• Title/Summary/Keyword: Hippocampal CA1 cells

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Biochemical Assessment of Deer Velvet Antler Extract and its Cytotoxic Effect including Acute Oral Toxicity using an ICR Mice Model (ICR 마우스 모델을 이용한 녹용 추출물의 생화학적 평가 및 급성 경구 독성을 포함한 세포 독성 효과)

  • Ramakrishna Chilakala;Hyeon Jeong Moon;Hwan Lee;Dong-Sung Lee;Sun Hee Cheong
    • Journal of Food Hygiene and Safety
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    • v.38 no.6
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    • pp.430-441
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    • 2023
  • Velvet antler is widely used as a traditional medicine, and numerous studies have demonstrated its tremendous nutritional and medicinal values including immunity-enhancing effects. This study aimed to investigate different deer velvet extracts (Sample 1: raw extract, Sample 2: dried extract, and Sample 3: freeze-dried extract) for proximate composition, uronic acid, sulfated glycosaminoglycan, sialic acid, collagen levels, and chemical components using ultra-performance liquid chromatography-quadrupole-time-of-light mass spectrometry. In addition, we evaluated the cytotoxic effect of the deer velvet extracts on BV2 microglia, HT22 hippocampal cells, HaCaT keratinocytes, and RAW264.7 macrophages using the cell viability MTT assay. Furthermore, we evaluated acute toxicity of the deer velvet extracts at different doses (0, 500, 1000, and 2000 mg/kg) administered orally to both male and female ICR mice for 14 d (five mice per group). After treatment, we evaluated general toxicity, survival rate, body weight changes, mortality, clinical signs, and necropsy findings in the experimental mice based on OECD guidelines. The results suggested that in vitro treatment with the evaluated extracts had no cytotoxic effect in HaCaT keratinocytes cells, whereas Sample-2 had a cytotoxic effect at 500 and 1000 ㎍/mL on HT22 hippocampal cells and RAW264.7 macrophages. Sample 3 was also cytotoxic at concentrations of 500 and 1000 ㎍/mL to RAW264.7 and BV2 microglial cells. However, the mice treated in vivo with the velvet extracts at doses of 500-2000 mg/kg BW showed no clinical signs, mortality, or necropsy findings, indicating that the LD50 is higher than this dosage. These findings indicate that there were no toxicological abnormalities connected with the deer velvet extract treatment in mice. However, further human and animal studies are needed before sufficient safety information is available to justify its use in humans.

Neuroprotective effects of herbal mixture HT070 on global cerebral ischemia in rats

  • Song, Jungbin;Lee, Donghun;Kim, Young-Sik;Lee, Hyun Jeong;Lee, Seunggyeong;Kim, Dong Kuk;Kang, Shin Ho;Shin, Yong Kook;Choi, Ho-Young;Kim, Hocheol
    • The Korea Journal of Herbology
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    • v.31 no.4
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    • pp.101-109
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    • 2016
  • Objectives : HT070 is a mixture of herbal extracts from root of Scutellaria baicalensis and stem bark of Eleutherococcus senticosus , which have long been used for stroke therapy in traditional Korean Medicine. The purpose of this study was to investigate the neuroprotective effects of HT070 on global cerebral ischemia and its potential mechanisms.Methods : Transient global cerebral ischemia was produced by 10 min of four-vessel occlusion (4-VO) in male Wistar rats. HT070 was administered orally at a dosage of 200 mg/kg twice at 0 and 90 min after reperfusion. Hippocampal neuronal damage was measured 7 days after reperfusion. To explore the potential mechanisms, we used hydrogen peroxide (H2O2)-induced rat pheochromocytoma (PC12) cells as an in vitro model. PC12 cells were pretreated with HT070 for 1 h and then exposed to 100 μM H2O2 for 6 h in the presence of HT070. Cell viability was measured by MTT assay and the mRNA expression of Bax, Bcl-2, iNOS and COX-2 were measured by quantitative RT-PCR.Results : Oral administration of HT070 at a dose of 200 mg/kg significantly reduced neuronal death in the hippocampal CA1 region by 13.4% as compared to the vehicle-treated group. HT070 increased cell viability, reversed the down-regulated Bcl-2 mRNA level, and suppressed the up-regulated mRNA expressions of Bax, iNOS, and COX-2 in H2O2-treated PC12 cells.Conclusions : HT070 protects against delayed neuronal death after global cerebral ischemia and its neuroprotection properties might be attributed to the inhibition of mitochondrial apoptosis and ROS-generating enzymes.

Effect of Saccharin Intake in Restraint-induced Stress Response Reduction in Rats (구속 스트레스 쥐 모델에서 스트레스 반응 감소에 대한 사카린 섭취의 효과)

  • Park, Jong Min;Song, Min Kyung;Kim, Yoon Ju;Kim, Youn Jung
    • Journal of Korean Biological Nursing Science
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    • v.18 no.1
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    • pp.36-42
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    • 2016
  • Purpose: Stress activates the sympathetic nervous system and hypothalamic-pituitary-adrenal (HPA) axis and induces the release of glucocorticoids. Saccharin is 300 times sweeter than sucrose, but does not increase blood insulin levels. Thus, this study was designed to evaluate the effect of saccharin intake in restraint-induced stress response reduction in rats. Methods: Adult male Sprague-Dawley (SD) rats had stress induced by restraint for 2 hours/day for 1 week. Saccharin was provided in sufficient amounts to allow them to intake it voluntarily at 0.1% diluted in water. The Y-maze test and forced swim test (FST) were performed to evaluate cognitive function and the depressive behavior of the rats. The protein expression of the glucocorticoid receptor (GR) in hippocampal cornu ammonis (CA) 1 was investigated by using immunohistochemistry. Results: It was found that, the percentage of alternation in the Y-maze test was significantly (p<.01) higher in the Stress + saccharin group than in the Stress group. Immobility time in the FST was significantly (p<.01) lower in the Stress + saccharin group than in the Stress group. Also, the positive cells of GR in hippocampus CA1 were significantly (p<.05) lower in the Stress + saccharin group than in the Stress group. Conclusion: This study showed that there was an effect of saccharin intake in restraint-induced stress response reduction in rats.

Neuroprotective and Anti-Neuroinflammatory Activities of Anthraquinones Isolated from Photorhabdus temperata Culture Broth

  • Yang, Eun-Ju;Kim, Seo-Hyun;Lee, Kyeong-Yeoll;Song, Kyung-Sik
    • Journal of Microbiology and Biotechnology
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    • v.28 no.1
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    • pp.12-21
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    • 2018
  • Photorhabdus temperata (PT), a gram-negative bacterium, lives symbiotically within entomopathogenic nematodes. The insecticidal compounds derived from Photorhabdus are used as biopesticides in agriculture. However, the physiological properties are not well characterized. In the course of our screening for neuroprotective and anti-neuroinflammatory substances from natural products, the culture broth of PT showed considerable activities. By activity-guided purification, five anthraquinones, namely, 3-methoxychrysazine (1), 1,3-dimethoxy-8-hydroxy-9,10-anthraquinone (2), 1,3,8-trihydroxy-9,10-anthraquinone (3), 3,8-dihydroxy-1-methoxy-9,10-anthraquinone (4), and 1,3,4-trimethoxy-8-hydroxy-9,10-anthraquinone (5), were isolated from the ethyl acetate fraction of the PT culture broth. Among the isolated compounds, $75{\mu}M$ 3 significantly protected mouse hippocampal neuronal cells (HT22) against 5 mM glutamate-induced cell death via the inhibition of reactive oxygen species production, $Ca^{2+}$ influx, and lipid peroxidation. Additionally, 3 and 4 effectively suppressed the interferon-${\gamma}$-induced neuroinflammation of mouse-derived microglial cells (BV2) at 10 ng/ml, via the reduction of nitric oxide, interleukin-6, and tumor necrosis factor-${\alpha}$. Anthraquinones 3 and 4 derived from the PT culture broth are a potential starting point to discover neuroprotective and anti-neuroinflammatory drug leads. The novel compound 5 is reported for the first time in this study.

Effects of ${\alpha}$-Asarone against Global Cerebral Ischemia in Rats

  • Kim, Young-Ock;Kwon, Yong-Soo;Lee, Seong-Eun;Lee, Sang-Won;Lee, Hee-Jae
    • Natural Product Sciences
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    • v.15 no.4
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    • pp.198-202
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    • 2009
  • Based on the use of Acorus gramineus SOLAND (AG) for the treatment of stroke in traditional Korean medicine, the present study was carried out to evaluate neuroprotective effects of ${\alpha}$-asarone after transient global cerebral ischemia using rat 4-vessel occlusion (4VO) model in rats. ${\alpha}$-Asarone (5 mg/kg) administered intraperitoneally significantly protected CA1 neurons against 10 min transient forebrain ischemia as demonstrated by measuring the density of neuronal cells stained with Cresyl violet. ${\alpha}$-Asarone significantly reduced hippocampal neuronal cell death by 85.2% where as its isolated single compounds from AG compared with a vehicle-treated group.

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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Neuroprotective Effect of Wild Radish Extract on Scopolamine Induced Memory Impairment (갯무 추출물의 스코폴라민 유도 기억력 저하 모델에서의 뇌신경 보호 효과)

  • Hur, Jinyoung;Choi, Sang Yoon;Yeom, Mijung
    • Journal of the Korean Society of Food Culture
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    • v.36 no.6
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    • pp.633-639
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    • 2021
  • Raphanus sativus var. hortensis f. raphanistroides Makino (Korean wild radish [WR]) are root vegetables belonging to the Brassicaceae family. These radish species mostly grow in sea areas in Asia, where they have been traditionally used as a medicinal food to treat various diseases. To investigate the effect of WR on neuronal cell death in SH-SY5Y cells, beta-amyloid was used to develop the cell death model. WR attenuated neuronal cell death in SH-SY5Y and regulated the mitogen-activated protein kinase (MAPK) signaling. WR extract also inhibited acetylcholinesterase inhibitor activity. Additionally, the WR treatment group ameliorated the behavior of the memory-impaired mice in a scopolamine-induced mouse model. In the behavior test, WR treated mice showed shorter escape latency and swimming distance and improved the platform-crossing number and the swimming time within the target quadrant. Furthermore, WR prevented histological loss of neurons in hippocampal CA1 regions induced by scopolamine. This study shows that WR can prevent memory impairment which may be a crucial way for the prevention and treatment of memory dysfunction and neuronal cell death.

Protective effects of Scutellariae Radix on impairments in learning and memory induced by brain ischemia in rats (뇌허혈로 인한 흰쥐에서의 기억력 및 학습효과 저해에 대한 황금의 보호효과)

  • Kim, Young-Ock;Lee, Se-Na;Kim, Myung-Gyou;Boo, Yung-Min;Kim, Sun-Yeou;Kim, Ho-Cheol;Leem, Kang-Hyun
    • The Korea Journal of Herbology
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    • v.21 no.4
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    • pp.189-195
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    • 2006
  • Objectives : It has been reported previously that the roots of Scutellaria baicalensis (known as Huang-Gum in Korean, henceforth referred to as S. baicalensis) could prevent neuronal cell death after global cerebral ischemia. In Genuine Korean medicine, S. baicalensis is known to relieve fever in upper body, and it was thus thought to be able to alleviate deteriorations in brain function. Methods : The protective effects of S. baicalensis against post-stroke memory retardation using 4-vessel occlusion model were examined in the present study. Results : S. baicalensis was shown to significantly alleviate the deficits in learning and memory by increasing the fraction of time spent in the quadrant in which the platform was initially placed ($34.9\;{\pm}\;3.2%$, p < 0.05) compared to that of the ischemia group ($28.0\;{\pm}\;2.5%$). The cytoprotective effect of S. baicalensis on CA1 hippocampal neurons was evaluated by measuring the neuronal cell density. Neuronal cell density in S. baicalensis extracts-treated ischemia group ($138.0\;{\pm}\;13.6\;cells/mm^2$) was significantly increased compared to saline-treated ischemia group ($22.1 \;{\pm}\;9.3\;cells/mm^2$, p < 0.05). In the study of OX-42 immunohistochemistry, S. baicalensis could decrease the micrgial activation in hippocampus after brain ischemia. Conclusion : These results may provide experimental support for the use of S. baicalensis in treating post-stroke memory impairment.

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Effects of Salvianolic Acid B Against Oxidative Stress in Skeletal Muscle and Brain Tissue following Exhaustive Exercise in Rats (Salvianolic acid B가 고강도 운동부하에 의한 흰쥐 골격근과 뇌조직의 Oxidative Stress에 미치는 영향)

  • Lee, Hyun-Joon;Kang, Sung-Han;Kweon, Su-Hyeon;Kim, Dae-Kyung;Kim, Jeeho;Moon, Ji-Hong;Shin, Jung-Won;Lee, Jong-Soo;Sohn, Nak-Won
    • The Korea Journal of Herbology
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    • v.31 no.5
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    • pp.99-106
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    • 2016
  • Objectives : Salvianolic acid B (SAB) is an active ingredient in Salvia miltiorrhiza frequently used for cardiovascular and cerebrovascular diseases. The present study investigated the antioxidant effects of SAB on the skeletal muscle and the brain tissue of rats following exhaustive exercise.Methods : The rats were treated with oral administration of SAB (30 mg/kg) daily for 5 days prior to the exhaustive exercise. The exhaustive exercise was performed as swimming for 150 min with 5% body weight attached to the tail on the 5th day. The antioxidant effects of SAB was evaluated by measuring the superoxide generation in the gastrocnemius and the 4-HNE expression in the hippocampal tissue. In addition, c-Fos-expressing cells in the brain tissue was observed using immunohistochemistry.Results : Histological features and muscle fiber type composition were not different between the SAB group and the exhaustive exercise group. SAB significantly reduced the upregulation of superoxide generation in the muscle tissue. SAB significantly reduced the increase of c-Fos-expressing cells in the cerebral cortex, paraventricular thalamic nucleus, dorsomedial hypothalamic nucleus, the CA1, CA3, and DG regions of hippocampus. SAB significantly reduced the upregulation of 4-HNE expression in the CA1 and DG regions of hippocampus caused by the exhaustive exercise.Conclusions : The results suggest that SAB exerts antioxidative effect against oxidative stress in the skeletal muscle and the brain tissue following exhaustive exercise, while SAB may has an anti-stress effect on stress responses in the brain.

The Morphologic Changes of Parvalbumin- Immunoreactive Interneurons of the Dentate Gyrus in Kainate-Treated Mouse Hippocampal Slice Culture Epilepsy Model (Kainic Acid로 처리한 해마박편배양 마우스 간질모델에서 치아이랑 Parvalbumin 면역 반응성 사이신경세포의 형태학적 변화)

  • Chung, Hee Sun;Shin, Mi-Young;Kim, Young-Hoon;Lee, In-Goo;Whang, Kyung-Tai;Kim, Myung-Suk
    • Clinical and Experimental Pediatrics
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    • v.45 no.12
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    • pp.1551-1558
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    • 2002
  • Purpose : Loss of hippocampal interneurons in dentate gyrus has been reported in patients with severe temporal lobe epilepsy and in animals treated with kainic acid(KA). Interneurons contain $Ca^{2+}$- binding protein parvalbumin(PV). The effects of kainic acid on parvalbumin-immunoreactive (PV-IR) interneurons in dentate gyrus were investigated in organotypic hippocampal slice cultures. Methods : Cultured hippocampal slices from postnatal day nine C57/BL6 mice were exposed to $10{\mu}M$ KA, and were observed at 0, 8, 24, 48, 72 hours after a one hour KA exposure. Neuronal injury was determined by morphologic changes of PV-IR interneuron in dentate gyrus. Results : Transient(1 hour) exposure of hippocampal explant cultures to KA produced marked varicosities in dendrites of PV-IR interneuron in dentate gyrus and the shaft of interbeaded dendrite is often much thinner than those in control. The presence of varicosities in dendrites was reversible with KA washout. The dendrites of KA treated explants were no longer beaded at 8, 24, 48 and 72 hours after KA exposure. The number of cells in PV-IR interneurons in dentate gyrus was decreased at 0, 8 hours after exposure. But there was no significant difference in 24, 48 and 72 hours recovery group compared with control group. Conclusion : The results suggested that loss of PV-IR interneurons in dentate gyrus is transient, and is not accompanied by PV-IR interneuronal cell death.