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Effect of Anti-inflammation on Oryeong-san Formulation for Mix Extract Tablet (오령산 정제 개발 및 항염증 효과)

  • Kim, Se Jin;Leem, Hyun Hee;Nam, Won Hee;Son, Su Mi;Choi, Hye Min;Kim, Myung Jin;Kim, Jung Ok;Lee, Hwa Dong
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.34 no.6
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    • pp.348-354
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    • 2020
  • Oryeong-san (ORS) is a traditional Korean herbal medicine widely used for renal associated diseases, composed of five medicine herbs; Atractylodes japonica Koidzumi, Cinnamomum cassia Presl, Polyporus umbellatus Fries, Poria cocos Wolf and Alisma orientale Juzepzuk. We studied to improve the convenience of intake and portability by developing modernized dosage forms, and examined the effect on anti-inflammation of ORS. In order to develop the tablet formulation of ORS (ORS-F), the tablets were evaluated on the basis of physical characteristics include diameter, thickness, weight variation, hardness, friability and disintegration. To analyze the marker components of ORS-F, eight index markers from five herbal medicines were chosen. And the method using high performance liquid chromatography (HPLC) with diode-array detector method was established for the simultaneous analysis. The biological activities were examined the effect of ORS-F on pro-inflammation mediated by LPS-stimulation. The production of nitric oxide (NO) and cytokines were determined by reacting cultured medium with griess reagent and enzyme-linked immunosorbent assay (ELISA). The expression of cyclooxygenase-2 (COX-2) and inducible NO synthase (iNOS) were investigated by Western blot and RT-PCR. The anti-oxidant activities of OJS-F increased markedly, in a dose-dependent manner. and, The total phenolic compound and flavonoids contents of OJS-F were 10.20±0.09 ㎍/㎎ and 12.86±0.86 ㎍/㎎. OJS-F which is LPS has diminished in the LPS-induced release of inflammatory mediators (NO, iNOS, COX2 and PGE2) and pro-inflammatory cytokines (TNF-α, IL-6 and IL-1β) from the RAW264.7 macrophages. Therefore, the developed formulation for tablet of ORS-F provide efficiency and usability, and indicated effect of anti-inflammation.

Immunostimulatory activity of hydrolyzed and fermented Platycodon grandiflorum extract occurs via the MAPK and NF-κB signaling pathway in RAW 264.7 cells

  • Jae In, Jung;Hyun Sook, Lee;So Mi, Kim;Soyeon, Kim;Jihoon, Lim;Moonjea, Woo;Eun Ji, Kim
    • Nutrition Research and Practice
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    • v.16 no.6
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    • pp.685-699
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    • 2022
  • BACKGROUND/OBJECTIVES: Platycodon grandiflorum (PG) has long been known as a medicinal herb effective in various diseases, including bronchitis and asthma, but is still more widely used for food. Fermentation methods are being applied to increase the pharmacological composition of PG extracts and commercialize them with high added value. This study examines the hydrolyzed and fermented PG extract (HFPGE) fermented with Lactobacillus casei in RAW 264.7 cells, and investigates the effect of amplifying the immune and the probable molecular mechanism. MATERIALS/METHODS: HFPGE's total phenolic, flavonoid, saponin, and platycodin D contents were analyzed by colorimetric analysis or high-performance liquid chromatography. Cell viability was measured by the MTT assay. Phagocytic activity was analyzed by a phagocytosis assay kit, nitric oxide (NO) production by a Griess reagent system, and cytokines by enzyme-linked immunosorbent assay kits. The mRNA expressions of inducible nitric oxide synthase (iNOS) and cytokines were analyzed by reverse transcription-polymerase chain reaction, whereas MAPK and nuclear factor (NF)-κB activation were analyzed by Western blots. RESULTS: Compared to PGE, HFPGE was determined to contain 13.76 times and 6.69 times higher contents of crude saponin and platycodin D, respectively. HFPGE promoted cell proliferation and phagocytosis in RAW 264.7 cells and regulated the NO production and iNOS expression. Treatment with HFPGE also resulted in increased production of interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α, C-X-C motif chemokine ligand10, granulocyte-colony-stimulating factor, granulocyte-macrophage colony-stimulating factor, and monocyte chemoattractant protein-1, and the mRNA expressions of these cytokines. HFPGE also resulted in significantly increasing the phosphorylation of NF-κB p65, extracellular signal-regulated kinase, and c-Jun N-terminal kinase. CONCLUSIONS: Taken together, our results imply that fermentation and hydrolysis result in the extraction of more active ingredients of PG. Furthermore, we determined that HFPGE exerts immunostimulatory activity via the MAPK and NF-κB signaling pathways.

Evaluation of Evodiae Fructus Extract on the Chronic Acid Reflux Esophagitis in Rats (오수유(吳茱萸) 추출물이 만성 역류성 식도염 흰쥐에 미치는 효능 평가)

  • Lee, Jin A;Park, Hae-Jin;Kim, Soo Hyun;Kim, Min Ju;Kim, Kyeong Jo;Shin, Mi-Rae;Roh, Seong-Soo
    • The Korea Journal of Herbology
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    • v.34 no.2
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    • pp.15-23
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    • 2019
  • Objective : Reflux esophagitis (RE) is a disease that caused gastric acid reflux and inflammation due to unstable gastroesophageal sphincter, as increasing worldwide respectively. This study was conducted to evaluate the effect of Evodiae Fructus (EF) extract on chronic reflux esophagitis in rats. Methods : The EF was measured antioxidant activity, such as total polyphenol and total flavonoid contents, 1, 1-diphenyl-2-picrylhydrazyl (DPPH) and 2, 2'-azinobis-3-ethyl-enzothiazoline-6-sulfonic acid (ABTS) radical scavenging activity. Rats were divided into 3 groups; Nor (normal group), Con (chronic acid reflux esophagitis rats treatment with water), EF (chronic acid reflux esophagitis rat treatment with EF 200 mg/kg body weight group). A surgically-induced chronic acid reflux esophagitis (CARE) model was established in SD rats, and treated with water or EF 200 mg/kg body weight for 14 consecutive days. Results : Administration of EF to rats of induction of chronic acid reflux esophagitis was found to reduce esophagus tissues injury. Reactive oxygen species (ROS) and produces peroxynitrite ($ONOO^-$) levels of esophagus tissues were significantly decreased in EF compared to Con group. As results of esophagus protein analyses, EF effectively reduce inflammatory-related factors ($NF-{\kappa}Bp65$, $p-I{\kappa}B{\alpha}$, iNOS, $TNF-{\alpha}$, IL-6), and increase anti-oxidant enzyme (Nrf2, HO-1, SOD, catalase, GPx-1/2). Conclusions : These results suggest that EF administration comfirmed that decreased esophagus tissues injury, oxidantive stress, anti-inflammation effect, and increased anti-oxidant effect. Therefore, EF was the potential to be used as a natural therapeutic drug.

The study of anti-inflammatory effect of Hyeonto-dan extract in RAW 264.7 macrophage (현토단(玄兎丹)의 RAW 264.7 대식 세포에서의 항염증 효과에 관한 연구)

  • Kim, Ma-Ryong;Kang, Ok-Hua;Kong, Ryong;Seo, Yun-Soo;Zhou, Tian;Kim, Sang-A;Kim, Eun-Su;Sin, Min-A;Lee, Young-Seob;Kwon, Dong-Yeul
    • The Korea Journal of Herbology
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    • v.32 no.2
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    • pp.77-85
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    • 2017
  • Objectives : This study aimed to investigate the unknown mechanisms behind the anti- inflammatory activity of Hyeonto-dan(HT) 70% ethanol extract on LPS-stimulated RAW 264.7 cells. Methods : Cells were treated with Hyeonto-dan 1 h prior to addition of 200 ng/mL of LPS. Cell viability was measured by the MTS assay. Nitric oxide levels were determined by the Griess assay. $PGE_2$ were measured using EIA kit. Pro-inflammatory cytokine production was measured by the enzyme-linked immunosorbent assay (ELISA). The expression of COX-2, iNOS, and MAPKs was investigated by Western blot, qRT-PCR. $NF-{\kappa}B$/p65 localization and interaction of the TLR-4 receptor with LPS was examined by immunofluorescence assays. Results : Hyeonto-dan had no cytotoxicity at the measured concentration. Hyeonto-dan inhibited NO production and pro-inflammatory cytokines such as IL-6, $TNF-{\alpha}$, and PGE2 as well as the protein and mRNA expression of iNOS and COX-2. Moreover, Hyeonto-dan inhibited the interaction between LPS and TLR-4 in murine macrophages. It suppressed phosphorylation of extracellular signal-regulated kinase (ERK 1/2), c-jun N-terminal kinase (JNK 1/2) and p38. Finally, it inhibited translocation of $NF-{\kappa}B$ in response to competitive LPS. Conclusions : Based on the results of this study, Hyeonto-dan inhibited the binding of TLR-4 receptor to LPS and inhibited the phosphorylation of extracellular signaling pathway MAPKs. These inhibitory effects are thought that the amount of $NF-{\kappa}B$ delivered to the nucleus was decreased and the inflammatory reaction was prevented by decreasing the production of LPS-induced $PGE_2$, NO, IL-6 and $TNF-{\alpha}$.

Anti-inflammatory effect of Crypsinus hastatus biorenovation extract (고란초 생물 전환 추출물의 항염증 효과)

  • Lee, Kyung-Mi;Choi, Byeong Min;Park, Tae-Jin;Hong, Hyehyun;Kim, Seung-Young
    • Journal of Applied Biological Chemistry
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    • v.65 no.1
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    • pp.49-55
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    • 2022
  • Biorenovation is a method for converting materials using the enzyme properties of microorganisms. Natural products converted by that method increase physiological activity or reduce cytotoxicity. In this study, we investigated the anti-inflammatory activity of crypsinus hastatus prothallium (CH) and biorenovated CH prothallium (CHB) using RAW 264.7 cells stimulated with lipopolysaccharide (LPS). CHB inhibited the production of nitric oxide, prostaglandin E2 and cytokines (interleukin-6, interleukin-1β, tumor necrosis factor-α) compared to CH at a concentration of 50-200 ㎍/mL. In addition, CHB concentration of 200 ㎍/mL inhibited the expression of inducible nitric oxide synthase and cyclooxygenase-2 protein by LPS stimulation to the level of the untreated control group. These results indicate that CHB could be a novel anti-inflammatory agent for cosmetic and pharmaceutical ingredients.It also suggests that the application of biorenovation has potential usefulness in developing anti-inflammatory materials. It also suggests that the application of bio-renovation has potential usefulness in the development of inflammatory material. We applied Biorenovation technology to Distylium racemosum extract (DR) to generate Distylium racemosum biorenovation product (DRB), and investigated the anti-inflammatory properties of DRB in lipopolysaccharide (LPS)-treated RAW264.7 macrophages. We are applying technology to Biorenovation Distylium racemosum extract (DR) Distylium racemosum was to create a biorenovation product (DRB), lipopolysaccharide (LPS) investigated the anti-inflammatory properties of DRB in RAW264.7 macrophages treated for.

Antioxidant Responses in Brackish Water Flea Diaphanosoma celebensis - Exposed to Mercury (수은 노출에 대한 기수산 물벼룩 Diaphnosoma celebensis의 항산화 반응)

  • Bae, Chulhee;Lee, Young-Mi
    • Journal of Marine Life Science
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    • v.3 no.2
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    • pp.74-80
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    • 2018
  • Mercury (Hg) poses a threat to marine ecosystem due to continuous inflow from various industries and bioaccumulation to higher trophic level via food web. Mercury can adversely affect growth, development, reproduction and metabolism to aquatic organisms. In the present study, acute toxicity and oxidative stress markers (total glutathione content, and activities of GST, GR and GPx) were investigated in brackish water flea Disphanosoma celebensis exposed to HgCl2 for 24 h. As results, Hg showed negative effect in survival of D. celebensis. 24 h-LC50 value was determined as 0.589 mg/l (95% C.I. 0.521~0.655 mg/l). After exposure to Hg (0.08 and 0.4 mg/l) for 24 h, total glutathione content was significantly decreased, whereas GST, GPx and GR activities were enhanced. These findings indicate that Hg induced oxidative stress in D. celebensis, and oxidative stress markers may be involved in cellular defense against Hg - mediated toxicity. This study provides a better understanding of molecular mode of action of Hg toxicity in this specie and potent of molecular markers for heavy metal monitoring in marine ecosystem.

Anti-inflammatory activities of carrot(Daucus carota) leaf Fermented by Weizmannia coagulans KK7 (김치에서 분리한 Weizmannia coagulans KK7으로 발효한 당근 잎 추출물의 함염증 활성 연구)

  • Yoonji Lee;Boram Ko;Hyejin Hyeon;Daeju Oh;Weon-Jong Yoon
    • Journal of the Korean Applied Science and Technology
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    • v.39 no.6
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    • pp.932-940
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    • 2022
  • In this study, the extracts of carrot (Daucus carota var. sativa) leaf fermented with Weizmannia coagulans KK7 strain were investigated for the anti-inflammatory activities and component changes. The KK7 strain was isolated from kimchi, a Korean fermented vegetable. The high-performance liquid chromatography was performed to analyze the changes in the components of the carrot leaf extracts before and after fermentation. It was confirmed that the content of luteolin, a kind of flavonoid, was significantly increased after fermentation. The anti-inflammatory activities of the carrot leaf extracts and the fermented carrot leaf extracts were evaluated by the inhibition of NO (nitric oxide) production in LPS-induced RAW 264.7 cells. The NO scavenging ability of the fermented carrot leaf extracts was higher than the other extracts. The protein expression of iNOS, an enzyme responsible for the NO production was significantly reduced in a concentration-dependent manner by treatment with the fermented carrot leaf extracts. In conclusion, we found that the anti-inflammatory effect of carrot leaf was increased by microbial fermentation, suggesting that carrot leaf generally discarded could be used as new food and cosmetic materials through fermentation.

The embryological studies on the interspecific hybrid of ginseng plant (Panax ginseng x P. Quiuquefolium) with special references to the seed abortion (인삼의 종간잡종 Panax ginseng x P Quinquefoilium의 발생학적 연구 특히 결실불능의 원인에 관하여)

  • Jong-Kyu Hwang
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.5 no.1
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    • pp.69-86
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    • 1969
  • On the growing of the interspecific hybrid ginseng plant, the phenomena of hybrid vigoures are observed in the root, stem, and leaf, but it can not produce seeds favorably since the ovary is abortive in most cases in interspecific hybrid plants. The present investigation was undertaken in an attempt to elucidate the embryological dses of the seed failure in the interspecific hybrid of ginseng (Panax Ginseng ${\times}$ P. Quinque folium). And the results obtained may be summarized as follows. 1). The vegetative growth of the interspecific hybrid ginseng plant is normal or rather vigorous, but the generative growth is extremely obstructed. 2). Even though the generative growth is interrupted the normal development of ovary tissue of flower can be shown until the stage prior to meiosis. 3). The division of the male gameto-genetic cell and the female gameto-genetic cell are exceedingly irregular and some of them are constricted prior to meiosis. 4). At meiosis in the microspore mother cell of the interspecific hybrid, abnormal division is observed in that the univalent chromosome and chromosome bridge occure. And in most cases, metaphasic configuration is principally presented as 23 II+2I, though rarely 22II+4I is also found. 5). Through the process of microspore and pollen formation of F1, the various developmental phases occur even in an anther loclus. 6). Macro, micro and empty pollen grains occur and the functional pollen is very rare. 7). After the megaspore mother cell stage, the rate of ovule development is, on the whole, delayed but the ovary wall enlargement is nearly normal. 8). Degenerating phenomena of ovules occur from the megaspore mother cell stage to 8-nucleate embryo sac stage, and their beginning time of constricting shape is variously different. 9). The megaspore arrangement in the parent is principally of the linear type, though rarely the intermediate type is also observed, whereas various types, viz, linear, intermediate, Tshape, and I shape can be observed in hybrid. 10). After meiosis, three or five megaspore are some times counted. 11). Charazal end megaspore is generally functional in the parents, whereas, in F1, very rarely one of the center megaspores (the second of the third megaspore) grows as an embryo sac mother cell. 12). In accordance with the extent of irregularity or abnormality in meiosis, division of embryo sac nuclei and embryo sac formation cause more nucellus tissue to remain within th, embryo sac. 13). Even if one reached the stage of embryo sac formation, the embryo sac nuclei are always precarious and they can not be disposed to theil proper, respective position. 14). Within the embryo sac, which is lacking the endospermcell, the 4-celled proembryo, linear arrangement, is observed. 15). Through the above respects, the cause of sterile or seed failure of interspecific hybrid would be presumably as follows, By interspecific crossing gene reassortments takes place and the gene system influences the metabolism by the interference of certain enzyme as media. In the F1 plant, the quantity and quality of chemicals produced by the enzyme system and reaction system are entirely different from the case of the parents. Generally, in order to grow, form, and develop naw parts it is necessary to change the materials and energy with reasonable balance, whereas in the F1 plant the metabolic process becomes abnormal or irregular because of the breakdown of the balancing. Thus the changing of the gene-reaction system causes the alteration of the environmental condition of the gameto-genetic cells in the anther and ovule; the produced chemicals cause changes of oxidatio-reduction potential, PH value, protein denaturation and the polarity, etc. Then, the abnormal tissue growing in the ovule and emdryo sac, inhibition of normal development and storage of some chemicals, especially inhibitor, finally lead to sterility or seed failure. Inconclusion, we may presume that the first cause of sterile or seed abortion in interspecific hybrids is the gene reassortment, and the second is the irregularity of the metabolic system, storage of chemicals, especially inhibitor, the growth of abnormal tissue and the change of the polarity etc, and they finally lead to sexual defect, sterility and seed failure.

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Effects of Hwangryunhaedok-Tang and Geongangbuja-Tang on the Change of Interleukin-6 and $TNF-{\alpha}$ Level Induced by LPS I.C.V. Injection in Mice (황연해독탕(黃連解毒湯)과 건강부자탕(乾薑附子湯)이 LPS유도에 의한 마우스 혈중 IL-6와 $TNF-{\alpha}$ 변화에 미치는 영향)

  • Park, Su-Hyun;Kwon, Yong-Uk;Lee, Tae-Hee
    • Herbal Formula Science
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    • v.15 no.1
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    • pp.185-197
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    • 2007
  • Objective : This study was conducted to investigate the effects of Hwangryunhaedok-Tang and Geongangbuja-Tang on the change of interleukin-6 (IL-6) and tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) level induced by LPS I.C.V. injection in mice. Method : We devided group into 6 mice and 6 mice were assingned to each group. In the normal group only saline was administered intragastrically, and in the control group LPS was injected intracerebroventricularly 1 hr after intragastric administration of saline. In the experiment groups Hwangryunhaedok-Tang(0.5g/kg, 1.0g/kg, 3.0g/kg) was administered intragastrically to mice 1 hr prior to LPS(100mg/mouse) I.C.V. injection.. Also Geongangbuja-Tang (0.5g/kg, 1.0g/kg, 3.0g/kg) was administered intragastrically to mice 1 hr prior to LPS(100mg/mouse) I.C.V. injection. To measure the plasma IL-6 and $TNF-{\alpha}$ level of mice, their blood samples were collected from retro-orbital plexus, immediately centrifuged at $4^{\circ}C$, and plasma was removed and stored frozen at $-83^{\circ}C$ for later determination of IL-6 and $TNF-{\alpha}$. The level of IL-6 and $TNF-{\alpha}$ production was measured by enzyme-linked immunosorbent assay in the plasma. Result : Regarding IL-6 level, The 0.5g/kg and the 1g/kg groups of Geongangbuja-Tang decreased IL-6 level. Especially the 3g/kg control group decreased IL-6 level significantly than the normal group(p<0.01). Regarding $TNF-{\alpha}$ level, the 3g/kg group of Geongangbuja-Tang decreased it significantly(p<0.05). Conclusion : These data revealed that Hwangryunhaedok-Tang might not have the anti imflammatory effect and Geongangbuja-Tang(3g/kg)might have the anti imflammatory effect by reducing the plasma IL-6 and $TNF-{\alpha}$ level in mice LPS Injection.EIM (Eighteen Incompatible Medicaments) is an important component in Oriental pharmacology and is directly related to clinical prescriptions. Medical practitioners argued that the definite cause and meaning of EIM was ambiguous and therefore debated the issue of clinical application of the EIM. This study conducted an in-depth literary research on the origin, meaning and contents of EIM with the purpose to contribute in its efforts to be used clinically. Even after thousands of years have past since establishment of Oriental medicine, EIM is still tabooed and was an obstacle that hindered ideologies. Modern herbal medicine texts claim that the use of EIM can reduce treatment effects and promote poisoning and side effects. However, since long ago, there has been medical practitioners who reject this as false. Recently, poisoning caused by EIM has been claimed to be from the toxicity of the drug itself, rather than the result of interaction between the drugs, and therefore they suggest that EIM is not a forbidden domain. In addition, EIM showed a difference in number depending on the era. However, this can be understood not as a definite number, but instead as a warning to be careful during combination of drugs for use as clinical medicine. Historically, there were very few cases in which EIM was used for clinical tests and thus, the clinical value is not, while others applied EIM directly to their bodies, which showed signs for the usefulness and potential of EIM for us. A more concrete and in-depth study must be made on EIM.

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Expression Analysis of Glutathione Peroxidase Genes in the Stage-Specific Seminiferous Tubules of Mice Excised by a Laser Capture Microdissection (Laser Capture Microdissection으로 절제된 마우스의 특정 단계별 정세관에서 Glutathione Peroxidase 유전자의 발현 분석)

  • Yon, Jung-Min;Lin, Chun-Mei;Park, Jung-Hoon;Hong, Min-Ki;Jung, A-Young;Kim, Mi-Ra;Baek, In-Jeoung;Lee, Beom-Jun;Nam, Sang-Yoon;Yun, Young-Won
    • Development and Reproduction
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    • v.14 no.2
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    • pp.99-105
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    • 2010
  • The seminiferous epithelium, with its division into 12 spermatogenic stages in the mouse, is a very complex tissue. Glutathione peroxidase (GPx) is a representative antioxidant enzyme that is capable of reducing organic hydroperoxides to their corresponding hydroxyl compounds utilizing glutathione and is related to the mammalian spermatogenesis. In this study, a real-time PCR was performed in the stage-specific seminiferous tubules of mouse testes excised by a laser capture microdissection (LCM) in order to quantitate the expression levels of a series of GPx genes including cytosolic GPx (cGPx), gastrointestinal GPx (GI-GPx), plasma GPx (pGPx), and phospholipid hydroperoxide GPx (PHGPx). Frozen sections (10 ${\mu}m$) were obtained from normal adult mouse testes. LCM was used to capture all the cells that were grouped into stages I-V, VII-VIII, and IX-XI in cross-sections of seminiferous tubules. The expression level of PHGPx mRNA was remarkably higher than those of other GPx mRNAs in mouse testes. During spermatogenesis, the expressions of GI-GPx, pGPx, and PHGPx mRNAs were highest on stages VII-VIII, began to decrease after stage XI, and showed a lowest level on stage I-V. However, the expressions of cGPx mRNA were highest on stages VII-VIII, and showed a lowest level on stage XI-XI. These findings indicate that GPx genes are expressed differentially on mouse spermatogenesis and also LCM can be an useful tool in cellular quantitative analysis of testes.