• The Journal of Pediatrics of Korean Medicine
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• v.25 no.1
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• pp.28-39
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• 2011

Objectives: SaengRyoSaMulTang is a herbal formula in Oriental Medicine, known anti-allergens. However, its mechanism and cellular targets have not been found yet. Thus the study has developed to investigate the suppressive effect of SaengRyoSaMulTang. Methods: In the study, cellular viability, IL-4, IL-13 mRNA expression, IL-4, IL-13 production, manifestations of GATA-1, GATA-2, NF-AT1, NF-AT2, AP-1 and NF-${\kappa}$B p65 transcription factors were examined by Real-Time PCR, ELISA analysis and western blotting. Results: As a result of treating with SaengRyoSaMulTang extract(SRSMT), the study has shown that the amount of Th2 cytokines, which include PI induced IL-4 and IL-13, plays a significant role in suppressing effect. RBL-2H3 mast cells significantly suppressed the PI-induced Th2 cytokine production including IL-4 and IL-13 in a dose dependent manner. PI-induced IL-4 and IL-13 production was significantly suppressed by SRSMT intervention. Western blot analysis of transcription factors involving IL-4 and IL-13 expression also revealed a prominent decreases of mast cell's specific transcription factors including GATA-1, GATA-2, NF-AT2, c-Jun and c-Fos, but NF-${\kappa}$B p65. Conclusions: The study suggests that the anti-allergenic activities of SRSMT may regulate the transcription factors GATA-1, GATA-2, NF-AT2, c-Jun and c-Fos inhibiting Th2 cytokines IL-4 and IL-13 in mast cells.

• The Korea Journal of Herbology
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• v.26 no.4
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• pp.31-37
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• 2011

Objectives : Chrysanthemum boreale flower is widely distributed in Korea, Japan, China, and Eastern countries. C. boreale flower is also one of the herbs used for the treatment of various inflammatory disease in Korean Medicine. So, this research was designed to study anti-inflammatory effect of C. boreale flower and its mechanism. Methods : We investigated nitro oxide (NO) and prostaglandin $E_2$ ($PGE_2$) production by ELISA. And expressions of inducible nitric oxide synthase (iNOS), Cyclooxygenase-2 (COX-2) and nuclear factor-${\kappa}B$ P50/65 (NF-${\kappa}B$ P50, NF-${\kappa}B$ P65) were measured in RAW 264.7 murine macrophage cells induced by LPS. Results : MeOH ex., EtOAc fr., $CHCl_3$ fr. and Water fr. of C. boreale flower showed anti-inflammatory effect through inhibition of NO and PGE expression respectively. Among them, EtOAc fr. and $CHCl_3$ fr. inhibited production of NO and $PGE_2$ through inhibition of iNOS and COX-2 expression. And MeOH ex., EtOAc fr. and $CHCl_3$ fr. inhibited translocation of NF-${\kappa}B$ P65, NF-${\kappa}B$ P50 by inhibiting phosphrylation of $I{\kappa}B$. Conclusions : MeOH ex. EtOAc fr, $CHCl_3$ fr., and Water fr. of the C. boreale flower have anti-inflammatory activity.

• The Korean Journal of Physiology and Pharmacology
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• v.14 no.4
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• pp.235-240
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• 2010

Toll-like receptors (TLRs) functionally expressed in salivary epithelial cells, but their roles remain elusive. Among TLRs family, TLR3 is activated by dsRNA, a byproduct of viral infection. The aim of this study was to investigate the role of TLR3 in the inflammatory immune responses using HSG cells. Reverse transcriptase-polymerase chain reaction (RT-PCR), real-time PCR and ELISA were performed to identify expression of TLRs and TLR3-mediated chemokine inductions. The chemotaxis assay of activated T lymphocytes was also performed. Treatment of HSG cells with polyinosinic: polycytidylic acid (poly(I:C)) significantly increased interferon-$\gamma$-inducible protein 10 (IP-10), interferoninducible T-cell $\alpha$ chemoattractant (I-TAC), and regulated on activation, normal T-cells expressed and secreted (RANTES) gene expressions in a concentration-dependent manner. Anti-TLR3 antibody blocked the increases of IP-10 and I-TAC genes. Poly(I:C)-induced increases of IP-10 and I-TAC were also confirmed at protein levels from cell lysates, but their release into extracellular medium was detected only in IP-10. We found that the culture media from HSG cells stimulated with poly(I:C) significantly increases T lymphocyte migration. Our results suggest that TLR3 plays an important role in chemokine induction, particularly IP-10, in salivary epithelial cells.

• Journal of Microbiology and Biotechnology
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• v.27 no.7
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• pp.1336-1344
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• 2017

Hepatitis B virus (HBV) is a major cause of liver cirrhosis and hepatocellular carcinoma. With recent identification of HBV receptor, inhibition of virus entry has become a promising concept in the development of new antiviral drugs. To date, 10 HBV genotypes (A-J) have been defined. We previously generated two murine anti-preS1 monoclonal antibodies (mAbs), KR359 and KR127, that recognize amino acids (aa) 19-26 and 37-45, respectively, in the receptor binding site (aa 13-58, genotype C). Each mAb exhibited virus neutralizing activity in vitro, and a humanized version of KR127 effectively neutralized HBV infection in chimpanzees. In the present study, we constructed a humanized version (HzKR359-1) of KR359 whose antigen binding activity is 4.4-fold higher than that of KR359, as assessed by competitive ELISA, and produced recombinant preS1 antigens (aa 1-60) of different genotypes to investigate the binding capacities of HzKR359-1 and a humanized version (HzKR127-3.2) of KR127 to the 10 HBV genotypes. The results indicate that HzKR359-1 can bind to five genotypes (A, B, C, H, and J), and HzKR127-3.2 can also bind to five genotypes (A, C, D, G, and I). The combination of these two antibodies can bind to eight genotypes (A-D, G-J), and to genotype C additively. Considering that genotypes A-D are common, whereas genotypes E and F are occasionally represented in small patient population, the combination of these two antibodies might block the entry of most virus genotypes and thus broadly neutralize HBV infection.

• Archives of Pharmacal Research
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• v.24 no.6
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• pp.557-563
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• 2001

Oral tolerance is thought to play a role in preventing allergic responses and immune-mediated diseases. An improved mouse model of the oral tolerance to Japanese cedar pollen (JCP) as antigen was developed in order to detect induction of the tolerance, and the immunological characteristics of this model were also elucidated. Oral tolerance was induced by C3H/ HeN mice given an oral administration of 10 mg JCP 7 days before immunization with an i.p. injection of 0.1 mg JCP in complete Freunds adjuvant (CFA). The effects of oral JCP on systemic immunity were assessed by enzyme-linked immunosorbent assay (ELISA) of immunoglobulin (Ig) levels in serum collected on day 7 or 14 after immunization. Oral tolerance to JCP was adequately induced on day 7 after immunization and was more effective in C3H/HeN mice than in BALB/c mice. The tolerance was primarily concerned with the decreased serum levels of antigen-specific IgG. In these mice, oral administration of JCP also suppressed various immune responses to the antigen including delayed-type hypersensitivity (DTH), total Igl level and anti-JCP IgGl level. The suppression of these immune responses by the oral antigen was associated with a significant reduction in interleukin-4 (IL-4) production. These findings therefore indicate that this C3H/HeN mice model has potential use in detecting the induction of oral tolerance by JCP and suggest that this tolerance model may be effective in the treatment and prevention of allergic responses caused by the antigen.

• Korean Journal of Agricultural Science
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• v.47 no.4
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• pp.815-827
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• 2020

The epidermal growth factor (EGF) transgenic soybean was developed and biosynthesis of human epidermal growth factor (hEGF) in soybean seeds was confirmed. Also, EGF transgenic soybean were found to contain a herbicide resistance selectable marker by introduction of phosphinothricin acetyltransferase (PAT) gene from the Streptomyces hygroscopicus. For biosafety assessment, the EGF transgenic soybean expressing the EGF biosynthesis gene EGF and herbicide resistant gene PAT was tested to determine effects on survival of Cyprinus carpio, commonly used as a model organism in ecotoxicological studies. C. carpio was fed 100% ground soybean suspension, EGF soybean or non-genetically modified (GM) counterpart soybean (Gwangan). Gene expression of EGF soybean was confirmed by PCR and ELISA to have EGF/PAT. Feeding test showed that no significant differences in cumulative immobility or abnormal response between C. carpio samples fed on EGF soybean and non-GM counterpart soybean. The 48 h-EC50 values of the EGF and non-GM soybean were 1,688 mg·L-1 (95% confidence limits: 1,585 - 1,798 mg·L-1) and 1,575 mg·L-1 (95% confidence limits: 1,433 - 1,731 mg·L-1), respectively. The soybean NOEC (no observed effect concentration) value for C. carpio was suggested to be 625 mg·L-1. We concluded that there was no significant difference in toxicity for non-target organisms (C. carpio) between the EGF soybean and non-GM counterparts.

• Journal of Veterinary Clinics
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• v.20 no.1
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• pp.66-73
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• 2003

Numerous physiological effects are attributed to conjugated linoleic acid(CLA). The purpose of this study is to consider these effects with respect to the cis-9, trans-11 and trans-10, cis-12 CLA isomer. Both isomers are natural products. The c9,t11-CLA isomer is the principal dietary form of CLA, but the concentrations of this isomer and the t10,c12-CLA Isomer in dairy products or beef vary depending on the diet fed to cows or steers, respectively. The influence of dietary CLA isomers on the immune response was examined, body weight and weight ratio of organ to body of Balb/C mice. Mice were divided into four groups of 8 mice. Balb/C mice were fed the experimental diets supplemented with 1% CLA (c9,t11-CLA isomer : t10,c12-CLA isomer = 2:3) (Group 1), 1% CLA (c9,t11-CLA isomer t10,c12-CLA isomer : 1:1) (Group 2), 1% safflower oil (Group 3) or nothing (Control) for 3 weeks. After 3 weeks, serum, gut lumen lavage, fat, liver, spleen and thymus were taken. Measurement of total immunoglobulin were executed using sandwich ELISA. Serum levels of IgA and IgM showed that group fed with t10,c12-CLA isomer significantly were higher than group fed with c9,tl1-CLA isomer. In addition serum level of IgG showed that group fed with t10,c9-CLA isomer significantly were lower than group fed with c9,tl1-CLA isomer. However, no significantly differences were observed in the serum IgE and secretory IgA. Weight ratio of spleen to body showed no significant differences. In weight ratio of liver and thymus to body, tl0,c9-CLA isomer significantly were respectively higher than group fed with c9,t11-CLA isomer. In weight ratio of fat to body, tl0,c9-CLA isomer significantly were respectively lower than group fed with c9,tl1-CLA isomer. In conclusion, t10,c12-CLA isomer produced a situation favorable for immunopotentiative effect and body composition. But it should be protected against hepatomegaly induced lipid accumulation in liver.

• Parasites, Hosts and Diseases
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• v.24 no.1
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• pp.25-41
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• 1986

The applicability of micro-ELISA was evaluated in human neuro-cysticercosis using paired samples of serum and CSF. A total of 355 cases who were mostly neurologic patients was subjected. Cystic fluid of C. cellulosae was used as antigen in protein concentration of $2.5{\;}{\mu}g/ml$. Serum was diluted to 1 : 100 and CSF was undiluted in the assay for the specific IgG antibody level. The differential criterion of the positive reaction was the abs. of o. 18 in both samples. The results were summarized as follows: 1. The overall sensitivity of the micro-ELISA in 71 confirmed neurocysticercosis was 90.1% ; the sensitivity by serum was 77.5% and that by CSF was 83.1%. CSF was a more sensitive and valuable material. Most of the false negative cases of neuro-cysticercosis showed far lower level of abs. rather than marginal. 2. The overall specificity of the micro-ELISA in 52 confirmed other neurologic diseases was 88.5%; the specificities by serum and by CSF were 94.2% respectively. Cases of other neurologic diseases did not show false positive reactions in both samples. 3. When serum was assayed, taeniasis(2/18), sparganosis(2/20), paragonimiasis(1/56), clonorchiasis(1/15) and fascioliasis(1/1) cases showed cross reactions. When CSF was assayed, 2 ot 10 neuro-sparganosis showed cross reactions while none of 9 neuro-paragonimiasis showed it. Out of 71 confirmed neuro-cysticercosis cases, 6 and 11 showed cross reactions by serum and CSF to crude extract antigen of sparganum; but no case did show it to crude extract antigen of Paragonimus westermani. 4. Ventricular CSF showed low or negative levels of IgG antibody than lumbar CSF unless the lesion was at the lateral ventricle itself. 5. Out of 4 racemose cysticercosis cases, 3 showed positive reaction in serum while all of 3 examined CSF were positive. The above results indicated that the serological test for detecting the specific IgG antibody by micro-ELISA using paired samples of serum and CSF was very helpful for clinical differentiation of neuro-cysticercosis from neurologic diseases of other causes.

• Journal of Korean Medicine Rehabilitation
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• v.34 no.2
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• pp.15-28
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• 2024

Objectives We used the D-galactose (D-gal) induced C2C12 myoblast senescence model to investigate whether ethanol extract of Perilla. fructescens leaves (EEPF) could delay cellular senescence and regulate related mechanisms. Methods C2C12 myogenic cells were cultured in an incubator under 37 ℃ and 5% CO₂ conditions. EEPF, dried perilla leaves were pulverized and extracted at 1:10 (v/v) at 50 ℃ for 4 hours. Cell counting kit-8 and western blot analysis was performed. Annexin V-FITC apoptosis detection kit and DAPI staining was applied. Catalase (CAT), glutathione peroxidase (GSH-Px), total antioxidant capacity (T-AOC), superoxide dismutase (SOD), and malondialdehyde analysis kits were used. To measure the level of reactive oxygen species generation, staining and flow cytometry was used. To analyze the mitochondrial activity, membrane potential changes were measured using JC-1. 𝛽-gal activity was analyzed using SA-𝛽-gal staining solution, and DNA damage was analyzed by using 𝛾-H2AX. Quantikine ELISA kit was used to analyze inflammatory cytokine production. Results According to the results of this study, EEPF significantly alleviated the decrease in cell viability in C2C12 cells treated with D-gal and suppressed the decrease in the expression of proliferating cell nuclear antigen. EEPF also markedly blocked D-gal-induced C2C12 cell apoptosis and restored reduced activity of CAT, GSH-Px, T-AOC, SOD. In addition, EEPF suppressed the decrease in 𝛽-galactosidase activity, the induction of DNA damage and the increase in expression of senescence-associated secretory phenotype proteins such as p16, p53 and p21 in D-gal-treated C2C12 cells. Furthermore, EEPF significantly attenuated D-gal-induced production and expression of inflammatory cytokines such as interleukin (IL)-6 and IL-18. Conclusions The results of this study indicate that EEPF can be used as a potential candidate for the prevention and treatment of muscle aging.

• The Korea Journal of Herbology
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• v.24 no.4
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• pp.215-224
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• 2009

Objectives : Atopic dermatitis is a chronic inflammatory disease characterized by typically distributed eczematous skin lesion with pruritus, lichenification and dry skin. In this study, we performed to assess the therapeutic effects of co-treatment of Chenilyeomgamibang (CGB) and Chenggihaedok-san (CHS, C&C) on the TNCB(trinitrochlorobenzene)-induced atopic dermatitis in NC/Nga mice, characterized by the onset of atopic dermatitis along with an increase the number of inflammatory cells and dysregulation of Th2 cytokines. Methods : Defined amount of CGB was sprayed on mice skin and CHS was simultaneously orally administrated to TNCB treated NC/Nga mice for 5 weeks. The immune cell types were caracterized by flow cytometry using each specific antibody. The amount of Th2 cytokines in serum and splenocytes culture supernatant were measured by ELISA. Results : Administration of C&C significantly reduced clinical dermatitis severity including pruritus, edema, eczematous and erythema. Histological findings indicated that the thickening of epidermis/dermis and dermal infiltration of inflammatory cells were dramatically reduced. Flow cytometry analysis showed that infiltrated immune cell numbers of CCR3+, B220+/IgE+, Gr-1+/CD11b+, and CD117+ were significantly reduced in C&C-treated dorsal skin lesion. Furthermore, T cell composition rate in PBMC was also dramatically decreased by the treatment. C&C greatly down-regulated production of Th2 cytokines including IL-4, IL-5, IL-13 in the serum. The down- regulatory effects of C&C on these Th2 cytokines production were also detected in CD3/ CD28 activated splenocytes. Conclusions : These results indicated that C&C is a plausible therapeutic agent for treatment of atopic dermatitis through regulating the Th2 skewed immune system.