• 동의생리병리학회지
• /
• 제20권6호
• /
• pp.1467-1476
• /
• 2006

This study was to evaluate the effect of Bulhwangeumjeonggi-san (BS) on mouse Th1 and Th2 cells' differentiation and ovalbumin (OVA)-induced allergic inflammation. The proliferation of mouse CD4 T cells and the secretion of Th1/Th2 cytokines under the influence of BS extract were measured as well as the amount ${\beta}$-hexosaminidase in RBL-wH3 cells and the levels of TNF-${\alpha}$ and IL-6 secretion in Raw264.7 cells. BALB/c mice were orally administered with BS extract and simultaneously inoculated with OVA to induce allergic reaction and measure the level of total lgE, OVA-specific lgE and the production of IFN- g, IL-4, IL-5 by the spleen cells. When mouse CD4- T cell were stimulated with anti-CD3 and anti-CD28 for 48 hours in various concentrations of BS extract, it increased proliferation of CD4 cells by 14% in 50 ${\mu}g/m{\ell}$ concentration but it showed an inhibition in higher concentrations. CD4 T cells under Th1/Th2 polarizing conditions for 3 days with BS resulted in mild decrease of IFN- g in Th1 cells and mild increase of IL-4 in Th2 cell at 50 ${\mu}g/m{\ell}$ but the level of IL-4 decreased by 18% at 100 ${\mu}g/m{\ell}$. BS extract had a dose-dependent inhibitory effect on antigen-induced release of ${\beta}$-hexosaminidase in RBL-2H3 cells. Treatment of BS extract on LPS stimulated Raw 264.7 cells showed dose-dependent decrease in TNF-${\alpha}$ production. Oral administration of BS extract on OVA-induced allergic mice showed an inhibitory effect on the levels of total serum lgE and OVA-specific lgE by 50% and 55%, respectively. Culture of spleen cells with OVA resulted in significant increase of IFN- g by 25% and significant decrease of IL-4 and IL-5 by 53%, and 38%, respectively. The results show that BS does not strongly induce mouse T cells to transform into Th1 or Th2 but it has an anti-allergic effect in vitro, and that it also corrects the unbalance between the reactions of Th cells in allergic diseases.

• 대한한방내과학회지
• /
• 제30권3호
• /
• pp.451-464
• /
• 2009

Background and Objective : Cyperus rotundus L. (CR) is a commonly used herbal medicine in Asian countries such as Korea, China and Japan. The present study was designated to evaluate the direct effects of CR on helper T cell activities and on Th1/Th2 lineage development in vitro. Materials and Methods : Spleen cells from 8 week BALB/c mice were cultured in CR extracts containing medium without activation for 24 hours and with activation for 48 hours. CD4+ T cells were isolated and analyzed for mRNA expression levels of INF-$\gamma$, IL-4, T-bet and GATA-3 by RT-PCR and secretion cytokines levels of INF-$\gamma$, IL-4, IL-5 and IL-10 by ELISA. Results : The results demonstrated that CR had no mitogenic effects on unstimulated CD4+ T cells, but augmented CD4+ T-cell proliferation upon activation with anti-CD3/anti-CD28 antibodies in a dose-dependent manner. CR treatment significantly increased CD4+ T cell population and the IFN-$\gamma$ expression was significantly enhanced, while IL-4 expression was significantly decreased. In addition, in vitro Th1/Th2 polarization experiments revealed that CR enhanced IFN-$\gamma$ secretion in Th1 cells, but reduced the IL-4 in Th2 cells in a dose-dependent manner. Conclusion : These results suggest that CR treatment could be a desirable alternative therapy for the prevention or correction of Th2 dominant pathological disorders, such as allergy and asthma.

• 대한한방내과학회지
• /
• 제28권4호
• /
• pp.681-693
• /
• 2007

Background and Objective : Atractylodes japonica (AJ) is a commonly-used herbal medicine in Asian countries such as Korea, China and Japan. The present study was designated to evaluate the direct effects of AJ on helper T cell activities and on Th1/Th2 lineage development in vitro. Materials and Methods : Spleen cells from 8-week BALB/c mice were cultured in CR extracts containing medium without activation for 24 hours and with activation for 48 hours. CD4+ T cells were isolated and analyzed for mRNA expression levels of INF-$\gamma$, IL-4, T-bet and GATA-3 by RT-PCR and secretion cytokines levels of INF-$\gamma$, IL-2, IL-4, IL-5 and IL-10 by ELISA. Results : The results demonstrated that AJ had no mitogenic effects on unstimulated CD4+ T cells, but augmented CD4+T-cell proliferation upon activation with anti-CD3/anti-CD28 antibodies in a dose-dependent manner. AJ treatment significantly increased CD4+ T cell population and IFN-$\gamma$ expression was significantly enhanced, while IL-4 expression significantly decreased. In addition, in vitro Th1/Th2 polarization experiments revealed that AJ enhanced IFN-$\gamma$ secretion in Th1 cells, but reduced the IL-4 in Th2 cells in dose-dependent manner. Conclusion : These results suggest that AJ treatment could be a desirable alternative therapy for the prevention or correction of Th2 dominant pathological disorders, such as allergy and asthma.

• 대한한방부인과학회지
• /
• 제28권2호
• /
• pp.1-14
• /
• 2015

Objectives : This study was designed to investigate the effects of Taraxaci Herba (TH) on cell death in breast cancer cells. Methods : In this experiment, the effects of TH on proliferation rates, cell morphology and growth pattern, intracellular reactive oxygen species (ROS) production. In addition, the effects on nuclear condensation, fragmentation and formation of acidic vesicular organelles (AVO) in MCF-7 cells were also investigated. Finally, autophagy related with protein was observed by using western blot method. Results : TH inhibited proliferation of MCF-7 cells, TH elevated intracellular ROS levels significantly. Treatment with TH did not affect nuclear morphologies such as condensation or fragmentation. On the other hand, TH treatment effectively induced AVO. Finally, one of autophagy related with protein, Microtubule-associated proteins 1A/1B light chain 3A (MAP1LC3A, LC3) level was elevated by treatment with TH. Conclusions : These data indicate that TH is able to be used for patient with breast cancer and mechanisms are involved in autophagy through ROS generation.

• Applied Microscopy
• /
• 제20권1호
• /
• pp.51-64
• /
• 1990

This experiment was performed to study the morphological changes of the goblet cells in the rabbit duodenal mucosa after common bile duct ligation. Healthy adult rabbits weighting about 2kg body weight were divided to normal and bile duct ligated groups. Common bile duct ligation was performed under ether anesthesia. Experimental animals were sacrificed on the 1st, 3rd, 5th, 7th and 14th day after the operation. Mucosal specimens from the upper part of duodenum were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde, followed by 1% osmium tetroxide, embedded in araldite mixture, cut with LKB-V ultratome, stained with uranyl acetate - lead citrate, and observed with JEM 100 CX II electron microscope. Observed results were as follow : 1. In the early stages(1st-5th day) of the experiments, the goblet cells showed apocrine and merocrine secretion. But those of the late stage(7th and 14th day) groups showed exocytotic merocrine secretion. 2. In the late stage of the experiments, there found than increase of newly formed goblet cells that contain electron lucent cytoplasms. 3. In the goblet cells of normal rabbit, mucous granules with higher or lower electron densities are found together in the cytoplasm, and electron lucent mucous granules occasionally fused together. But in the early stage of the common bile duct ligation, goblet cells contained granules of higher electron densities. 4. Considering the above findings, common bile duct ligation probably initiates the hypersecretion of mucous granules of goblet cells in the early stage, and may facilitate the differentiation of goblet cells in the later stage.

• 동의생리병리학회지
• /
• 제18권5호
• /
• pp.1397-1403
• /
• 2004

By recently study, GM (Gamipaemo-tang) treatment have worked well on the allergic asthma. The purpose of this study was effect of GM extract on helper T cell, major regulator of immune system. Splenic cells from 8-week BALB/c mice were cultured in GM containing media without activation for 48 hours. The MTS assay and flow cytometry study revealed that lymphocyte treated with GM were not effective on CD4+ T cells. Subsequently CD4+ T cells were isolated and cultured in GM containing media. Either GM were not effective on CD4+ T cell without APCs. By FACS scan analysis, the expression of INF-γ, IL-4 were down-regulated in the condition skewed Th1 and Th2 cells respectively, Using ELISA analysis, the expression of INF-γ is up-regulated and IL-4 is down-regulated in the condition skewed Th1, Th2 cells respectively. With RT-PCR analysis, the expression of mRNA for INF-γ is down-regulated and IL-4 is down-regulated in the condition skewed Th1 and Th2 cells respectively. The result suggests that GM inhibited the differetiation of Th2 cells significantly and indicates GM could enhance anti-allergic immune system.

• Restorative Dentistry and Endodontics
• /
• 제27권1호
• /
• pp.1-11
• /
• 2002

Immune responses associated with bacterial infection involve various inflammatory cells. Clinical symptoms and pathologic features are particularly influenced by the predominant cells Among inflammatory cells, T cells have the heterogenity. T cells may develop into the mature cells expressing the cell surface markers with different functions and T helper cells are categorized into Th1 and Th2 cells based on their different patterns of cytokine production. The objective of this study was to investigate the change of expression of surface markers on T cells and the Th1/Th2 immune response in pulpal inflammation associated with specific bacteria. We experimentally induced pulpal inflammation in rat incisors by drilling without coolant and innoculated with Streptococcus mutans (S.M. group), Porphyromonas endodontalis (P.E. group), or only sterile cotton (control group). After 1, 2, and 5 days, mandibular incisors were extracted and the pulp tissues were extirpated The expressions of IL-2 recepters (CD25) and ICAM-1 (CD54) on CD4+ and CD8+ cells in the pulps were determined using a flow cytometer, and the concentration of IL-2, IFN-$\gamma$ and IL-4 was measured by enzyme-linked immunosorbent assay. The results were as follows: 1 In the S.M. group, CD4+ cells were more increased at 2nd day than 1st day and in the P.E. group, CD8+ cells were more increased at 2nd day than 1st day. 2. The percentages of CD4+, CD4+25+ and CD4+54+ cells were decreased in the pulp tissues at 5th day after irritation in all groups. 3. The ratios of CD4+/CD8+, CD4+/CD4+25+ and CD4+/CD4+54+ in the pulps at 2nd day after irritation by P. endodontalis were significantly lower than the other groups. 4. The higher concentrations of IFN-$\gamma$ than IL-4 in the pulps at 2nd day after irritation by P. endodontalis showed that T helper 1 reaction were predominant in the early stage of the pulpal inflammation induced by P. endodontalis. 5. The higher concentrations of IL-4 than IFN-$\gamma$ in the pulps at 1st day and 5th day after irritation by S. mutans were measured but the differences were not significant.

• 한국동물번식학회:학술대회논문집
• /
• 한국동물번식학회 2004년도 춘계학술발표대회
• /
• pp.272-272
• /
• 2004

The objective of this study is to test whether human embryonic stem cells expressing Nurr1 (Nurr1-transfected hES cells) could be expressed TH according to neuronal differentiation. As an effort to direct differentiation of hES (MB03 registered in NIH) cells to dopamine-producing neuronal cells, Nurr1 was transfected using conventional transfection protocol into MB03 cell and examined the expression of tyrosine hydroxylase (TH) after differentiation induced by retinoic acid (RA) and ascorbic acid (AA). (omitted)

• Clinical and Experimental Reproductive Medicine
• /
• 제37권3호
• /
• pp.231-237
• /
• 2010

목 적: 반복적 유산 또는 반복적 착상실패 환자의 T 림프구의 Th1 면역반응 정도를 알아보고, T 림프구 활성 정도를 분석하고자 하였다. 연구방법: 반복적 유산 및 반복적 착상실패를 경험한 37명의 환자를 연구군으로 설정하고, 유산이나 불임의 병력이 없이 정상분만의 경력이 있는 11명의 가임 여성을 대조군으로 모집하였다. 유세포분석기를 이용하여, 이들의 말초혈액 중 T helper 세포 내 TNF-$\alpha$와 INF-$\gamma$ 및 IL-10의 발현도를 측정하고 Th1/Th2 세포 비율 (TNF-$\alpha$/IL-10 및 INF-$gamma$/IL-10 발현도)을 계산하여 Th1 면역반응의 우세 정도 및 T 림프구의 활성도를 분석하였으며, 활성 표지자인 CD154와 CD69 발현 정도를 비교하였다. 결 과: 연구군의 평균연령은 $35.3{\pm}4.3$세였으며, 이들에서 T helper cell 내 TNF-$\alpha$/IL-10의 발현 비율이 대조군에 비해 유의하게 높았고 ($42.1{\pm}2.3$ vs. $28.7{\pm}2.7$, p=0.002), CD154와 CD69의 발현율 또한 대조군에 비해 전반적으로 높았다. CD154의 경우, T helper cell ($1.7{\pm}0.5$ vs. $0.3{\pm}0.2$, p=0.038)과 T suppressor cell ($0.6{\pm}0.2$ vs. $0.1{\pm}0.0$, p=0.024) 모두에서 유의하게 높은 발현을 보인 반면, CD69의 경우, 전체 T 림프구 ($5.6{\pm}1.9$ vs. $1.3{\pm}5.4$, p=0.046)와 T suppressor cell ($4.8{\pm}1.3$ vs. $1.8{\pm}0.2$, p=0.035)에서 통계적으로 높은 발현을 확인할 수 있었다. 결 론: 반복적 유산 및 반복적 착상실패를 보이는 여성에서 T 림프구의 활성도와 Th1 면역반응이 증가하였으며 이는, 이들 여성에서 활성화된 T 림프구가 Th1 면역반응을 유도하여 초기 임신의 유지와 착상에 좋지 않은 영향을 미치기 때문으로 생각된다.

• Biomolecules & Therapeutics
• /
• 제23권1호
• /
• pp.71-76
• /
• 2015

Peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) was identified as a cell-intrinsic regulator of Th17 cell differentiation. Th17 cells have been associated with several autoimmune diseases, including experimental autoimmune encephalomyelitis (EAE), inflammatory bowel disease (IBD), and collagen-induced arthritis. In this study, we confirmed $PPAR{\gamma}$-mediated inhibition of Th17 cell differentiation and cytokine production at an early stage. Treatment with ciglitazone, a $PPAR{\gamma}$ ligand, reduced both IL-$1{\beta}$-mediated enhancement of Th17 differentiation and activation of Th17 cells after polarization. For Th17 cell differentiation, we found that ciglitazone-treated cells had a relatively low proliferative activity and produced a lower amount of cytokines, regardless of the presence of IL-$1{\beta}$. The inhibitory activity of ciglitazone might be due to decrease of CCNB1 expression, which regulates the cell cycle in T cells. Hence, we postulate that a pharmaceutical $PPAR{\gamma}$ activator might be a potent candidate for treatment of Th17-mediated autoimmune disease patients.