• Journal of Acupuncture Research
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• v.24 no.5
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• pp.171-183
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• 2007

Objectives : The purpose of this study was to analyze the effects of Erythrinae Cortex herbal-acupuncture solution(EC-HAS) at the Joksamni($ST_{36}$) of mice with collagen II-induced arthritis(CIA). Methods : The author performed several experimental procedures to observe the effects of the EC-HAS at the arthritis. The severity of arthritis, changes of cytokine level and antibody level, histological changes of the CIA mouse joint were analyzed. Results : 1. The incidence of arthritis and arthritis index were significantly decreased in the cases which were treated with the EC-HA. 2. Cartilage destruction and synovial cell proliferation were reduced, and the expression of the collagen fibers was similar with that of the normal group. 3. The levels of IL-6, $IFN-{\gamma}$, $TNF-{\alpha}$, $IL-1{\beta}$ in serum of the CIA mice which were treated with the EC-HA were significantly decreased compared with those of the normal group. 4. The levels of IgG, IgM and anti-collagen II in serum of CIA mice when they treated with the EC-HA were significantly decreased compared with those of the normal group. 5. The expression ratio of $CD4^+$ to $CD8^+$ cells in the EC-HA treated mice were maintained as much as the normal group of the lymph nodes in the CIA mice. 6. The $CD3e^+CD69^+$ and $CD11b^+Gr-1^+$ cell populations in the knee joint were significantly decreased in the EC-HA treated group. Conclusions : These results suggest that the EC-HA at the ST36 may be responsible roles to control on the synovial cell proliferation and to prevent the cartilage destruction in rheumatoid arthritis. These results will be important supporting evidence for the practical use of the EC-HA at rheumatoid arthritis clinic in the future.

• The Korea Journal of Herbology
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• v.35 no.4
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• pp.51-60
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• 2020

Objective : The objective of this study was to demonstrate the effect of Persicae Semen (PS) in DNCB-induced atopic dermatitis mouse and HaCaT cell. Methods : The BALB/c mice were divided into four groups. To develop atopic dermatitis, 200 ㎕ of 1 and 0.5% DNCB solution was put on the back of mice in the Control group, the PS-Low group and the PS-High group once a day. After application of DNCB, 200 ㎕ of the PS extract was also treated. The Normal group was given PBS. The mice dorsal skin was stained with Masson's trichrome, H&E, and toluidine blue to evaluate the thickness of the epidermis and dermis, infiltration of eosinophils and mast cells respectively. ELISA was applied to measure the serum level of IgE and IL-6. Toxicity of PS was measured by MTS assay in HaCaT cell. To investigate the effects of PS on HaCaT cells, cells were pre-treated with PS for 1h, and then stimulated with TNF-α and IFN-γ. After 24 hours, the expression of TARC was analyzed using RT-PCR. Results : PS not only significantly diminished the thickness of the epidermis and dermis, but also reduced the infiltration of eosinophil and mast cell in skin lesion. PS also reduced the serum IgE and IL-6 level which plated important roles in the atopic dermatitis. The expression of TARC was decreased significantly in TNF-α/IFN-γ stimulated HaCaT cell. Conclusion : These results suggest that PS may be effective in alleviating the atopic dermatitis induced by DNCB and inflammation by TNF-α/IFN-γ.

• The Journal of Pediatrics of Korean Medicine
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• v.30 no.4
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• pp.29-59
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• 2016

Objectives PRAL (Prunus armeniaca Linne Var) has been known to suppress allergic reaction. However, the cellular target and its mechanism of action were unclear. This study was designed to investigate the effect of PRAL on RBL-2H3 mast cell, which is PMA-Ionomycin-induced activated in vitro and the effect of PRAL on the MNC/Nga mice that are DNCB-induced activated in vivo. Methods In this study, IL-4, IL-13 production were examined by ELISA analysis; IL-4, IL-13, IL-31, IL-31Ra, $TNF-{\alpha}$ and GM-CSF mRNA expression were examined by Real-time PCR; manifestations of AP-1 and MAPKs transcription factors were examined by western blotting in vitro. Then skin rashes have been evaluated and verified the distribution of mast cells by H&E and toluidine blue. Also, WBC, eosinophil and neutrophil, IgE level in serum, $IFN-{\gamma}$, IL-4, IL-5 in the splenocyte culture supernatant, the absolute cell numbers of $CD4^+$, $CD8^+$, $Gr-1^+CD11b^+$, $B220^+CD23^+$, $CD3^+CD69^+$ in the Axillary Lymph Node (ALN), PBMCs and dorsal skin and IL-5, IL-13, IL-31, IL-31Ra in the dorsal skin by Real-time PCR were all evaluated from the NC/BNga mice. Results As a result of this study, the mRNA expression of IL-4, IL-13, IL-31, IL-31Ra and $TNF-{\alpha}$ and IL-4, IL-13 production, shown in ELISA analysis, were suppressed by PRAL. Results from the western blot analysis showed decrease on the expression of mast-cell-specific transcription factors, including AP-1 and p-JNK, p-ERK. Histological examination showed that infiltration levels of immune cells in the skin of the AD-induced NC/Nga mice were improved by PRAL orally adminstration. Orally- administered PRAL group also showred decreased level of IgE in the serum. This group has shown decreased the level of IL-4, IL-5, but shown elevated $IFN-{\gamma}$ level in the splenocyte culture supernatant. The same group also has shown decreased cell numbers of $CD4^+$, $CD8^+$, $CD3^+CD69^+$ in the ALN, and $CD4^+$, $Gr-1^+CD11b^+$ in the dorsal skin. PRAL oral adminstration increased cell numbers of $CD4^+$, but decreased cell numbers of $CD8^+$, $Gr-1^+CD11b^+$, $B220^+CD23^+$ in the PBMCs. Conclusions Obtained results suggest that PRAL can regulate molecular mediators and immune cells that are functionally associated with atopic dermatitis (AD) induced in the NC/Nga mice. This may play an important role in recovering AD symptoms and suppressing pruritus.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.14 no.1
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• pp.129-153
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• 2001

Background: SMG (升麻葛根湯加味方) is an herbal medicine which has been used in oriental medicine as a traditional therapeutic agent of pruritus and skin disease. Objective: This study was performed to investigate the effect of SMG on the anti-hypersensitivity and immune response in the murine of type I hypersensitivity induced by the experiment. Materials and Methods: Laboratory rats were primary sensitized with OA (ovalbumin); on day 1, rats of a Control group and Sample group (SMG group) were systemically immunized by subcutaneous injection of 1mg OA and 300mg of Al(OH)3 in a total volume of 2ml saline. The rats of the sample group were orally administered with an SMG water extract for 14 days after primary immunization. On day 14 after the systemic immunization, rats received local immunization by inhaling $0.9\%$ saline aerosol containing $2\%$(wt/vol) OA. A day after local immunization, BAL fluid and peripheral blood were collected from the rats. Total cell, lymphocyte, $CD4^+\;T\;cell,\;CD8^+\;T\;cell,\;CD4^+/CD8^+$ ratio in the BALF, and IgE, $CD4^+\;T\;cell,\;CD8^+$ T cell in the peripheral blood were measured and evaluated. Results: SMG showed a suppressive effect on the immune response in the rats. 1. Total Cells in the BALF decreased in the SMG treated group in comparison group, but statistic differences were not observed. 2. Total lymphocytes in the BALF were statistically decreased in SMG treated group in comparison to the control group. 3. CD4+ T cells in the BALF were statistically decreased in SMG treated group in comparison to the control group. 4. CD8+ T cells in the BALF were decreased in SMG treated group in comparison to the control group, but statistic differences were not observed. 5. The ratio of CD4+/CD8+ in the BALF was statistically decreased in SMG treated group in comparison to the control group. 6. The IgE level in serum was statistically decreased in SMG treated group in comparison to the control group. 7. The ratio of CD4+ and CD8+ in peripheral blood showed undetectable differences between each group of rats. From the experiment cited above, this study shows that SMG has both anti-hypersensitivity effects and immunoregulatory effects when administered to rats. Based on this experiment, it is suggested that SMG could be a useful immunomodulator and anti-allergy agent.

• Journal of Environmental Health Sciences
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• v.38 no.6
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• pp.541-549
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• 2012

Objectives: This study was undertaken in order to evaluate a potential mechanism involved in gastro-intestinal problems observed in autistic subjects and uses an animal model of autism investigation. Methods: BTBR T+tf/J, a mouse strain with typical socio-behavioral characteristics of autistic subjects and FVB mice with highly social behaviors as the control strain were used. Both genders of mice aged three weeks and six months were used from four separate litters for each strain. Serum was prepared following cardiac puncture, and mesenteric lymph nodes were collected for in vitro stimulation and enumeration of major immune cell proportion. Results: The level of serum IgA was significantly enhanced in six-month-old BTBR mice compared with three-week-old BTBR, which was not observed with the FVB control mice. The serum IgE level was also higher among BTBR mice than among age-sex matched FVB mice, respectively. Considering the ratio of interleukin-4 vs interferon-gamma production from mesenteric lymph node T cells, skewedness toward type-2 reactivities was observed. In addition, the proportion of B cells in mesenteric lymph nodes was significantly higher in BTBR mice than in FVB mice. Conclusion: Upregulation of mucosal immunity related with enhanced type-2 immune reactivity observed in BTBR mice could be involved with the etiology of gastro-intestinal abnormalities in autism.

• Journal of Acupuncture Research
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• v.33 no.2
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• pp.21-34
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• 2016

Objectives : The aim of the present study is to examine the effect and mechanism of Erycibae Caulis and Corydalis Tuber Pharmacopuncture (ECP) on a mouse model with collagen induced rheumatoid arthritis (CIA). Methods : We evaluated the Aspartate aminotransferase (AST), Alanine aminotransferase (ALT), Creatinine, and the Blood urea nitrogen (BUN) of serum to examine the safety of this study. In vivo, we compared the results of the non-treated group, the normal saline pharmacopuncture treated control group, the indomethacin treated group and the ECP group. We evaluated rheumatoid arthritis manifestation and the Rheumatoid Arthritis Index (AI). Also, immune cells in blood affected by ECP were evaluated by calculating the level of white blood cells (WBC), neutrophil, lympocytes and monocytes. Next, the level of Immunoglobulin M (IgM), Immunoglobulin G (IgG), Interleukin (IL)-$1{\beta}$, IL-6, IL-17, Tumor Necrosis Factor (TNF)-${\alpha}$ and Granulocyte-macrophage Stimulating Factor (GM-CSF)in serum were measured. We examined the imaging of cartilage degeneration using micro CT-arthrography of the hind paw. Additionally, we examined the effects of reducing bone volume (BV) ratio and bone surface/bone volume (BS/BV) ratio with 3D Micro-CT. Finally, we did a histopathologic examination analysis. Results : The absence of liver and kidney toxicity was evident. In vivo, edema of the joints of the ECP group decreased greatly in macroscopic observation. AI measurement of the ECP group also decreased significantly compared to the control group. The level of WBC, neutrophil, lympocytes, and monocytes in the blood decreased but there was no statistical significance of this data. IgM of the ECP group decreased significantly compared to the control group. IL-$1{\beta}$, IL-6, TNF-${\alpha}$, and GM-CSF production of the ECP group decreased significantly compared to the control group. As a result of examining joint condition with 3D micro CT, deformation and destruction of the joint was shown to have decreased. Bone density of ECP group increased at a statistically significant level compared to the control group. Degree of joint inflammation of ECP group decreased significantly compared to the control group. After H&E and M-T staining, infiltration of immune cells, subsidence of the cartilage, damage to the synovial cells and joint erosion decreased. Conclusion : This study showed that ECP hindered the process of rheumatoid arthritis and protected joints and cartilage.

• Journal of Life Science
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• v.20 no.4
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• pp.503-510
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• 2010

Most allergens have protease activities, suggesting that proteases may be a key link between Th2-type immune reactions in allergic responses. Protease activated receptor (PAR) 2 is activated via the proteolytic cleavage of its N-terminal domain by proteinases. To know the role of PAR2 in Aspergillus protease allergen activated Th2 immune responses in airway epithelial cells, we investigated and compared immune cell recruitment and level of chemokines and cytokines between PAR2 knock out (KO) mice and wild type (WT) mice. There were evident immune cell infiltrations into the bronchial alveolar lavage fluid (BALF) of WT mice, but the infiltrations in PAR2 KO mice were significantly lowered than those of WT mice. The IL-25, TSLP, and eotaxin gene expressions were profoundly increased after Aspergillus protease, but their expression was significantly lowered in PAR2 KO mice in this study. Compared to PAR2 KO mice, OVA specific IgE concentrations in serum of WT mice were quite increased; moreover, the IgE level of PAR2 KO mice was lower than in WT mice. The IL-25 expression by Aspergillus protease stimulation was significantly reduced by p38 specific inhibitor treatment. In this study, we determined that Th2 response was initiated with IL-25 and TSLP mRNA up-regulation in lung epithelial cells via PAR2 after Aspergillus protease allergen treatment.

• Korean Journal of Acupuncture
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• v.24 no.2
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• pp.163-184
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• 2007

Objectives & Methods : The purpose of this study is to observe the effects of Phellodendri Cortex Herbal-acupuncture solution (PC-HAS) at Joksamni (ST36) on collagen II induced arthritis in DBA-1J mice. The author performed several experimental items to analyze arthritis evaluation, change of weight, spleen size and adhesion rate, change of cytokine level, IgG, IgM and anti-collagen II, chang of immunocyte count, histological change of CIA mouse joint. Results : 1. In the PC-HA group, arthritis index, the incidence of arthritis and joint edema were significantly decreased. 2. In the PC-HA group, the change of spleen size, spleen adhesion rate and the knee joint were significantly decreased. 3. The levels of $IL-1{\beta}$, IL-6 and INF- in serum of the CIA mouse were significantly decreased by PC-HA. 4. The levels of IgG, IgM and anti-collagen II in serum of the CIA mouse were significantly decreased by PC-HA. 5. In the CIA mouse spleen cell culture, the levels of IFN- , IFN- / IL-4, IL-10 were significantly decreased by PC-HA, but the level of IL-4 was significantly increased by PC-HA. 6. In the PC-HA group, the ratios of $CD3e^+$ to $CD45R^+$ cell, $CD4^+$ to $CD8^+$ cell and $CD4^+/CD25^+$ cell were similarly maintained as normal group in the CIA mouse spleen cell. 7. In the PC-HA group, $CD4^+CD25^+$ and $CD45R^+/CD69^+$ cell were significantly decreased in the lymph nodes. 8. In the PC-HA group, $CD3^+/CD69^+$ and $CD11b^+/Gr-1^+$ cell were significantly decreased in knee joint. 9. In histology, the cartilage destruction and synovial cell proliferation in the PC-HA group were similar with that of the normal group and the collagen fiber expressions in the PC-HA group were similar with that of the normal group. Conclusions : Form the result above, the results suggest that the PC-HA at ST36 has significant effect on collagen-induced arthritis, and can be put to practical use in the future rheumatoid arthritis clinic.

• Journal of Acupuncture Research
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• v.24 no.6
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• pp.45-61
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• 2007

Objectives : The purpose of this study is to observe the effects of Carthami Flos herbal-acupuncture (CF-HA) at Joksamni($ST_{36}$) on arthritis in mice induced by Collagen II. Methods : The author performed several experimental items, including arthritis evaluation, change in weight, spleen size and stenosis rate, change in cytokine level, IgG, IgM and anti-collagen II, change of immunocyte count and histological change of the CIA mouse joint. Conclusions are as follows: Results : 1. In the CF-HA, the arthritis index and rate and the incidence of arthritis were decreased as the experiment proceeded. 2. In the CF-HA, spleen swell and stenosis, joint edema and change were decreased. 3. In the CF-HA, the level of $IL-1{\beta}$, IL-6, $TNF-{\alpha}$ and $IFN-{\gamma}$ in blood serum were significantly decreased. 4. In the CF-HA, the level of IgG, IgM and anti-collagen II were decreased. 5. In the CF- HA, $IFN-{\gamma}$, $IFN-{\gamma}/IL-4$, IL-10 of the culture fluid was decreased. 6. In the CF-HA, the cell rate of $CD3e^+$ and $CD45R^+$, $CD4^+$ and $CD8^+$, $CD4^+/CD25^+$ in spleen was similar to the cell rate of the normal group. 7. In the CF-HA, the cell rate of $CD4^+/CD25^+$, $CD45R^+/CD69^+$ in a lymph node was decreased as in the normal group. 8. In the CF-HA, the cell rate of $CD3^+/CD69^+$, $CD11b^+/Gr-1^+$ in joints was decreased as in the normal group. 9. In the CF-HA, the cartilage destruction and the inflammation cell growth in the H&E stain were decreased. The collagen fiber in the M&T stain were less destructed, therefore the result was similar to the normal group. Conclusions : These results suggest that CH-HA at $ST_{36}$ has an effect in controlling immune reaction and suppressing inflammation in rheumatoid arthritis therefore, the continuous flow of the following study is expected.

• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.5
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• pp.1168-1177
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• 2008

Atopic dermatitis(AD) is a chronic inflammatory skin disease. AD has increased gradually, many people are tortured with AD. Chunggi-san(CG) and Samhwangseze-gamibang(SG) has been used for many kinds of skin disease in the Oriental medicine. But reports about the effect of CG and SG are insufficient. So, author investigated the effect of CG and SG on NC/Nga atopic mice. Major findings are summarized as follows: The clinical skin severity scores of experimental group in 13 and 16 week were decreased by 42% and 50% compared to the control group. Serum IgE, IL-4, IL-5, IL-6, IgM, IgGI levels of experimental group were significantly decreased compared to the control group. Serum $IFN-\nu$ was significantly increased in the experimental group compared to the control group. mRNA expression levels of IL-4, IL-5, and CCR3 in the skin tissues of experimental group were significantly decreased, and expression level of IL-6 in the skin tissues of experimental group was significantly decreased compared to the control group. $IFN-\nu$ mRNA expression levels was increased compared to the control group. According to biopsy reports of the ear and skin tissues showed that the tissue damage, experimental group were highly reduced compared to the control group. Judging from that $IL-1{\beta}$, $TNF-{\alpha}$, IL-6 express of gene, the effects of inflammatory cytokines revelation were significantly decreased compared to the control group. Depending on the density of CG, inflammatory RAW 264.7 in the serum of CG were significantly inhibited compared to the control serum that leaded a COX-2 activity model.