cDNA microarray를 이용한 침자극에 대한 반응군과 비반응군의 유전자적 특이성 연구

노삼웅,이기석,이진우,홍무창,신민규,민병일,배현수 대한동의병리학회 2001 동의생리병리학회지 Vol.15 No.4

Numerous studies show that acupuncture stimulation increases experimental pain threshold in various animal species. It has been suggested that the acupuncture analgesic effects could be mediated by endogenous opioid system. However, the analgesic effect on the rat in response to electroacupuncture(EA) shows marked individual variations. Individual variations in effectiveness(IVE) of acupuncture analgesia has frequently provoked clinical troubles. The intrinsic mechanisms that cause IVE are an interesting subject for the study. The purpose of this study is to identify and characterize genes that cause differences between the responder and non-responder upon EA treatments using cDNA microarray. Sprague-Dawley male rats were stimulated at the zusanli(ST36) acupoint, then the tail flick latency (TFL) test was performed to classify the responder and non-responder group. Among the 19 rats tested, the ratio of responder group to non-responder group was 6:4. The hypothalamus of each group was dissected and mRNA was further purified. After synthesizing cDNA using oligo-dT primer, the cDNA were applied to Incyte rat GEMTM 2 microarray chips. The microarray results show that genes such as Brain factor(BF)-1, a transcription factor essential for the proliferation of the progenitor cell of the cerebral cortex, Dopamine beta-hydroxylase and Phosphotidylethanolamine N-methyltransferase which play a major role in catecholamine synthesis, are expressed abundantly in the responder group, whereas tau and cathepsin B, which are neurodegeration markers are abundant in the non-responder group. These results suggest that the sensitivity of EA mediated analgesic effects are closely related with neurodegeneration of the brain.

足三里 전침자극에 의한 흰쥐 hypothalamus의 유전자 발현 profile 분석

노삼웅(Sam Woong Rho),이기석(Gi Seog Lee),최기순(Gi Soon Choi),나영인(Young In Na),홍무창(Moo Chang Hong),신민규(Min Kyu Shin),민병일(Byung il Min),배현수(Hyun Su Bae) 한의병리학회 2004 동의생리병리학회지 Vol.18 No.4

The association of cholecystokinin-A receptor expression with the responsiveness of electropacupuncture analgesic effects in rat

Lee, Giseong,Rho, Samwoong,Shin, Minkyu,Hong, Moochang,Min, Byung-il,Bae, Hyunsu WHO COLLABORATING CENTRE FOR TRADITIONAL MEDICINE 2002 東西醫學硏究所 論文集 Vol.2002 No.-

The purpose of this study is to determine whether the level of cholecystokinin (CCK) receptor expression causes the differences between the responder and non-responder to electroacupuncture mediated analgesic effects. Male Sprague-Dawley rats were stimulated at the Zusanli (ST36) acupoint in the absence of any anesthetics and holders. The tail flick latency test was performed to quantify analgesic effects and then the responder and non-responder groups were classified. The hypothalamus of each group was dissected and RNA was purified. The amount of mRNA expression of CCK-A and CCK-B receptors was determined by reverse transcription-polymerase chain reaction. The results show that CCK-A receptors are significantly more expressed in non-responders than responders, whereas CCK-B receptor expresson is similar in both groups. ⓒ 2002 Elsevier Science lreland Ltd. All rights reserved.

육미지황탕가미방에 의한 흰쥐 기억력 향상과 관련된 Hippocampus 부위의 특이 유전자 발현에 대한 연구

심대식,노삼웅,이진우,이은아,조종운,배현수,신민규,홍무창 대한동의병리학회 2003 동의생리병리학회지 Vol.17 No.3

The herbal extract(YMT_02) is a modified extracts from Yukmijihwang-tang(YMJ) to promote memory-enhancing. The YMJ extracts has been widely used as replenishing yin and tonifying the kidneys herbal medicine for hundred years ia Asian countries. The purpose of this study is to: 1) quantitatively evaluate the memory-enhancing effect of YMT_02 by passive avoidance test, 2) statistical evaluation of candidate gene expression (pentraxin, PEP-19, transthyretin) in rat hippocampus. The hippocampi of YMT_02 and control group were dissected and mRNA was further purified. After synthesizing cDNA using oligo-dT primer, the cDNA were applied to Real Time PCR. The results were as follows : 1) passive avoidance test showed enhancing memory retentin by YMT_02 treatment, 2) expression of pentraxin, that accelerate degenerating of neuronal cell, was significantly decreased, 3) the mRNA of genes that has been known to be associated with protecting neuronal cell degeneration, such as PEP-19 and transthyretin, were significantly increased upon YMT_02 treatment. From above results, the administration of YMT_02 which tonify the function of Kidneys could enhance the ability of memory and learning. In addition, the administration of YMT_02 enhance memory retention through modulating particular gene (pentraxin, PEP-19, transthyretin) expressions in hippocampu

전침이 자연살해세포 활성에 미치는 유전자 발현 profile에 대한 연구

최기순,노삼웅,오상덕,배현수,안현종,하윤문,김강호,민병일 대한침구의학회 2002 대한침구의학회지 Vol.19 No.6

牡丹皮의 PC12 cell 酸化抑制 효과 및 neuronal 유전자 발현 profile 분석에 대한 연구

김현희,노삼웅,나영인,배현수,신민규,김정숙,홍무창 대한동의생리학회,대한동의병리학회 2003 동의생리병리학회지 Vol.17 No.2

Yukmijihwang-tang has been widely used as an anti-aging herbal medicine for hundred years in Asian countries. Numerous studies show that Yukmijihwangtang has anti-oxidative effect both in vivo and in vitro. It has been reported that Moutan Cortex Radicis extract (MCR) was the most effective herb in Yukmijihwang-tang on undifferentiated PC12 cells upon oxidative-stressed with hydrogen peroxide. The purpose of this study is to; 1) evaluate the recovery of neuronal damage by assessing the anti-oxidant effect of MCR on PC12 cells differentiated with nerve growth factor (NGF), 2) identify candidate genes responsible for anti-oxidative effect on differentiated PC12 cells by oligonucleotide chip microarray. PC12 cells, which were differentiated by treating with NGF, were treated without or with hydrogen peroxide in the presence or absence of various concentration of MCR. Cell survival was determined by using MTS assay. Measurement of intracellular reactive oxygen species (ROS) generation was determined using the H2DCFDA assay The Viability of cells treated with MCR was significantly recovered from stressed PC12 cell. In addition, wide rage of concentrations of MCR shows dose-dependent inhibitory effect on ROS production in oxidative-stressed cells. Total RNAs of cells without treatment(Control group), only treated with H_2O_2 (stressed group) and treated with both H2O2 and of MCR (MCR group) were isolated, and cDNAs was synthesized using oligoT7(dT) primer. The fragmented cRNAs, synthesized from cDNAs, were applied to Affymetrix GeneChip Rat Neurobiology U34 Array. mRNA of Calcium/calmodulin-dependent protein kinase Ⅱ delta subunit(CaMKⅡ), neuron glucose transporter (GLUT3) and myelin/oligodendrocyte glycoprotein(MOG) were downregulated in Stressed group comparing to Control group. P2X2-5 receptor (P2X2R-5), P2X2-4 receptor (P2X2R-4), c-fos, 25 kDa synaptosomal attachment protein(SNAP-25a) and GLUT3 were downregulated, whereas A2 adenosine receptor (A2AR), cathechol-O-methyltransferase(COMT), glucose transporter 1 (GLUT1), EST223333, heme oxygenase (HO), VGF, UI-R-C0-ja-a-07-0-UI.s1 and macrophage migration inhibitory factor (MIF) were upregulated in MCR group comparing to Control group. Expression of Putative potassium channel subunit protein (RCK4), P2X2R-5, P2X2R-4, Interferon-gamma inducing factor isoform alpha precursor (IL-18α), EST199031, P2XR, P2X2 purinoceptor isoform e (P2X2R-e), Precursor interleukin 18 (IL-18) were downregulated, whereas MOG, EST223333, GLUT-1, MIF, Neuronatin alpha, UI-R-C0-ja-a-07-0-UI.s1, A2 adenosine receptor, COMT, neuron-specific enolase (NSE), HO, VGF, A rat novel protein which is expressed with nerve injury (El2625) were upregulated in MCR group comparing to Stressed group. The results suggest that decreased viability and ROS production of PC12 cell by H_2O_2 may be, at lease, mediated by impaired glucose transporter expression. It is implicated that the MCR treatment protect PC12 cell from oxidative stress via following mechanisms; improving glucose transport into the cell, enhancing expression of anti-oxidative genes and protecting from dopamine cytotoxicity by increment of COMT and MIF expression. The list of differentially expressed genes may implicate further insight on the action and mechanism behind the anti-oxidative effects of herbal extract Moutan Cortex Radicis.

牡丹皮의 PC12 cell 항산화 효과와 관련 HO, MIF, COMT 유전자 발현에 미치는 영향

손무성,노삼웅,고은정,나영인,조종운,배현수,홍무창,신민규 대한동의생리학회,대한동의병리학회 2003 동의생리병리학회지 Vol.17 No.4

Oriental medicine explains aging as the weakening of Kidney-Qi, and Kidney-strengthening herbal medicines such as Yukmijihwang-tang have been studied for anti-aging effects. In Western Medicine, the hypothesis that reactive oxidant species(ROS) contribute to the aging process is generally accepted. It has been reported that Moutan Cortex Radicis extract (MCR) was the most effective constituent of Yukmijihwang-tang in decreasing ROS production in oxidative-stressed cells. The purpose of this study is to confirm the anti-oxidant effect of MCR on PC12 cells, the expression of Heme oxygenase (HO), Macrophage migration inhibitory factor (MIF), Catechol-0-methyltransferase (COMT) using real time RT PCR. PC12 cells were treated without or with hydrogen peroxide in the presence or absence of MCR using MTS assay. Hydrogen peroxide decreased the viability of PC12 cells by 53% and MCR did not influence that of stressed PC 12 cells irrespective of dose or incubation period. However, MCR showed an inhibitory effect on production of ROS in stressed cells, both dose and incubation time dependently. In particular, 1 mg/ml of MCR for 24 h culture almost returned to normal level. In the quantiation of anti-aging related gene expression, MCR at 1 mg/ml increased the expression of HO by 370%, MlF by 180% and COMT by 280% through real time RT PCR. In conclusion, MCR treatment protected PC12 cells from hydrogen peroxide and decreased ROS production and enhanced anti-oxidative gene expression such as HO, COMT and MlF, which suggests that MCR is involved in controlling anti-aging of nerve cells through elimination of cytotoxic stimuli..

鍼刺戟에 대한 반응과 비반응군 흰쥐의 특이 유전자 발현 연구

서영찬,노삼웅,이기석,고은정,홍무창,신민규,민병일,배현수 대한동의병리학회 2003 동의생리병리학회지 Vol.17 No.3

A pain is the symptom defends against noxious stimulus about a human body, it is known that if the periphery of perceptive nerve were stimulated by a physical or chemical factors, the stimulation is induced by transmission to pain center in the cerebral cortex according to pain conduction tract. The treatment of pain is to decrease a stimulus that causes a pain or block off a nerve transmitting a stimulus or puts on a way to calm down pain center, but It is for adjustment of a pain to be the most representative in acupuncture among various ways to cure a pain in Oriental medicine. However, the analgesic effect of an individual response to acupuncture stimulation shows marked individual variations, so these days genetic a few approach is attempted. On this the author determined that the responding group was appointed those whose tail flick latency (TFL) responding time delayed the minimum of 30% comparing with basal reaction time. For those whose TFL time had shorter than 30% was grouped as a non-responding group. And then the hypothalamus of each group was dissected and RNA was further purified. After synthesizing cDNA using oligo dT primer, products were finally applied to the PCR. The results were as follows ; The ratio of responding group to non-responding group was 6:4. Ach T (acetylcholinesterase T subunit), BF-1 (Brain factor-1), DBH (Dopamine β-hydroxylase) and PNM (Phosphotidylethanolamine N-Methyltransferase) were revealed significantly in the responding group. Cathepsin B and Tau were revealed significantly in the non-responding group. The PCR results show that Ach T, BF-1, DBH and PNM are expressed abundantly in the responding group, where as cathepsin B and tau are abundant in the non-responding group. These results suggest that the analgesic effect on acupuncture stimulation is related to regulation of neurotransmitter as well as neurodegeration of cerebrum.

한국인의 G Protein ß3 Subunit 유전자의 多形性과 腦血管질환과의 상관성에 대한 연구

최민희,이진우,이경진,이효정,노삼웅,최현,조기호,홍무창,신민규,김영석,배현수 대한동의병리학회 2003 동의생리병리학회지 Vol.17 No.3

Background and purpose: Hypertension and obesity has been implicated in the most important risk factors for stroke. The original finding that the G-protein beta3 subunit (GNB3) C825T allele associates with essential hypertension and obesity has been confirmed in several different populations. Hence, our objective was to determine whether the GNB3 C825T polymorphism predicts interindividual variation in stroke. Method: We recruited 361 stroke patients (cerebral infarction, n=278; intracerebral hemorrhage (ICH), n=83) and 199 healthy control subjects. Subjects were genotyped for GNB3 C825T mutation and findings were investigated for association with stroke. Result: The GNB3 T/T type was significantly associated with cerebral infarction prevalence (OR, 1.98; 95% CI, 1.14-3.46; p=0.015). While, ICH was not found to be significantly associated with GNB3 T/T type (OR, 1.63; 95% ICH, 0.74-3.56; p=0.219). Similarly, no significant association was determined between GNB T/C type, and cerebral infarction (OR, 1.09; 95% CI, 0.68-1.74; p=0.76), and ICH (OR, 1.14; 95% CI, 0.59-2.21; p=0.697). Conclusion: In clinical characteristics, this study shows no differences among GNB3 genotypes, that are BMI, WH ratio, hypertension rate, and ischemic heart disease rate, total lipid level, triglycerides level, total cholesterol level, HDL cholesterol level, prothrombine time, with the exception of LDL cholesterol concentrations. However, our subjects showed an inverse relationship between LDL cholesterol level and the risk of cerebral infarction. We have shown that the GNB3 T/T genotype is strongly associated with cerebral infarction. (OR, 1.98; 95% CI, 1.14-3.46; p=0.015)

한국인의 G Protein β3 Subunit유전자의 다형성과 뇌혈관 질환과의 상관성에 대한 연구

최민희,이진우,이경진,이효정,노삼웅,최현,조기호,홍무창,신민규,김영석,배현수 한의병리학회 2003 동의생리병리학회지 Vol.17 No.3