• Biomolecules & Therapeutics
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• 제21권3호
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• pp.229-233
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• 2013

This study was aimed at investigating the possible effects of phytoceramide (Pcer) on learning and memory and their underlying mechanisms. Phytoceramide was orally administered to ICR mice for 7 days. Memory performances were assessed using the passive avoidance test and Y-maze task. The expressions of phosphorylated cAMP response element binding protein (pCREB), brain-derived neurotrophic factor (BDNF) were measured with immunoblot. The incorporation of 5-bromo-2-deoxyuridine (BrdU) in hippocampal regions was investigated by using immunohistochemical methods. Treatment of Pcer enhanced cognitive performances in the passive avoidance test and Y-maze task. Immunoblotting studies revealed that the phosphorylated CREB and BDNF were significantly increased on hippocampus in the Pcer-treated mice. Immunohistochemical studies showed that the number of immunopositive cells to BrdU was significantly increased in the hippocampal dentate gyrus regions after Pcer-treatment for 7 days. These results suggest that Pcer contribute to enhancing memory and BDNF expression and it could be secondary to the elevation of neurogenesis.

• 한방재활의학과학회지
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• 제29권2호
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• pp.91-99
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• 2019

Objectives The loss of maternal care during early postnatal period may increase development of mood-related disorders, such as depression, anxiety, and personality disorders. In this study, the effect of acupuncture on depression in relation with cell proliferation in the hippocampal gyrus dentatus was investigated using maternal-separated rat pups. Methods On the postnatal 14th day, rat pups from six dams were grouped into following groups: maternal care group, maternal separation group, maternal separation and non-acupoint-acupunctured group, maternal separation and Zusanli-acupunctured group, and maternal separation and fluoxetine-treated group. Acupuncture was performed from postnatal 28th day to postnatal 37th day. The rat pups that belong in the maternal separation and fluoxetine-treated group were injected subcutaneously with 5 mg/kg fluoxetine hydrochloride once a day for the same period of time. To evaluate activity of the rat pups, open field test was performed. Immunohistochemistry for serotonin (5-hydroxytryptamine, 5-HT) and tryptophan hydroxylase (TPH) in the dorsal raphe and for 5-bromo-2'-deoxyuridine (BrdU) in the hippocampal gyrus dentatus was conducted. Results The present results reveal that the activity was decreased by maternal separation. In contrast, acupuncture at Zusanli overcame maternal separation-induced hypoactivity. Maternal separation suppressed TPH expression and 5-HT synthesis in the dorsal raphe and decreased cell proliferation in the hippocampal gyrus dentatus of rat pups. In contrast, acupuncture at Zusanli alleviated maternal separation-induced decrease of 5-HT synthesisand TPH expression. Conclusions The present results demonstrate that acupuncture at Zusanli ameliorated depressive state through increasing cell proliferation and enhancing 5-HT synthesis.

• 동의신경정신과학회지
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• 제21권2호
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• pp.29-44
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• 2010

Objectives : The purpose of this study was to investigate the effect of Gentianae Radix on neurogenesis and apoptosis in ethanol- induced newborn rats hippocampus dentate gyrus. Methods : In vivo, laboratory animals were divided into three groups; Normal group(N), Control group(C) and Treated group (TG)(n=7 for each group). N were treated saline daily for five days. C were treated 1.5 g/kg ethanol and saline daily for five days. TG were treated 1.5 g/kg ethanol and 300 mg/kg Gentianae Radix daily for five days. BrdU(5-bromo-2-deoxyuridine) assay was used to test neurogenesis in the dentate gyrus. And TUNEL(Terminal deoxynucleotidyl transferase dUTP nick end labeling) assay was used to test apoptosis in the dentate gyrus. Three groups were measured body weight, serum ethanol concentration, BrdU-positive cells and TUNEL-positive cells in the dentate gyrus. In vitro, MTT(3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was used to test viability in SK-N-MC cells. BrdU assay was used to test neurogenesis in SK-N-MC cells. DNA fragmentation and caspase-3 enzyme activity assay were used to test apoptosis in SK-N-MC cells. And treated ethanol and Gentianae Radix of all in vitro tests were made various concentration. Results : In vivo, Gentianae Radix modulated ethanol-induced neurogenesis and apoptosis in newborn rats hippocampus dentate gyrus. In vitro, TG 100 ${\mu}g/ml$ have significantly modulated ethanol-induced neurogenesis and apoptosis in SK-N-MC cells. And only TG 100 ${\mu}g/ml$ have significantly protected SK-N-MC cells from ethanol-induced cytotoxicity. Conclusions : Gentianae Radix may have the effect that modulated ethanol-induced neurogenesis and apoptosis in SK-N-MC cells.

• 대한한의학회지
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• 제39권4호
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• pp.86-113
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• 2018

Objectives: We examined the effects of a mixed formula consisting of dried pomegranate concentrate powder (PCP) and the aqueous extracts of Eucommiae cortex (EC) and Achyranthis radix (AR) in rats with surgically induced osteoarthritis (OA). Methods: Two weeks after OA-inducing surgery, a PCP:EC:AR 5:4:1 (g/g) combination or single formula was orally administered. Changes in body weight, knee thickness, maximum knee extension angle, bone mineral density of the knee joints, femoral and tibial articular surfaces, and compressive strength of the femoral and tibial articular cartilage (AC) were assessed, along with the prostaglandin E2 level, 5-lipoxygenase, matrix metalloproteinase (MMP)-2 and MMP-9 activity, and chondrogenic gene mRNA expression in the femoral and tibial AC with the synovial membrane (SM). In addition, the number of cleaved poly(ADP-ribose) polymerase, cyclooxygenase and tumor necrosis factor-${\alpha}$-immunoreactive cells in the femoral and tibial AC with SM were monitored, and the rate of cell proliferation was determined with a 5-bromo-2'-deoxyuridine uptake assay. Results : The signs of surgically induced OA in rats were significantly inhibited by both PCP, EC and AR combined and single formulas. In particular, the combination formula-treated OA model rats showed dose-dependent, significantly increased inhibitory activity against all tested criteria compared with single formula-treated rats. Conclusions: Taken together, our results suggest that the combination formula synergistically increased the anti-OA effects of its components through anti-inflammatory and chondrogenic activity in rats with surgically induced OA. In addition, 200, 100 and 50 mg/kg combination formula treatments showed dose-dependent inhibitory activity against all of the tested criteria.

• Parasites, Hosts and Diseases
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• 제31권2호
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• pp.83-90
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• 1993

간흡충에 감염된 담관은 확장되면서 상피세포의 수가 크게 증식하여 점막층이 두꺼워지고 과식증에 의해 점막의 단면이 유두상으로 관찰된다. Bromodeoxyuridine (BrdU)이 세포가 핵산을 복제할 때에 끼어 들어가는 성질을 이용하여, 이 연구에서는 담관 상피세포 중에서 합성기(Sphase)에 있는 세포의 분율을 파악하고자 하였다. 흰쥐에 간흡충의 피낭유충을 100 개씩 경구 감염시키고, 15주까지 간의 각 엽에 있는 담관의 조직절편을 만들어서 다세포군항체와 ABC complex를 이용한 염색을 실시하였다. 그 결과 충제가 들어 있는 담관에서는 감염 1주에 상피 세포의 2.9-10.2%가 염색에 양성이었고, 2주에 7.3-12.8%가, 5주에는 7.3-13.4%가, 15주에 8.4-14.8%가 BrdU를 함유하였다. 대조군 흰쥐나 실험군 동물의 감염되지 않은 담관의 경우에는 0-2.7%가 염색에 양성으로 나타났다. 염색된 상피세포는 주로 점막 상피세포층의 기저부에서 관찰되었다. 이 결과에 의하면 간흡충과 직접 접촉하는 담관의 상피세포가 뚜렷하게 증식하고 있어 충체의 어떠한 물리적인 또는 화학적인 자극이 있더라도 가까운 부위에서 주로 작용하고 있다고 판단하였다.

• 대한감각통합치료학회지
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• 제2권1호
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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.

• 대한한의학회지
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• 제31권2호
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• pp.36-47
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• 2010

Objective: The purpose of this study was to investigate the effects of skin application with Gelidium amansii extract on skin with deep second degree burns in mice. Methods: BALB/c mice were divided into four groups: normal (NOR) group; burn-elicited mice (CON) group, Silmazine-treated mice after burn elicitation (ST) group, and Gelidium amansii-extract treated mice after burn elicitation (GT) group. To examine the skin recovery effect after burn, changes of burn area, angiogenesis and histologic structure were analyzed. To measure effect of edema regulation, matrix metalloproteinase-9 (MMP-9) was analyzed. To estimate the skin regenerative & stable effect, 5-bromo-2'-deoxyuridine (BrdU) and substance P were analyzed. Results: 2 weeks later, 1. The size of burn area decreased in the GT and ST groups more than the CON group. 2. Alleviation of angiogenesis appeared in the GT and ST groups more than in the CON group. 3. Blood clot, epithelial cell hyperplasia, and inflammatory cell infiltration declined in the GT and ST groups more than in the CON group. 4. MMP-9, BrdU, and substance P positive reaction decreased in the GT and ST groups more than in the CON group 5. In the comparative study, the GT group was superior to the ST group. Conclusion: The skin application of Gelidium amansii extract could lessen skin damage by the medium of regulation MMP-9 activation. This skin stabilization was induced in mice with deep second degree burns.

• Radiation Oncology Journal
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• 제23권3호
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• pp.157-160
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• 2005

목적: 저선량 방사선이 당뇨쥐의 해마에서 신경 세포의 증식에 미치는 영향을 확인하기 위해서이다. 대상 및 방법: 정상 쥐를 대상으로 방사선을 조사하지 않은 군, 0.1 Gy 방사선 조사군, 10 Gy 방사선 조사군으로 나눠 방사선이 정상 신경 세포의 증식에 미치는 영향을 확인하였다. 당뇨쥐를 대상으로 한 이차 실험은 방사선을 조사하지 않은 정상 쥐, 방사선을 조사하지 않은 당뇨쥐, 0.01 Gy 방사선 조사한 당뇨쥐, 0.1 Gy를 조사한 당뇨쥐, 1 Gy를 조사한 당뇨쥐, 10 Gy를 조사한 당뇨쥐로 나눠 시행하였다. 5-bromo-2'-deoxyuridine (BrdU)를 이용한 면역 염색을 시행하여 쥐 해마의 치아이랑 세포의 수를 세어 비교하였다. 결과: BrdU 양성 세포의 수는 대상으로 방사선을 조사하지 않은 정상쥐, 0.1 Gy 방사선 조사한 정상쥐, 10 Gy 방사선 조사한 정상쥐에서 각각 $45.95{\pm}3.42/mm^2,\;59.34{\pm}5.20/mm^2,\;19.26{\pm}2.98/mm^2$로 나타났다. 당뇨쥐를 대상으로 한 이차 실험에서 BrdU 양성 세포의 수는 방사선을 조사하지 않은 정상 쥐, 방사선을 조사하지 않은 당뇨쥐, 0.01 Gy 방사선 조사한 당뇨쥐, 0.1 Gy를 조사한 당뇨쥐, 1 Gy를 조사한 당뇨쥐, 10 Gy를 조사한 당뇨쥐에서 각각 $55.44{\pm}8.57/mm^2,\;33.33{\pm}6.46/mm^2,\;67.75{\pm}10.54/mm^2,\;66.63{\pm}10.05/mm^2,\;23.59{\pm}6.37/mm^2,\;14.34{\pm}7.22/mm^2$로 나타났다. 결론: 0.1 Gy 이하의 저선량 방사선은 당뇨쥐의 해마 치아이랑세포의 수를 증가시켜 세포 증식효과를 보이는 것을 확인하였다.

• 한방재활의학과학회지
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• 제21권2호
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• pp.15-30
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• 2011

Objectives : The present study investigated the effects of Achyranthis Radix on short-term memory, apoptotic neuronal cell death in the hippocampus following transient global ischemia in gerbils. Methods : The gerbils were divided into 5 groups(n=10); Sham operation group, ischemia-induced group, ischemia-induced and 50 mg/kg Achyranthis Radix-treated group, ischemia-induced and 100 mg/kg Achyranthis Radix-treated group, ischemia-induced and 200 mg/kg Achyranthis Radix-treated group. For this study, a step-down avoidance task, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL) assay, immunohistochemistry for caspase-3 and BrdU(5-Bromo-2'-deoxyuridine), and western blotting for bax, bcl-2 were performed. Results : The results revealed that ischemic injury impaired short-term memory and increased apoototic neuronal cell death in the hippocampal CA1(cornu ammonis area 1) region. Ischemic injury enhanced cell proliferation in the hippocampal CA1 region, the compensatory and adaptive process for excessive apoptosis. Achyranthis Radix treatment improved short-term memory by suppressing ischemia-induced apoptotic neuronal cell death in the hippocampal CA1 region. Also, Achyranthis Radix suppressed the ischemia-induced increase in cell proliferation in the hippocampal CA1 region. Conclusions : We showed that Achyranthis Radix alleviates ischemia-induced apoptotic neuronal cell death, thus facilitates the recovery of short-term memory impairment induced by ischemic cerebral injury.

• 운동영양학회지
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• 제23권3호
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• pp.13-21
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• 2019

[Purpose] Chronic stress is a precipitating factor for depression, whereas exercise is beneficial for both the mood and cognitive process. The current study demonstrates the anti-depressive effects of regular exercise and the mechanisms linked to hippocampal neurogenesis. [Methods] Mice were subjected to 14 consecutive days of restraint, followed by 3 weeks of treadmill running, and were then subjected to behavioral tests that included the forced swimming and Y-maze tests. Protein levels were assessed using western blot analysis and newborn cells were detected using 5-bromo-2'-deoxyuridine (BrdU). [Results] Three weeks of treadmill running ameliorated the behavioral depression caused by 14 days of continuous restraint stress. The exercise regimen enhanced BrdU-labeled cells and class III β-tubulin levels in the hippocampal dentate gyrus, as well as those of thioredoxin-1 (TRX-1) and synaptosomal β2-adrenergic receptors (β2-AR) under stress. In vitro experiments involving treatment with recombinant human TRX-1 (rhTRX-1) augmented the levels of phospho-extracellular signal-regulated kinases 1 and 2 (ERK1/2), nuclear β-catenin, and proliferating cell nuclear antigens, which were previously inhibited by U0216 and FH535 (inhibitors of ERK1/2 and β-catenin/T cell factor-mediated transcription, respectively). The hippocampal neurogenesis elicited by a 7-day exercise regimen was abolished by a selective inhibitor of β2-AR, butoxamine. [Conclusion] These results suggest that TRX-1-mediated hippocampal neurogenesis by β2-AR function is a potential mechanism underlying the psychotropic effect of exercise.