• Title/Summary/Keyword: Cellular activity

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EFFECT OF CURCUMIN AND RESVERATROL ON THE CELL CYCLE REGULATION, APOPTOSIS AND INHIBITION OF METASTASIS RELATED PROTEINS IN HN-4 CELLS (Curcumin과 resveratrol에 의한 두경부암 유래의 HN-4 세포의 세포주기, 세포사 및 전이관련 단백질의 발현 조절)

  • Kim, Sa-Yub;Lee, Sang-Han;Kwon, Taeg-Kyu
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.29 no.5
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    • pp.272-281
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    • 2003
  • Nontraditional or alternative medicine is becoming an increasingly attractive approach for the treatment of various inflammatory disorders and cancers. Curcumin is the major constitute of turmoric powder extracted from the rhizomes of the plant Curcuma longa. Resveratrol is a phytoalexin present in grapes and a variety of medicinal plants. In this report, We investigated the effect of curcumin and resveratrol on regulatory protein of cell cycle, induction of apoptosis and MMP activity. Treatment with 75 M curcumin for 24 hrs produced morphological changing in HN-4 cells. Curcumin and resveratrol inhibited the cellular growth in HN-4 cells. Inhibition of cell growth was associated with down-regulation of cell cycle regulatory proteins. Curcumin-induced caspase-3 activation and Bax degradation were dose-dependent with a maximal effect at a concentration of 100 M. The elevated caspase-3 activity in curcumin treated HN-4 cells are correlated with down-regulation of survivin and cIAP1, but not cIAP2. Curcumin induced a dose-dependent increase of cytochrome c in the cytosol. Curcumin induced-apoptosis was mediated through the release of cytochrome c. In addition, curcumin-induced apoptosis was caused by the generation of reactive oxygen species, which was prevented by antioxidant N-acetyl-cysteine (NAC). Cotreatment with NAC markedly prevented cytochrome c release, Bax cleavage and cell death. Also resveratrol-induced apoptosis was preceded by down-regulation of the anti-apoptotic Bcl-2, cIAP1, and caspase-3 activity. However, resveratrol-induced apoptosis was not prevented by antioxidant NAC. In addition, HN-4 cells release basal levels of MMP2 when cultured in serum-free medium. Treatment of the cells with various concentrations of PMA for 24 hr induced the expression and secretion of latent MMP9 as determined by gelatin zymography. HN-4 cells were treated with various concentrations of curcumin and resveratrol in the presence of 75 nM PMA, and MMP2 and 9 activities were inhibited by curcumin and resveratrol. These findings have implications for developing curcumin-based anticancer and anti-inflammation therapies.

Antioxidative and Protective Effects of Corn Silk (Zea mays L.) Extract on Human HaCaT Keratinocyte (옥수수수염 추출물의 항산화효과 및 피부각질세포 보호효과)

  • Kim, Hyun Young;Seo, Woo Duck;Seo, Kyung Hye;Lee, Mi-Ja;Choi, Sik-Won;Lee, Kwang-Sik;Kim, Sun Lim;Kang, Hyeon Jung
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.61 no.3
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    • pp.184-190
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    • 2016
  • We investigated the antioxidative and protective effects of corn silk (Zea mays L.) ethanol extracts on human HaCaT cells and erythrocytes. The NICS-2 fraction, extracted from corn silk, exhibited favorable 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) radical scavenging activities with $IC_{50}$valuesof$13.3{\pm}0.3{\mu}g/mL$ and $14.2{\pm}0.1{\mu}g/mL$ when compared with those of ${\alpha}$-tocopherol, a positive control, with $IC_{50}=10.4{\pm}02.2$ and $22.2{\pm}3.6{\mu}g/mL$, respectively. In addition, we investigated skin protection effects of NICS extracts of corn silk in HaCaT keratinocytes. To investigate the pharmacological potential of NICS-1 and NICS-2 extracts of corn silk on UV-B-induced damage in HaCaT cells, we measured the activity of interleukin (IL) 1a. Our results showed that all the corn silk extracts inhibited the UV-B-induced activity of IL-1a. In particular, NICS-1 extracts of corn silk significantly suppressed IL-1a activity in a dose-dependent manner without inducing cytotoxicity. These results indicate that the ethanol extracts of corn silk (Zea mays L.) could function as natural cytoprotective agents and antioxidants in biological systems, particularly the skin exposed to UV radiation, by protecting cellular membrane against reactive oxygen species (ROS).

The Antioxidant Effect of Hot Water Extract from the Dried Radish (Raphanus sativus L.) with Pressurized Roasting (가압볶음 무말랭이 열수 추출물의 항산화 효과)

  • Song, Yeong-Bok;Choi, Jeong-Sun;Lee, Ji-Eun;Noh, Jeong-Sook;Kim, Mi-Jeong;Cho, Eun-Ju;Song, Yeong-Ok
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.8
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    • pp.1179-1186
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    • 2010
  • The antiradical property of hot water extract from dried radish (DR) or dried radish roasted with pressure (DRRP) was investigated in vitro and in LLC-PK1 cell system. The contents of total free amino acid and reducing sugar in DR were decreased by 72.86% and 3.17%, respectively, after pressurized roasting. In vitro test, $IC_{50}$ for DR and DRRP for DPPH radical scavenging activity were 646.70 and $135.45\;{\mu}g/mL$, 896.10 and $566.98\;{\mu}g/mL$ for superoxide anion radical, and 722.26 and $531.84\;{\mu}g/mL$ for hydroxy radical, respectively. The radical scavenging effects of DRRP was significantly greater than those for DR (p<0.001). These radical scavenging effects of DR and DRRP were confirmed in LLC-$PK_1$ at which oxidative stresses were induced by superoxide, nitric oxide and peroxynitrite generated in the treatment of pyrogallol, SNP, and SIN-1, respectively. Cell viability was increased in the presence of DR or DRRP, dose dependently (p<0.05), and TBARS formation was decreased. The protective effects of DRRP against oxidative damage in LLC-$PK_1$ were greater than those of DR at the same concentration tested (p<0.05). This superior antiradical activity of DRRP might be due to the products produced during the pressurized roasting in addition to the antioxidative compounds originally present in the radish. 5-hydroxyl methyl furfural (5-HMF) known as an intermediate product of the maillard reaction was detected in DRRP (0.57 mg/g), but not from DR. In conclusion, daily consumption of DRRP may prevent oxidative damage by retarding oxidative stress.

Antioxidative, and Inhibitory Activities on Melanogenesis of Vitex negundo L. Leaf Extract (목형 잎 추출물의 항산화 활성과 멜라닌 생합성에 대한 저해활성)

  • Kim, A Reum;Park, Su Ah;Ha, Ji Hoon;Park, Soo Nam
    • Microbiology and Biotechnology Letters
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    • v.41 no.1
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    • pp.135-144
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    • 2013
  • The aim of this study was to evaluate various aspects of Vitex negundo L. leaf extract, such as the antioxidative activity, tyrosinase inhibitory effects, and inhibitory activities on ${\alpha}$-MSH induced melanogenesis, and active component analysis. The DPPH (1, 1-diphenyl-2-picrylhydrazyl) scavenging activities ($FSC_{50}$) of the ethyl acetate fraction and aglycone fraction of V. negundo L. leaf extract were $14.51{\mu}g/ml$ and $13.96{\mu}g/ml$, respectively. A luminol-dependent chemiluminescence assay revealed that the reactive oxygen species (ROS) scavenging activity ($OSC_{50}$) of the aglycone fraction of V. negundo L. leaf extract on ROS generated in an $Fe^{3+}$-$EDTA/H_2O_2$ system was the most prominent at $0.22{\mu}g/ml$. The protective effects of the extracts fractions of V. negundo L. leaf against the rose-bengal sensitized photohemolysis of human erythrocytes were increased in a concentration dependent manner ($1{\sim}50{\mu}g/ml$). In particular, there were greater protective effects of the aglycone fraction on the cellular membrane than that of the fat-soluble antioxidant (+)-${\alpha}$-tocopherol. The inhibitory effects ($IC_{50}$) on mushroom tyrosinase were the highest for the ethyl acetate fraction ($IC_{50}$ = $48.58{\mu}g/ml$). The inhibitory effect on ${\alpha}$-MSH induced melanogenesis in B16 melanoma cells was 41.80% at $50{\mu}g/ml$ of ethyl acetate fraction. Active component analyses by TLC, HPLC and LC/ESI-MS revealed luteolin and isoorientin. These results indicate that V. negundo L. leaf extract can be used as an antioxidant for ROS scavenging. Particularly, the luteolin and isoorientin of the ethyl acetate fraction may be applicable to new whitening cosmetics because of its inhibitory effect on mushroom tyrosinase and ${\alpha}$-MSH induced melanogenesis in B16 melanoma cells.

A Study on Anti-Bone Resorption & Osteoporosis by Taeyoungion-Jahage Extracts

  • Bae, Hyo-Sang;Cho, Hyung-Lae;Kim, Dong-Il;Lee, Tae-Kyun;Kim, Jun-Ki;Shin, Jung-Sik
    • The Journal of Korean Obstetrics and Gynecology
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    • v.15 no.4
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    • pp.61-75
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    • 2002
  • Recombinant human $interleukin-1{\beta}$ $(rhIL-1{\beta})$ regulates several activities of the osteoblast cells derived from mouse calvarial bone explants in vitro. $rhIL-1{\beta}$ stimulated cellular proliferation and the synthesis of prostaglandin $E_2(PGE_2)$ and plasminogen activator activity in the cultured cells in a dose-dependent manner. However, the induction of osteocalcin synthesis and alkaine phosphatase activity in response to vitamine D, two characteristics of the osteoblast phenotype, were antagonized by $rhIL-1{\beta}$ over a similar dose range. This study supports the role of $IL-1{\beta}$ in the pathological modulation of bone cell metabolism, with regard to implication in the pathogenesis of osteoporosis by $IL-1{\beta}$. When the mouse calvarial bone cells were used, the bone resorption induced by $IL-1{\beta}$ was strongly inhibited by calcitonin treatment, indicating osteoclast-mediated bone resorption. On the other hand, the medicinal extracts of Taeyoungjon-Jahage (T.Y.J-J.H.G extracts) was tested for whether they could inhibit $IL-1{\beta}-induced$ $PGE_2$ production. Cell viability was not significantly affected by treatment with the indicated concentration of the extracts. The T.Y.J.-J.H.G. extracts were shown to have the inhibitory effects against the synthesis of $PGE_2$. We also examined the effect of the pretreatment with a various concentrations of the T.Y.J.-J.H.G. extracts then treated the $PGE_2-induction$ agents. Pretreatment of the T.Y.J.-J.H.G. extracts for 1 h, which by itself had little effect on cell survival, did not enhance the synthesis of $PGE_2$. Furthermore, the T.Y.J-J.H.G. extracts were shown to have the protective effects against plasminogen dependent fibrinolysis induced by the bone resorption agents of $IL-1{\beta}$. Pretreatment of the T.Y.J.-J.H.G. extracts for 1 h did not enhance the plasminogen dependent fibrinolysis. Finally, calcitonin showed the inhibitory activity the $IL-1{\beta}-stimulated$ bone resorption in the mouse calvarial bone cells having both of the osteoblast and osteoclast cells. Seemingly, pretreatment of the T.Y.J.-J.H.G. extracts for 1 h reduced the bone resorption. These results clearly indicated that calcitonin and T.Y.J.-J.H.G. extracts play key roles in inhibition of the osteoclast-mediated bone resorption.

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Antioxidative Activity and Component Analysis of Cayratia japonica Extract (오렴매 추출물의 항산화 활성, 성분 분석)

  • Yang, Hee-Jung;Kim, Eun-Hee;Park, Soo-Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.34 no.2
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    • pp.117-127
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    • 2008
  • In this study, the antioxidative effects, inhibitory effects on elastase, and components of Cayratia japonica extracts were investigated. The free radical(1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activities($FSC_{50}$) of extract/fractions of Cayratia japonica were in the order: 50% ethanol extract(114.3 ${\mu}g/mL$)${\mu}g/mL$)${\mu}g/mL$). Reactive oxygen species(ROS) scavenging activities($OSC_{50}$) of some Cayratia japonica extracts in $Fe^{3+}-EDTA/H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The order of ROS scavenging activities were deglycosylated flavonoid aglycone fraction($OSC_{50},\;3.30{\mu}g/mL$)<50% ethanol extract(1.21 ${\mu}g/mL$)${\mu}g/mL$). Ethyl acetate fraction showed the most prominent scavenging activity. The protective effects of extract/fractions of Cayratia japonica on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The Cayratia japonica extracts suppressed photohemolysis in a concentration dependent manner($1{\sim}25{\mu}g/mL$), particularly deglycosylated flavonoid aglycone fraction exhibited the most prominent celluar protective effect(${\tau}_{50}$, 175.05min at 25 ${\mu}g/mL$). Aglycone fractions obtained from the deglycosylation reaction of ethyl acetate fraction among the Cayratia japonica extracts, showed 2 bands in TLC and 2 peaks in HPLC experiments(360 nm). Two components were identified as luteolin(composition ratio, 47.50%), apigenin(52.50). TLC chromatogram of ethyl acetate fraction of Cayratia japonica extract revealed 3 bands and HPLC chromatogram showed 4 peaks, which were identified as luteolin-7-O-${\beta}$-D-glucopyranoside(composition ratio, 11.14%), apigenin-7-O-${\beta}$-D-glucuronopyranoside(15.38%), luteolin(23.55%) and apigenin(49.92%) in the order of elution time. The inhibitory effect of aglycone fraction on elastase($IC_{50},\;70.5{\mu}g/mL$) was very high. These results indicate that extract/fractions of Cayratia japonica can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging $^1O_2$ and other ROS, and protect cellular membranes against ROS. And component analysis of Cayratia japonica extract and antioxidative effects could be applicable to new cosmetics.

Antioxidative Activities and Antiaging Effects of Geum aleppicum Jacq. Extracts (큰뱀무 추출물의 항산화 활성 및 항노화에 관한 연구)

  • Kim, Min-Ji;Yang, Hyun-Gab;Park, Soo-Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.37 no.2
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    • pp.191-198
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    • 2011
  • In this study, antioxidative effects and inhibitory effects of Geum aleppicum Jacq. extracts on tyrosinase and elastase were investigated. The ethyl acetate fraction of G. aleppicum Jacq. extract ($4.70\;{\mu}g$/mL) showed the most prominent free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity (FSC50). Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of some G. aleppicum Jacq. extracts on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The ethyl acetate fraction showed the most prominent ROS scavenging activity ($0.22 \;{\mu}g$/mL). The protective effects of extract/fraction of G. aleppicum Jacq. against the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The G. aleppicum Jacq. extracts suppressed photohemolysis in a concentration dependent manner ($1{\sim}25{\mu}g$/mL), particularly the ethyl acetate fraction exhibited the most prominent celluar protective effect (${\tau}_{50}$, 416.20 min at $10 \;{\mu}g$/mL). The inhibitory effect of G. aleppicum Jacq. extracts on tyrosinase and elastase were investigated to assess their whitening and anti-winkle efficacy. The half maximal inhibitory concentration ($IC_{50}$) of the ethyl acetate fraction on tyrosinase was $95.23\;{\mu}g$/mL. The $IC_{50}$ of 50 % ethanol extract and the ethyl acetate fraction on elastase were $6.27 \;{\mu}g$/mL and $4.31 \;{\mu}g$/mL, respectively. These results indicate that extract/fraction of G. aleppicum Jacq. can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging $^1O_2$ and other ROS, and protect cellular membranes against ROS. Especially the ethyl acetate fraction of G. aleppicum Jacq. extracts could be applicable to new functional cosmetics for antioxidant, antiaging.

The Effect of the IGF-I treated Gingival and Periodontal Ligament Fibroblast on Osteoblasts (IGF-I으로 처리한 치은 및 치주인대 섬유모세포가 골모세포에 미치는 영향)

  • Kim, Mi-Jeong;Yang, Won-Sik
    • The korean journal of orthodontics
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    • v.31 no.6 s.89
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    • pp.589-600
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    • 2001
  • Insulin-like growth factor I (IGF-I) has the local tissue regulating actions. In bone, IGF-I increases the replication of osteoblastic lineage, probably preosteoblasts, and enhances osteoblastic collagen synthesis and matrix composition rates. The purpose of this study was to investigate the local regulatory effect of IGF-I on periodontium totally, both in an autocrine and paracrine manner. To examine the effect of IGF-I directly on osteoblast (OB) of test rats, and indirectlv on OB via periodontal ligament fibroblast (PDLF), and the effect of gingival fibroblast (GF) on OB via cellular paracrine manner for the understanding of humoral action of adjacent tissue, GF and PDLF were obtained from male Sprague-Dawley rats of six to eight weeks of age. OB was obtained iron frontal and parietal calvarial bone of Sprague-Dawley 21day-old-fetus. After each tell was Incubated 24 hours, for collecting conditioned medium, different concentrations of IGF-I (1,10,100 ng/ml,1ml/well) was adding in the GF, PDLF cells, and the supernatant from these cultures was put into the primary OB culture with $1{\times}10^4$cell/ml/well. The experimental group was divided into six groups control OB, IGF-I treated OB, OB culture with conditioned medium from PDLF, OB culture with conditioned medium from IGF-I treated PDLF, OB culture with conditioned medium from GF, OB culture with conditioned medium from IGF-I treated GF. After final IGF-I treatment, OB was Incubated for 24 hours, and alkaline phosphatase activity assay, BMP expression, cell proliferation measurement using MTT assay, total protein measurement, Collagen synthesis assay using western blot, and examination of bone nodule synthesis were done. Alkaline phosphatase expressions were increased in the group of PDLF-IGF-I supernatant treatment. Direct IGF-I treatment with concentrations of 100ng/m1 showed increased viable tell number measured by MTT assay. And IGF-I treatment did not increase total protein amount. The entire experimental group showed BMP2, 4 expression in western blot, and there was no significant difference between control and experimental groups. These results suggested that supernatant from PDLF effects on increasing cellular activities of OB regardless of IGF-I, and at high concentration, IGF-I increases OB tell proliferation.

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Carbohydrate Metabolism in Preimplantation Stage Embryos and the Role of Metabolites (착상전 초기 배아에서 탄수화물 대사와 그 대사물의 역할)

  • Cheon, Yong-Pil
    • Development and Reproduction
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    • v.12 no.1
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    • pp.19-30
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    • 2008
  • Proper development of fertilized oocyte to blastocyst is a key step in mammalian development to implantation. During development of preimplantation embryos, the mammalian embryo needs supply the energy substrate for keep viability. Usually mammalian oocyte get substrate especially energy substrate from oviduct and uterus, because it does not store much substrate into cytoplasm during oogenesis. Carbohydrates are known as a main energy substrate for preimplantation stage embryos. Glucose, lactate and pyruvate are essential component in preimplantation embryo culture media and there are stage specific preferences to them. Glucose transporter and $H^+$-monocarboxylate cotransporter are a main mediator for carbohydrate transport and those expression levels are primarily under the control of intrinsic or extrinsic factors like insulin and glucose. Other organic substances, amino acids, lipids and nucleotides are used as energy substance and cellular regulation factor. Though since 1960s, successful development of fertilized embryo to blastocyst has been accomplished with chemically defined medium for example BWW and give rise to normal offspring in mammals, the role of metabolites and the regulation of intermediary metabolism are still poorly understood. Glucose may permit expression of metabolic enzymes and transporters in compacting morula, capable of generating the energy required for blastocyst formation. In addition, it has been suggested that the cytokines can modulate the metabolic rate of carbohydrate in embryos and regulate the preimplantation embryonic development through control the metabolic rate. Recently we showed that lactate can be used as a mediator for preimplantation embryonic development. Those observations indicate that metabolites of carbohydrate are required by the early embryo, not only as an energy source, but also as a key substrate for other regulatory and biosynthetic pathways. In addition metabolites of carbohydrate may involve in cellular activity during development of preimplantation embryos. It is suggested that through these regulation and with other regulation mechanisms, embryo and uterus can prepare the embryo implantation and further development, properly.

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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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