• Title/Summary/Keyword: TH-immunohistochemistry

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The comparison of histological effects of musk containing and civet containing WooHwangChungSimWon on the cerebral ischemia (Mongolian Gerbil의 뇌허혈 유발에 따른 뇌조직 손상에 있어서 사향 함유 우황청심원액과 영묘향 함유 우황청심원액의 조직학적 효능 비교)

  • 최은욱;이장헌;신상덕;마응천
    • Biomolecules & Therapeutics
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    • v.8 no.3
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    • pp.255-261
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    • 2000
  • WooHwangChungSimWon is a traditional prescription for treating with hypertension, arteriosclerosis, coma, and stroke in China, Korea, and Japan. In the new presciption of WooHwangChungSimWon, the covet is substituted for the musk, the major component of WooHwangChungSimwon, because of the prohibition law about the musk. We have made a comparative study of the effects on the ischemic damage between the musk containing and civet containing WooHwangChungSimWon. In order to investigate the effects of WooHwangChungSimWon on the ischemic damage, each samples were administrated for 12 days, ischemia was induced for 10 minutes at 7th day, and immunohistochemistry was performed in the region of hippocampus of mongolian gerbils. According to the result of immunohistochemistry, the survival rates of neuroal cells in the hippocampal CA1 region are 37.8% in the high dose of musk containing WooHwangChungSimWon (HM-WHCSW) administrated group, 27.8% in low dose of civet containing WooHwangChungSimWon (LC-WHCSW), and 35.5% in high dose of civet containing WooHwangChungSimWon (HC-WHCSW) administrated group. These survial rates were significantly different from the survival rate of sham control group (14.4%). The results suggest that all the samples except the low dose of musk containing WooHwangChungSimWon (LM-WHCSW) have protective or preventive effects on cerebral ischemia.

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Immunohistochemical studies on the relationship between pineal body and superior cervical ganglia of the Korean native goat (한국재래산양 송과체와 앞쪽목신경절의 관계규명을 위한 면역조직화학적 연구)

  • Lee, Heungshik S.;Lee, In-Se;Song, Seung-hoon;Yoon, Sung-tae;Hwang, In-koo;Lee, Choong-hyun
    • Korean Journal of Veterinary Research
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    • v.40 no.2
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    • pp.197-211
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    • 2000
  • The pineal body have been known to be affected by superior cervical ganglia, and most of its nerve fibers containing peptidergic neurotransmitters have been considered to be originated from this ganglia. To confirm this relationships, some peptidergic neurotransmitters were identified in both of pineal body and superior cervical ganglia of the Korean native goat, which were divided into two group; breeding season and non-breeding season. The localizations of two catecholamine-synthesizing enzymes; tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DBH), were investigated by immunohistochemistry in the superior cervical ganglia and the pineal body of adult Korean native goats. Substance P (SP), calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY) and galanin (GAL) were also identified in these organs by immunohistochemical and double immunofluorescent methods. In superior cervical ganglia, immunoreactivities for TH and DBH were confirmed in the same ganglion cells. The immunoreactivites for SP, VIP(only in male), NPY and GAL were identified in both of ganglion cell bodies and nerve fibers in the ganglia. CGRP immunoreactivity, however, was observed only in nerve fibers. Most NPY- and VIP-immunoreactive(IR) ganglion cells also contained TH. SP and TH were colocalized in the cell bodies, but not in the nerve fibers. TH immunoreactivity was shown in almost all of ganglion cells in the superior cervical ganglia. The immunoreactivity for NPY had some seasonal variation and was stronger in breeding season than in non-breeding season. In pineal body, lots of TH-IR fibers were observed throughout the parenchyma including the pineal stalk and most of them also contained DBH. SP- and NPY-IR fibers were also immunostained with TH or DBH. But a few SP- and NPY-IR fibers were not colocalized with TH or DBH. Exceptionally, a bipolar neuron-like cell was observed to be immunostained with NPY in the pineal body. A few CGRP and GAL-IR fibers were observed, while VIP-IR fibers were not present. It is concluded that most TH- and DBH-IR fibers as well as the peptidergic immunoreactive fibers of the pineal body might be originated from the superior cervical ganglia. Some peptidergic immunoreactive fibers, however, might be come from other regions of brain. We also suggest that NPY in pineal body plays a important role for pineal function. The seasonal variation of NPY immunoreactivity indicates that the synthesis and use of NPY may be different between in breeding and non-breeding seasons.

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Acupuncture inhibits microglial activation in the rat model of Parkinson's disease (파킨슨병 모델 흰쥐에서 침치료에 의한 microglia 활성화 억제에 관한 연구)

  • Hwang, Jeong-Yeon;Choi, Il-Hwan;Park, Jae-Hyun;Kang, Jun-Mo;Park, Hi-Joon;Lim, Sa-Bi-Na
    • Korean Journal of Acupuncture
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    • v.24 no.1
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    • pp.131-144
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    • 2007
  • Objectives : Although the cause of neuronal death of Parkinson's disease remains unclear, increasing evidence points to the role of inflammatory processes. And the hallmark of brain inflammation is the activation of microglia. This study was performed to prove the effect of acupuncture on inhibiting microglial activation. Methods : The rat models which were injected with 6-hydroxydopamine were treated with acupuncture once a day on LR3 (太衝) and GB34 (陽陵泉). To prove the effect of inhibiting microglial activation, we examined the tyrosine hydroxylase (TH) immunopositive neurons and CD11b immunohistochemistry in the substantia nigra. Results : There were 18% (third day), 32% (seventh day) loss of TH-positive cell bodies in the control group and 23% (third day), 26% (seventh day) in the acupuncture group, whereas 3% (third day), 10% (seventh day) in vehicle group. The difference of optical density in substantia nigra was evaluated by subtracting log inverse gray value of contralateral side from that of ipsilateral side. With regards to the result of CD11b immunohistochemistry, acupuncture group showed significantly inhibited microglial activation compared with control group (p<0.01) on the seventh day. Conclusions : Acupuncture showed the effect of inhibition of microglial activation in seventh day. However, the effect of protection of TH positive cell bodies was not shown. So we need longer investigation of the effect of acupuncture on Parkinson's disease.

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Dopaminergic neurons of the substantia nigra and ventral tegmentum in the stripped field mouse(apodemus agrarius coreae) (야생등줄쥐 흑색질 및 배쪽피개의 dopamine성 신경세포)

  • Jeong, Young-gil;Kim, Kil-soo;Lee, Chul-ho;Yoon, Won-kee;Hyun, Byung-hwa;Oh, Yang-seok;Won, Moo-ho;Kim, Moo-kang
    • Korean Journal of Veterinary Research
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    • v.37 no.3
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    • pp.489-497
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    • 1997
  • The distributions characteristics of neurons displaying immunoreactivity to the catecholamine synthetic enzymes, tyrosine hydroxylase(TH), dopamine-${\beta}$-hydroxylase(DBH), and phenylethanolamine-N-methyltransferase(PNMT) were examined in the adjacent sections of the substantia nigra & ventral tegmentum of the Striped Field Mouse(Apodemus agrarius coreae). None of these cell groups displayed either DBH or PNMT immunoreactivity. Many TH-immunoreactive neurons were present in the substantia nigra & ventral tegmentum. The major dopaminergic cell(TH-positive, DBH- & PNMT-negative) group in the midbrain was present in the pars compacta of substantia nigra and adjacent ventral tegmentum. And smaller dopaminergic cell groups Were found in the pars reticulata of the substantia nigra and central liner nucleus.

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Tyrosine hydroxylase immunoreactive neurons of the olfactory bulb in the stripped field mouse(apodemus agrarius coreae) (야생등줄쥐 후각망울의 Tyrosine hydroxylase 면역반응신경세포)

  • Jeong, Young-gil;Lee, Nam-seob;Kim, Kil-soo;Lee, Chul-ho;Hyun, Byung-hwa;Won, Moo-ho;Kim, Moo-kang
    • Korean Journal of Veterinary Research
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    • v.37 no.3
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    • pp.499-508
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    • 1997
  • The distributions and morphological characteristics of neurons displaying immunoreactivity to the catecholamine synthetic enzymes, tyrosine hydroxylase(TH), dopamine-${\beta}$-hydroxylase(DBH), and phenylethanolamine-N-methyltransferase (PNMT) were examined in the adjacent sections of the olfactory bulb of the Striped Field Mouse(Apodemus agrarius coreae). None of these cell groups displayed either DBH or PNMT immunoreactivity. Many TH-immunoreactive neurons were present in the olfactory bulb. The vast majority of such cells occurred in the glomerular layer as periglomerular cells surrounding the glomeruli. Numerous addtional cells were present in the external plexiform layer, and scattered in the mitral cell layer and internal plexiform layer. Also TH-immunoreactive neurons were found in the glomerular layer and granular layer of the accessory olfactory bulb.

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Dopaminergic neuronal development in the embryonic mesencephalon of mouse

  • Kim, Mun-Ki;Lee, Si-Joon;Won, Chung-Kil
    • Korean Journal of Veterinary Research
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    • v.60 no.4
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    • pp.203-207
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    • 2020
  • This study presents neuronal migration pattern of dopamine (DA) neurons generated in separate regions occupying the ventral mesencephalic territory. A single pulse 5-bromodeoxyuridine (BrdU) was administered at embryonic day (E)10-E15. Distribution of tyrosine hydroxylase (TH) positive cells was determined at E13-postnatal day 0 (P0) by immunohistochemistry. BrdU positive cells labeled at E10 were spread out uniformly in the mesencephalon from E13 to E15, migrating through dorsal and ventral routes at E17 and P0. TH expression labeled at E10 was observed at E13 in the ventromedial region and clearly formed in the ventral tegmental area (VTA) at E15. At E17, TH expression in the substantia nigra (SN) was observed in the ventrolateral region, spreading more outward of the mesencephalon at P0. Generation of TH-positive cells labeled at E13 was also observed in VTA and SN of the mesencephalon at E17 and P0. The expression of these cells labeled after E15 was markedly decreased. These results demonstrated that an almost complete primary structure of DA neuron was formed at the early embryonic stage in the ventral mesencephalon, showing the most active neuronal migration was occurred at E13-E17.

Post-cancer Treatment with Condurango 30C Shows Amelioration of Benzo[a]pyrene-induced Lung Cancer in Rats Through the Molecular Pathway of Caspase-3-mediated Apoptosis Induction -Anti-lung cancer potential of Condurango 30C in rats-

  • Sikdar, Sourav;Mukherjee, Avinaba;Bishayee, Kausik;Paul, Avijit;Saha, Santu Kumar;Ghosh, Samrat;Khuda-Bukhsh, Anisur Rahman
    • Journal of Pharmacopuncture
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    • v.16 no.3
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    • pp.11-22
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    • 2013
  • Objectives: The present investigation aimed at examining if post-cancer treatment with a potentized homeopathic drug, Condurango 30C, which is generally used to treat oesophageal cancer, could also show an ameliorating effect through apoptosis induction on lung cancer induced by benzo[a]pyrene (BaP) in white rats (Rattus norvegicus). Methods: Lung cancer was induced after four months by chronic feeding of BaP to rats through gavage at a dose of 50 mg/kg body weight for one month. After four months, the lung-cancer-bearing rats were treated with Condurango 30C for the next one ($5^{th}$), two ($5^{th}-6^{th}$) and three ($5^{th}-7^{th}$) months, respectively, and were sacrificed at the corresponding time-points. The ameliorating effect, if any, after Condurango 30C treatment for the various periods was evaluated by using protocols such as histology, scanning electron microscopy (SEM), annexinV-FITC/PI assay, flow cytometry of the apoptosis marker, DNA fragmentation, reverse transcriptase-polymerase chain reaction (RT-PCR), immunohistochemistry, and western blot analyses of lung tissue samples. Results: Striking recovery of lung tissue to a near normal status was noticed after post-cancerous drug treatment, as evidenced by SEM and histology, especially after one and two months of drug treatment. Data from the annexinV-FITC/PI and DNA fragmentation assays revealed that Condurango 30C could induce apoptosis in cancer cells after post-cancer treatment. A critical analysis of signalling cascade, evidenced through a RT-PCR study, demonstrated up-regulation and down-regulation of different pro- and anti-apoptotic genes, respectively, related to a caspase-3-mediated apoptotic pathway, which was especially discernible after one-month and two-month drug treatments. Correspondingly, Western blot and immunohistochemistry studies confirmed the ameliorative potential of Condurango 30C by its ability to down-regulate the elevated epidermal growth factor receptor (EGFR) expression, a hallmark of lung cancer. Conclusion: The overall result validated a positive effect of Condurango 30C in ameliorating lung cancer through caspase-3-mediated apoptosis induction and EGFR down-regulation.

The Characteristics of Supramammillary Cells Projecting to the Hippocampus in Stress Response in the Rat

  • Choi, Woong-Ki;Wirtshafter, David;Park, Hyun-Jung;Lee, Mi-Sook;Her, Song;Shim, In-Sop
    • The Korean Journal of Physiology and Pharmacology
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    • v.16 no.1
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    • pp.17-24
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    • 2012
  • The hypothalamus-pituitary-adrenocortex (HPA) axis is the central mediator of the stress response. The supramammillary (SuM) region is relatively unique among the hypothalamic structures in that it sends a large, direct projection to the hippocampal formation. It has been shown that mild stress could activate the SuM cells that project to the hippocampus. However, the role of these cell populations in modulating the stress response is not known. The present study examined the effect of stress on different populations of SuM cells that project to the hippocampus by injecting the fluorescent retrograde tracer, fluorogold (FG), into the hippocampus and utilizing the immunohistochemistry of choline acetyltransferase (ChAT), corticotrophin releasing factor (CRF), serotonin (5-HT), glutamate decarboxylase (GAD), tyrosine hydroxylase (TH) and NADPH-d reactivity. Immobilization (IMO) stress (2 hr) produced an increase in the expression of ChAT- immunoreactivity, and tended to increase in CRF, 5-HT, GAD, TH-immunoreactivity and nitric oxide (NO)-reactivity in the SuM cells. Fifty-three percent of 5-HT, 31% of ChAT and 56% of CRF cells were double stained with retrograde cells from the hippocampus. By contrast, a few retrogradely labeled cells projecting to the hippocampus were immunoreactive for dopamine, ${\gamma}$-aminobutyric acid (GABA) and NO. These results suggest that the SuM region contains distinct cell populations that differentially respond to stress. In addition, the findings suggest that serotonergic, cholinergic and corticotropin releasing cells projecting to the hippocampus within the SuM nucleus may play an important role in modulating stress-related behaviors.

Effect of Kamicheonggan-tang on Pre-hepatocarcinogenesis Induced by N-nitrosomorpholine (가미청간탕이 N-nitrosomorpholine으로 유도된 전암성 간병변에 미치는 영향)

  • Kim Dong Hee;Choi Jeung Mok;Jo Dong il
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.4
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    • pp.734-744
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    • 2002
  • The purpose of this study was to evaluate the protective effect of Kamicheonggan-tang(KCGT) on pre-hepatocarcinogenesis induced by N-nitrosomorpholine. The studied using blood chemistry, lipidperoxidation, antioxidant, immunohistochemistry and morphological change. The results were obtained as follows. In the pre-hepatocarcinogenesis induced by NMP, serum AST, ALT, ALP and total bilirubin were not changed in NMP and NK treated group after 1 st week, but desreased in NK treated group after 4th week as compared with NMP treated 4th week group. The content of GSH was similary to in NK treated groups as compares with data of normal group. The content of MDA was increased in NK treated group after 1st and 4th week, and more increased in the NMP treated group than those of their NK treated group. The immunohistochemically, stain of GST-p, positive lesions of KCGT were significantly decreased than those of NMP treated group. The histopathologically, fat changes, nucleotic changes, oval cell and inflammatory cells in periportal were observed in NMP treated fater 4th week, but those were significantly decreased from 4th week in the NK treated group. And the enlarged nucleus was not changed in KCGT treated group, but increased in NMP treated group after 1st and 4th week. The ultrastructurally, nucleotic changes, glycogen degeneration, lipid droplet and rER fragmentation were observed in NMP treated group after 4th week, but those changes were significantly decreased from 4th week in the NK treated group. These results suggested that KCGT extracts has protective effect on prehepatocarcinogenesis by NMP, might be usefully applied for clinical treatment of hapatic disease and also it was necessary to do more studies about its mechanisms.

Induction of Apoptosis in Glioma Cells and Upregulation of Fas Expression Using the Human Interferon-β Gene

  • Guo, Yan;Wang, Gan;Gao, Wen-Wei;Cheng, Shi-Wen;Wang, Ren;Ju, Shi-Ming;Cao, He-Li;Tian, Heng-Li
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.6
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    • pp.2837-2840
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    • 2012
  • We investigated whether IFN-${\beta}$ inhibits the growth of human malignant glioma and induces glioma cell apoptosis using the human IFN-${\beta}$ gene transfected into glioma cells. A eukaryonic expression vector ($pSV2IFN{\beta}$) for IFN-${\beta}$ was transfected into the glioma cell line SHG44 using liposome transfection. Stable transfection and IFN-${\beta}$ expression were confirmed using an enzyme-linked immunosorbent assay (ELISA). Cell apoptosis was also assessed by Hoechst staining and electron microscopy. In vivo experiments were used to establish a SHG44 glioma model in nude mice. Liposomes containing the human IFN-${\beta}$ gene were injected into the SHG44 glioma of nude mice to observe glioma growth and calculate tumor size. Fas expression was evaluated using immunohistochemistry. The IFN-${\beta}$ gene was successfully transfected and expressed in the SHG44 glioma cells in vitro. A significant difference in the number of apoptotic cells was observed between transfected and non-transfected cells. Glioma growth in nude mice was inhibited in vivo, with significant induction of apoptosis. Fas expression was also elevated. The IFN-${\beta}$ gene induces apoptosis in glioma cells, possibly through upregulation of Fas. The IFN-${\beta}$ gene modulation in the Fas pathway and apoptosis in glioma cells may be important for the treatment of gliomas.