• Restorative Dentistry and Endodontics
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• 제20권2호
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• pp.527-537
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• 1995

The effect of calcitonin gene-related peptide (CGRP), substance P (SP) and electrical stimulation of the tooth on the intradental nerve activtiy (INA) was investigated in anesthetized cats. The INA was recorded from single pulp nerve units dissected from the inferior alveolar nerve under stereomicroscope. The INA elicited by 3 minute application of 4M NaCl in deep dentinal cavity was compared before and after stimulation at 10 minute intervals. The magnitude of INA was calculated as the total number of nerve impulses produced in given period, and the changes of INA are expressed as % of control INA. The results obtained were as follows. 1. 16 single pulp nerve units were classified as 14 $A{\delta}$-fibers (3.4~19.4m/sec) and 2-fibers (1.5~1.7m/sec) according to the conduction velocity. 2. 4M NaCl evoked an irregular bursts of spikes which continued until washing out. Isotonic saline did not affect INA to subsequent applications of the hypertonic NaCl solution (P>0.05). 3. Local application of CGRP ($200{\mu}g$/ml) in deep dentinal cavity reduced the INA induced by 4M NaCl in $A{\delta}$-fiber units (P<0.01) and some units of those responded to CGRP during application. 4. Local application of SP ($100{\mu}g$/ml) in deep dentinal cavity reduced the INA induced by 4M NaCl in AS-fiber units (p<0.05), but increased the INA in C-fiber unit coincided with large reduction of the INA of $A{\delta}$-fiber units. 5. Monopolar electrical stimulation applied to the crown at intensities high enough to excite C-fibers (12V, 5ms, 10Hz, 10~30min) decreased the INA in $A{\delta}$-fiber units (P<0.01) and systemic pretreatment with phenoxybenzamine (3mg/kg, i.v.) enhanced this inhibitory effect (P<0.01). On the contrary, electrical stimulation increased the INA in C-fiber unit.

• Archives of Plastic Surgery
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• 제34권2호
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• pp.149-155
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• 2007

Purpose: This study was to investigate how the heparin, which has been known to induce neovascularization by MMP in the infarcted tissue of the myocardium, had influence on the expression of mRNA of MMP 1,2,9 of the skin wound of rat. Methods: Full depth skin wounds were created on the dorsum of Sprague-Dawley 60 rats. The experimental rats were divided into two groups according to the concentration of heparin($100{\mu}g/ml$ in 20, $300{\mu}g/ml$ in 20). Heparin soaked gelatin sponges in different concentration were inserted into the pocket of experimental rats and the wounds were closed. Normal saline soaked gelatin sponges were used in control rats. Wounds were harvested at 48 and 72 hours after closure. We performed histologic study in H-E stain. RNA was isolated from the harvested tissue and then real time polymerase chain reaction was performed to determine the gene expression of MMP-1,2,9. Results: We observed that inflammatory cell decreased in heparin soaked group and heparin increased the expression of MMP-1,9 mRNA of dorsal wound of rat at 72 hours (p < 0.05). Conclusion: This result suggest that heparin may be used inducing another factor inducing scarless wound healing by increasing MMP.

• 대한본초학회지
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• 제24권1호
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• pp.79-88
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• 2009

Objectives : It has been recently shown that Piperis Longi Fructus (PLF) is involved in the reduction of eosinophil recruitment and production of Th2 cytokines in vivo. However, the main therapeutic mechanisms of PLF remains a matter of considerable debate. To investigate the therapeutic mechanisms of PLF, we examined the influence of PLF on regulatory T cells number, IgE, histamine production in vivo and Th1/Th2 cytokine balance in vitro. Methods : All mice were immunized on two different days (21 days and 7 days before inhalational exposure) by i.p. injections of 0.2 $m\ell$ alum-precipitated Ag containing 100 ${\mu}g$ of OVA bound to 4 mg of aluminum hydroxide in PBS. Seven days after the second sensitization, mice were exposed to aerosolized ovalbumin for 30 min/day on 3 days/week for 12 weeks(at a flow rate of 250 L/min, 2.5％ ovalbumin in normal saline) and PLF (150 mg/kg) were orally administered 3 times a week for 8 weeks. Splenocytes from C57BL/6 mice at 8 weeks of age were stimulated with anti-CD3 (1 mg/ml) plus anti-CD28 (1 mg/ml) antibody for 48hrs. IL-4 and IFN-$\gamma$ in the culture supernatants were measured by ELISA Results : The suppressive effects of PLF on asthma model were demonstrated by the increase the number of regulatory T cells and by reducing IgE, histamine production in vivo and modulation of Th1/Th2 cytokine balance. Conclusions : These results indicate that PLF has a deep inhibitory effects on asthma model mice by increase the number of regulatory T cells, and by reducing IgE, histamine production.

• The Korean Journal of Pain
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• 제9권2호
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• pp.311-317
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• 1996

Background: Following peripheral nerve injury, rats will show a tactile allodynia and hyperalgesia. But the mechanism of allodynia is still obscure. Previous studies have shown this allodynia was reversed by intrathecal alpha-2 agonists and NMDA antagonists, but not by morphine. In formalin test, either the pretreatment of NMDA antagonist or morphine prevents the hyperalgesia. The present studies, using rats rendered allodynic by ligation of the left L5 and L6 nerves, aimed to investigate the effects of pretreatment of MK-801 and morphine on the development of tactile allodynia. Methods and Material: Male Sprague-Dawley rats (100~150g) were anesthetized with halothane, the left L5 and L6 spinal nerves were ligated tightly by 6-0 black silk. For sham operation muscle dissection was performed but the spinal nerve was not ligated. For pretreatment of drugs, MK-801 (NMDA antagonist; 0.3 mg/kg). CNQX (non-NMDA) antagonist; 0.3 mg/kg), morphine (1 mg/kg) or saline (placebo) was administered subcutaneously 30 minutes before operation. A second dose was administered subcutaneously 24 hours after operation and further doses were given daily for 2 days further. The volume of injection was 5 ml/kg. To assess the mechanical allodynia, paw withdrawal thresholds of ipsilateral limb were determined using 8 von Frey hairs. Results: Within 2 days saline, CNQX or morphine injected rats developed tactile allodynia (paw withdrawal threshold was about 2g), and persisted for over 2 weeks. Pretreatment of MK-801 delayed the development of tactile allodynia for 3 days comparing to that of saline injected rat. Conclusion: NMDA receptor in the central nerve system plays an important role in the development of tactile allodynia induced by peripheral nerve injury. But the mechanism may be different from hyperalgesia developed in formalin test.

• 치위생과학회지
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• 제15권2호
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• pp.98-105
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• 2015

본 연구는 외과적으로 형성된 흰쥐 하악골 결손부에 S.mutan에서 분리 정제된 mutan을 주사하고, ${\mu}CT$로 촬영하여 골결손부의 치유에 미치는 영향을 3차원 골 미세구조 지표로 평가하기 위하여 실시하였다. 흰쥐 치조골에 임계 크기 결손부인 1.5 mm 지름의 원형 결손부를 형성하여 대조군에는 동일량의 saline을 투여하고, 실험군으로 LPS와, mutan을 주사하여 골치유에 미치는 영향을 관찰하였다. 수술 후 4주에 희생하고 치유 결과를 3차원 미세구조의 형태 계측학적 지표값을 비교 분석하여 다음과 같은 결론을 얻었다. 실험동물의 체중변화는 총 12회 투여 기간 동안 일정한 증가를 나타냈다. 8회차부터 mutan 처리군은 대조군과 비교하여 유의한 차이를 보였으며(p<0.05), 9회차에는 mutan 처리군이 대조군과 LPS 처리군과 비교하여(p<0.05), 10~12회차에는 LPS 처리군과 비교하여 유의한 체중증가를 나타냈다(p<0.01). Mutan이 생체 내에서 하악의 골치유 효과를 알아보기 위해 ${\mu}CT$를 통해 형태계측학적 지표 분석 결과, 대조군에 비해 LPS 처리군에서 뚜렷한 골결손부 치유 지연 및 골흡수 양상을 보였으며, mutan 처리군 은 LPS의 작용보다 미약하지만 골치유를 지연시키는 것으로 확인되었다. ${\mu}CT$를 통해 골밀도 분석 결과, 대조군과 비교하여 LPS와 muatn 처리군에서 낮은 골밀도치를 나타냈고, mutan은 LPS 처리군보다 유의하게 높은 골밀도치를 보였다(p<0.01).

• 대한소아치과학회지
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• 제38권2호
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• pp.129-136
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• 2011

치수 및 치근단의 감염은 미생물에 의해 야기되며, 성공적인 근관치료는 근관 내의 미생물의 수를 감소시키는 것에 의해 좌우된다. 서양산 고추냉이(Horseradish, Armoracia ruticana)의 주성분인 Allyl isothiocyanate (AIT)는 여러 연구에서 항균효과가 있음이 보고되었다. 이에 본 연구에서는 근관세척제로써 서양산 고추냉이 추출물의 항균효과를 알아보고자 발거된 사람의 치아에 Enterococcus faecalis를 접종하여, 대표적인 근관세척제인 차아염소산나트륨(NaOCl)과 항균력을 비교하여 다음과 같은 결론을얻었다. 1. 치큰관 내를 21일간 E. faecalis에 감염시키고, 근관세척제로써 각 실험군을 1분간 적용한 후 1차 표본(S1)을 채취하여 배양한 결과, 서양산 고추냉이 뿌리 추출물은 $5.815{\times}10^3$ CFU/ml, NaOCl은 $3.465{\times}10^3$ CFU/ml이었다(p=0.086). 2. 1차 표본을 채취한 후 실험 치아를 7일 동안 더 배양하여 2차 표본(S2)을 채취한 결과 서양산 고추냉이 뿌리 추출물은 $3.100{\times}10^3$ CFU/ml, NaOCl은 $5.252{\times}10^5$ CFU/ml로, 서양산 고추냉이 뿌리 추출물을 적용한 실험 치아에서 더 적은 수의 세균수가 측정되었다(p<.05). 3. NaOCl로 근관세척을 시행한 치아의 1차 표본(S1)과 2차 표본(S2)을 채취하여 배양한 결과 2차 표본에서 CFU 값이 더 증가하였다(p<.05). 반면에 고추냉이 추출물로 근관세척을 시행한 치아의 1차 표본(S1)과 2차 표본(S2)을 채취하여 배양한 결과 유의한 차이는 없는 것(p=0.076)으로 보아 근관세척후 7일 후에도 향균효과가 지속됨을 알 수 있었다.

• 생명과학회지
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• 제25권2호
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• pp.164-173
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• 2015

유산 균주들을 신생아 및 성인의 분변으로부터 분리하여 그 중에서도 가장 중금속 흡착능이 크고 중금속에 내성이 있다고 사료되는 균주 KP-3를 선별하여 동정한 결과 Lactobacillus sp.으로 밝혀졌다. 동물 실험에서는 생후 4주령의 Sprague dawley male rat에게 선별한 균주 KP-3를 투여하여 혈액 및 장기 내의 중금속 축적 저해효과가 어느 정도인지 조사하였다. 실험군으로는 SD rat 한 마리당 Lactobacillus sp. KP-3와 중금속 10 ppm을 혼합하여 약 100 ml/day씩 7일간 투여하였다. 대조군으로 중금속과 멸균 생리 식염수를 혼합하여 투여한 군과 Lactobacillus sp. KP-3와 멸균 생리 식염수를 혼합하여 투여한 군, 멸균 생리 식염수 만을 투여한 군으로 분리하여 SD rat 한 마리당 약 100 ml/day씩 7일간 투여하였다. 혈액, 간장 및 신장의 시료를 채취하여 카드뮴(Cd), 크롬(Cr), 납(Pb) 등의 중금속 축적량을 측정하였다. 측정 결과 크롬(Cr)은 신장에 주로 축적되었고, 납(Pb)은 간장과 신장에 주로 많은 축적을 보였고, 실험군인 Lactobacillus sp. KP-3와 중금속을 혼합하여 투여한 군에서는 중금속만을 투여한 군보다 중금속 축적량이 낮은 것으로 나타났다. 카드뮴(Cd), 크롬(Cr), 납(Pb)의 평균 중금속 축적 저해율은 각각 41.8%, 33.4% 44.2%로 나타났다. 특히, 혈액에서의 중금속 축적 저해율은 약 70%로 매우 높게 나타났고 혈액에서의 카드뮴 축적 저해율은 100%로 나타났다. 본 실험에서 탐색하여 최적화된 유산균이라고 생각되는 Lactobacillus sp. KP-3균주는 실험 결과에 의하면 중금속 축적 저해에 효과가 있는 것으로 여겨지며, 또한 유산균이 함유된 식품을 지속적으로 섭취함으로써 체내의 중금속의 축적 억제에 도움이 크게 되리라 생각된다.

• 대한미생물학회지
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• 제21권1호
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• pp.133-144
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• 1986

This study was undertaken to assess the effect of ginseng administration on T lymphocyte induced local xenogenic graft-versus-host(GVM) reactions which were induced with thymocyte, spleen cell and lymph node cell of ICR mice. Mice received daily 10mg of 70% alcohol ginseng extract oral1y for 100days and control mice remained untreated for the same period of time. The cells from donor mice were injected intradermally into the closely shaven abdominal skin of Sprague-Dawley rats for GVH tests. The thymocyte from control(ginseng-untreated) mice showed a negative local GVH reaction, whereas thymocyte from experimental(ginseng-treated) mice showed a positive reaction with the rate of 17.4%. When spleen cells were injected, the incidence of positive local GVH reaction was 66.7% among ginseng-treated mice, as opposed to incidence of 45.5% of positive local GVH reaction among control mice. The incidence of positive local GVH reaction of the lymph node cells when injected into a recipient was 71.4% among ginseng-treated mice as compared with that of 18.9% among control mice. The relationship between spleen cell inoculum and intensity of the local GVH reaction was assessed in ginseng-untreated mice. The intensity of GVH reaction clearly appears to be dose related. In ginseng-treated mice, a minimum of $1{\times}10^7$ spleen cell was required for production of positive local GVH reaction with almost linear relationship up to an inoculum of $5{\times}10^8$ cells. In control mice, however, a minimum of $1{\times}10^8$ spleen cells was required for positive GVH reaction. These results strongly suggest that the ginseng administration augments significantly the local xenogenic GVH reaction which was used to assess T lymphocyte function and immunocompetence of mice and in addition to this, these results appear to support previous suggestions that the local GVH reaction consitutes a qualitative test of the functional activity of T lymphocytes. These results may be the first to induce local GVH reaction, employing rats as recipient and mice as donor. This study was also desingned to investigate some of the effects of ginseng extract on lymphocyte-macrophage interactions. This was accomplished by in vitro quantification of 1) migratory inhibitory factor(MIF) synthetic capacity of splenic lymphocytes in mice previously primed with ginseng 2) MIF responsiveness of mouse peritoneal macrophages or chicken peripheral leucocytes under the presence of ginseng extract 3) migration ability of chicken peripheral leucocytes by direct stimulation of ginseng extract or ginseng saponin and 4) immunosuppressive effects of immunosuppressants such as cyclophosphamide, cyclosporin A or dexamethasone. Mice divided equally into the ginseng and the saline groups, which received intraperitoneally daily 0.2ml of ginseng absolute alcohol-extract(5mg/ml) and same amount of saline for 15 days, respectively. The cellular immune responsiveness of these mice was assayed 15 days after ginseng pretreatment. Splenic lymphocytes of mice treated with ginseng, when stimulated with sensitized specific-antigen such as sheep red blood cells or toxoplasmin, or with polyclonal activator concanavalin A, produced significantly more MIF than those of control saline group. MIF responsiveness of normal mouse macrophages was significantly augmented when assayed under the presence of ginseng extract (1mg/ml). The migratory ability of normal chicken leucocytes in the absence of MIF was significantly decreased by the stimulation of ginseng extract alone. MIF response was significantly decreased by immunosuppressants and this impaired response was not restored by ginseng pretreatment. This study was additionally performed to evaluate the effect of ginseng on the expulsion of adult Trichinella spiralis in mice. ICR mice were infected experimentally by esophageal incubation of 300 T. spiralis infective muscle larvae prepared by acid-pepsin digestion of infected mice. and received oral administration of 70% alcohol ginseng extract(10mg/mouse/day) for the indicated days plus 4 days before infection. At various times after infection, the number of adult T. spiralis worms in small intestines was determined. Interestingly, ginseng-treatment was accompanied by accelerated expulson of T. spiralis. These results led to the conclusion that Panax ginseng caused some enhancing effect on GVH reaction, macrophage migration inhibition reaction and expulsion of T. spiralis. In addition these results suggested that the mechanisms responsible for this enhancement of ginseng may be chiefly or partially due to nonspecific stimulation of cell-mediated immune response.

• Journal of Periodontal and Implant Science
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• 제25권2호
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• pp.210-221
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• 1995

The purpose of this study was performed to investigate the mineralization and differentiation of osteobalsts for bone regeneration in vitro and the effect of rate of the composition in periodontal cells on mineralization. For this study, healthy gingival tissues were surgically obtained from the patients during 1st premolar extraction for the purposes of orthodontic treament. Gingival tissue was washed several time with Phosphate buffered saline contained high concentration of antibiotics and antifungal agent, and cultured in Dulbecco's Modified Eagle's Medium(DMEM, Gibco, U.S.A.). Every cell were cultured in state at $37^{\circ}C$, 100% of humidity, 5% of $CO_2$ incubator. Bone marrow stromal cells were isolated from 5-clay-old rat femur with using medium irrigation mathod by syringe. Cell suspension medium were centrifuged at 1500 rpm for 5 min and then cultured in the petri dish. Two kinds of cell were freezed and stocked in the liquid nitrogen tank until experiment. Cell were incubated into the 24 multi-well plate with $5{\times}10^4$cell/well of medium at $37^{\circ}C$, 100% of humidity 5% $CO_2$ incubator for 24 hours. After discarded of the supernatent of medium, O.5ml of medium were reapplied and incubated. And counted the number of cell using the hemocytometer and inverted light microscope. We have measured the number of mineralized nodule with using Alizarin red S. staining in microscope. Furthermore every cell were observed the morphological change between every rate of co-culture of the two kinds of cell. The results were as follows; The rate of proliferation of co-culture cell revealed high rate tendency compared the bone marrow stromal cell only and low growth rate to compared with gingival fibroblast only. The tendency of formation of the mineralized nodule were observed dose-depend pattern of bone marrow stromal cell. It is concluded that the gingival fibroblast may inhibit the formation of mineralized nodule in the culture of the bone marrow stromal cell.

• 한방비만학회지
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• 제17권2호
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• pp.101-110
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• 2017

Objectives: In this study, the lipolytic effects of Eriobotrya folium extract (EFE) on local fat was investigated in high fat diet (HFD)-induced obesity mouse and 3T3-L1 adipocytes. Methods: C57BL/6J mice (5 weeks) were fed HFD for 6 weeks to induce obesity. EFE (20 mg/ml, $100{\mu}l$) or saline ($100{\mu}l$) as a normal control was injected into left inguinal fat pad region, 3 times per a week for last 2 weeks. After sacrifice, body weight, and histological changes of the inguinal fat pad were evaluated. The expressions of hormone-sensitive lipase (HSL) and adipose triglyceride lipase (ATGL) in inguinal fat pad were analyzed by Western blotting. Also, lipid accumulation and lipases release were determined in 3T3-L1 adipocytes by oil red o staining. Results: EFE significantly reduced the weight of inguinal fat pad and the size of adipocytes in HFD-induced obesity mice compared to control. The treatment of EFE up-regulated the expressions of HSL and ATGL in inguinal fat pads of obesity mice, as well as 3T3-L1 adipocytes. In addition, EFE inhibited the lipid accumulation in 3T3-L1 adipocytes in a dose dependent manner. Conclusions: EFE showed lipolytic effect on local fat of HFD-induced obesity mice by up-regulation of the lipases secretion. This suggests that EFE could be considered as anti-obese substance with lipolytic property on local fat.