• Archives of Pharmacal Research
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• v.29 no.10
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• pp.849-858
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• 2006

Previous screening of the pharmacological action of Gastrodia elata (GE) root (Orchidaceae) showed that methanol (MeOH) extracts have significant anti-inflammatory properties. The antiinflammatory agents of GE, however, remain unclear. In this experiment, MeOH extracts of GE were fractionated with organic solvents for the anti-inflammatory activity-guided separation of GE. Eight phenolic compounds from the ether (EtOEt) and ethyl acetate (EtOAc) fractions were isolated by column chromatography: 4-hydroxybenzaldehyde (I), 4-hydroxybenzyl alcohol (II), benzyl alcohol (III), bis-(4-hydroxyphenyl) methane (IV), 4(4'-hydroxybenzyloxy)benzyl-methylether (V), 4-hydroxy-3-methoxybenzyl alcohol (VI), 4-hydroxy-3-methoxybenzaldehyde (VII), and 4-hydroxy-3-methoxybenzoic acid (VIII). To investigate the anti-inflammatory and anti-oxidant activity of these compounds, their effects on carrageenan-induced paw edema, arachidonic acid (AA)-induced ear edema and analgesic activity in acetic acid (HAc)-induced writhing response were carried out in vivo; cyclooxygenase (COX) activity, reactive oxygen species (ROS) generation in rat basophilic leukemia (RBL 2H3) cells and 1,1-diphenyl-2-picryl-hydroazyl (DPPH) scavenging activity were determined in vitro. These phenolic compounds not only had anti-inflammatory and analgesic properties in vivo, but also inhibited COX activity and silica-induced ROS generation in a dose-dependent manner. Among these phenolic compounds, compound VII was the most potent anti-inflammatory and analgesic. Compound VII significantly inhibited silica-induced ROS generation and compound VI significantly increased DPPH radical scavenging activity. Compounds I, II and III significantly inhibited the activity of COX-I and II. These results indicate that phenolic compounds of GE are anti-inflammatory, which may be related to inhibition of COX activity and to anti-oxidant activity. Consideration of the structure-activity relationship of the phenolic derivatives from GE on the anti-inflammatory action revealed that both C-4 hydroxy and C-3 methoxy radicals of benzyl aldehyde play an important role in anti-inflammatory activities.

• Archives of Pharmacal Research
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• v.29 no.10
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• pp.898-903
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• 2006

The antiplatelet and antithrombotic activities of Korean Red Ginseng (KRG) were examined on rat carotid artery thrombosis in vivo, and platelet aggregation in vitro and ex vivo. Administration of KRG to rats not only prevented carotid artery thrombosis in vivo in a dose-dependent manner, but also significantly inhibited ADP- and collagen-induced platelet aggregation ex vivo, while failed to prolong coagulation times such as activated partial thromboplastin time (APTT) and prothrombin time (PT), indicating the antithrombotic effect of KRG might be due to its anti platelet aggregation rather than anticoagulation effect. In line with the above observations, KRG inhibited U46619-, arachidonic acid-, collagen- and thrombin-induced rabbit platelet aggregation in vitro in a concentration-dependent manner, with $IC_{50}$ values of $620{\pm}12$, $823{\pm}22$, $722{\pm}21$ and $650{\pm}14\;{\mu}g/mL$, respectively. Accordingly, KRG also inhibited various agonists-induced platelet serotonin secretions as it suppressed platelet aggregation. These results suggest that KRG has a potent antithrombotic effect in vivo, which may be due to antiplatelet rather than anticoagulation activity, and KRG intake may be beneficial to the individuals with high risks of thrombotic and cardiovascular diseases.

• Journal of Acupuncture Research
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• v.23 no.5
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• pp.23-29
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• 2006

Objectives : The present study was conducted to evaluate the effects of automatically controlled rotating acupuncture (ACRA) on thermal allodynia in neuropathic pain rats, and to examine whether the endogenous opioid system mediates the effects of ACRA. Methods : For the neuropathic surgery, the right superior caudal trunk was resected at the level between S1 and S2 spinal nerves innervating the tail. Two weeks after the nerve injury, ACRA stimulation with 4 different stimulation conditions (i.e., angle and frequency of rotation: 90o+1Hz, 90o+1/4Hz, 360o+/1Hz, and 360o+1/4Hz) was delivered to the Zusanli (ST36) acupoint for 15 min. The behavioral signs of thermal allodynia were evaluated by the tail immersion test (i.e., immersing the tail in cold $(4^{\circ}C)$ or warm $(4^{\circ}C)$ water and measuring the latency to an abrupt tail movement) before and after the stimulation. In an additional set of experiments, we examined the effects of naloxone (opioid Results : ACRA stimulations under all of the conditions above significantly relieved thermal antagonist, 2mg/kg, i.p.) on the action of ACRA stimulation. allodynia. There is no difference in the anti-allodynic effects among the 4 stimulation conditions. In addition, the effect of ACRA on thermal allodynia was reversed by naloxone pretreatment. Conclusion : These results indicate that ACRA stimulations have relieving effects on thermal allodynia in neuropathic pain rats, irrespective of stimulation parameters, and that this is mediated by the endogenous opioid system.

• Journal of Acupuncture Research
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• v.22 no.1
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• pp.145-153
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• 2005

Objective : It was well knownth at electro-acupuncture(EA) of ST36 in the hindimb affected a gastrointestinal disease. This study was designed to compare the factors affected EA`s effect such as stimulation duration, operating time and inserted needle`s length. Meyhods and Results : The lightly anaesthetized rat was used and serum gastrin level was observed in this study. Standard stimulation of ST36 consisted of a 5 mA at 2 Hz for a duration of 30 min, applied at 5 times the threshold to evoke muscle twitch. In the study of comparing stimulation duration, 60 min of EA produced no effect, whereas both of 10 and 30 min of EA increased gastrin level. When operating time was altered, both of 5 and 10 days only produced effects of increasing gastrin level. When comparing the depth of inserting needle, stimulation of the outer layer increased gastrin serum level as well as stimulation of full length. Conclusion : These data suggest that the difference of factors affected EA`s effect such as stimulation duration, operating time and inserted needle`s length produced difference effects.

• Journal of Acupuncture Research
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• v.30 no.4
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• pp.69-78
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• 2013

Objectives : The purpose of this study is to examine whether Eucommiae Cortex pharmacopuncture may affect to the neuropathic pain in a rat model. Methods : To produce the model of neuropathic pain, under isoflurane 2.5 % anesthesia, underwent tight ligation by 6.0 silk thread and transection of the tibial and sural nerves, leaving the common peroneal nerve intact. After neuropathic surgery, the author examined if the exhibited the behavioral sign of allodynia. The allodynia was assessed by stimulating the plantar with Dynamic Plantar Aesthesiometer. Three days after the neuropathic surgery, Eucommiae Cortex pharmacopuncture was injected at Sinsu($BL_{23}$) once every week for 6 weeks. After that, the author examined the withdrawal response of neuropathic rats' leg by Dynamic Plantar Aesthesiometer. And also the author examined Bax, Bcl-2, Bax/Bcl-2 ratio in the spinal cord of neuropathic rats and the change of WBC, RBC, HGB, HCT count in the blood of neuropathic rats. Results : 1. The Eucommiae Cortex pharmacopuncture decreased the withdrawal response of mechanical allodynia that assessed with Dynamic Plantar Aesthesiometer in EC2-$BL_{23}$ group as compared with control group. 2. The Eucommiae Cortex pharmacopuncture decreased Bax/Bcl-2 ratio in EC1-$BL_{23}$, EC2-$BL_{23}$ group. But The Eucommiae Cortex pharmacopuncture injected at Sinsu($BL_{23}$) didn't change Bax, Bcl-2 expression level in the all group. 3. The Eucommiae Cortex pharmacopuncture decreased WBC count in EC1-$BL_{23}$, EC2-$BL_{23}$ group. Conclusions : We have noticed that Eucommiae Cortex pharmacopuncture decreased mechanical allodynia in the model of neuropathic pain compared with the control group. Bax/Bcl-2 ratio in spinal cord of that group was also decreased compared with the control group. This study can be used as a basic resource on a study and a treatment of neuropathic pain.

• Physical Therapy Korea
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• v.18 no.1
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• pp.83-92
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• 2011

Osteoarthritis is a degenerative joint disease and is led to physical disability. Yet the development of effective disease-modifying treatments has lagged. In this study, I examined the effect of physical therapeutic intervention through microcurrent stimulation and attempt to find which degree of intensity, either 25 ${\mu}A$ or 500 ${\mu}A$ with a regular 5 pps pulse, is more effective in the osteoarthritis. Osteoarthritis was induced with a mixture of 2% carrageenan and 2% kaolin in 26 male Sprague-Dawley rats. The mixture (0.1 $m{\ell}$) was injected into the intra-articular capsule of knee joint once a week for three weeks. Five animals did not show degenerative changes by radiological findings and excluded in the following experiment. Osteoarthritic animals were randomly divided into 3 groups ($n_1$, $n_2$, $n_3$=7/each): untreated, treated with 25 ${\mu}A$, treated with 500 ${\mu}A$. All experimental groups received microcurrent stimulation for four weeks (15 min/day, 5 days/week). The ethological inspection of foot print analysis on the walking corridor was accomplished every week. Histological preparations and immunohistochemical staining with insulin-like growth factor-1 were also done in the articular cartilages. All of these parameters were compared with those of osteoarthritic control group (n=7). The ethological inspection of foot print analysis revealed that changes of walking track (paw width) and stride length was significantly increased in both experimental groups. The better results were observed in experimental group treated with 25 ${\mu}A$ intensity without significance than group treated with 500 ${\mu}A$. Histological preparations disclosed that routine hyaline cartilage of articular surface were altered to fibrous cartilage in untreated group and experimental group treated with 500 ${\mu}A$ intensity. But a little changes were seen in experimental group treated with 25 ${\mu}A$ intensity. Immunolocalization of insulin-like growth factor-1 was simultaneously decreased according to the duration of osteoarthritis, and did not show significant difference among the groups. In this study discovered that the microcurrent stimulation, especially 25 ${\mu}A$ intensity, had a positive effect by the ethological inspection, histological and immunohistochemical stainings. These results suggest that microcurrent stimulation with low-intensity might be effective in the promotion of healing process for the osteoarthritis.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.19 no.3
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• pp.265-286
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• 1997

Bone morphogenetic proteins(BMPs) are a group of transforming growth factor beta(TGF-${\beta}$)-related factors and multifunctional proteins, especially the only known biologic factors capable of inducing endochondral bone formation at an extraskeletal site. This study was performed to investigate the effect of the partially purified porcine BMP(pBMP) at an ectopic site. PBMP was partially purified from porcine bone matrix and its activity was monitored by an in vivo bioassay. The purification method utilized extraction of the bone-inducing activity with 4M guanidine, followed by chromatography on heparin-Sepharose. Active fractions were assayed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. And the fractions were reconstituted with inactive insoluble collagenous bone matrix from rats, acid soluble type I collagen from rat tail and chondroitin-6-sulfate sodium salt and implanted into the pectroralis muscle pouches of Sprague-Dawley rats. And the carrier complex was implanted on the opposite side as control. The rats were sacrificed at the day of 1st, 3rd, 5th, 7th, 11th, 14th and 21st after implantation and examined histologically, radiologically and biochemically. And alkaline phosphatase activity and calcium content were used as indices of bone formation. The results were as follows ; 1. Active fractions were localized in a zone between 31 and 40 KDa on SDS-PAGE. 2. The implanted 3.0mg of the partially purified pBMP induced cartilage and bone in the muscle tissue of rats through an endochondral ossification process. 3. Inactive insoluble bone matrix, type I collagen and chondroitin-6-sulfate have functioned as carriers for pBMP, but revealed some foreign body reactions. 4. Soft X-ray didn't reveal significant change between the experimental and the control group. 5. The alkaline phosphatase activities in the experimental group of 5th, 7th, 11th, 14th and 21st were increased significantly compared with control (p<0.01) with the peak in the group of 11th day. 6. With time, the calcium content of the experimental group increased. And the calcium contents in the experimental group of 11th, 14th and 21st were increased significantly compared with control (p<0.01).

• The korean journal of orthodontics
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• v.20 no.3 s.32
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• pp.463-498
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• 1990

전기적 자극에 의한 골성장기전의 개념을 이용하여 임상적 효율성을 증진시키기 위한 연구는 현재 교정학을 비롯한 치과영역에서 활발히 진행되고 있는 분야 중의 하나이다. 전기적 자극의 여러 형태 중의 하나인 전자기장과 하악의 기능적 전방 이동을 유도하는 악기능교정장치가 백서의 하악과두 성장에 미치는 영향을 구명하기 위하여 본 연구를 시행하였다. 생후 4주된 Sprague Dawley계 백서 48마리를 대조군 12마리, 실험군 36마리로 나누고, 실험군은 다시 전자기장을 가한 군, 하악골 전방이동 장치를 장착시킨 군, 전자기장과 하악골 전방이동 장치를 병용시킨 군으로 분류하여 각각 12마리씩 실험동물을 배정하였다. 각 군의 실험동물은 15 HZ의 특수 전자기장이나 하악전방이동 자치가 하루10시간씩 작용되도록 특별히 제작한 실험장치 속에 넣어 1주간, 4주간씩 사육하여 희생시킨 후 하악골을 분리하고 연조직을 박리한 후 $10\%$ formalin에 보관하였다. 하악골 길이를 측정하기 위해 0.05mm까지 계측 가능한 캘리퍼를 이용하여 하악과두의 후연에서 이공까지의 거리를 계측하였고, 하악과두를 절제하여 0.5M EDTA에 탈회시켜 파라핀 포매를 하였다. 표본의 절단방향은 시상평면에 평행하게 하여 $6{\mu}m$두께로 연속절단 하였으며, 그 중 중심의 3절편을 취하여 통법에 의한 H-E 중염색을 시행하였다. 하악골 계측과 H-E 중염색 표본을 통한 조직학적 관찰을 통해 다음과 같은 결론을 얻었다. 1. 4주군에서 전자기장만에 노출된 실험군은 대조군에 비해 하악골 길이가 유익성 있게 증가되었다. 2. 전자기장과 하악골 전방이동 장치를 병용한 실험군은 하악골 전방이동장치만을 사용한 실험군에 비하여 하악골 길이가 증가되었다. 3. 전자기장에 노출된 실험군은 전구 연골아세포(prechondroblast)의 증식, 비대연골 세포층의 세포간질 및 연골내 골화층의 석회화가 모두 증가되었다. 4. 본 실험에 사용한 15 HZ전자기장의 주요작용부위는 백서의 하악과두 성장지역 중 연골내 골화의 석회화 지역이며, 또한 이는 하악골 전방이동 장치와 병용시 하악과두 성장을 촉진시킬 수 있음이 관찰되었다.

• International Journal of Oral Biology
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• v.31 no.4
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• pp.141-148
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• 2006

The effects of adenosine triphosphate(ATP) on salivary glands have been recognized since 1982. The presence of purinergic recepetors(P2Rs) that mediate the effects of ATP in various tissues, including parotid and submandibular salivary gland, has been supported by the cloning of receptor cDNAs and the expression of the receptor proteins. P2Rs have many subtypes, and the activation of these receptor subtypes increase intracellular $Ca^{2+}$, a key ion in the regulation of the secretion in the salivary gland. The apical pores of taste buds in circumvallate and foliate papillae are surrounded by the saliva from von Ebner salivary gland(vEG). Thus, it is important how the secretion of vEG is controlled. This study was designed to elucidate the roles of P2Rs on salivary secretion of vEG. Male Sprague-Dawley rats (about 200 g) were used for this experiment. vEG-rich tissues were obtained from dissecting $500-1,000\;{\mu}m$ thick posterior tongue slices under stereomicroscope view. P2Rs mRNA in vEG acinar cells were identified with RT-PCR. To observe the change in intracellular $Ca^{2+}$ activity, we employed $Ca^{2+}-ion$ specific fluorescence analysis with fura-2. Single acinar cells and cell clusters were isolated by a sequential trypsin/collagenase treatment and were loaded with $10\;{\mu}M$ fura -2 AM for 60 minutes at room temperature. Several agonists and antagonists were used to test a receptor specificity. RT-PCR revealed that the mRNAs of $P2X_4$, $P2Y_1$, $P2Y_2$ and $P2Y_3$ are expressed in vEG acinar cells. The intracellular calcium activity was increased in response to $10\;{\mu}M$ ATP, a P2Rs agonist, and 2-MeSATP, a $P2Y_1$ and $P2Y_2R$ agonist. However, $300\;{\mu}M\;{\alpha}{\beta}-MeATP$, a $P2X_1$ and $P2X_3R$ agonist, did not elicit the response. The responses elicited by $10\;{\mu}M$ ATP and UTP, a $P2Y_2R$ agonists, were maintained when extracellular calcium was removed. $10\;{\mu}M$ suramin, a P2XR antagonist, and reactive blue 2, a P2YR antagonist, partially blocked ATP-induced response. However, when extracellular calciums were removed, suramin did not abolish the responses elicited by ATP. These results suggest that P2Rs play an important role in salivary secretion of vEG acinar cells and the effects of ATP on vEG salivary secretion may be mediated by $P2X_4$, $P2Y_1$, $P2Y_2$, and/or $P2Y_3$.

• International Journal of Oral Biology
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• v.41 no.3
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• pp.133-139
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• 2016

The ultrastructural parameters related to synaptic release of endings which are presynaptic to tooth pulp afferent terminals (p-endings) were analyzed to understand the underlying mechanism for presynaptic modulation of tooth pulp afferents. Tooth pulp afferents were labelled by applying wheat-germ agglutinin conjugated horseradish peroxidase to the rat right lower incisor, whereafter electron microscopic morphometric analysis with serial section and reconstruction of p-endings in the trigeminal oral nucleus was performed. The results obtained from 15 p-endings presynaptic to 11 labeled tooth pulp afferent terminals were as follows. P-endings contained pleomorphic vesicles and made symmetrical synaptic contacts with labeled terminals. The p-endings showed small synaptic release-related ultrastructural parameters: volume, $0.82{\pm}0.45{\mu}m^3$ ($mean{\pm}SD$); surface area, $4.50{\pm}1.76{\mu}m^2$; mitochondrial volume, $0.15{\pm}0.07{\mu}m^3$; total apposed surface area, $0.69{\pm}0.24{\mu}m^2$; active zone area, $0.10{\pm}0.04{\mu}m^2$; total vesicle number, $1045{\pm}668.86$; and vesicle density, $1677{\pm}684/{\mu}m^2$. The volume of the p-endings showed strong positive correlation with the following parameters: surface area (r=0.97, P<0.01), mitochondrial volume (r=0.56, P<0.05), and total vesicle number (r=0.73, P<0.05). However, the volume of p-endings did not positively correlate or was very weakly correlated with the apposed surface area (r=-0.12, P=0.675) and active zone area (r=0.46, P=0.084). These results show that some synaptic release-related ultrastructural parameters of p-endings on the tooth pulp afferent terminals follow the "size principle" of Pierce and Mendell (1993) in the trigeminal nucleus oralis, but other parameters do not. Our findings may demonstrate a characteristic feature of synaptic release associated with p-endings.