• 한국약용작물학회지
• /
• 제26권1호
• /
• pp.8-18
• /
• 2018

Background: This present study was conducted to evaluate the anti-inflammatory and immune regulatory effects of Aucklandia lappa Decne (AL). Methods and Results: We measured cytotoxicity, nitric oxide (NO) content, mRNA expression (iNOS, IL-1${\alpha}$, IL-$1{\beta}$, and TNF-${\alpha}$), protein expression (iNOS, COX-2, and $I{\kappa}B$) and phagocytic activity in RAW264.7 cells. Male BALB/c mice were fed 100 mg/kg AL (Aucklandia lappa Decneon 70% ethanol extract) and 250 mg/kg AL for 4 weeks; thereafter, we observed B/T or $CD4^+/CD8^+$ lymphocyte subpopulation change, and expression patterns of $CD4^+$ and $CD8^+$ lymphocytes by immunohistochemical staining in mouse splenocytes and/or thymocytes. To determine the experimental concentration of AL, cell viability was measured by MTT assay and tested at $12.5{\mu}g/m{\ell}$ or less. AL inhibited the levels of NO, lymphokine production (IL-$1{\beta}$, and TNF-${\alpha}$), and mRNA (iNOS, IL-1${\alpha}$, IL-$1{\beta}$, and TNF-${\alpha}$) and protein (iNOS, and COX-2) expression. Additionally, the levels of $I{\kappa}B$, phagocytic activity, and splenic and thymic T lymphocytes, especially $T_H$ and $T_C$ cells were significantly increased in AL administered mice. The immuno-reactive density of $CD4^+$ and $CD8^+$ lymphocytes was stronger in AL groups than in the normal group. AL stimulated NO, iNOS, and COX-2, and regulated IL-1${\alpha}$, IL-$1{\beta}$, TNF-${\alpha}$, and $I{\kappa}B$ in macrophages treated with LPS (lipopolysaccharide). In addition, AL increased the phagocytic activity of macrophages and the immunity of mouse T ($T_H$, and $T_C$) cells. Conclusions: These results suggested that AL might show anti-inflammatory activity via the suppression of various inflammatory markers and immuno-regulatory activity.

• 대한약리학회지
• /
• 제30권1호
• /
• pp.39-48
• /
• 1994

포만중추 (satiety center)를 자극하여 음식섭취를 억제한다고 알려진 CCK-8을 흰쥐 복강에 투여하여, 흰쥐 뇌의 도파민 변화에 대한 CCK-8의 효과를 관찰하였다. 흰쥐 뇌의 부위별 도파민 함량은 HPLC-ECD방법으로 측정하였으며, 시상하부와 흑질에서의 TH-immunoreactive neuron은 면역조직화학법과 영상분석법을 시행하였다. 굶긴 쥐에서는 정상 쥐에 비하여, 도파민 함량이 전두 피질, 해마, 시상하부 및 편도체에서 각각 감소하였다. CCK-8을 투여한 쥐는 정상 쥐와 굶긴 쥐에 비하여, 도파민 함량이 시상하부에서 의미있게 감소하였다. 또한 굶긴 쥐는 정상 쥐와 비교하여, TH-positive neuron의 분포와 수가 뇌실옆핵, 깔때기핵, 정중융기 및 혹질에서 현저히 감소하였다. CCK-8투여시, 시상하부와 흑질에서의 TH-immunoreactive neuron의 수는 굶긴 쥐에 비하여 증가하였다. 이상의 실험 결과로 보아 음식물 섭취를 억제하는 작용이 있는 CCK-8은 시상하부의 도파민 신경계와 일부분 관련되어 있으며, 또한 시상하부와 흑질에 존재하는 TH-positive neuron은 음식물 섭취 행위에 중요한 역할이 있음을 시사하고 있다.

• Journal of Acupuncture Research
• /
• 제23권6호
• /
• pp.117-132
• /
• 2006

Objectives : The purpose of this study is to investigate the effect of Ulmus davidiana Planch herbal acupuncture solution in LPS-stimulated RAW 264.7 cells and mouse knee joints, perfom1ed several experimental items: those are MIF mRNA, MIF, $TNF-{\alpha}$, $NF-{\kappa}B$ p65, iNOS mRNA, iNOS, NO, synovial hyperplasia, angiogenesis and fibrosis. Methods : In order to observe mRAN expression of MIF and iNOS in LPS-stimulated RAW 264.7 cells, RT-PCR was used. NO production in LPS-stimulated RAW 264.7 cells was measured by nitrite assay. All the female BALB/c mice were bred and maintained in pathogen-free mouse colonies and were 6 weeks of age on beginning of the experiment. The experimental model of RA was induced by injection of $50{\mu}g/kg$ LPS. Ulmus davidiana Planch herbal acupuncture solution was injected into either S 35 (犢鼻) or EX-LE 202 (內膝眼) of mice in turn daily for 19 days. Immunohistochemical staining was carried out to assess $TNF-{\alpha}$, $NF-{\kappa}B$ p65 and iNOS expression in synovial membrane. Synovial hyperplasia, angiogenesis and fibrosis in synovial membrane was observed with a microscope. Results : 1. Ulmus davidiana Planch herbal acupuncture solution inhibited mRNA expression of MIF and iNOS in dependence on a density of it in LPS-stimulated RAW 264.7 cells. 2. Ulmus davidiana Planch herbal acupuncture solution decreased synovial hyperplasia, angiogenesis and fibrosis in LPS-stimulated mouse knee joints. 3. Ulmus davidiana Planch herbal acupuncture solution curtailed production of MIF, $TNF-{\alpha}$, $NF-{\kappa}B$ p65, iNOS in LPS-stimulated mouse knee joints. Conclusion : On the basis of these results, It was shown that Ulmus davidiana Planch herbal acupuncture solution is significantly able to inhibit the production of MIF as a top in cytokines related to inflammatory or irrlll1une responses. Our results may provide that Ulmus davidiana Planch herbal acupuncture solution has beneficial effect in not only RA but other inflammatory or immune deases.

• 대한두경부종양학회지
• /
• 제20권2호
• /
• pp.135-142
• /
• 2004

Objectives: The purpose of this study was to evaluate for prognostic significance of VEGF expression and tumor angiogenesis in papillary carcinomas of the thyroid. Materials and Methods: The materials were 79 cases of papillary thyroid carcinomas, and age, sex, tumor size, multiplicity of tumor, capsular invasion, lymph node metastasis, recurrence, TNM stage, DeGroot stage and AMES scale were evaluated. An immunohistochemical stains for CD 34 to estimate microvessel density (MVD), and VEGF were done. MVD was defined as an average count of vessels per ${\times}400$ power field in the most vascularized area. VEGF expression was interpreted as 1+ and 2+ according to staining intensity and percentages of positive cells. Results: Mean score of MVD was $39.7{\pm}16.9.$ MVD were significantly higher in cases with capsular invasion (p=0.0001), lymph node metastasis (p=0.0001), TNM stage III (p=0.0022), DeGroot stage III (p=0.0163) and high risk group by AMES scale (p=0.0001). VEGF 2+ expression rate was significantly increased in cases with capsular invasion and lymph node metastasis (p=0.0006, p=0.0013), and in cases with TNM stage III, DeGroot stage III and high risk group by AMES scale (p=0.0236, p=0.0003, p=0.0293). In VEGF 2 + expression group, MVD was significantly higher than in VEGF 1 + group (p=0.0008), and MVD showed positive relation to VEGF 2 + expression (r=0.4616). Conclusion: VEGF expression and high MVD were significantly correlated to capsular invasion, lymph node metastasis, TNM stage III, DeGroot stage III and high risk group by AMES scale. The expression of VEGF and high MVD could be considered to be one of prognostic factor in papillary thyroid carcinomas.

• Asian Pacific Journal of Cancer Prevention
• /
• 제15권5호
• /
• pp.2353-2358
• /
• 2014

Purpose: We aimed to detect the expression of HIF-1${\alpha}$, VEGF, HPSE-1 and CD31 in SKOV3 xenografts in nude mice treated with different doses of ionizing radiation, trying to explore the possible mechanism of hypoxia and radioresistance. Methods: Nude mice bearing SKOV3 xenografts were randomly divided into 4 groups: Group A (control group, no ionizing radiation), Group B (treated with low dose of ionizing radiation: 50cGy), Group C (treated with high dose of ionizing radiation: 300cGy), Group D ( combined ionizing radiation, treated with ionizing radiation from low dose to high dose : 50cGy first and 300cGy after 6h interval). The mRNA levels of HIF-1 and VEGF in each group were detected by real time polymerase chain reaction, while HPSE-1 expression was measured by ELISA. The microvessel density (MVD) and hypoxic cells were determined through immunohistochemical (IHC) staining of CD31 and HIF-1a. Results: Significant differences of HIF-1${\alpha}$ mRNA level could be found among the 4 groups (F=74.164, P<0.001): Group C>Group A>Group D> Group B. The mRNA level of VEGF in Group C was significantly higher than in the other three groups (t=-5.267, P=0.000), while no significant difference was observed among Group A, B and D (t=1.528, 1.588; P=0.205, 0.222). In addition, the MVD was shown to be the highest in Group C (t=6.253, P=0.000), whereas the HPSE-1 level in Group A was lower than in Group B (t=14.066, P=0.000) and higher than in Group C (t=-21.919, P=0.000), and similar with Group D (t=-2.066, P=0.058). Through IHC staining of HIF-1a, the expression of hypoxic cells in Group A was (++), Group B was (+), Group C was (+++) and Group D was (+). Conclusion: Ionizing radiation with lowerdoses might improve tumor hypoxia through inhibiting the expression of HIF-1 and HPSE-1, whereas higherdoses worsen tumor hypoxic conditions by up-regulating HIF-1${\alpha}$, HPSE-1, VEGF and CD31 levels. A protocol of low-dose ionizing radiation followed by a high-dose irradiation might at least partly improve tumor hypoxia and enhance radiosensitivity.

• Asian-Australasian Journal of Animal Sciences
• /
• 제21권9호
• /
• pp.1244-1251
• /
• 2008

The aim of the present study was to delineate the expression and secretion of insulin-like growth factor (IGF) system components by pig liver cells. Hepatocytes were prepared from 3-wk-old weanling piglets following a two-step collagenase perfusion procedure, after which the cells were incubated for 24 or 48 h at a density of $2{\pm}10^5$ cells per 35-mm dish in 2-ml Williams' medium E. The cells were found to express the genes encoding IGF-I, IGF-binding proteins (IGFBPs)-2 and -3 and acid-labile subunit (ALS) by reverse transcription-polymerase chain reaction (RT-PCR) following the culture. However, IGF-I was localized to hepatocytes by immunohistochemical analysis, whereas IGFBP-3 was localized to endothelial cells, but not to hepatocytes. This indicated that the IGFBP-3 gene expression detected by RT-PCR was likely to have been contributed by unidentified non-parenchymal cells that had not been removed during the hepatocyte preparation. The conditioned culture medium (CCM) of the cells contained immunoreactive IGF-I and IGF-II, with the latter being seven-fold more abundant than the former. The CCM also contained 43-, 40-, 34-, 31-kDa doublet and 26-kDa IGFBPs as examined by Western ligand blotting. The 40-, 34- and 31-kDa doublet IGFBPs were approximately three-fold as abundant as the 43- and 26-kDa IGFBPs. Moreover, the 43- and 40-kDa doublet and the 34-kDa IGFBPs were immunoprecipitable with IGFBP-3 and IGFBP-2 antibodies, respectively. Overall, these results are similar to those known in the rat, which suggests that the IGF system components are likely to be expressed and secreted in pig liver in a manner similar to that in rat liver.

• Asian Pacific Journal of Cancer Prevention
• /
• 제16권10호
• /
• pp.4177-4183
• /
• 2015

Background: Gastric cancer is the second leading cause of cancer-related death in Asia, and the majority type is gastric adenocarcinoma (GAC). Most GAC patients die of recurrence and metastasis. Cancer stem cells (CSCs) have been thought to be responsible for the initiation, development, metastasis, and ultimately recurrence of cancer. In this study, we aimed to investigate expression and clinical significance of CSCs markers, CD133 and Lgr5, and vasculogenic mimicry (VM) in primary GAC. Materials and Methods: Specimens from 261 Chinese patients with follow-up were analyzed for CD133, Lgr5 protein expression and VM by immunohistochemical and histochemical staining. The Pearson Chi's square test was used to assess the associations among the positive staining of these markers and clinicopathological characteristics. Postoperative overall survival time was were studied by univariate and multivariate analyses. Results: In GAC tissues, positive rates of 49.0%, 38.7%, and 26.8% were obtained for CD133, Lgr5, and VM, respectively. The mean score of microvessel density (MVD) was $21.7{\pm}11.1$ in GAC tissues. There was a significantly difference between the positive and negative groups. There was a positive relationship between the VM, the expression of CD133 and Lgr5, and the score of MVD and the grades of tumor, lymph node metastasis, TNM stages (all p<0.05). The overall mean survival time of the patients with CD133, Lgr5, VM, and MVD (${\geq}22$) positive expression was lower than that of patients with negative expression. The score of MVD, positive expression of CD133 and VM were independent prognostic factors of GAC (p<0.05). Conclusions: VM, and expression of CD133, Lgr5, and the score of MVD are related to grades of tumor, lymph node metastasis, TNM stages, and overall mean survival time. It is suggested that CSCs and VM could play an important role in the evolution of GAC.

• 동의생리병리학회지
• /
• 제23권3호
• /
• pp.619-630
• /
• 2009

To investigate the hair growth activity of fractions and extract of Arisaematis Rhizoma in the hair removed skin of normal and spontaneous alopecia areata model in C57B/6N mice. These experiments were performed with the macroscopic, microscopic, immunohistochemical(VEGF, c-kit, PKC-${\alpha}$, TGF and FGF) and RT-PCR(TGF-${\beta}$, IGF, prolactin and placenta lactogen) methods. The results were as follows: Macroscopic observation after topical application of vehicle, 50% EtOH as control and extract of Arisaematis Rhizoma to the hair removed skin of C57BL/6N mice on the 9th, 11th and 15th day. Extensive hair growth activity was observed in treated group with extract of Arisaematis Rhizoma on the 9th, 11th and 15th day. In Arisaematis Rhizoma extracts treated group, hair follicles of middle stage of anagen was observed and it were grown down to subcutaneous tissue of skin in all the normal mice on 15th day. But in control group, most of hair follicles of telogen phase was observed in skin. The treatment of extract of Arisaematis Rhizoma increased expression of IGF(145%) and placenta lactogen(108%) in the skin of normal C57BL/6N mice on the 11th day compared to control group(100%). But expression of TGF-${\beta}$(90%) and prolactin(91%) decreased in the skin of normal C57B/6N mice on the 11th day compared to control group(100%). After application of fractions(chloroform, ethyl acetate and water fractions) of Arisaematis Rhizoma extract for 9th day, hair growth effect was observed in whole skin area in 50% of normal mice. But in control group, hair growth effect was not observed in whole skin area of normal mice. Immunoreactive density of VEGF, c-kit, PKC-${\alpha$ and FGF in skin of fractions of Arisaematis Rhizoma extracts was strongly stained in epidermis, bulge, secondary hair germ cells, cutaneous trunci m., subcutaneous tissue, root sheath compare to control group on the 9th day. In spontaneous alopecia areata model, The hair growth activity of Arisaematis Rhizoma extrat treated group(75%) was observed to be strong compared to control group(O%) on 7th day. These experiments suggest that fractions and extracts of Arisaematis Rhizoma may stimulate the topical hair growth activity. Thus it can be useful for treatment of alopecia areata.

• 대한수의학회지
• /
• 제40권1호
• /
• pp.1-16
• /
• 2000

Nowadays, mongolian gerbil is notably utilized for the research of brain and water deprivation because of a congenital incomplete willis circle structure in the brain, audiogenic seizure in low noise, and special cholesterol metabolism without water absorption for a long time. In this study, we are intend to identify the morphological changes of the catecholaminergic neuron of brain according to the time lapse in the condition of long term water deprivation. 55 mongolian gerbil were divided 10 groups(control, 1, 2, 3, 4, 5, 10, 15, 20, 42th day water deprivation group), of which each group include 5 mongolian gerbils and 5 normal mongolian gerbils in control group were also used for brain atlas as a control. The brains were observed by the immunohistochemical stain using the TH, DBH and PMNT antibody. The results were as followings; 1. The nerve fibers of the TH-immunoreactive neuron were observed only in the and corpus striatum of the telencephalon. 2. Intensity of the immunostain of the nerve fiber in the cerebral cortex and corpus striatum was decreased gradually day by day after water deprivation. 3. The TH-immunoreactive nerve cells were observed in the paraventricular and periventricular nucleus of the 3rd ventricular in the hypothalamus of mongolian gerbil but the number of nerve cells were decreased from the first day of the water deprivation to the 10th day and increased until the 20th day, after than redecreased from the 20th day by the continuous water deprivation. The number of nerve fibers in this area were increased in the first day, but decreased from the 2nd day of water deprivation. The shape and density of the dopamine secreting cells in the brain of mongolian gerbil by the immunoreactive stain were changed in the continuous water deprivation. In this results, we can conclude that dopamine concerned in the water metabolism of mongolian gerbil, and mongolian gerbil could be used as an animal model for the research of water deprivation.

• 대한두경부종양학회지
• /
• 제13권2호
• /
• pp.206-212
• /
• 1997

H$\"{u}$rthle cell neoplasm of the thyroid gland is an uncommon, but potentially malignant lesion. However, in many instances, the malignant potential of the H$\"{u}$rthle cell neoplasm is very difficult to judge histologically. For this reason, the biologic behavior of this tumor and its optimal treatment have come under considerable debate in recent years. In order to review the clinicopathologic features of the H$\"{u}$rthle cell neoplasm and to determine its optimal treatment modalities, we studied 26 patients with path logical proof of H$\"{u}$rthle cell tumor from January 1987 to September 1997. We also performed an immunohistochemical study using the monoclonal antibodies against antigen CD34 for the angiogenic activity of this tumor and evaluated the differences of microvessel density(MVD) between benign and malignant tumors. The age of the patients ranged from 1 to 71 years with a mean of 44.2 years. There were 6 males and 20 females(M : F= 1 : 3.3). The accuracies of fine needle aspiration biopsy and frozen section were very low; 6.3% and 34.8%, respectively. There were 20 benign tumors and 6 malignant tumors(23.1%). All the malignant tumors were microinvasive(intermediate) type which had minimal capsular invasion and most of them(5 cases) were diagnosed postoperatively. Any specific clinicopathologic differences were not seen between benign and intermediate groups. Most of the cases had conservative surgeries(15 ipsilateral lobectomy-isthmusectomy, 7 subtotal thyroidectomy) while total thyroidectomy was performed in 4 cases. Of the cases with malignant tumor, 2 had ipsilateral lobectomy-isthmusectomy, 3 had subtotal thyroidectomy and the remaining 1 had total thyroidectomy. Mean size of the tumors was 3.0 cm(0.1- 8.5 cm) in the greatest diameter and multiple tumors were seen in 6 cases(23.1 %). During the follow-up period, only one recurrence(3.8%) of benign tumor occurred but distant metastasis or cause-specific death was seen in the benign or intermediate groups. Mean MVDs of the benign(n=13) and intermediate(n=6) groups were $121.7{\pm}35.3$ and $114.3{\pm}31.7$, respectively and there was no statistical significance between them. In conclusion, because of the low accuracies of fine needle aspiration biopsy and frozen section for the H$\"{u}$rthle cell neoplasm, the extent of surgery could be individualized based on permanent pathologic examination; Conservative surgery would be adequate for patients with benign or intermediate H$\"{u}$rthle cell neoplasm and total or near-total thyroidectomy for those with definite malignancy.