간질(癎疾) 동물(動物) 모델을 이용한 소부혈(少府穴)의 간질발작(癎疾發作) 및 해마(海馬)의 염증(炎症) 억제 효과(效果) 검증(檢證)

두아름 ( Ah Reum Doo ),김승남 ( Seung Nam Kim ),인창식 ( Chang Shik Yin ),김연정 ( Yeon Jung Kim ),이혜정 ( Hye Jung Lee ),김승태 ( Seung Tae Kim ),박히준 ( Hi Joon Park ) 경락경혈학회 2012 Korean Journal of Acupuncture Vol.29 No.3

Objectives : To confirm whether acupuncture can suppress the degree of seizure and the activation of astrocytes in hippocampi using kainic acid(KA)-induced epilepsy mouse model. Methods : 8 weeks C57BL/6 mice(20~25 g) were given acupuncture once a day at acupoint HT8(Shaofu) bilaterally during 2 days before KA injection. After an intraperitoneal injection of 30 mg/kg KA, acupuncture treatment was subsequently administered once more(total 3 times), and the degree of seizure was observed for 120 min. The neuronal cell death, pERK expression, and astrocyte activation confirmed 1 hour and 24 hours after KA injection. Results : Acupuncture treatment at HT8 suppressed KA-induced epileptic seizure. One hour after KA injection, the pERK expression was increased, which was reduced by the acupuncture treatment. Twenty four hours after injection, the treatment decreased the KA-induced neuronal cell death, the interleukin-1β expression and the astrocyte activation in the CA3 region of the mouse hippocampus. Conclusions : Acupuncture treatment at HT8 decreases the KA-induced epileptic seizure, the neural cell inflammation and death.

$MPP^+$로 유도된 SH-SY5Y신경세포 사멸에 대한 고분자성분제거 봉독약침액의 신경보호 효과 연구

배광록,두아름,김승남,박지연,박히준,이혜정,권기록,Bae, Kwang-Rok,Doo, Ah-Reum,Kim, Seung-Nam,Park, Ji-Yeon,Park, Hi-Joon,Lee, Hye-Jung,Kwon, Ki-Rok 대한한방내과학회 2010 大韓韓方內科學會誌 Vol.31 No.2

Objectives : The neuroprotective effects of bee venom (BV) have been demonstrated in many studies, but bee venom has many side effects. So we used sweet bee venom (SBV), which has high molecular elements removed to reduce the side effects. I examined the neuroprotective effect of sweet bee venom in 1-methyl-4-phenylpyridine ($MPP^+$)-induced human neuroblastoma SH-SY5Y cells. Methods : To observe the possible toxicity of SBV itself, SH-SY5Y cells were treated with SBV in various concentrations for 3 h and $MPP^+$ in concentrations (1 and 5mM) for 24h. To investigate the protective effect of SBV against $MPP^+$ toxicity, SH-SY5Y cells were pretreated with vehicle or nontoxic concentrations of SBV for 3h and the cells were not washed, followed by incubation with respective concentrations of SBV and 1 mM $MPP^+$ for 24h. To investigate the protective effect of SBV against $MPP^+$ toxicity, SH-SY5Y cells were pretreated with vehicle or nontoxic concentrations of SBV for 3h and the cells were not washed, followed by incubation with respective of SBV(0.5%), 1 mM $MPP^+$, 5uM AKT inhibitor(LY984002) and 10uM ERK inhibitor(PD98059) for 24 h. The protective effect was measured by cell viability assay. To investigate the degree of apoptosis, caspase-3 enzyme activity was measured in control, $MPP^+$, SBV+$MPP^+$. Results : SBV (0.5%) pretreatment protected the SH-SY5Y cells against $MPP^+$-induced apoptotic cell death. The cell viability was higher in the SH-SY5Y cells that were pretreated with vehicle or nontoxic concentrations of SBV than those not pretreated. The caspase-3 activity was lower in the pretreated groups than these not pretreated. ERK and AKT enzymes have a role in the neuroprotective effects of the sweet bee venom. Conclusions : The results demonstrate that SBV has a protective effect on dopaminergic neurons against $MPP^+$ toxicity. This data suggest that SBV could be a potential therapeutic tool for neurodegenerative diseases such as Parkinson's disease(PD).

MTR and In−vivo 1H-MRS Studies on Mouse Brain with Parkinson’s Disease

윤문현,김현진,정진영,두아름,박히준,김승남,최보영 한국물리학회 2012 THE JOURNAL OF THE KOREAN PHYSICAL SOCIETY Vol.61 No.11

The aim of this study was to investigate whether the changes in the magnetization transfer ratio (MTR) histogram are related to specific characteristics of Parkinson’s disease (PD) and to investigate whether the MTR histogram parameters are associated with neurochemical dysfunction by performing in vivo proton magnetic resonance spectroscopy (¹H-MRS). MTR and <i>In−vivo</i> ¹HMRS studies were performed on control mice (n = 10) and 1-methyl-1,2,3,6-tetrahydropyridine intoxicated mice (n = 10). All the MTR and in vivo ¹H-MRS experiments were performed on a 9.4 T MRI/MRS system (Bruker Biospin, Germany) using a standard head coil. The protondensity fast spin echo (FSE) images and the T2-weighted spin echo (SE) images were acquired with no gap. Outer volume suppression (OVS), combined with the ultra-short echo-time stimulated echo acquisition mode (STEAM), was used for the localized in-vivo ¹H-MRS. The quantitative analysis of metabolites was performed from the 1H spectra obtained in vivo on the striatum (ST) by using jMRUI (Lyon, France). The peak height of the MTR histograms in the PD model group was significantly lower than that in the control group (p < 0.05). The midbrain MTR values for volume were lower in the PD group than the control group(p < 0.05). The complex peak (Glx: glutamine+glutamate+ GABA)/creatine (Cr) ratio of the right ST in the PD group was significantly increased as compared to that of the control group. The present study revealed that the peak height of the MTR histogram was significantly decreased in the ST and substantia nigra, and a significant increase in the Gl_x/Cr ratio was found in the ST of the PD group, as compared with that of the control group. These findings could reflect the early phase of neuronal dysfunction of neurotransmitters.

Acupuncture Stimulation to HT8 Enhances Cell Proliferation in Hippocampus on an Epilepsy Mouse Model

김승태,박해정,홍미숙,김승남,두아름,인창식,이혜정,정주호,박히준,Kim, Seung-Tae,Park, Hae-Jeong,Hong, Mee-Sook,Kim, Seung-Nam,Doo, Ah-Reum,Yin, Chang-Shik,Lee, Hye-Jung,Chung, Joo-Ho,Park, Hi-Joon Society for Meridian and Acupoint 2010 Korean Journal of Acupuncture Vol.28 No.2

목적 : 뇌의 신경세포 증식은 해마 치상회와 뇌실하영역에서만 나타나는 현상이다. Kainic acid(KA)를 이용한 간질 동물모델을 연구하던 중 침이 해마 치상회의 신경세포증식을 촉진하는 현상을 발견하여 이를 보고하고자 한다. 방법 : 수컷 ICR계 생쥐를 Saline(n=8), KA(n=8), KA+Acu(n=8)의 세 군으로 나누고, 모든 생쥐들에게 KA 주입 3일 전부터 1일 1회씩 5'-bromodeoxyuridine(BrdU)을 3일간 주입하였다. Saline군에는 멸균된 생리식염수를 뇌실 내에 주입하였고, KA군 및 KA+Acu군에는 $0.1{\mu}g$의 KA를 뇌실 내에 주입하였으며, KA+Acu군에 속한 쥐들에게는 KA 주입 2일전, 1일전, 주입 직후에 양쪽 소부(少府)(HT8)에 자침하였다. KA 주입 3시간 후 쥐의 뇌를 적출하고 해마 치상회부위의 BrdU 및 neuropeptide Y (NPY)의 발현을 측정하였다. 결과 : 소부(少府) 자침이 KA의 독성으로 인한 신경세포의 파괴를 줄여주었으며, BrdU 양성 세포 및 NPY를 유의하게 증가시켰다. KA 주입시 세포증식이 일어나긴 하나, 3시간 안에는 거의 일어나지 않는다. 결론 : 소부(少府) 자침이 해마 치상회의 신경세포증식을 촉진하며, 이는 KA의 효과가 아닌 KA 투여 전 소부(少府) 자침으로 인한 것으로 사료된다.

Investigation of the Neuroprotective Effects of Bee-venom Acupuncture in a Mouse Model of Parkinson’s Disease by Using Immunohistochemistry and <i>In−vivo</i> ¹H Magnetic Resonance Spectroscopy at 9.4 T

윤문현,이도완,김현진,정진영,두아름,박히준,김승남,최보영 한국물리학회 2013 THE JOURNAL OF THE KOREAN PHYSICAL SOCIETY Vol.62 No.2

Neuroprotective therapeutics slows down the degeneration process in animal models of Parkinson’s disease (PD). The neuronal survival in PD animal models is often measured by using immunohistochemistry. However, dynamic changes in the pathology of the brain cannot be explored with this technique. Application of <i>in-vivo</i> ¹H magnetic resonance spectroscopy (¹H MRS) can cover this shortcoming, as these techniques are non-invasive and can be repeated over time in the same animal. Thus, the sensitivity of both techniques to measure changes in the PD pathology was explored in an experiment studying the neuroprotective effects of the vigilance enhancer bee-venom (BV) in a mouse model of PD. The mice were pre-treated with 0.02-ml BV administered to the acupuncture point GB34 (Yangneungcheon) once every 3 days for 2 weeks. Three groups were classified as control, MPTP-intoxicated PD model and BV-treated mice. Outer volume suppression combined with the ultra-short echo-time STEAM (TE = 2.2 ms, TM = 20 ms, TR = 5000 ms) was used for localized <i>in-vivo</i> ¹H MRS. Based on the ¹H MRS spectral analysis, substantial changes of the neurochemical profiles were evaluated in the three investigated groups. In particular, the glutamate complex (Glx)/creatine (Cr) ratio (7.72 ± 1.25) in the PD group was significantly increased compared to that in the control group (3.93 ± 2.21, P = 0.001). Compared to the baseline values, the Glx/Cr ratio of the BV-treated group was significantly decreased 2 weeks after MPTP intoxication (one-way ANOVA, p < 0.05). In conclusion, the present study demonstrated that neurochemical alterations occurred in the three groups and that the neuroprotective effects of the BV acupuncture in a mouse model of PD could be quantified by using immunohistochemistry and ¹H MRS.

마우스 간질 동물모델에서 소부혈 자침이 해마 치상회의 신경세포증식에 미치는 영향

김승태 ( Seung Tae Kim ),박해정 ( Hae Jeong Park ),홍미숙 ( Mee Sook Hong ),김승남 ( Seung Nam Kim ),두아름 ( Ah Reum Doo ),인창식 ( Chang Shik Yin ),이혜정 ( Hye Jung Lee ),정주호 ( Joo Ho Chung ),박히준 ( Hi Joon Park ) 대한경락경혈학회 2010 Korean Journal of Acupuncture Vol.27 No.2

목적: 뇌의 신경세포 증식은 해마 치상회와 뇌실하영역에서만 나타나는 현상이다. Kainic acid(KA)를 이용한 간질동물모델을 연구하던 중 침이 해마 치상회의 신경세포증식을 촉진하는 현상을 발견하여 이를 보고하고자 한다. 방법: 수컷 ICR계 생쥐를 Saline(n=8), KA(n=8), KA+Acu(n=8)의 세 군으로 나누고, 모든 생쥐들에게 KA 주입 3일 전부터 1일 1회씩 5`-bromodeoxyuridine(BrdU)을 3일간 주입하였다. Saline군에는 멸균된 생리식염수를 뇌실 내에 주입하였고, KA군 및 KA+Acu군에는 0.1μg의 KA를 뇌실 내에 주입하였으며, KA+Acu군에 속한 쥐들에게는 KA 주입 2일전, 1일전, 주입 직후에 양쪽 少府(HT8)에 자침하였다. KA 주입 3시간 후 쥐의 뇌를 적출하고 해마 치상회부위의 BrdU 및 neuropeptide Y (NPY)의 발현을 측정하였다. 결과: 少府 자침이 KA의 독성으로 인한 신경세포의 파괴를 줄여주었으며, BrdU 양성 세포 및 NPY를 유의하게 증가시켰다. KA 주입시 세포증식이 일어나긴 하나, 3시간 안에는 거의 일어나지 않는다. 결론: 少府 자침이 해마 치상회의 신경세포증식을 촉진하며, 이는 KA의 효과가 아닌 KA 투여 전 少府 자침으로 인한 것으로 사료된다.