• The Korean Journal of Pain
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• 제21권2호
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• pp.112-118
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• 2008

Background: This study was conducted to quantitatively analyze proteins associated with the calcitonin gene-related peptide (CGRP) in cerebrospinal fluid (CSF) that was obtained from a rat model of chronic neuropathic pain following administration of intrathecal $CGRP_{8-37}$. Methods: Male Sprague-Dawley rats (100-150 g, 5-6 wks) were divided into two groups, sham controls and neuropathic pain models. At the time of operation for neuropathic pain model, an intrathecal catheter was threaded through the intrathecal space. At 1 or 2 wks after the operation (maximum pain state), a test dose of 1, 5, 10, or 50 nM of $CGRP_{8-37}$ was injected into the intrathecal catheter and the CSF was then aspirated. Conventional proteomics to evaluate the CSF were then performed using high resolution 2-D, gel electrophoresis followed by computational image analysis and protein identification by mass spectrometry. Results: Treatment with $CGRP_{8-37}$ effectively alleviated mechanical allodynia in a dose dependent manner. The most effective response was obtained when a dose of 50 nM was administered, but significant differences were obtained following administration of only 5 nM $CGRP_{8-37}$. Furthermore, the results of the proteomic analysis were consistent with the experimental results. Specially we detected 30 differentially expressed spots in 7 images when 2-D gel electrophoresis was conducted. The intensity of 6 of these spots (spot number: 20 and 26-30) was found decrease the $CGRP_{8-37}$ dose increased; therefore, these spots were evaluated by mass spectrometry. This analysis identified 2 different proteins, CGRP (spot numbers: 26-30) and neurotensin-related peptide (spot number: 20). Conclusions: The results of this study suggest that CGRP plays a role in chronic central neuropathic pain and is a major target of chronic neuropathic pain management.

• 대한치과교정학회지
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• 제26권5호
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• pp.581-590
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• 1996

교정력에 의하여 치주인대내에 생긴 압박측에서는 국소적 염증반응과 골 흡수가 활발히 일어나며 신장측은 골생성이 활발하므로 위 두 부위는 서로 다른 조직반응이 일어난다. 치아이동에 관여하는 것으로 알려진 CGRP 면역 양성 신경섬유도 압박 및 신장 부위에 따라 다를 것으로 생각된다. 저자등은 CGRP면역 양성 신경섬유의 교정력 적용시간에 따른 변화를 관찰하고 압박 및 신장력을 받는 부위에 따른 변화를 관찰하기 위하여 200gm내외의 생후 9주령 백서 상악 제1 대구치에 Ni-Ti coil spring으로 80gm내외의 교정력을 12시간, 1일, 3일, 7일, 12일동안 가한 후 희생하여 면역 조직화학법으로 염색하여 관찰하였다. $\cdot$ 교정력 적용 12시간 및 1일군에서 압박 및 신장측 모두에서 CGRP 면역 양성 신경섬유가 감소되었다. $\cdot$ 교정력 적용 3일군에서 CGRP면역 양성 신경섬유는 많은 가는 분지가 관찰되며 증가되었고 특히 신장측에서 현저하였다. $\cdot$ 교정력 적용 7일군에서는 비교적 굵은 CGRP 면역 양성 신경섬유도 증가 되었으며 신장측에서 현저하였다. $\cdot$ 교정력 적용 12일군에서 압박측 치주인대에서는 거의 정상과 유사하였으나 신장측치주인대에서는 증가된 채 유지되었다. $\cdot$ CGRP면역 양성 신경섬유는 압박측보다 신장측에서 더 많이 증가하여 치아이동시 CGRP가 말초조직에서 염증반응에 관여함과 동시에 골 개조에도 상당한 연관성을 가지고 있을 것으로 생각된다.

• 대한소아치과학회지
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• 제24권2호
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• pp.339-351
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• 1997

출생 후 성장 발육에 따른 흰쥐 유곽유두의 신경분포 및 변화를 관찰하기 위하여 생후 1, 5, 10, 15, 20, 25, 35일 및 완전히 성장한 흰쥐를 대상으로 calcitonin gene-related peptide (CGRP)를 이용한 면역조직화학적 방법으로 관찰하여 다음과 같은 결과를 얻었다. CGRP-면역양성 신경섬유들이 혀의 양측에서부터 유두 내로 진입하여 성장발육에 따라 그 분포밀도가 증가하다가 10일군부터는 상피의 직하방에 특히 풍부하게 분포하여 subepithelial plexus를 형성하고 있었다. 1일군에서는 CGRP-면역양성 신경섬유의 특징적인 양상인 염주알 모양이 명확히 관찰되었으나 그 이후로는 염주알 모양은 다소 감소하였고 하나의 선으로 보이는 경우도 있었다. 상피 내로 진입하는 CGRP-면역양성 신경섬유들은 5일군에서 드물게 관찰되었으며 taste pore를 가진 성숙한 미뢰가 처음으로 관찰되는 10일군에서 그 수가 증가하여 출생 후 15일군에서는 상피 전반에 걸쳐 풍부하게 관찰되었다. 그러나 그 이후 20일군부터는 미뢰가 존재하는 trench wall의 하방 2/3부위의 CGRP-면역양성 신경섬유는 감소하고 유두의 top surface와 trench wall의 상방 1/3에 CGRP-면역양성 신경섬유가 제한적으로 관찰되었다.

• Restorative Dentistry and Endodontics
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• 제24권2호
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• pp.372-380
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• 1999

The purpose of this study was to investigate the distribution of calcitonin gene-related peptide(CGRP) containing nerve fivers after pulp exposure in rats. The Spague-Dawley rats weighing about 250 - 300g were used. The animals were devided into normal control group and experimental groups. Experimental animals were sacrified on 2, 4, 7, 10 days after pulp exposure. The maxillary teeth and alveolar bone were removed and immersed in the 4% paraformaldehyde plus 0.1M phosphate buffer (pH 7.4). Serial frozen $50{\mu}m$ thick sections were cut with a cryostat. In the immunohistochemical staining procedure, the rabbit CGRP antibody was used as a primary antibody. The sections were incubated for 48 hours at $4^{\circ}C$, and placed into biotinylated anti-rabbit IgG as a secondary antibody and incubated in ABC (avidin-biotin complex), The sections were visualized by 0.05% 3.3 diaminobenzidine tetrahydrochloride. The results of this study were as follows: 1. In control group, CGRP containing nerve fibers ran parallel to the long axis of root and reached the coronal pulp. They were distributed on Raschkow plexus under the odontoblastic layer. 2. In 2 day group after pulp exposure, tissue necrosis and acute inflammation occurred and CGRP containing nerve fibers increased. In 4 day group, the necrotic tissue extended to the pulp and CGRP containing nerve fibers were distributed around the inflammation zone. 3. In 7 day group after pulp exposure, pulp necrosis occurred, and in 10 day group, the abscess under the necrotic pulp extended to the root apex area and CGRP containing nerve fibers were not observed in root canals. 4.The sprouting of CGRP nerve fibers was most remarkable at the pulp chamber under injury in 4 day group, and it was found at inflammation zone under the necrotic tissue in 7 day group and the remaining root pulp tissue in 10 day group. As mentioned above, CGRP nerve fibers had a tendency to increase around the inflammatory zone, especially around the acute inflammation tissue, when compared with control group. It is suggested that CGRP nerve fibers maybe related to the control of inflammatory response of pulp tissue.

• 동의생리병리학회지
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• 제16권1호
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• pp.117-123
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• 2002

To investigate the antiinflammatory and analgesic effects of Sophorae radix extracts administered to the arthritic rat model, immunohistochemical stains for CGRP in the L4, L5 and L6 spinal cord and ganglia were done, and paw swelling thickness were measured. Complete Freund,s Adjuvant(CFA) were injected to subcutaneous tissue of left foot paw of rats to induce arthritis. Sophorae radix extracts was administered immediately after CFA injection for 10 days. The spinal cord and ganglia were frozen sectioned(30㎛). These sections were stained by CGRP immunohistochemical staining method, and observed with light microscope. The results were as follows : 1. The change of paw swelling thickness of experimental group decreased from 4 day to 10day after CFA injection compared to control group. 2. The change of differential leukocytes counts of experimental group increased the ratio of lymphocytes. and decreased the ratio of neutrophils compared to control group. 3. The change of CGRP immunoreactive nerve fiber of dorsal horn of experimental group was dense stained compared to control group. 4. The number of CGRP immunoreactive neurons of L4 and L5 spinal cord of experimental group was less than in those control group. These results suggested that Sophorae radix extracts reduces the number of CGRP immunoreactive neurons and nerve fibers of spinal cord and ganglia, and decrease paw swelling thickness in arthritic rat model, which may be closely related to analgesic and antiinflammatory effects of Sophorae radix.

• 대한기관식도과학회지
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• 제2권1호
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• pp.97-102
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• 1996

The larynx has three major functions such as protective reflex, respiration and phonation, and is richly innervated by sensory, sympathetic and parasympathetic nerves. The sensory innervation of the laryngeal mucosa, which is involved in the perception of pain, mechanical and chemical irritation, prtects the airway via various laryngeal reflexes. We studied the distribution of Substance P (SP) and Calcitonin Gene-Related Peptide (CGRP) sensory fibers in the rat's larynx using the immuno-histochemical methods. Many SP and CGRP immunoreactive fibers were found in all regions of the laryngeal mucosa except the vocal cords. SP immunoreactive fibers showed a very similar distribution to the CGRP fibers in the epithelium and submucosa. But SP immunoreactive fibers were sparser than CGRP immunoreactive fibers in distribution density. Both reactive fibers were denser in the supraglottic region than subglottic region. Especially, intraepithelial fibers displayed the densest innervation to the laryngeal surface of the epiglottis. h the subepithelium, SP and CGRP immunoreactive fibers were distributed along the wall of vessels and around the glands. The present results suggest that the regional distribution of SP and CGRP immunoreactivity may be responsible for the protective reflex function of the laryngeal inlet.

• 대한소아치과학회지
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• 제26권1호
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• pp.44-52
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• 1999

The purpose of this study was to investigate the distribution of CGRP immunoreactive nerve fibers in rat squamosomandibular joint with postnatal development. Squamosomandibular joint with adjacent tissue of 8 groups of rats(1, 5, 10, 15, 20, 25, 35-postnatal day and adult groups) were removed on bloc and processed for immunohistochemistry and were subjected to light microscopic examination. The results obtained were as follows; 1. The anterior portion of the articular disk was most densely innervated, followed by the posterior, lateral, and medial portions in each group. 2. Increase of CGRP immunoreactive nerve fibers was evident in 10, 15, 20 postnatal day groups. 3. Almost no CGRP immunoreactive nerve fibers were observed in articular surface of disk proper, mandibular fossa and condyle head of each group. These result suggest that CGRP immunoreactive nerve fibers increased in synovial membrane and peripheral portion of articular disk during 10-20 postnatal day may play a important role in squamosomandibular joint function after weaning period.

• 대한수의학회지
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• 제37권4호
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• pp.719-726
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• 1997

Dorsal root ganglion(DRG) cells are primary sensory neurons which contain some biologically active neuropeptides which play a role as neurotransmitters or neuromodulators. This study was performed to observe normal distribution of calcitonin gene-related peptide (CGRP) and substance P (SP) immunoreactive cells and colocalization of CGRP and SP in a single DRG cell of the lumbar DRGs($L_1{\sim}L_6$) in the Wistar Kyoto(WKY) rat by immunohistochemistry. About 55.8% of DRG cells contained CGRP-immunoreactivity, while about 12.7% of DRG cells showed SP-immunoreactivity. There was no significant difference in percentage of each neuropeptied-immunoreactive cells between each neuropeptide-immunoreactive cells between each levels of DRGs ($L_1{\sim}L_6$) (p>0.01). In size distribution, CGRP-immunoreactive cells were identified below $1,500{\mu}m^2$; SP-immunoreactive cells below $600{\mu}m^2$. In serial sections, about 86.7% of the SP immunoreactive cells contained CGRP immunoreactivity.

• 대한수의학회지
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• 제36권2호
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• pp.265-276
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• 1996

This study was performed to identify the localization of several neuropeptides; calcitonin gene-related peptide(CGRP), substance P(SP), vasoactive intestinal polypeptide(VIP), neuropeptide Y(NPY), serotonin(5-HT) and neurotensin in the tongue of Korean native goat(Capra hircus) by immunohistochemical method. The results were summarized as follows: CGRP- and SP-immunoreactive fibers were observed as moderate immunoreactivity at the subepithelial plexus and subgemmal fibers in lamina propria of lingual papillae, but not seen in intragemmal, intergemmal, perigemmal fibers as well as in the supporting, basal and taste cells. Fibers around the acinus of the von Ebner's gland and blood vessels showed weak immunoreactivities against CGRP and SP. In the intrinsic ganglion cells, CGRP- and SP-immunoreactivities were not observed. The distribution patterns of VIP- and NPY-immunoreactive fibers were similar to CGRP-or SP-immunoreactive fibers, but their immunoreactivities were stronger than those of CGRP- or SP immunoreactive fibers. The immunoreactivities to VIP or NPY were seen in the intrinsic ganglion. Only a few serotonin immunoreactive fibers were seen in some filiform or fungiform papillae. Neurotensin immunoreactivity was not observed in the tongue of Korean native goat.

• 대한소아치과학회지
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• 제35권1호
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• pp.11-17
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• 2008

본 연구는 Er:YAG laser에 의한 치수의 초기 조직반응과 calcitonin gene-related peptide (CGRP) 함유 신경섬유의 발현을 비교 평가하고자 시행하였다. Er:YAG laser와 conventional bur를 이용하여 상악 제1대구치의 교합면에 와동을 형성하였다. 와동형성 후 48시간 후에 조직을 처리하여 hematoxylin-eosin 염색과 CGRP 면역조직화학을 시행한 후 다음과 같은 결과를 얻었다. 1. Conventional bur를 이용하여 삭제한 경우, 상아질 바닥면은 균열이 없는 평면의 균일한 모양을 보였으며, Er:YAG laser의 경우에는 상아질의 면은 bur에 비해서 파동형의 불규칙적인 형태와 바닥면에서 상아세관의 주행방향을 따라 미세한 균열을 보였다. 2. Er:YAG laser에 의해 손상 받은 치수에서는 혈관확장과 노출된 치수의 손상부위를 따라 bur에서 손상 받은 치수보다 더 많은 염증세포의 침윤과 상아모세포층의 파괴를 보였다. 3. 노출손상 후에 치수신경 말단에서 CGRP 발현은 bur와 laser군 모두에서 손상을 받지 않은 정상 치수에 비해서 증가된 양상을 보였으며, 그 증가된 정도는 laser의 경우에서 더 강하였다. 위의 결과들을 종합하면, Er:YAG laser가 치수손상을 최소화하여 적용되면 치아우식 제거와 와동형성과 같은 여러 수복치료 시 유용하게 사용될 수 있으리라 사료된다.