• Title/Summary/Keyword: BrdU immunohistochemistry

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Evaluation of Skin Sensitization Potential of Skin Whitening Agents by Local Lymph Node Assay (국소 림프절 시험법(Local lymph node assay)을 이용한 피부미백화장품 원료의 피부감작성 연구)

  • 이종권;박승희;박재현;김형수;엄준호;남기택;한범석;장동덕;최광식
    • Toxicological Research
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    • v.19 no.4
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    • pp.285-291
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    • 2003
  • The use of skin whitening agents has been recently increased in various kinds of cosmetic products, although there were some reports that whitening agents might cause allergic contact dermatitis. A murine local lymph node assay (LLNA) has been developed as an alternative to guinea pigs for contact sensitization potential. This study was carried out to investigate the skin sensitization potential of three whitening agents, arbutin, azelaic acid, and kojic acid, by LLNA using a non-radiois-topic endpoint. Female Balb/c mice were exposed topically to a weak allergen, $\alpha$-hexylcinnamalde-hyde (HCA), and three whitening agents following LLNA protocol. Lymph node (LN) weight and cell proliferation in ears and auricular lymph node using bromodeoxyuridine (BrdU) immunohistochemistry were evaluated. LN weights were significantly increased at the HCA group compared to the vehicle control. A weak allergen, HCA elicited 3-fold or greater increase in cell proliferation of lymph nodes as well as increase in cell proliferation of ear as measured by BrdU immunohistochemistry. However, in the case of skin whitening agent groups, there were no significant changes in LN weight and cell proliferation in the ear and lymph node of mice treated with 5, 10 and 20% of three whitening agents compared to the vehicle control. These results show that these three skin whitening agents may not have contact sensitization potentials at tested concentrations in Balb/c mice by LLNA.

EVALUATION OF CELL PROLIFERATION IN EAR AND LYMPH NODE USING BRDU IMMUNOHISTOCHEMISTRY FOR MOUSE EAR SWELLING TEST

  • Lee, Jong-Kwon;Park, Jae-Hyun;Kim, Hyung-Soo;Jung, Seung-Tae
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2002.05a
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    • pp.97-97
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    • 2002
  • A mouse ear swelling test (MEST) has been developed as an alternative to guinea pig models for contact sensitization potential. However, the MEST relies on a quantitative measurement of ear swelling as an endpoint by micrometer. In this study, we aimed to investigate the cell proliferation in ear and lymph node using Bromodeoxyuridine (BrdU) immunohistochemistry as a possible reliable marker for MEST.(omitted)

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Effects of Compression Stimulation Application on Cell Proliferation in the Hippocampal Dentate Gyrus of the Sprague-Dawley Rats (흰쥐 해마의 치상회에서 압박자극 적용이 뇌 신경세포 증식에 미치는 영향)

  • Yu, Byong-Kyu;Kim, Kyeong-Mi;Kim, Chang-Ju
    • The Journal of Korean Academy of Sensory Integration
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    • v.2 no.1
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    • pp.21-32
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    • 2004
  • Objective : Effect of treadmill exercise on hippocampal neural cell proliferation under normal conditions and alcohol intoxication conditions has been recently studied; however, this effect under sensory stimulation application has not clarified yet. In the present study, the effect of compression stimulation application on hippocampal neural cell proliferation in the dentate gyrus in normal and alcohol intoxicated rats was investigated. Methods : Experimental design: comparative investigation on number of 5-Bromo-2'-deoxyuridine(BrdU)B-positive cells in dentate gyrus 5 days after commencement. Setting: animal laboratory. Participants: male Sprague-Dawley rats of 3weeks old in age weighing $80{\pm}10gm$. Intervention: animals were randomly assigned into 4 groups; control-rest group(n=8), control-compression group(n=8), alcohol intoxication-rest group(n=8) and alcohol intoxication-compression group(n=8). Animals of the alcohol intoxicated groups were injected intraperitoneally with alcohol(2g/kg) twice per day for 3 days. All animals were injected BrdU(50mg/kg) intraperitoneally, and rats compression stimulation application groups were compressed using sphygmomanometer cuff times per day, for 5 days following alcohol administration. Measures: mean number of BrdU-positive cells in dentate gyrus was observed via immunohistochemistry. Results : Compression stimulation application significantly increased the number of BrdU-positive cells in the dentate gyrus. Also, treatment with alcohol for 3 days inhibited cell proliferation, and compression stimulation application alleviated alcohol-induced inhibition of new cell formation. Conclusion : These results suggest the possibility that compression stimulation application may help in improvement following alcohol-induced brain damaged.

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Shenqi-wan Increases Cell Proliferation of Cultured Hippocampal Cell Line HiB5 and Dentate Gyrus of Young Sprague-Dawley Rats (신기환 약침이 HiB5 해마세포와 어린 Sprague-Dawley계 흰쥐의 치상회 세포생성에 미치는 영향)

  • Kim Youn-Jung;Jang Mi-Hyun;Shin Min-Chul;Lim Baek-Vin;Chung Joo-Ho;Bahn Gyun-Ho;Paik Eun-Kyung;Park Jae-Hyung;Kim Ee-Hwa;Kim Chang-Ju
    • Korean Journal of Acupuncture
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    • v.19 no.1
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    • pp.77-84
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    • 2002
  • 목적 : 소아의 정신적 및 신체적 발달지연에 사용되고 있는 신기환이 해마의 신경세포 생성에 미치는 영향을 관찰하고자 하였다. 방법 : 신기환이 세포생성에 미치는 영향을 알아보기 위하여 해마 세포인 HiB5에 신기환을 처리, 배양하여 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay를 통해서 세포생성을 관찰하였다. 또한 3주령의 Sprague-Dawley (S-D)계 흰쥐의 중완혈에 신기환 약침을 자침후 해마의 치상회에서 bromodeoxyuridine (BrdU) immunohistochemistry를 시행하여 세포생성을 관찰하였다. 결과 : HiB5 배양세포에서는 신기환을 처리시 배양시간과 농도별로 세포생성율이 증가하였다. 또한, 대조군의 치상회에서 BrdU 양성세포수는 $128.50{\pm}9.53$, 1 mg/kg 신기환 자침군은 $157.08{\pm}10.82$ 및 10 mg/kg 신기환 자침군은 $204.80{\pm}17.68$로 나타났다. 이러한 결과로 신기환은 어린 S-D계 흰쥐의 치상회에서 세포생성을 증가시키는 것으로 사려된다.

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Effects of Acanthopanax senticosus herb-acupuncture on cell proliferation and nitric oxide synthase expression in dentate gyrus of alcohol-intoxicated rats (오가피(五加皮) 약침(藥針)이 알코올 중독(中毒) 흰쥐의 치상회(齒狀回)에서 신경세포생성(神經細胞生成) 및 NOS 발현(發顯)에 미치는 영향(影響))

  • Kim Jong-Chul;Lee Eun-Yong;Kim Ho-Hyun;Kim Ee-Hwa
    • Korean Journal of Acupuncture
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    • v.20 no.2
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    • pp.67-76
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    • 2003
  • 목적 : 오가피 약침이 알코올에 의해서 중독된 Sprague-Dawley(S-D)계 흰쥐의 해마 치상회에서 새로운 신경세포 생성 및 NOS발현에 미치는 영향을 조사하였다. 방법 : S-D계 흰쥐에 알코올을(2g/kg) 3일간 연속으로 투여한후, 5일간 인체의 중완혈에 상응하는 부위에 오가피 약침(30mg/kg) 치료를 시행하였다. 치료효과를 관찰하기 위해서 BrdU-면역조직화학 염색법 및 NADPH-d-조직화학염색법을 이용하였다. 결과 : 알코올 처치군에서는 BrdU-양성세포수 및 NADPH-양성세포수가 모두 정상군에 비해서 감소한 반면에 알코올 처치후 오가피 약침으로 치료한 군에서는 BrdU-양성세포수 및 NADPH-양성세포수 모두 알코올 처치군에 비해서 증가하였다. 결론 : 오가피 약침치료가 알코올에 의해서 중독된 S-D계 흰쥐 해마 치상회에서 새로운 신경세포의 생성을 증가시키는 것을 확인하였으며, 그 기전으로 산화질소가 관여할 것으로 사려된다.

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Effect of Postnatal Angelicae Gigantis Radix Herb-acupuncture on Cell Proliferation in Offspring Rats with Prenatal Noise Stress during Pregnancy (당귀약침(當歸藥鍼)이 소음 Stress를 받은 새끼 쥐의 신경세포 생성에 미치는 영향)

  • Chang, So-Young;Kim, Ee-Hwa;Lee, Eun-Yong
    • Journal of Acupuncture Research
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    • v.23 no.3
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    • pp.47-56
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    • 2006
  • Methods : 40 of Female rats were allowed to mate with 40 of male rats. Then, female rats were delivered of offspring rats, After birth 28 days, offspring rats were divided 8 groups, The normal group(Group A), the 10 mg/kg Angelicae gig antis radix~treated group(Group B), the 50 mg/kg Angelicae gigantis radix-treated group(Group C), the 100 mg/kg Angelicae gig antis radix-treated group(Group D), The control (noise-treated) group(Group E). the noise -10 mg/kg Angelicae gigantis radix-treated group(Group F), the noise-50 mg/kg Angelicae gigantis radix~treated group(Group G), and the noise-100 mg/kg Angelicae gigantis radix-treated group(Group H)(n = 5 in each group), From the 15th day of pregnancy, all rats were subcutaneously injected with 50 mg/kg BrdU once a day 30 min before the starting of experimental treatment. Rats of the prenatal noise-treated group were applied with 95 decibel supersonic machine sound for 1h once a day until delivery, After birth 28 days, offspring rats intraperitoneally injected with 50 mg/kg of BrdU and offspring rats were treated Angelicae gigantis radix Herb-acupunture on chungwan(CV12) for 7 consecutive days. For the detection of BrdU-positive cells and Ki-67 positive cells in hippocampus, immunohistochemistry was performed. Results : 1. The number of BrdU-positive cells in the dentate gyrus of noise-treated group was significantly decreased to normal group, and the Group F, G, H were significantly increased to control group. 2. The number of Ki-67 positive cells in the dentate gyrus of noise-treated group was significantly decreased to control group, and the Group G, H were significantly increased to control group. Conclusion : We concluded that postnatal Angelicae gigantis radix administration has effect on cell proliferation in offspring rats with prenatal noise stress during pregnancy.

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The Effects of Swim Training on Neurogenesis in the Hippocampal Dentate Gyrus and Functional Ability After Focal Ischemic Stroke in Rats (수영 훈련이 뇌허혈 유발 흰쥐의 해마 치아이랑에서 뇌신경생성과 기능적 능력에 미치는 영향)

  • Kim, Ho-Sung;Kim, Deuk-Ho;Lee, Jeong-Pil;Kim, Young-Joo;Shin, Young-Oh;Kim, Sang-Hoon;Kwon, Ki-Wook;Oh, Jae-Keun
    • Physical Therapy Korea
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    • v.12 no.3
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    • pp.11-21
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    • 2005
  • The present study was aimed at investigating the effect of swimming training on brain function after focal cerebral ischemia in rats. Therefore, this study was examined on neurogenesis in dentate gyrus of hippocampus using 5-bromo-2'-deoxyuridine (BrdU) to label proliferating cells and assessed the neurological response following focal cerebral ischemia in rats using neurological motor behavioral test. In an observer-blinded fashion, twenty male Sprague-Dawley (280~310 g, 7 weeks old) rats were divided into four groups: MCAO plus swimming group (ME, $n_1$=5), MCAO plus control group (MC, $n_2$=5), SHAM plus swimming group (SE, $n_3$=5), SHAM plus control group (SC, $n_4$=5). The results of this study were as follows: 1) The limb placing time before and after swimming in the ME group were significantly longer than the MC group (p<.05), the SE group were significantly longer than the SC group (p<.01). 2) The balance beam scores before and after swimming in the ME group was higher than the SE group, the MC group was higher than the SC group but was not significantly different (p>.001). 3) The foot fault index before and after swimming training in ME group was significantly lower (i.e., improved) than the MC group (p<.001) and the SE group (p<.001), the SE group was significantly lower (i.e., improved) than the SC group (p<.001). 4) The mean number of BrdU-positive cells in the dentate gyrus in the ME group was significantly higher than the MC group (p<.001) and the SE group (p<.01). The MC group and the SE group was significantly higher than the SC group (p<.001). 5) There was significantly correlation between limb placing time and number of BrdU-positive cells on swimming training, there was positive correlation (r=.807, p<.0001) and between foot fault index and BrdU-positive cells number, there was negative correlation (r=-.503, p<.05). However, between balance beam scores and BrdU-positive cells number, there was no correlation. In conclusion, the present study demonstrates that the role of swimming training improves behavioral motor function probably by enhancing cell proliferation in that hippocampus. This study provides a model for investigating the stroke rehabilitation that underlies neurogenesis and functional ability.

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Effects of Puerariae flos herb-acupuncture on cell proliferation and neurogenesis in the dentate gyrus of ethanol-induced Sprague-Dawley rats (갈화 약침이 알콜 중독 흰쥐의 치상회에서 신경세포 생성에 미치는 영향)

  • Kim Youn-Hee;Kim Ee-Hwa;Jang Mi-Hyun;Lim Back-Vin;Kim Youn-Jung;Chung Joo-Ho;Seo Jung-Chul;Kim Chang-Ju
    • Journal of Acupuncture Research
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    • v.18 no.6
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    • pp.206-214
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    • 2001
  • The purpose of this study was to determine the effects of Puerariae flos herb-acupuncture on hippocampal neural cell proliferation. Sprague-Dawley rats were randomly assigned into 4 groups; control group, control with herb-acupuncture group, alcohol group, alcohol with herb-acupuncture group group. Control groups were received with NaCl, while alcohol intoxication groups were injected intraperitoneally with alcohol (2 g/㎏) twice per day for 3 days. Herb-acupuncture groups were injected on Zhongwan (CV 12) for 5 consecutive days. Bromo-deoxyuridine (BrdU) was injected into all animal per day for 5 days. For the detection of BrdU-positive cells in dentate gyrus of hippocampus, immunohistochemistry was performed. In alcohol group, a significant decrease in BrdU-positive cells was observed compared to control group. In alcohol with herb-acupuncture group, BruU-positive cells increased significantly compared to alcohol group. In conclusion, the present results revealed that new cell proliferation is enhanced in the dentate gyros of young Sprague-Dawley rats through Puerariae flos herb-acpuncuture in an acute alcoholic intoxication condition.

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Effect of the Water Extract of Cultivated Wild Ginseng on the Toxicities Induced by Cyclophosphamide in Mice (산양삼이 Cyclophosphamide의 독성에 미치는 영향)

  • Lee, Sung-Woo;Kim, Ki-Tae;Ko, Heung
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.25 no.5
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    • pp.849-856
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    • 2011
  • This study was investigated the protective effect of Cultivated Wild Ginseng(WG) on the toxicities induced by cyclophosphamide(CP) in mice. Twenty-four male BALB/c mice were divided into non-treated normal group(n=6), CP treated control group(n=6), CP+WG treated CP+WG group(n=6), WG treated WG group(n=6). CP(100 mg/kg of b.w., i.p) was injected at 0, 7 th, 14 th, 21 th, and 28 th day of the experiment respectively. WG(4.4 g/kg, i.p.) was administrated for 35days. Body and organ(heart, liver, kidney, testis) weight were measured. Histopathological examination on the organ(heart, liver, kidney, testis), morphometric analysis, and BrdU immunohistochemistry on the testis were performed. Body weight was decreased following CP administration. In contrast, such a decrease was significantly attenuated by WG administration. CPK and AST of CP+WG group were significantly decreased compared with CP group. Histopathologically, cross sectional area of testis and diameter of seminiferous tubule were significantly increased in CP+WG group compared with CP group. BrdU labelled cells in the seminiferous tubules were remarkably decreased in CP group. Whereas the number of seminiferous tubules labelled with BrdU in spermatogonia was increased by CP+WG administration. The obtained results suggest that WG has protective effect on CP-induced toxicity. This effect might be mediated through the supplementation of vital energy.