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A Simple, Sensitive, and Specific HPLC Analysis of Tissue Polyamines using FNBT Derivatization: Its Application on the Study of Polyamine Metabolism in Regenerating Rat Liver (생체의 Polyamine-분석을 위하여 FNBT-유도체를 이용하는 간편하고 특이적이며 예민한 Isocratic RP-HPLC 분석법과 재생성 흰쥐-간의 Polyamine-대사의 변동에 관한연구)

  • Choi, Sang-Hyun;Kim, Hyung-Gun;Park, Hong-Ik;Chun, Boe-Gwun
    • The Korean Journal of Pharmacology
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    • v.24 no.2
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    • pp.233-240
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    • 1988
  • A simple and selective isocratic HPLC method for the analysis of tissue polyamine contents is described and applied on the study of the changes of the hepatic polyamine contents after partial hepatectomy in male rats. The hepatic polyamines are extracted with 0.4 M perchloric acid containing 2 mM disodium EDTA, and then the extract is redissolved in 100 ul of 1 M sodium carbonate and incubated with 300 ul of FNBT-dimethylsulfoxide (1: 100) mixture. The N-2'-nitro-4'-trifluoromethylphenyl drivatives of polyamines are separated through a ERC-ODS column in an isocratic mode with an acetonitrile-water (80:20) mobile phase within 20 min. per a sample, while monitoring the effluent at 242 nm. This improved method which could detect subnanogram of each polyamines is highly specific and reproducible as evidenced by the application of it on the study of the changes of polyamine contents in the regenerating rat liver after partial hepatectomy.

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Effect of the Paclitaxel and Radiation on the Large Bowel Mucosa of the Rat (횐쥐의 대장점막에 Paclitaxel(Taxol)과 방사선조사의 효과)

  • Lee Kyung-Ja
    • Radiation Oncology Journal
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    • v.17 no.1
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    • pp.57-64
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    • 1999
  • Purpose : Paclitaxel is a chemotherapeutic agent with a potent microtubule stabilizing activity that arrests mitosis at G2-M phase of cell cycle which is the most radiosensitive period. Therefore paclitaxel is considered as a cell cycle-specific radiosensitizer. This study investigates the effect of paclitaxel on the radiation response of the normal large bowel mucosa of the rat. Materials and Methods: The rats were divided into the three groups i.e., single intraperitoneal infusion of paclitaxel (10 mg/kg), a single fraction of irradiation (8 Gy, x-ray) to the whole abdomen, and a combination of irradiation (8 Gy, x-ray) given 24 hours after paclitaxel infusion. The histological changes as well as kinetics of mitotic arrest and apoptosis were evaluated on the large bowel mucosa at 6 hours, 1 day, 3 days and 5 days after treatment with paclitaxel alone, radiation alone and combination of paclitaxel and radiation. Results : The incidence of the mitotic arrest was not increased by paclitaxel infusion. The apoptosis appeared in 24 hours after paclitaxel infusion, and the histopathologic changes such as vesiculation, atypia and reduction of the goblet cell of the mucosa of the large bowel were demonstrated during the period from 6 hours to 3 days after, and returned to normal in 5 days after paclitaxel infusion. In irradiated group, the apoptosis was increased in 6 and 24 hours after irradiation, and the histopathologic changes of the mucosa were appeared in 24 hours and markedly increased in 3 days and returned to normal in 5 days. In combined group of irradiation and paclitaxel infusion, the apoptosis was appeared in 3 days and the histopathologic changes appeared during the period from 6 hours to 3 days after infusion. On the basis of the incidence of apoptosis and the degree of the histopathologic changes of the large bowel mucosa, there seemed to be additive effect by paclitaxel on radiation rather than sensitizing effect. Conclusions: The histopathological changes of large bowel mucosa in combined group compared to radiation alone group suggested an additive effect of paclitaxel on radiation response in the large bowel of rat.

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Effect of the Paclitaxel and Radiation on the Gastric Mucosa of the Rat (흰쥐의 위점막에 Paclitaxel (Taxol)과 방사선조사의 효과)

  • Lee Kyung-ja;Koo Heasoo
    • Radiation Oncology Journal
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    • v.17 no.4
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    • pp.314-320
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    • 1999
  • Purpose : Paclitaxel is a chemotherapeutic agent with potent microtubule stabilizing activity that arrests cells in $G_2$-M phase. Because $G_2$ and M are the most radiosensitive phase of the cell cycle, paclitaxel has potential role as a cell-cycle specific radiosensitizer. This study was peformed to see the effects of paclitaxel on the radiation-induced damage of gastric mucosa of the rat. Materials and Methods : The rats were divided into the three groups i.e., paclitaxel alone group, radiation alone group and, a combination of paclitaxel and radiation in combined group. A single intraperitoneal infusion of paclitaxel (10 mg/kg) was done in paclitaxel alone group. In radiation alone group, a single fraction of irradiation (8 Gy, x-ray) to the whole abdomen and, a combination of a single fraction of irradiation (8 Gy, x-ray) to the whole abdomen was given 24 hrs after paclitaxel infusion In combined group of paclitaxel and radiation. The incidence of mitosis and apoptosis as well as histologic changes of the gastric mucosa were evaluated at 6 hrs, 24 hrs, 3 days and 5 days after treatment. Results : The number of the mitosis was not increased by paclitaxel infusion. The incidence of the apoptosis was similar from 6 hrs to 3 days after paclitaxel infusion and was decreased at 5 days. Paclitaxel induced minimal glandular dilatation and cellular atypia of gastric mucosa at 24 hrs and 3 days. In irradiation group, the incidence of apoptosis was $6.0\%$ in 6 hrs and $1.25\%$ in 24 hrs after irradiation and minimal glandular dilatation and cellular atypia were noted throughout the experimental period. The incidence of apoptosis in the combined group of paclitaxel and irradiation ($4.5\%$) was significantly higher than irradiation alone group ($1.25\%$) at 3 days (p<0.05). Conclusion : Paclitaxel had no mitotic on mitotic arrest in gastric mucosa of the rat. Increased number of apoptosis in combined paclitaxel and irradiation group suggested the additive effects of paclitaxel on irradiation.

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Radiation-induced Apoptosis in Developing Fetal Rat Cerebral Cortex (발육 중의 백서 태아 대뇌 피질에서 방사선에 의한 아포토시스)

  • Chung Woong-Ki;Nam Taek-Kehn;Lee Min-Cheol;Ahn Sung-Ja;Song Ju-Young;Park Seung-Jin;Nah Byung-Sik
    • Radiation Oncology Journal
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    • v.21 no.4
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    • pp.315-321
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    • 2003
  • Purpose: This study was peformed to Investigate apoptosis by radiation In the developing fetal rat brain. Materials and Methods: Fetal blains were Irradiated In utero between the 17th and 19th days of fetal life (El7-19) by linear accelerator. A dose of Irradiation ranging from 1 Gy to 4 Gy was used to evaluate dose dependency. To test time dependency the ra)s were Irradiated with 2 Gy and then the fetal brain specimens were removed at variable 41me course; 1, 3, 5, 12 and 24 hours after the onset of irradiation. Immunohistochemlcal staining using in situ 707-mediated dUTP nick end labelling (TUNEL) technlfue was used for apoptotic cells. The cerebral cortex, including three zones on coriicai zone (Cf). Intermediate zone (if), and ventricular zone (VZ), was examined. Results : TUNEL positive cells revealed typical features of apoptotic cells under light microscope In the fetal rat cerebral cortex. Apoptotic cells were not found In the cerebral cortex of non-Irradiated fetal rats, but did appear In the entire cerebral cortex after 1 Gy Irradiation, and were more expensive at the ventricular and Intermediate zones than at the cortical zone. The extent of apoptosis was Increased with Increasing doses of radiation. Apoptosis reached the peak at S hours after the onset of 2 Gy Irradiation and persisted until 24 hours. Conclusion: Typical morphological features of apoplosis by irradiation were observed In the developing fetal rat cerebral cortex. It was more extensive at the ventricular and Intermediate zones than at the cortical zone, which suggested that stem cells or early differentiated cells are more radiosensitive than differentiated cells of the cortical zone.

A Comparison Study on Animal Models for Osteoarthritis in Temporomandibular Joint (측두하악관절에서의 골관절염 유도 동물모델 비교연구)

  • Yu, Sun-Nyoung;Yi, Young-Chul;Park, Hae-Ryoun;Ryu, Mi-Heon;Jeon, Hye-Mi;Kim, Kwang-Youn;Kim, Sang-Hun;Ok, Soo-Min;Ko, Myung-Yun;Ahn, Yong-Woo;Ahn, Soon-Cheol;Jeong, Sung-Hee
    • Journal of Oral Medicine and Pain
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    • v.36 no.4
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    • pp.261-271
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    • 2011
  • Osteoarthritis in patients with temporomandibular disorders(TMDs) induces pain, limitation of mouth opening, occlusal problems, and most commonly affects their life quality. Control method and progressive process of osteoarthritis are being extensively researched. The researchers focus on histologic changes, synovial changes, muscular and ligamental changes and observed reaction to pain. Therefore most of them developed the animal model for osteoarthritis in TMD patients. In this study, we applied several methods which induces osteoarthritis of temporomandibular joint(TMJ) in rats or mice. For locally induce osteoarthritis in TMJ, Monosodium iodoacetate(MIA) or interleukin-$1{\alpha}$(IL-$1{\alpha}$) were injected into TMJ joint space for 5 or 3 weeks. Other groups are chosen for osteoarthritis under systemic control including hormonal changes and aging. To observe cellular change, increased collagen, degenerative bony destruction and distribution of proteoglycans (PGs), safranin-O staining and Masson's trichrome staining were used.

The Effect of Platelet Derived Growth Factor - BB Loaded Chitosan/Calcium Metaphosphate on Bone Regeneration (혈소판유래성장인자를 함유한 Chitosan/Calcium Metaphosphate의 골조직재생효과에 관한 연구)

  • Lee, Seung-Yeol;Seol, Yang-Jo;Lee, Yong-Moo;Lee, Ju-Yeon;Lee, Seung-Jin;Kim, Suk-Young;Ku, Young;Rhyu, In-Chul;Han, Soo-Boo;Choi, Sang-Mook;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.31 no.1
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    • pp.1-23
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    • 2001
  • Chitosan is biodegradable natural polymer that has been demonstrated its ability to improve wound healing, and calcium metaphosphate(CMP) is a unique class of phosphate minerals having a polymeric structure. In this study, chitosan/CMP and platelet derived growth factor(PDGF-BB) loaded chitosan/CMP sponges were developed, and the effect of the sponges on bone regeneration and their possibility as scaffolds for bone formation by three-dimensional osteoblast culture were examined. PDGF-BB loaded chitosan/CMP sponges were prepared by freeze-drying of a mixture of chitosan solution and CMP powder, and soaking in a PDGF-BB solution. Fabricated sponge retained its 3-dimensional porous structure with $100-200\;{\mu}m$ pores. The release kinetics of PDGF-BB loaded onto the sponge were measured in vitro with $^{125}I-labeled$ PDGF-BB. In order to examine their possibility as scaffolds for bone formation, fetal rat calvarial osteoblastic cells were isolated, cultured, and seeded into the sponges. The cell-sponge constructs were cultured for 28 days. Cell proliferation, alkaline phosphatase activity were measured at 1, 7, 14 and 28 days, and histologic examination was performed. In order to examine the effect on the healing of bone defect, the sponges were implanted into rat calvarial defects. Rats were sacrificed 2 and 4 weeks after implantation and histologic and histomorphometrical examination were performed. An effective therapeutic concentration of PDGF-BB following a high initial burst release was maintained throughout the examination period. PDGF-BB loaded chitosan/CMP sponges supported the proliferation of seeded osteoblastic cells as well as their differentiation as indicated by high alkaline phosphatase activities. Histologic findings indicated that seeded osteoblastic cells well attached to sponge matrices and proliferated in a multi-layer fashion. In the experiments of implantation in rat calvarial defects, histologic and histomorphometric examination revealed that chitosan/CMP sponge promoted osseous healing as compared to controls. PDGF-BB loaded chitosan/CMP sponge further echanced bone regeneration. These results suggested that PDGF-BB loaded chitosan/CMP sponge was a feasable scaffolding material to grow osteoblast in a three-dimentional structure for transplantation into a site for bone regeneration.

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Effect of Platelet-Activating Factor on Cyclic Nucleotide Level in Rat Uterine tissue during Preimplantation Period (흰쥐의 임신초기에 있어서 자궁 조직중 Cyclic Nucleotide의 변화 및 Platelet-Activating Factor의 영향에 관한 연구)

  • Park, Kyoung-Sik;Kwun, Jong-Kuk
    • Clinical and Experimental Reproductive Medicine
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    • v.18 no.2
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    • pp.133-142
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    • 1991
  • This study was carried out to observe the change in uterine cyclic nucleotide level and the effect of PAF on cyclic nucleotides in uterine tissue in early pregnany in order to understand reciprocal relation ship between PAF and cyclic nucleotides in pregnancy in the rat. The test groups were injected intramuscularly with $1{\mu}g$ of PAF or 1.25mg of BN-52021 on day 0, 1, 2, 3, 4 and 5 of pregnancy. The level of cyclic nucleotide in removed uterine tissue was assayed by using cyclic nucleotides test kits. The results showed that the cyclic AMP content in uterine tissue of non-pregnant at pro-oestrus rat was $2.91{\pm}0.33$ pmol/mg protein which was lower than those of pregnant rat. The cyclic GMP content in uterine tissue of non-pregnant rat was $0.39{\pm}0.20$ pmol/mg pro-tein which was also lower than those of pregnant rats. The maximum level in cAMP was $5.92{\pm}1.72$ pmol/mg protein on day 3 and cGMP, $1.03{\pm}0.22$ pmol/mg protein on day 4. On each day of pregnancy, PAF induced the increased cAMP level ompared with that of intact rat. That was significant on day 0, 2 and 4 of pregnancy, p<0.05, on the other hand PAF receptor antagonist, BN-52021 ecreased cAMP level in uterine tisssue. PAF as well as BN-52021 had not an consistent effect on changes in cGMP level. These results suggest that cyclic nucleotide levels in uterine tissue ware increased during early pregnancy and PAF influences cAMP level in uterine rather than cGMP level during peri-implantation period, accordingly demonstrating a possible involvement of PAF in the regulation of implantation-related events through cAMP-mediated process.

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The Effects of Sea Cucumber as an Anti-gastritis, Anti-gastric Ulcer, and Anti-Helicobater (해삼의 항위염, 항위궤양 및 항헬리코박터 효과)

  • Oh, Hong-Geun;Moon, Dae-In;Kim, Jung-Hoon;Kang, Young-Rye;Park, Jung-Woo;Seo, Min-Young;Park, Sang-Hoon;Kang, Yang-Gyu;Choe, Chung-Hyeon;Park, In-Sun;Kim, Ju;Yu, Kang-Yeol;Seol, Eu-Ddeum;Kim, Ok-Jin;Lee, Hak-Yong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.5
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    • pp.605-611
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    • 2012
  • Sea cucumber, $Stichopus$ $japonicus$, is used not only as an outstanding tonic food but also as a traditional medicine for the treatment of asthma, hypertension, rheumatism, anemia, and sinus congestion. The purpose of this study was to examine sea cucumber as an anti-gastritis and anti-gastric ulcer in HCl-ethanol-induced gastric and $H.$ $pylori$-infected animal models. Thirty 7-week-old SD rats and Mongolian gerbils were divided into normal (Nor, n=6), control (Con, 60% HCl-ethanol+water, n=6), groupI (DSCI, 60% HCl-ethanol+sea cucumber 30 mg/kg, n=6), groupII (DSCII, 60% HCl-ethanol+sea cucumber 100 mg/kg, n=6), and group III (DSCIII, 60% HCl-ethanol+sea cucumber 300 mg/kg, n=6). Sea cucumber significantly suppressed gastric lesions and ulcers in the 60% HCl-ethanol-induced gastric model. Especially, 100 mg/kg of sea cucumber showed significantly inhibitory effects. In histopathological analysis of the $H.$ $pylori$ model, we found that sea cucumber augmented the eradication rates of $H.$ $pylori$ and attenuated gastric ulcer formation. Our results suggest that sea cucumber has inhibitory effects on gastritis and gastric ulcers. In addition, sea cucumber can be applied for the treatment of $H.$ $pylori$.

Effect of Dietary Fish Oil on Lipid Peroxidation and Antiperoxidative System in Rat Liver and Brain -Sex-related Differences- (어유(魚油)섭취가 흰쥐의 간과 뇌조직의 지질과산화물 형성과 항산화계에 미치는 영향 -성(性)의 차이를 중심으로-)

  • Choi, Kyung-Won;Park, Myungg-Hee;Chang, Kyung-Sook;Cho, Sung-Hee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.16 no.2
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    • pp.147-155
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    • 1987
  • In order to examine the effect of dietary fish oil on lipid peroxide formation and antiperoxidative efficiency in liver and brain, a group of male and female rats weighing about 70 grams were fed for three months, diet containing mackerel oil(MO) at the level of 10% (w/w). Results were compared, according to sex and source of dietary fat, i.e., in addition to MO, perilla oil(PO), soybean oil(SO), rapeseed oil(RO) or beef tallow(BT). Liver lipid peroxide level was significantly higher and levels of ${\alpha}-tocopherol$ and reduced glutathione(GSH) were lower in MO group than in other groups. This phenomenon was less clear in male than in female. Liver GSH level was lower in male, compared to female, but oxidized glutathione (GSSG) level did not vary, depending on either sex or dietary fat source. Brain lipid peroxide and ${\alpha}-tocopherol$ levels were not different among five experimental groups. Activities of liver and brain glutathione peroxidase and superoxide dismutase were not changed by dietary fat source, but glutathione peroxidase activity was higher in female than in male. The present study shows (a) that there is sex-related difference in antiperoxidatiye activity and (b) that fish oil containing $C_{20-22}({\omega}3)$ fatty acids, increases body lipid peroxide level and consumes more of cellular antioxidant, although it has lower total PUFA content than perilla or soybean oils.

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Inhibitory Effects of Dropwort (Oenanthe javanica) Extracts on Memory Impairment and Oxidative Stress and the Qualitative Analysis of Isorhamnetin in the Extracts (미나리 추출물의 기억력 손상 억제와 산화스트레스 억제 효과 및 Isorhamnetin 분석)

  • Won, Beom Young;Shin, Ki Young;Ha, Hyun Jee;Wee, Ji-Hyang;Yun, Yeo Sang;Kim, Ye Ri;Park, Yong Jin;Jung, Kyoung Ok;Sung, Hea Mi;Lee, Hyung Gun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.1
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    • pp.1-11
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    • 2016
  • This study was conducted to investigate effect of the acetylcholinesterase inhibitor activity, the protective effect of the extract on SH-SY5Y cell death by $H_2O_2$, the memory improvement from scopolamine-induced rat. Moreover, the antioxidant activity of isorhamnetin from the dropwort (Oenanthe javanica) was investigated. Acetylcholinesterase inhibitor activity was highest (28.59%) in Hwasun O. javanica extract (H-OJE). H-OJE and Naju O. javanica extract (N-OJE) were not significantly different. SH-SY5Y cell death deceased to 37.23% and 36.68% for H-OJE and N-OJE, respectively, following treatment with the extracts. O. javanica extracts showed a protective effect against $H_2O_2$-induced neurotoxicity. Treatment with O. javanica extracts slightly improved scopolamine-induced (1 mg/kg, i.p.) memory impairment in rats. H-OJE contained the highest total phenolic and flavonoid contents of 117 mg/g and 30 mg gallic acid equivalents/g, respectively, and had a DPPH radical scavenging activity ($SC_{50}$) of $113.8{\mu}g/mL$ and ABTS radical scavenging activity of $48.2{\mu}g/mL$, which was higher than the other extracts. The thiobarbituric acid reactive substances value was highest (50.2%) in H-OJE. Antioxidant activity differed significantly among dropwort extracts. Isorhamnetin was known as one of the flavonoid and for having neuroprotective effect. So we analyzed acid-hydrolyzed O. javanica extract HPLC. The results were that peak at 14 min and spectrum of the extracts was consistent with standard solution. The results of LC/MS/MS analysis were that the extract and standard solution were confirmed total ion chromatogram at identical time, precursor ion was 317 $[M-H]^+$ m/z, product ion was 302 $[M-H]^+$ m/z. Overall, the results showed that the dropwort extract led to memory improvement and had antioxidant activity. Based on these finding, further research to investigate the production of ethanol extract of dropwort as a processed food is warranted.