• Korean Journal of Clinical Laboratory Science
• /
• v.51 no.2
• /
• pp.245-251
• /
• 2019

This study examined the effects of 2-methoxy-1,4-naphthoquinone (MQ) on the monocyte chemoattractant protein-1 (MCP-1)-induced migration of monocytes, which is an important phenomenon for the body defense and immune response. MQ is a major component extracted from Impatiens balsamina leaves, which have been used for many years in Asian medicine for the treatment of a range of diseases and pain. The cytotoxicity of MQ began to appear at a concentration of $10{\mu}M$, and approximately 50% cytotoxicity was confirmed at $100{\mu}M$. The MCP-1 induced migration of the THP-1 monocyte cell line increased after MQ treatment in a dose dependent manner and the largest increase was observed at $0.1{\mu}M$. The level of cAMP expression decreased after a treatment with $0.1{\mu}M$ MQ. The phosphorylation of extracellular signal-regulated kinases 1/2 (Erk1/2), a key signaling protein involved in the signaling pathway of C-C motif chemokine receptor 2 (CCR2), a receptor for MCP-1, was increased by the simultaneous treatment of $0.1{\mu}M$ MQ. These results show that MQ increases the MCP-1-induced migration of THP-1, decreases the level of cAMP expression, and increases the level of Erk1/2 phosphorylation.

• Journal of Life Science
• /
• v.32 no.2
• /
• pp.135-141
• /
• 2022

Thymosin β-4 (TB4) is a small peptide composed of 43 amino acids. To obtain sufficient biologically active mouse TB4 economically, we cloned and overexpressed this gene in an Escherichia coli system. With the isopropyl β-D-1-thiogalactopyranoside induction of the E. coli transformant, TB4 fusion protein with intein- and chitin-binding domain was successfully expressed in the soluble fraction within the E. coli cell. The TB4-intein - chitin-binding domain fusion protein was purified from the soluble fraction of E. coli cell lysate. The affinity chromatography with chitin beads and dithiothreitol-mediated intein self-cleavage reaction releases the TB4 peptide into the stripping solution. Sodium dodecyl sulphate - polyacrylamide gel electrophoresis and Western blot analyses were used to confirm that the recombinant TB4 peptide was produced with the expected size of 5 kDa. We found that the recombinant TB4 stimulated cell migration in the transwell plate chamber assay. After 18 hr of the treatment of the recombinant TB4 with 1 ng/ml concentration, the migration of the HT1080 cell was increased by 20% compared with that of the chemically synthesized TB4. The recombinant TB4 was also observed to promote the healing of a wound area in C57BL/6 mice by as high as 35% compared with that of the chemically synthesized TB4. These results suggest that the recombinant TB4 has better biological activity for cell migration and wound healing than that of the chemically synthesized TB4 peptide.

• Development and Reproduction
• /
• v.5 no.1
• /
• pp.23-33
• /
• 2001

When mammalian oocytes ovulate into the oviduct, associating follicular fluid components are exposed to the oviductal environment, possibly resulting in the mutual interaction between fillicu1ar and oviductal fluids. In the Present study, we have demonstrated for the first time that components of fallicular fluid could be modified by the oviductal fluid. Gelatin zymographic analyses of human follicular fluid (hFF) obtained from IVF patients showed consistently the presence of 110 kDa gelatinase (GA110) in addition to many bands among which 62 kDa gelatinase was predominant. Addition of EDTA or phenanfhroline to the gelatinase substrate buffer during gel incubation abolished GA110 band whereas phenylmethylsulffnyl fluoride (PMSF) did not. In contrast, bovine oviductal fluid(bOF) exhibited only 62 kDa gelatinase. Surprisingly, when bOF was added to hFF in 1:1 ratio and then the mixture was incubated for 3 h at 37$^{\circ}$C, GA110 of hFF disappeared. Disappearance of GA110 by bOF was observed even within 30 min after mixing with hFF. Addition of aminophenylmercuric acetate (APMA) to hFF also abolished enzymatic activity of GA110 but increased the activityof 62 kDa gelatinase. However, APMA abolished many other gelatinases as well unlike bOF. Interestingly, treatment of hFF with EDTA for 3 h remarkably increased the enzymatic activity of GA110 but not that of other gelatinases. Addition of phenanthroline, PMSF or soybean trypsin inhibitor (SBTI) did not affect overall gelatinase activities. Again, addition of bOF to the hFF pretreated with any of the above proteinase inhibitors abolished the appearance of GA110. Human serum also showed GAI 10 of which activity was greatlyenhanced by EDTA treatment. Similar to hFF, serum GA110 also disappeared by the addition of bOF. Human granulosa cell homogenate did not reveal any appreciable gelatinase activity except 92 kDa gelatinase. Anti-human gelatinase A antibody reacted with 62 kDa gelatinase of hFF. Based upon these results, it is concluded that bOF could selectively degrade an isoform of gelatinase A present in hFF and human serum.

• Journal of Acupuncture Research
• /
• v.22 no.3
• /
• pp.155-167
• /
• 2005

Objectives : The purpose of this study was to investigate the anti-inflammatory effect of Cobrotoxin on binding affinity of cobrotoxin with P50, $IKK{\alpa}$ and $IKK{\beta}$, activities of NF-${\kappa}B$, Cell viability of astrocyte, expressions of protein molecules of NF-${\kappa}B$ such as P50, P-$1{kappa}B$, $1{\kappa}B$ and iflammation related genes such as Cox-2, iNOS, cPLA2 in the SNP or LPS induced Inflammatory pathway of Rats' astrocytes. Methods : In this study, The expression of cytosolic phospholipase A2, Nitric oxcide, Cyclooxygenase-2 and inducible nitrogen oxide synthase was determined by western blotting with corresponding antibodies, and the generation of NF-${\kappa}B$ was assayed by EMSA method in astrocytes of rats. The Cell viability of astrocytes was determined by MTT assay, and Binding affinity of Cobrotoxin with P50, $IKK{\alpha}$ and $IKK{\beta}$ was assayed by Surface plasmon resonance analysis, and NF-${\kappa}B$ dependent luciferase activity was determined by luciferase analysis, and Uptake of cobrotoxin in astrocytes was identified by Confocal laser scanning microscope Results : 1. Compared with control, LPS-induced NF-${\kappa}B$ DNA binding activity was decreased significantly by 0.1, $0.5{\mu}g/m{\ell}$ of Cobrotoxin in Astrocyte. 2. Compared with control, LPS-induced NF-kB dependent luciferase expression was decreased significantly by 0.1, 0.5 and $1{\mu}g/m{\ell}$ of Cobrotoxin in Astrocyte. 3. Compared with control, SNP induced P50, $I{\kappa}B$ expressions in astrocyte were decreased significantly by 0.1, 0.5 and $1{\mu}g/m{\ell}$ of Cobrotoxin and P-$1{\kappa}B$ expression was decreased significantly by 0.5 and $1{\mu}g/m{\ell}$ of Cobrotoxin. 4. Compared with control, LPS induced P50, $1{\kappa}B$ expressions in astrocyte were decreased significantly by 0.5 and $1{\mu}g/m{\ell}$ of Cobrotoxin. 5. Compared with control, SNP induced Cox-2, iNOS, CPLA2 expressions in astrocyte were decreased significantly by $1{\mu}g/m{\ell}$ of Cobrotoxin. 6. Compared with control, LPS induced Cox-2, cPLA2 expressions in astrocyte were decreased significantly by 0.1, 0.5, $1{\mu}g/m{\ell}$ of Cobrotoxin and iNOS expression was decreased significantly by 0.5, $1{\mu}g/m{\ell}$ of Cobrotoxin. 7. Compared with $0.5{\mu}g/m{\ell}$ of Cobrotoxin, SNP-induced NF-${\kappa}B$ DNA bindins activity in astrocyte was increased significantly by Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 1mM and Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 5mM. 8. Compared with $0.5{\mu}g/m{\ell}$ of Cobrotoxin, LPS-induced NF-${\kappa}B$ DNA binding activity in astrocyte was increased significantly by Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 1mM, Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 5mM, Cobrotoxin $0.5{\mu}g/m{\ell}$with GSH 1mM and Cobrotoxin $0.5{\mu}g/m{\ell}$ with GSH 5mM 9. Compared with $0.1{\mu}g/m{\ell}$ of cobrotoxin, SNP induced P50 expressions in astrocyte were increased significantly by Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 1mM, Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 5mM Cobrotoxin $0.5{\mu}g/m{\ell}$ with GSH 1mM and Cobrotoxin $0.5{\mu}g/m{\ell}$ with GSH 5mM. 10. The uptake of the labeled cobrotoxin into the cells was shown under a confocal laser scanning microscope. cobrotoxin was uptaken into the membrane and nucleus of astrocytes. Conclusions : In summary, the present results demonstrate that cobrotoxin directly binds to sulfhydryl group of p50 and IKKS resulting In the reduction of translocation of p50 and IkB release, thereby inhibits activation of NF-${\kappa}B$, and suggest that pico to nanomolar range of cobrotoxin could inhibit the expression of genes in the NF-${\kappa}B$ signal pathway.

• Journal of Korean Medical Ki-Gong Academy
• /
• v.7 no.2
• /
• pp.40-94
• /
• 2004

Recently, meditation, Zen meditation and Qigong became popular in western. For that reason, Value of Korea traditional methods for health promotion have been evaluated but theoretical evidence about that is insufficient. ${\ll}$The Yellow Emperor's Inner Classic${\gg}$ (黃帝內經). So, I reviewed history of Korea Qigong according to period, in order to produce theoretical evidence of Korea medical Qigong and investigated ${\ulcorner}$Han${\lrcorner}$ thought (${\ulcorner}$한${\lrcorner}$ 思想) as theoretical evidence of Korea Qigong. I have obtained following results by comparing meaning of god, a view of human body and practice method of the ${\ll}$Samilshingo${\gg}$ (三一神誥) with ${\ll}$The Yellow Emperor's Inner Classic${\gg}$ (黃帝內經). Sinsundo(神仙道) of native to race became active during the period of the Three Kingdoms but more disappeared due to under the influence of Taoism. So only has been remained in existence since the period of the unity silla. There could accept positively the foreign ideas belonging to Confucianism, Buddhism and Taoism have been brought since the period of the Three Kingdoms because ${\ll}$Samilshingo${\gg}$ (三一神誥), one of three the scriptures as the origin of ${\ulcorner}$Han${\lrcorner}$ thought(${\ulcorner}$한${\lrcorner}$ 思想) included the original form of three religion belonging to Confucianism, Buddhism, Taoism. Three common results as theoretical evidence of Qigong were found out by comparing ${\ll}$Samilshingo${\gg}$ (三一神誥) with ${\ll}$The Yellow Emperor's Inner Classic${\gg}$ (黃帝內經). First, in meaning of god, there is not only different from the gods of heaven and the gods of human but also keep understanding with each other and there was common feature of logical structure that function of god was divided into three. Second, In a view of human body, there were in common with ${\ll}$Samilshingo${\gg}$ (三一神誥) as theory of only as energy(氣一元論), theory of bisection as truth false(眞妄二分論), theory of trisection as spirit(性) destiny(命) nature(精) and mind(心) energy(氣) body(身)(性 命 精, 心 氣 身의 三分論) and ${\ll}$The Yellow Emperor's Inner Classic${\gg}$ (黃帝內經) as theory of only as energy(氣一元論), theory of bisection as shape and god(形神二分論), theory of trisection as nature(精) energy(氣) god(神) or shape(形) energy(氣) god(神)(精 氣 神, 形 氣 神의 三分論) according to formal part. Also, spirit(性) destiny(命) nature(精) as truths of three(三眞) were understand as nature(精) energy(氣) god(神) as treasure of three(三寶) by part of reason and mind(心) energy(氣) body(身) as false of three(三妄) were understand as nature(精) energy(氣) god(神) as treasure of three(三寶) by part of function. Third, In practice method, pause of thinking(止感) modulation of breath(調息). prohibition of sensibility(禁觸) mentioned in ${\ll}$Samilshingo${\gg}$ (三一神誥) as practice method each were understand regulation of an etiological cause as an internal cause, an external cause and not internal and external cause in oriental medicine. Namely, pause of thinking(止感) was understand as regulation method of emotion, mind and nature. modulation of breath(調息) was not only as modulation of energy from the inner parts of the body but also that from the internal and external parts of the body, prohibition of sensibility(禁觸) was understand as regulation method of ear, eye, mouth, and nose and posture, life style. These results suggest that ${\ll}$Samilshingo${\gg}$ (三一神誥) is worth meaning of Korea medical Qigong because meaning of god, a view of human body, practice method of mentioned in ${\ll}$Samilshingo${\gg}$ (三一神誥) is common with that of ${\ll}$The Yellow Emperor's Inner Classic${\gg}$ (黃帝內經) as theoretical evidence of Qigong.