Objectives : Nitric oxide (NO) plays an important role in normal and pathophysiological cells as a messenger molecule, neurotransmitter, microbiological agent, or dilator of blood vessels and arteriosclerosis, respectively. This study was undertaken to understand the mechanism of NO production and effect of Hyeolbuchukeo-tang (Xiefuzhuyu-tang) on NO production in cultured vascular smooth muscle cell (VSMC). Methods and Results : VSMC was isolated from aorta and cultured. Cultured primary cells were identified as VSMC with anti--smooth muscle actin antibody. A large amount of NO was produced in cultured VSMC treated with $IFN-{\gamma}$ plus TNF in a time- and dose-dependent manner. $TNF-{\alpha}$ was a more efficient stimulator than $IFN-{\gamma}$ in NO production of cultured VSMC. iNOS protein wasdetected within 3 hrs and it increased up to 12 hrs in a time-dependent manner. However, accumulated NO in cytokine-treated VSMC was not detected within 3 hrs. NO production in cytokine-treated VSMC showed the dose- and time-dependent manner, and increased up to 48 hrs. The activated VSMC produced a large amount of NO (about 60 uM). Hyeolbuchukeo-tang (Xiefuzhuyu-tang) alone did not induceNO production, but it potentiated the effect of $TNF-{\alpha}$ on NO production and increased NO production by about 20%. Hyeolbuchukeo-tang (Xiefuzhuyu-tang) did not affect the transcriptional activity of iNOS gene, but increased the accumulation of iNOS. These results indicate that Hyeolbuchukeo-tang (Xiefuzhuyu-tang) could modulate the translational level of iNOS. PKC did not modulate NO production, but calcium ionophore A23187 decreased NO production. However, Hyeolbuchukeo-tang (Xiefuzhuyu-tang) elevated the decreased NO production in A23187-treated VSMC by modulating the stability of iNOS transcripts. Half-life of the synthesized transcripts appeared to have about 6 hrs. PDTC, an $NF-{\kappa}B$ inhibitor, blocked the accumulation of iNOS mRNA, indicating that $NF-{\kappa}B$ served as an important modulator in the transcriptional regulation of iNOS. As Hyeolbuchukeo-tang (Xiefuzhuyu-tang) potentiated the effect of the $TNF-{\alpha}$ on NO production but had no additional effect on PDTC-modulated NO production, it is suggested that Hyeolbuchukeo-tang (Xiefuzhuyu-tang) enhances the $TNF-{\alpha}-mediated$ NO production of VSMC by modulating the iNOS activity and the stability of iNOS transcripts in activated VSMC having the elevated intracellular calcium ion. Conclusions : This study suggests that Hyeolbuchukeo-tang (Xiefuzhuyu-tang) has a potential capacity for preventing and treating diseases of the circulation system, including arteriosclerosis.
Background: Membranous nephropathy (MN) is the most common cause of adult nephrotic syndrome worldwide and has been defined as granular subepithelial deposition of immune complexes along the glomerular basement membrane (GBM). MN has few known treatments and gives rise to side effects under treatment with steroids and immunosuppressives. Objective: The purpose of this experimental study was to demonstrate the effects of Scutellariae Radix extract (SRE) treatment on MN mouse model induced by cBSA. Methods: We divided mice into 4 groups. The Normal group had no treatment. We induced MN mouse model to the other 3 groups by injecting cBSA into the abdominal cavity. The control group was treated with cBSA (10 mg/kg, i.p.) only. The second group, 'SRE-250', was treated with cBSA (10 mg/kg, i.p.) and SRE (250 mg/kg, p.o.). The third group, 'SRE-500', was treated with cBSA (10 mg/kg, i.p.) and SRE (500 mg/kg, p.o.). After cBSA and SRE treatment for 4 weeks, gain in body weight, 24hrs proteinuria, serum albumin, total cholesterol, triglyceride, BUN and creatinine of all groups were measured. TNF-$\alpha$, IL-6, IL-1$\beta$, IL-10, IFN-$\gamma$, IgA, IgM and IgG levels of all groups were gauged. H&E staining and electron microscopy of the kidney were observed. Results: SRE showed significant decrease in the 24hrs proteinuria, serum triglyceride, BUN, TNF-$\alpha$, IL-6, serum IgA, IgM and IgG levels compared with the control group. SRE showed increase in the serum IL-10 and IFN-$\gamma$ levels compared with control on RT-PCR. SRE considerably decreased in the thickening of the GBM on H&E staining and deposition of electron-density on electron microscopy of the kidney compared with the control. Conclusions: According to the above results, it is suggested that SRE decreases the symptoms of MN induced by cBSA in mouse model. Therefore, SRE seems to be applicable to MN in clinical practice.
Bamboo salt has been used for the purpose of prevention and treatment of various diseases in Korea. Present study was carried out to ascertain the effects of purple bamboo salt upon anti-allergic effect, anti-inflammatory activity and immune-enhance effect as well. Purple bamboo salt significantly inhibited the ear swelling response and histamine release induced by compound 48/80 in mice and rat peritoneal mast cells. Purple bamboo salt (0.01 ∼ lg/kg) also dose-dependently inhibited the passive cutaneous anaphylaxis by oral administration. Purple bamboo salt (1 mg/mL) in hibited phorbol 12-myristate 13-acetate plus calcium ionophore A23187-stimulated tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-1${\beta}$ and IL-6 secretion, by 67.04${\pm}$0.08%, 68.01${\pm}$1.85%, 69.48${\pm}$0.54%, respectively. In addition, purple bamboo salt inhibited the expression of TNF-${\alpha}$ mRNA in HMC-1 cells. Finally, we investigated the effect of purple bamboo salt in the forced swimming test (FST) and the change of purple bamboo salt-mediated cytokine production from MOLT-4 cells. At the 7th, immobility time was significantly decreased in the purple bamboo salt-administration group (35.4 ${\pm}$5.9 s for 1 g/kg) in comparison with the control group (93.2 ${\pm}$ 15.45). After FST, the content of glucose in the blood serum was increased and the levels of blood urea nitrogen, lactic dehydrogenase was decreased in purple bamboo salt-administration group. However, it had no effect on the elevation of CK and TP level. Purple bamboo salt (1 mg/mL) significantly increased the interferon (IFN)-${\gamma}$ and IL-2 level compared with media control (about 3.7-fold for IFN-${\gamma}$, about 3.5-fold for IL-2, p〈0.05) but did not affect the IL-4.
This study was conducted to determine the effects of dietary supplementation of either 100 mg/kg chito-oligosaccharide (COS) or chlortetracycline (CTC) with corn-soybean-fish meal on immunity in broiler chickens. A total of 147 one-day old male broiler chicks were randomly allocated to 3 treatments with 7 replicate pens per treatment and 7 birds per pen. The experimental diets consisted of a control diet based on corn, soybean and fish meal without COS and any antibiotic supplement and similar diets supplemented with either CTC (80 mg/kg from d 1 to 21 and 50 mg/kg from d 22 to 42) or COS (100 mg/kg from d 1 to 42). During the entire experimental period, all birds had ad libitum access to diets and water. The main immune organ indices, T-lymphocyte proliferation, serum cytokine concentrations, serum NO level and serum iNOS activity were measured on d 21 and d 42. On d 21, broilers fed 100 mg/kg COS had improved (p<0.01) indices of spleen, thymus, and bursa of Fabricius compared with the control and CTC birds. Birds receiving 100 mg/kg COS had higher (p<0.05) serum concentrations of $IL-1{\beta}$, IL-6, IgM, NO and iNOS than birds on the control treatment. Serum $Ca^{2+}$ level of birds fed 100 mg/kg COS tended to be higher (p = 0.049) than in birds fed CTC. On d 42, the birds fed 100 mg/kg COS had higher (p<0.05) concentrations of TNF-${\alpha}$ and IgM in serum than birds in both the CTC and control treatments. Birds fed 100 mg/kg COS had a higher concentration of IFN-$\gamma$ than the control group. In conclusion, dietary supplementation of COS appeared to improve the immunity of broilers by promoting the weight of the main immune organs, increasing IgM secretion, stimulating microphages to release $TNF-{\alpha}$, $IL-1{\beta}$, IL-6 and IFN-$\gamma$, and activating iNOS to induce NO.
In order to reveal immunopathogenesis of periodontal tissue destruction, it is important to clarify the molecular mechanism of trafficking and retention of activated leukocytes, including monocytes/macrophages. Gingival fibroblasts may be involved in the regulation of inflammatory cell accumulation in the extravascular periodontal connective tissues via cytokine production and surface expression of adhesion molecules. In this study, it was investigated the molecular basis for the adhesive interactions between monocytes and fibroblasts such as peri-odontal ligament fibroblast(PDLF), human gingival fibroblast(HGF), and human dermal fibroblast(HDF). First, it was examined the evidence whether monocyte-fibroblast cell contact may cause signal transduction in fibroblasts. Being directly in contact with fixed human monocyte cell line THP-1, or U937, upregulation of IL-6 production, $TNF-{\alpha}$ mRNA expression and increased cell proliferation could be seen for fibroblasts. IL-6 production induced by monocyte- fibroblast coculture were further increased when fibroblasts had been pretreated with $IFN-{\gamma}$ or $IL-1{\beta}$ , and monocytes with LPS. Next, it was examined the expression of ICAM-1 which has been known to be involved in accumulation and activation of leukocytes in inflammatory diseases such as periodontitis. ICAM-1 was upregulated up to 10-fold on PDLF, HGF, and HDF by exposure to $IFN-{\gamma}$ or $IL-1{\beta}$. Furthermore, anti-ICAM-1 monoclonal antibody clearly blocked cocultureinduced IL-6 production by fibroblasts, suggesting that $ICAM-1/{\beta}_2$integrin pathway is involved in periodontal fibroblastmonocyte interaction. Overall, these findings provide evidence that periodontal fibroblasts could be involved in the accumulation and retention of monocytes/macrophages in periodontal inflammatory lesion at least in part by ICAM-1 expression. In addition, periodontal fibroblast-monocyte interaction could cause activation signals in fibroblasts intracellularly which result in cytokine production and cell proliferation. Thus, periodontal fibroblasts are speculated to play an important role in immunoregulation and tissue destruction in chronic periodontal diseases by interaction with monocytes/macrophages.
This study was conducted to compare the general composition and immunomodulatory activity of breast and thigh meats from four lines of Korean native chickens: Yeonsan Ogye, Hyunin Black, Hwangbong, and Hoengseong Yakdak. White Leghorn was used as a control. Fifteen male chickens (three chickens in each line) were grown under the same conditions and slaughtered at 13 weeks old. The four lines of Korean native chickens, regardless of the part, had higher contents of crude fat (p<0.05) than White Leghorn. The cholesterol contents were significantly higher in Hyunin Black and significantly lower in Hoengseong Yakdak than those of other chickens (p<0.05). The immunomodulatory effect, assessed by macrophage cell proliferation and nitric oxide production, was only observed in the breast meat of the four lines of Korean native chickens. The phagocytic activity of macrophage cells was significantly augmented by the breast meat of Hyunin Black and Hoengseong Yakdak. Furthermore, the mRNA expressions of pro-inflammatory cytokines, such as IL-4, IL-10 and $IFN-{\gamma}$, was significantly suppressed by Korean native chickens compared with White Leghorn. These results suggested that the four lines of Korean native chickens exhibited greater immune-enhancing activity than White Leghorn.
High levels of inflammatory cytokines were proposed contributors to the pathogenesis of a various inflammatory skin disorders. Therefore, investigating the immune response of the inflammatory skin disorder allows a better understanding of pathogenesis of a various inflammatory skin disorders and therapeutic approaches. The aim of this study was to analyze of the immune response in dogs with chronic inflammatory skin disease. To this aim, the present study evaluated relative mRNA expression of canine $IFN-{\gamma}$, IL-4, $TGF-{\beta}$ and IL-10 using TaqMan realtime PCR assays and semi-quantitative RT-PCR in freshly isolated peripheral blood mononuclear cells from twenty dogs with chronic inflammatory skin disease and ten normal dogs. The relative mRNA expression levels of IL-4 mRNA were significantly higher in dogs with chronic inflammatory skin disease than those in normal dogs (P < 0.01). The results of present study also showed a tendency towards increased expression of IL-10 transcripts in dogs with chronic inflammatory skin disease. However, there were no significant differences in the levels $IFN-{\gamma},\;TGF-{\beta}$ between normal and chronically inflammed dogs. In addition, the concentration of serum IgE was significantly increased in dogs with chronic inflammatory skin disease compared with those in normal dogs (P < 0.01). In histopathological examination, we found that there were markedly increased mast cell counts in chronically inflammed dogs (P < 0.05). These results suggest that the pathogenesis of chronic inflammatory skin disease might be associated with a T-cell mediated inflammatory responses characterized by a Th2-skewed immune response. Based on these results, the modulation of Th1/Th2 balance may be an effective therapeutic strategy for the treatment of chronic inflammatory skin disease.
In this experiment, the effects of Pueraria thunbergiana extracts on the activation of immune cells were studied. An immune cell-activating factor was partially purified from P. thunbergiana by means of physiological saline extraction, acetone precipitation, and heating inactivation. P. thunbergiana extracts increased the proliferation of spleen cells and induced the production of IL-2, IL-6, TNF-${\alpha}$, and IFN-${\gamma}$ by spleen cells. Also, they increased the proliferation of purified B cells and the production of IgM antibody in a dose-dependent fashion. The extract self-induced NO synthesis in a mouse macrophage cell line (RAW264.7). When cell lines were treated with extracts, the cytokines' (IL-$1{\beta}$, IL-6, and TNF-${\alpha}$) production was markedly increased. Therefore, P. thunbergiana extract can self-activate spleen cells, B cells, and macrophages. These results might be useful in further studies into a possible immune-activating agent derived from P. thunbergiana for the development of functional foods and drugs.
Kim, Hyang Suk;Chung, Kyung Tae;Lee, In Hwan;Choi, Woo Bong;Lee, Jong Hwan;Hyun, Sook Kyung;Kim, Byung Woo;Hwang, Hye Jin
Journal of Life Science
/
v.24
no.1
/
pp.61-66
/
2014
The purpose of this study was to investigate the immunomodulatory effects of Alpina officinarum (AO) ethanol extract on immunocompromised mice. The mice were injected intraperitoneally with an immunosuppressive drug, cyclophosphamide, and then administrated orally with 30, 100, and 300 mg/kg of ethanol extract of AO (AO 30, AO 100, and AO 300, respectively). The concentrations of cytokines and immunoglobulins (IgM, IgA, IgG) in serum were measured. The body weight of the mice and spleen cell number of the AO-fed group showed no significant difference compared to a control group. The concentrations of several cytokines, including IL-2, IFN-${\gamma}$, and TGF-${\beta}$, in serum showed a significant increase in the AO 100 group compared to the control and other groups (p<0.05). The IL-4 level showed no significant difference in the experimental groups. The supplementation of AO (30, 100, 300 mg/kg) significantly increased the concentration of IgM (p<0.05). The concentration of IgA was significantly increased in the AO 100 group (p<0.05) compared to the control group. It can be concluded that AO ethanol extract enhances immune function by promoting the production of cytokines and immunoglobulins.
Bartonella henselae is the causative agent of cat scratch disease. Although cats are the main zoonotic reservoirs of Bartonella spp., unusual cases of cat scratch disease caused by a domestic dog scratch have been recently reported. For the in vivo B. henselae infection, eight dogs were inoculated intradermally with $2{\times}10^8CFU$ of B. henselae Houston-1 suspended in 1 ml of phosphate buffered saline on day 0 and subsequent injections of the same amount given intradermally on days 21, 28, 36, 58 and 64. After in vivo canine B. henselae infection was confirmed by nested PCR, the IFN-$\gamma$ levels of the culture supernatant of PBMC stimulated with B. henselae was significantly higher in the B. henselae-PCR positive group than the B. henselae-PCR negative group. Our results showed that the canine immune responses against B. henselae were different from those of cats. Th1 activation by B. henselae stimulation was characterized in dog peripheral blood mononuclear cells, whereas Th2 activation was reported in B. henselae-infected cats.
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