothelial changes based on plasma TXB$_2$and vWf occurs in diabetic rats. and olive oil with L-arginine supplementation contributes to a better control of the hyperglycemia, endothelial changes and hypercholesterolemia accompanying diabetes as compared with beef tallow or soy bean oil in this rat model.

Immunohistological expression of cytochrome P450 1A2 (CYP1A2) in the ovarian follicles of prepubertal and pubertal rat

  • Hwang, Jong-Chan;Park, Byung-Joon;Kim, Hwan-Deuk;Baek, Su-Min;Lee, Seoung-Woo;Jeon, Ryoung-Hoon;Jang, Min;Bae, Seul-Gi;Yun, Sung-Ho;Park, Jin-Kyu;Kwon, Young-Sam;Kim, Seung-Joon;Lee, Won-Jae
    • Journal of Animal Reproduction and Biotechnology
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    • v.35 no.4
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    • pp.329-337
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    • 2020
  • Cytochrome P450 1A2 (CYP1A2) is a member of the cytochrome P450 superfamily enzymes in mammals and plays a major role in metabolizing endogenous hormones in the liver. In recent days, CYP1A2 expression has been found in not only the liver but also other tissues including the pancreas and lung. However, little information is available regarding the expression of CYP1A2 in the ovary, in spite of the facts that the ovarian follicle growth and atresia are tightly associated with controls of endocrine hormonal networks. Therefore, the expression of CYP1A2 in the ovaries of prepubertal and pubertal rats was investigated to assess its expression pattern and puberty-related alteration. It was demonstrated that the expression level of CYP1A2 was significantly (p < 0.01) higher in the pubertal ovaries than prepubertal counterparts. At the ovarian follicle level in both groups, whereas CYP1A2 expression was less detectable in the primordial, primary and secondary follicles, the strongly positive expression of CYP1A2 was localized in the granulosa cell layers in the antral and pre-ovulatory follicles. However, the ratio of CYP1A2-positive ovarian follicle was significantly (p < 0.01) higher in the ovary of pubertal group (73.1 ± 3.1%) than prepubertal one (41.0 ± 10.5%). During the Immunofluorescence, expression of CYP1A2 was mainly localized in Fas-positive follicles, indicating the atretic follicles. In conclusion, these results suggested that CYP1A2 expression was mainly localized at the atretic follicular cells and affected by the onset of puberty. Further study is still necessary but we hypothesize that CYP1A2 expresses in the atretic follicles to metabolize residue of the reproductive hormones. These findings may have important implications for the fields of reproductive biology of animals.

Effect of blended protein nutritional support on reducing burn-induced inflammation and organ injury

  • Yu, Yonghui;Zhang, Jingjie;Wang, Jing;Wang, Jing;Chai, Jiake
    • Nutrition Research and Practice
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    • v.16 no.5
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    • pp.589-603
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    • 2022
  • BACKGROUND/OBJECTIVES: Previous studies have reported that protein supplementation contributes to the attenuation of inflammation. Serious trauma such as burn injury usually results in the excessive release of inflammatory factors and organs dysfunction. However, a few reports continued to focus on the function of protein ingestion in regulating burn-induced inflammation and organ dysfunction. MATERIALS/METHODS: This study established the rat model of 30% total body surface area burn injury, and evaluated the function of blended protein (mixture of whey and soybean proteins). Blood routine examination, inflammatory factors, blood biochemistry, and immunohistochemical assays were employed to analyze the samples from different treatment groups. RESULTS: Our results indicated a decrease in the numbers of white blood cells, monocytes, and neutrophils in the burn injury group administered with the blended protein nutritional support (Burn+BP), as compared to the burn injury group administered normal saline supplementation (Burn+S). Expressions of the pro-inflammatory factors (tumor necrosis factor-α and interleukin-6 [IL-6]) and chemokines (macrophage chemoattractant protein-1, regulated upon activation normal T cell expressed and secreted factor, and C-C motif chemokine 11) were dramatically decreased, whereas anti-inflammatory factors (IL-4, IL-10, and IL-13) were significantly increased in the Burn+BP group. Kidney function related markers blood urea nitrogen and serum creatinine, and the liver function related markers alanine transaminase, aspartate aminotransferase, alkaline phosphatase, and lactate dehydrogenase were remarkably reduced, whereas albumin levels were elevated in the Burn+BP group as compared to levels obtained in the Burn+S group. Furthermore, inflammatory cells infiltration of the kidney and liver was also attenuated after burn injury administered with blended protein supplementation. CONCLUSIONS: In summary, nutritional support with blended proteins dramatically attenuates the burn-induced inflammatory reaction and protects organ functions. We believe this is a new insight into a potential therapeutic strategy for nutritional support of burn patients.

Effects of a Combined Diet of Jerusalem Artichoke's Inulin, Lotus Leaf and Herb Extracts in Obesity-induced White Rat with Fat Diet (돼지감자의 이눌린, 연잎, 허브의 병합식이가 고지방식이로 유도된 비만흰쥐에 미치는 영향)

  • Lee, Eun-Hye;Lee, Ye-Jin;Choi, Ok-Byung;Kang, Sang-Mo
    • Applied Biological Chemistry
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    • v.50 no.4
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    • pp.295-303
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    • 2007
  • A preliminary diet experiment utilizing Jerusalem artichoke's inulin, lotus leaf powder, nettle powder and eucalyptus powder extract indicated that combining all four elements gave the most effective result. Therefore, a study was conducted to evaluate the effectiveness of combined diet for weight loss. In this study, Sprague-Dawley, male white rats about 200 g in weight was fed with high fat diet for 8 weeks in order to induce obesity followed by 4 week administration of combined diet to look into the effect of the diet. After a total of 12 weeks of feeding, factors relevant to weight, blood, and lipid metabolism by liver in the body were researched and histologic change was examined with optical microscope. In terms of weight change, both high fat diet group and regular diet group gained weight from high fat diet for 8 weeks compared to normal group. Then, for another 4 weeks, while normal group and high fat diet group kept gaining weight, combined diet group which was provided with high fat diet for 8 weeks, lost weight to the normal group level after 3 week administration of diet. However, after the 4th week of administration, the group weighted significantly less than the normal group and the efficiency of diet also significantly dropped. In the biochemical analysis of blood, total cholesterol, LDL-cholesterol, triglyceride, glucose, GOT, GPT, ${\gamma}-GTP$ and creatine showed significant increase in high fat diet group and there was no significant difference between diet group and normal group except for GPT, ${\gamma}-GTP$ and creatine. In the biochemical analysis of liver, there was significant increase in LDL-cholesterol, triglyceride of high fat diet group compared to normal group, while there was no significant difference in term of total cholesterol and HDL-cholesterol. Compared to normal group, diet group had higher HDL-cholesterol, while total cholesterol dropped significantly. There was no significant difference in terms of LDL-cholesterol and triglyceride. Besides, in high fat diet group, observation of histologic change in liver and change in ultrastructure showed volume increase of hepatic cell and severe fatty degeneration in hepatic cell around hepatic vein. However in diet group, like normal group, no pathological change was observed in terms of cytoplasm, nucleus and capillary in hepatocyte and the alignment of hepatocyte had regularity thanks to the administration of combined diet. Therefore, combined diet utilizing Jerusalem artichoke's inulin, lotus leaf powder, nettle powder and eucalyptus powder was proven to be an effective measure to prevent and improve obesity as a result of abnormal adipose deposition.