• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.5
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• pp.1251-1256
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• 2003

To determine the effect of Zibachunggan-tang(ZCT) on the process of lipopolysaccharide (LPS)-induced nuclear factor-kBp65 (NF-kBp65) activation, and LPS-induced expression of pro-inflammatory proteins including tumor necrosis factor-α (TNF-α), nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), in HepG2 cell. Immunoblot analysis showed that the level of nucleic NF-kBp65 was rapidly up-regulated and cytosolic inhibitory I-kBα was down-regulated by LPS challenge. While ZCT inhibited an increase of NF-kBp65 and degradation of I-kBα in HepG2 cell. Beside LPS-induced expression of a group of genes, such as TNF-α, inducible iNOS and COX-2, are repressed by ZCT. It may be concluded that ZCT attenuates the progress of LPS-induced inflammation by reduction of NF-kBp65 activation. The ZCT would be useful as a therapeutic agent for endotoxin-induced liver disease.

• Journal of Life Science
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• v.10 no.2
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• pp.55-60
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• 2000

We examined the effects of Kamgil-Tang on the process of lipopolysaccharide (LPS)-induced unclear factor (NF)-${\kappa}$ Bp65 and inhibitory (I)-${\kappa}$ B${\alpha}$ alteration in RAW 264.7 cell and acute lung injury in rats. Immunoblot analysis showed that LPS-induced degradation of I-${\kappa}$ B${\alpha}$ in RAW 264.7 was inhibited by pretreatment of Kamgil-Tang. The total cells of bronchoalveolar lavage fluid by LPS challenge markedly decreased in the Kamgil-Tang pretreatment rats. Kamgil-Tang pretreatment caused also a decline in neutrophils infiltration into interstitium of the lung. In the alveolar macrophages and neutrophils, decreased NF-${\kappa}$ Bp65 and inducible nitric oxide synthase and increased I-${\kappa}$ B${\alpha}$ immunoreaction were detected in Kamgil-Tang pretreated rats compared with LPS alone treated ones. It may be concluded that Kamgil-Tang attenuates the development of LPS-induced inflammation by reduction of NF-${\kappa}$ Bp65 activation and neutrophil-mediated acute lung injury. Kamgil-Tang would be useful as a therapeutic agent for endotoxin-induced lung disease.

• The Journal of Korean Medicine
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• v.26 no.1 s.61
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• pp.18-25
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• 2005

Objective & Methods: We examined the effects of Platycodi radix on the process of lipopolysaccharide (LPS)-induced nuclear factor $NF-{\kappa}Bp65$ and inhibitory $(I)-{\kappa}B{\alpha}$ alteration in RAW 264.7 cells and acute lung injury in rats. Results: Immunoblot analysis showed that LPS-induced degradation of $I-{\kappa}B{\alpha}$ in RAW 264.7 was inhibited by pretreatment of Platycodi radix. The total cells of bronchoalveolar lavage fluid by LPS challenge markedly decreased in the Platycodi radix pretreatment rats. Platycodi radix pretreatment also caused a decline in neutrophils infiltration into interstitium of the lung. In the alveolar macrophages and neutrophils, decreased $NF-{\kappa}Bp65$ and inducible nitric oxide synthase and increased $I-{\kappa}B{\alpha}$ immunoreaction were detected in Platycodi radix pretreated rats compared with LPS alone treated ones. Conclusion : It may be concluded that Platycodi radix attenuates the development of LPS-induced inflammation by reduction of $NF-{\kappa}Bp65$ activation and neutrophil-mediated acute lung injury. Platycodi radix would be useful as a therapeutic agent for endotoxin-induced lung disease.

• Journal of Life Science
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• v.14 no.2
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• pp.215-220
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• 2004

We investigated the anti-inflammatory effects of aqueous extract of Artemisia capillaris Thunb. (AEAC), a traditional Korean herb for remedying liver disease, for suppression in the process of lipopolysaccharide (LPS)-induced inflammation in the liver of rat. Level of glutamate oxaloacetate transaminase (GOT), glutamate pyruvate transaminase (GPT) and lactate dehydrogenase (LDH) was increased in the serum of LPS-treated rats compared to normal, however, in the rats pretreated with AEAC, the increase of GOT, GPT and LDH value was arrested. More severe histological changes of liver such as cloudy swelling, hydropic degeneration, Kupffer cell reaction and inflammatory cells infiltration were demonstrated in the rats challenged with LPS compared with normal. Fewer scores of these changes were observed in rats pretreated with AEAC. Immunohistochemical analysis showed that while the expression of the nuclear factor (NF)-kBp65, inducible nitric oxide synthase (iNOS), tumor necrosis factor (TNF)-$\alpha$ and COX (cyclooxygenase)-2 tended to increase, that of inhibitory (I)-kBa was decreased in the hepatocytes of rats challenged with LPS. A slight decline of NF-kBp65, TNF-$\alpha$ and COX-2, but increase of I-kB$\alpha$ were observed in the hepatocytes of the rats pretreated with AEAC. These results suggest that AEAC may act as a therapeutic agent for liver disease through a regulation of inflammation-related proteins.

• Journal of Physiology & Pathology in Korean Medicine
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• v.26 no.5
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• pp.738-744
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• 2012

Gastrodia elata Blume is used for a variety of purposes including treatment of inflammation in the Korean medicine. The present study investigated whether the G. elata extracts have the anti-inflammatory effect on lipopolysaccharide(LPS)-stimulated BV-2 microglia cells. G. elata extracts showed an anti-inflammatory effects in the morphological and nitric oxide(NO) analysis, especially in hexane extract. So we investigated the hexane extract from G. elata in the following experiments. Hexane extract significantly inhibited the secretion of NO with protein level of inducible nitric oxide synthase in LPS-stimulated BV2 microglia cells. Hexane extract also inhibited LPS-stimulated inflammatory responses involving the degradation of cytosolic inhibitory(I)-${\kappa}B{\alpha}$ and the translocation of nuclear factor(NF)-${\kappa}Bp65$ to nucleus in LPS-stimulated BV-2 microglia cells by morphological analysis. Western blot analyse confirmed that I-${\kappa}B{\alpha}$ and NF-${\kappa}Bp65$ showed a similar pattern as morphological analysis. Our results suggest that G. elata extracts, especially hexane extract, may act as a therapeutic agent for inflammatory disease in the central nervous system through a selective regulation of NO production and NF-${\kappa}B$ activation.

• Biomedical Science Letters
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• v.10 no.2
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• pp.65-73
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• 2004

I report that single-stranded antisense as a part of large circular (LC-) genomic DNA of recombinant M13 phage exhibits enhanced stability, sequence specific antisense activity, and no need for target site search. A cDNA fragment (708 bp) of rat TNF-$\alpha$ was inserted into a phagemid vector, and TNF-$\alpha$ antisense molecules (TNF$\alpha$-LCAS) were produced as single-stranded circular DNA. When introduced into a rat monocyte/macrophage cell line, WRT7/P2, TNF$\alpha$-LCAS was able to ablate LPS-induced TNF-$\alpha$ mRNA to completion. The antisense effect of TNF$\alpha$-LCAS was shown to be sequence-specific because expressions of three control genes ($\beta$-actin, GAPDH and IL-1$\beta$) were not significantly altered by the antisense treatment. Further, TNF$\alpha$-LCAS was found to be highly efficacious as only 0.1 $\mu$g (0.24 nM) of TNF$\alpha$-LCAS was sufficient to block TNF-$\alpha$ expression in 1$\times10^5$ WRT7/P2 cells. I have also observed specific antisense activity in reduction of NF-$\kappa$B gene expression. The results suggest that an antisense sequence as a part of single-stranded circular genomic DNA has a specific antisense activity.

• Journal of Microbiology and Biotechnology
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• v.26 no.2
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• pp.421-431
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• 2016

Recently, poly-γ-glutamic acid synthetase A (pgsA) has been applied to display exogenous proteins on the surface of Lactobacillus casei or Lactococcus lactis, which results in a surface-displayed component of bacteria. However, the ability of carrying genes encoded by plasmids and the expression efficiency of recombinant bacteria can be somewhat affected by the longer gene length of pgsA (1,143 bp); therefore, a truncated gene, pgsA, was generated based on the characteristics of pgsA by computational analysis. Using murine IL-10 as an exogenous gene, recombinant Lactobacillus plantarum was constructed and the capacity of the surface-displayed protein and functional differences between exogenous proteins expressed by these strains were evaluated. Surface expression of IL-10 on both recombinant bacteria with anchorins and the higher expression levels in L. plantarum-pgsA'-IL-10 were confirmed by western blot assay. Most importantly, up-regulation of IL-1β, IL-6, TNF-α, IFN-γ, and the nuclear transcription factor NF-κB p65 in RAW264.7 cells after stimulation with Poly(I:C) or LPS was exacerbated after co-culture with L. plantarum-pgsA. By contrast, IL-10 expressed by these recombinant strains could reduce these factors, and the expression of these factors was associated with recombinant strains that expressed anchorin (especially in L. plantarum-pgsA'-IL-10) and was significantly lower compared with the anchorin-free strains. These findings indicated that exogenous proteins could be successfully displayed on the surface of L. plantarum by pgsA or pgsA', and the expression of recombinant bacteria with pgsA' was superior compared with bacteria with pgsA.