• 대한한방내과학회지
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• 제29권1호
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• pp.265-277
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• 2008

In order to investigate the protective effects of Gami Yugan-tang on liver damage in rats induced by $CCl_{4}$ and d-galactosamine, the serum transaminase(ALT & AST), alkaline phosphatase(ALP), lactic dehydrogenase(LDH), superoxide dismutase(SOD), catalase, glutathione S-transferase(GST), glutathione peroxidase(GPX) for enzyme activities, and lipid peroxidation were measured. All animals were divided into 4 groups: normal group (untreated), control group (treated with 0.9% saline solution), sample I group (treated with 740mg/kg Gami Yugan-tang), and sample II group (treated with 1,480mg/kg Gami Yugan-tang). The results were as follows : 1. The results of liver damage in rats induced by $CCl_4$ : The protective effects of ALT were displayed in sample I and sample II, and AST, ALP, LDH, SOD, catalase, GST, GPX, and lipid peroxidation were noted in sample II group. It showed slight necrosis of hepatic cell and pathologic changes, for example, inflammatory cells infiltration were improved in sample II group compared to the control group. 2. The results of liver damage in rats induced by d-galactosamine : The inhibitory effects of AST, ALT, LDH, and ALP activities were noted in both sample I and sample II groups. The findings from this experiment suggests that Gami Yugan-tang has protective effects against liver damage in rats induced by $CCl_{4}$ and d-galactosamine.

• 동의생리병리학회지
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• 제30권2호
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• pp.109-115
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• 2016

Hataedock is a Korean medical treatment that administers herbal extracts orally to newborn infants. This method is used for alleviating harmful heat and excreting fetal wastes by meconium. The purpose of this study was to evaluate anti-inflammatory effect of Hataedock method with Douchi on 2,4-dinitrofluorobenzene (DNFB)-induced atopic dermatitis (AD). The 3-week-old NC/Nga mice were divided into 3 groups: the control group (Ctrl), the AD-induced group (AE), and the Hataedock-treated group (GT). Only the GT group was treated with Hataedock at the 3rd week. After 28 days from Hataedock treatment, we induced AD-like dermatitis to the AE and GT group by DNFB. The effects of Hataedock were evaluated by immunohistochemical method. In the epithelium, PKC-positive reaction of the GT group was decreased by 57%. In the dermal papillae, IL-4-positive reaction was decreased by 34%. In the dermis, the distribution of degranulated mast cells was decreased and substance P-positive reaction was decreased by 49%. In the skin tissue, edema was decreased and MMP-9-positive reaction was decreased by 71%. Tissue damage such as epithelial cell hyperplasia, infiltration of granulocyte and lymphocyte, and capillary distribution were also decreased. The Hataedock method with Douchi maintained skin barrier and inhibited skin-damaging factors via regulating Th2 differentiation. In conclusion, Hataedock has a potential for preventative treatment of AD. Further studies are necessary to investigate the immune-regulating mechanism and verify the safety and efficacy of the Hataedock method.

• 동의생리병리학회지
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• 제23권3호
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• pp.590-598
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• 2009

The purpose of this research is to evaluate the effect of Mahaenggamseok-tang-gagambang (MGTG) on airway hyper- responsiveness (AHR), immune cells, cytokines and lung tissue in OVA-induced asthmatic mice. C578L/6 mice were injected, inhaled and sprayed with OVA for 12 weeks (3times a week) for asthma sensitization and challenge. Two experimental groups were treated with different concentrations of MGTG (400 mg/kg and 200 mg/kg) extract and cyclosporin A (10 mg/kg) for the later 8 weeks. Enhanced pause (Penh) levels were measured by whole body plethysmography. Immune cells were analyzed by flow cytometer in peripheral blood monocyte cell (PBMC) and lung cells. The IL-1b, IL-12, IFN-${\gamma}$, OVA-lgE, IL-4, IL-5, TNF-${\alpha}$ were analyzed by ELISA kit in serum and splenocyte+a-cCDS/a-CD28. Enhanced pause (Penh) levels of the MGTG groups (400 mg/kg and 200 mg/kg) were decreased significantly compared with that of control group. The numbers of MGTG groups (400 mg/kg and 200 mg/kg) on lung total cells were decreased significantly compared with that of control group. The numbers of MGTG groups (400 mg/kg and 200 mg/kg) on $CD3^+/CD69^+$, $B220^+/CD22^+$, $B220^+/CD23^+$, $B220^+/lgE^+$, $CCR3^+$ cells were decreased significantly compared with that of control group. The number of MGTG group (400 mg/kg) on $CD3^+/CD49b^+$ cells was decreased significantly compared with that of control group. The level of MGTG groups (400 mg/kg and 200 mg/kg) on IL-4, IL-5, IL-12, TNF-${\alpha}$, OVA-lgE were decreased significantly compared with that of control group. The level of MGTG group (400 mg/kg) on IL-1b, IL-1S, OVA-lgE were decreased significantly compared with that of control group. These results demonstrate that MGTG could be a desirable alternative therapy for allergic asthma by inhibiting the expression of immune cells, the activation of inflammatory mediator.

• 동의생리병리학회지
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• 제24권4호
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• pp.661-667
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• 2010

Astragali Radix (AR) has been used in traditional medicinal herb, which is known to tonify the qi and blood. It has been asserted to be a tonic that can improve the function of the lungs, adrenal glands and the gastrointestinal tract, increase metabolism, promote healing and reduce fatigue. Also, Plantago asiatica (PA) has the anti-inflammatory, antiseptic, antitussive, cardiac, diuretic, expectorant, haemostatic effects, though it is considered to be less powerful than the seed. Recent research is showing that it is promising in lowering cholesterol and controlling diabetes. Obesity is characterized at the cellular level by an increase in the number and size of adipocytes differentiated from fibroblastic preadipocytes in adipose tissue. In the present study, we investigated the effects of 3 to 7 extracts of mixed medicinal herbs (water and ethanol extract of Astragali Radix; ARW and ARE, water and ethanol extract of Plantago asitica; PAW and PAE, and those mixed extracts; ARW+PAE, ARW+PAW and ARE+PAE) on adipogenesis and apoptosis in 3T3-L1 adipocytes by MTT assay, Oil-Red-O staining and TUNEL assay. And then, we determined total contents of phenolic compounds and flavonoid compounds in mixed herbal extracts. These result showed that herbal extracts (ARW, PAE and ARW+PAE) affected on adipogenesis and apoptosis in 3T3-L1 adipocytes among 3 to 7 extracts tested. Especially herbal mixed extracts (ARW+PAE) had the highest inhibition on differentiation of preadipocytes and lipoid accumulation of maturing adipocytes. And it did not affect cell viability of mature adipocytes, but herbal mixed extracts (ARW+PAE) increased apoptosis in mature adipocytes, as proved by highest concentration $200\;{\mu}g/m{\ell}$ using TUNEL assay. In addition, contents of total phenoilc compounds and flavonoids showed the highest level in ARE+PAE and PAE than ARW. These results suggest that this main target for adipocyte functions could be effective in improving the metabolic syndrome including obesity.

• 생명과학회지
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• 제18권6호
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• pp.814-825
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• 2008

아스코크로린(Ascochlorin, ASC)은 Ascochyta viciae로부터 추출된 프레닐페놀 물질로, 혈청 콜레스테롤과 트리글리세라이드 수치를 감소시키고 종양 성장을 억제한다는 연구 결과가 보고되어 있다. 본 논문에서는 아스코크로린이 생리학적 혹은 병리학적인 작용과 염증반응에서 약리학적으로 유도되는 반응을 어떻게 조절하며, 이러한 메커니즘에 대해 이해하기 위해 mouse macrophage Raw264.7 세포에 아스코크로린을 처리하여 이에 대한 프로테옴의 특이적인 발현에 대해 분석하였다. 따라서 본 연구는 LPS를 처리한 mouse macrophage Raw264.7 세포에 아스코크로린을 처리하여 염증과정에 관련된 단백질의 발현 양상을 확인하기 위해 프로테오믹스를 시행하였다. Mouse macrophage RAW264.7 세포에 아스코크로린을 처리한 조건과 무처리한 조건으로 나누어 two-dimensional electrophoresis (2-D SDS-PAGE), matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF-MS)와 bioinformatics 방법으로 아스코크로린을 처리한 mouse macrophage Raw264.6 세포의 프로테옴을 분석하였다. 그 결과 mouse macrophage Raw264.7 세포에 아스코크로린 처리 시 Calreticulin이 4배 감소, ${\beta}-actin$도 4배 감소 그리고 vimentin이 1.5배 감소함을 확인 할 수 있었다. 그러나 rabaptin 아스코크로린 처리에 의해 3배 증가함을 확인 할 수 있었다. 이러한 단백질 발현은 RT-PCR을 수행하여 결과에 대해 재확인 하였으며, 프로테오믹스와 동일한 결과를 얻을 수 있었다. 따라서 본 연구를 통해 LPS 처리에 의해 활성화된 mouse macrophage RAW264.7 세포에 ASC를 처리한 후 이차원 전기영동법을 이용하여, 단백질의 발현 변화 및 양상을 규명하고 단백질 지도를 확립 하였으며, RAW264.7 세포를 이용한 면역세포 모델에서 ASC의 항염증 작용을 중심으로 생리활성 조절기능을 확인 할 수 있었다. 향후 분자 기능 조절 연구와 전 임상 연구를 통해 ASC의 생리활성 조절 기능을 규명한다면 ASC는 항염증 및 항암활성을 갖는 약물로 개발될 것으로 기대된다.

• Archives of Plastic Surgery
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• 제35권4호
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• pp.385-392
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• 2008

Purpose: The most effective methods of harvesting, preparing, and injecting autologous fat grafts have been inconsistent and conflicting. With its limitation as resorption in fat grafting, handling various techniques affect adipocyte survival, and is crucial to optimizing its long-term survival. To improve graft survival, re-implantation of cryopreserved adipocytes was developed. In addition, adipocytes do not induce immune rejection in response to non-self lymphocytes in a mixed lymphocyte reaction. The purpose of this study is to analyze the changes in cryopreserved adipocytes so as to determine the most efficient long-term storage period, and to analyze the changes in cryopreserved allografted adipocytes so as to determine the efficacy of cryopreserved adipocytes allografting. Methods: Fat tissues were harvested from the inguinal and retroperitoneal fat pad of mice. After the centrifugation of the harvested fat tissues, they were disintegrated with collagenase. The adipocytes were obtained by centrifugation of the disintegrated fat tissues. The adipocytes were treated as follows: (1) They were examined for weight and then frozen at $-20^{\circ}C$(n=25). For four months, each five frozen samples were taken and examined for weight and histologic changes in the 1st week, the 1st month, the 2nd month, the 3rd month, and the 4th month, respectively. (2) The adipocytes were immediately frozen at $-20^{\circ}C$(n=125). For four months, five frozen samples were taken, and allografted in the same time period as above. Finally, for four months, five cryopreserved allografted adipocytes were taken and examined for histologic changes in the same time period as above. Results: (1) Significant weight changes and histologic findings with inflammatory and destructive changes were observed in the cryopreserved adipocytes in three months. (2) Significant fat necrotic changes in the histologic changes with Hematoxylin and eosin stain were observed in the cryopreserved allografted adipocytes since the first week, independent of the freezing period. Conclusion: The study results show that the adipocytes that were cryopreserved for more than three months underwent obvious weight reductions and necrotic changes, and the adipocytes that were allografted without freezing were viable for four months, but the cryopreserved allografted adipocytes had obvious necrotic changes since the first week regardless of the freezing period.

• Annals of Clinical Neurophysiology
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• 제1권2호
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• pp.91-98
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• 1999

Background : The pathogenesis of acute inflammatory demyelinating polyradiculoneuropathy (AIDP), Guillain Barre syndrome (GBS) is not clear, but it has been known that the immune mechanisms play an important role. Authors performed this study to establish an animal model of experimental allergic neuritis (EAN) by immunizing the myelin components of peripheral nerves and to understand the electrophysiological and histopathological features as well as the ${CD_5}^+$ B-lymphocyte changes in peripheral bloods in the EAN models. Methods : Lewis rats weighing 150-200 gm were injected subcutaneously in soles two times with total myelin, P0, P1, or P2 proteins purified from the bovine cauda eguina. The EAN induction was assessed by evaluating clinical manifestations. The electrophysiological and histopathological features were studied as routine methods. The ${CD_5}^+$ Blymphocytes were double stained using monoclonal FITC conjugated anti-rat CD45RA and R-PE conjugated anti-rat ${CD_5}^+$ antibodies and calculated using a fluorescence activated cell sorter (FACS). Results : The EAN animal models were established. In two out of five, in one out of two, in none out of three, and in none out of one Lewis rats injected with purified total myelin, P0, P1, P2 proteins respectively, They showed slow spontaneous motor activity and weak resistance against pulling back by tails. The typical electrophysiological and histologic findings in total protein and P0 induced EAN animal models were the decreased conduction velocity, the decreased compound muscle action potential (CMAP) amplitude and the dispersion phenomenon. The perivascular infiltrates of lymphocytes with focal demyelinating process were found in light microscopy. The ${CD_5}^+$ B-lymphocyte expression in three EANs were 2.38%, 3.50% 2.50%, which were not significantly increased, compared with those in normal controls. Conclusion : The EAN animal models were successfully established by injecting the total myelin and P0 myelin and they showed electrophysiological and histological features typical of demyelinating process. However they did not show an increased expression of ${CD_5}^+$ B-lymphocyte in peripheral bloods which could be indirect evidence of humoral autoimmunity.

• Applied Microscopy
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• 제41권1호
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• pp.21-29
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• 2011

피부질환의 개선을 위한 수치료요법은 예로부터 세계 각지에서 널리 이용되어 왔다. 본 연구에서는 국내 알칼리환원온천수를 이용한 수치료요법이 손상된 피부에 어떠한 영향을 미치는지 확인하고자 하였다. 무모생쥐(hairless mice)의 등 부위에 자외선을 조사하여 피부 손상을 인위적으로 유발한 후 매일 실온에서 40분간 침적시켜 21일간 수치료를 실시한 후 생쥐의 피부조직을 분리하여 검경하였다. 수치료를 하는 동안 실험군의 피부 회복속도가 대조군에 비해 더 빠른 경향으로 관찰되었다. 조직학적으로 관찰한 결과, 대조군에 비해 실험군에서 표피층 비후 및 염증세포 침착 현상이 상대적으로 적게 나타났으며, 비만세포의 증가 및 탈과립 현상 또한 적게 나타났다. 이는 온천수에 함유된 황성분의 박피효과 및 항균작용과 함께 온천수의 항산화능이 피부조직의 치유에 도움을 준 것으로 사료된다.

• 생명과학회지
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• 제25권5호
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• pp.515-522
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• 2015

Treculia africana Decne는 빵나무종으로 뽕나무과, Treculia 속에 속하는 식물로서, 이 식물의 다양한 부위에서 추출한 물질은 항염증, 항균등의 효과를 가지고 있어 백일해의 치료등 다양한 질환에서 민간요법으로 사용되어 왔다. 하지만 정확한 생리활성에 관한 연구는 보고된 바가 없다. 따라서 본 연구에서는 T. africana Decne 메탄올추출물(META)을 사용하여 항산화능 및 인체 대장암 세포주인 HT29에 대한 항암활성에 관하여 분석하였다. DPPH radical scavenging activity를 통해 분석한 결과, META의 IC₅₀가 4.53 μg/ml로 강력한 항산화능을 보유하 였음을 확인하였다. 또한 META 처리에 의해 HT29의 생존율이 감소함과 더불어 IC₅₀가 66.41 μg/ml로 강력한 세포사멸효과를 나타냈다. META 처리에 의해 HT29의 subG1 세포비율 및 Annexin V⁺ 세포의 비율이 증가하고, DAPI로 염색된 apoptotic body가 증가하였다. 또한 apoptosis 관련 단백질인 Fas, Bax, cytochrome c의 발현이 증가하였으며, 이는 caspase 3, 8, 9를 활성화 시켜 최종적으로 PARP가 분해되어 apoptosis가 유도되었음을 확인하였다. 이러한 결과들을 통해 META는 강한 항산화 활성과, 대장암세포에서 apoptosis 유도에 의한 높은 항암활성을 보유한 물질임을 확인하였다.

• 한방재활의학과학회지
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• 제21권2호
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• pp.63-85
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• 2011

Objectives : This study was carried out to know the effects of Danggwisayeokgaohsuyusaenggang-tang(hereinafter referred to DST) on arthritis induced by collagen on DBA/1 OlaHsd mice. Methods : For this purpose, DST was orally administered to mouse with arthritis induced by collagen II. Cytotoxicity, high performance liquid chromatograph(HPLC) analysis, arthritis index, value of immunocyte in draining lymph node and paw joint, cytokine were measured in vivo. Results : 1. The cytotoxicity against human fibroblast cells(hFCs) was not measured in any concentration. 2. In HPLC analysis, There are high peak patterns at 8 minute(min), 12 min, 35 min, 45 min. 3. The arthritis index was decreased significantly. 4. The degree of arthritis induced damage of joint of DST group is slight compared with control group in histopathologic observation(Hematoxylin and eosin stain(H&E), Masson's trichrome(M-T) staining). 5. In total cell counts of draining lymph node(DLN) and paw joint, the cells in DLN decreased significantly on DST 200 mg/kg and the cells in paw joint decreased significantly on 200 mg/kg and 50 mg/kg. 6. In DLN, $CD4^+/CD25^+$, $CD3^+/CD69^+$, major histocompatibility complex(MHC), class-II/$CD11c^+$ cells decreased significantly on DST 200 mg/kg and 50 mg/kg $CD3^+/CD8^+$ cells decreased significantly on DST 200 200 mg/kg, $CD4^+$, $CD3^+/CD44^+$ cells decreased. 7. In paw joints, $CD4^+$, $CD11b^+/Gr-1^+$ cells decreased significantly on DST 200 mg/kg and 50 mg/kg. 8. In joints, levels of $IL-1{\beta}$, IL-6, $TNF-{\alpha}$, cyclo-oxygenase-2(COX-2), NOS-II were decreased on DST 200 mg/kg and DST 50 mg/kg. 9. In analysing of cytokine in CD3/CD28 activated spleen, IL-17 was decreased significantly, IL-4 was increased significantly $INF-{\gamma}$ was decreased on DST 200 mg/kg. 10. In analysing of cytokine in collagen activated spleen, IL-17 were decreased significantly, IL-4 was increased significantly. Conclusions : This results demonstrated that DST suppressed the inflammatory progression of collagen-induced arthritis(CIA) mice and supported further studies are required to survey continuously in looking for the effective substance and mechanism in the future.