• Journal of Physiology & Pathology in Korean Medicine
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• v.24 no.2
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• pp.220-227
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• 2010

Puerariae radix (PR) is a popular natural herb and a traditional food in Asia, which has antithrombotic and anti-allergic properties and stimulates estrogenic activity. One of the major side effects of cisplatin is nephrotoxicity, leading to acute renal failure. Recent study has suggested a role of ROS and p53 in renal cell injury by cisplatin. We studied that protective effects of PR on cisplatin-induced apoptosis in rat mesangial cell. Rat mesangial cell was preincubated with PR (50, 100, 150 and 200 ${\mu}g/m{\ell}$) for 12 hr and then treated with 30 ${\mu}M$ cisplatin for 24 hr. Protective effect of PR on cisplatin-induced apoptosis in ECV304 cells was determined using MTT assay, FDA-PI staining, flow cytometric analysis, caspase-3 activity assay, ROS assay and western blot. Our results showed that PR inhibited in cisplatin-induced apoptosis and ROS production in ECV304 cells. Moreover, PR reduced ERK, p38 and JNK activation that increased in cisplatin-treated rat mesangial cell. Furthermore, activation of p53 by cisplatin in rat mesangial cell was inhibited by PR treatment. These results suggest that protective effect of PR on cisplatin-induced apoptosis in rat mesangial cell may be associated with reduction of ERK, p38, JNK, p53 activation.

• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.7 no.1
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• pp.39-60
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• 2001

Fuzhengfangaitang is a prescript for inhibiting recurrence and metastasis of cancer. We had examined the anti-metatstastic effect of Fuzhengfangaitang. Furthermore, we performed the following experiments with ingredients of Fuzhengfangaitang. The purpose of this thesis is to study what ingredients of Fuzhengfangaitang have more valuable anti-cancer effects. And the results are listed below: 1. Cell Viability assay At the dose of 400$\mu$g/ml, most ingredients of Fuzhengfangaitang depressed viability of ECV-304. And especially, Scutellaria barbata D. DON$50{\mu}g/ml$ : 53.118%, $100{\mu}g/ml$: 49.092%, $200{\mu}g/ml$ : 43.765%, $400{\mu}g/ml$ : 12.747%), Polygonum bistorta L.($50{\mu}g/ml$ : 45.554%, $100{\mu}g/ml$ : 45.554%, $200{\mu}g/ml$ : 0.0%, $400{\mu}g/ml$ : 0.0%) and Psoralea corylifolia L.($50{\mu}g/ml$ : 86.591%, $100{\mu}g/ml$ : 81.307%, $200{\mu}g/ml$ : 24.801%, $400{\mu}g/ml$ : 3.111%) highly depressed cell viability more than the other ingredients. (${\alpha}$<0.05) 2. Cell Proliferation assay Proliferation assay with ingredients of Fuzhengfangaitang on ECV-304 showed that Crataegus pinnatifuda BGE ($50{\mu}g/ml$: 63.276%, $100{\mu}g/ml$ : 64.092%, $200{\mu}g/ml$ : 68.966% $400{\mu}g/ml$ : 38.517%, ED50=$296.974{\mu}g/ml$), Polygonum bistorta L.($50{\mu}g/ml$ : 83.981%, $100{\mu}g/ml$ : 86.997%, $200{\mu}g/ml$ : 58.780%, $400{\mu}g/ml$ : 26.408%), ED50=$266.725{\mu}g/ml$) and Psoralea corylifolia L.($50{\mu}g/ml$ : 103.037%, $100{\mu}g/ml$ : 82.529%, $200{\mu}g/ml$ : 2.829%, $400{\mu}g/ml$ : 0.998%), ED50=$177.369{\mu}g/ml$) depressed cell proliferation more than the other ingredients. 3. Tube Formation assay Compared with the control group, most ingredients of Fuzhengfangaitang did not remarkably inhibited the Tube Formation assay of ECV-304 at the dose of $100{\mu}g/ml$. But, Polygonum bistorta L. highly inhibited the tube formation of ECV -304 at the lower dose of $50{\mu}g/ml$. 4. Rat Aortic Ring assay In comparison with the control group, Scutellaria barbata D. DON., root of Polygonum bistorta L. and Psoralea corylifolia L. restricted the angiogenesis of the rat aortic ring at the dose of $100{\mu}g/ml$. And the other ingredients of Fuzhengfangaitang did not restricted the angiogenesis of the rat aortic ring at that dose. Especially, Polygonum bistorta L. highly inhibited the angiogenesis of the rat aortic ring at the lower dose of $50{\mu}g/ml$. From our research, the anti-angiogenic effects of the ingredients of Fuzhengfangaitang was proven. Moreover, it will be helpful for designing more effective prescription for anti angiogenesis.

• Proceedings of the PSK Conference
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• 2003.04a
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• pp.129.1-129.1
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• 2003

Alcohol, Helicobacter pylori, stress and NSAIDs-activated neutrophils all produce reactive oxygen species (ROS), which play an important role in gastric mucosal damage. Eupatilin is an active component of Artemisia asiatica possessing cytoprotective effect. The effect of eupatilin on the production of ROS and cellular damage in AGS and ECV304 cells were evaluated to prove the cytoprotective action against the above mentioned gastric mucosal cell damages. (omitted)

• BMB Reports
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• v.38 no.5
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• pp.577-583
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• 2005

Subtilisin QK, which is newly identified as a fibrinolytic enzyme from Bacillus subtilis QK02, has the ability of preventing nitrotyrosine formation in bovine serum albumin induced by nitrite, hydrogen peroxide and hemoglobin in vitro verified by ELISA, Western-blot and spectrophotometer assay. Subtilisin QK also attenuates the fluorescence emission spectra of bovine serum albumin in the course of oxidation caused by nitrite, hydrogen peroxide and hemoglobin. Furthermore, subtilisin QK could suppress the transformation of oxy-hemoglobin to met-hemoglobin caused by sodium nitrite, but not the heat-treated subtilisn QK. Compared with some other fibrinolytic enzymes and inactivated subtilisin QK treated by phenylmethylsulfonylfluoride, the ability of inhibiting met-hemoglobin formation of subtilisin QK reveals that the anti-oxidative ability of subtilisin QK is not concerned with its fibrinolytic function. Additionally, nitrotyrosine formation in proteins from brain, heart, liver, kidney, and muscle of mice that is intramuscular injected the mixture of nitrite, hydrogen peroxide and hemoglobin is attenuated by subtilisin QK. Subtilisin QK can also protect Human umbilical vein endothelial cell (ECV-304) from the damage caused by nitrite and hydrogen peroxide.

• BMB Reports
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• v.40 no.5
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• pp.832-838
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• 2007

A novel inhibitory protein against blood coagulation factor Va (FVa) was purified from muscle protein of granulated ark (Tegillarca granosa, order Arcoida, marine bivalvia) by consecutive FPLC method using anion exchange and gel permeation chromatography. In the results of ESI-QTOF tandem mass analysis and database research, it was revealed that the purified T. granosa anticoagulant protein (TGAP) has 7.7 kDa of molecular mass and its partial sequence, HTHLQRAPHPNALGYHGK, has a high identity (64%) with serine/threonine kinase derived from Rhodopirellula baltica (order Planctomycetales, marine bacteria). TGAP could potently prolong thrombin time (TT), corresponding to inhibition of thrombin (FIIa) formation. Specific factor inhibitory assay showed that TGAP inhibits FVa among the major components of prothrombinase complex. In vitro assay for direct-binding affinity using surface plasmon resonance (SPR) spectrometer indicated that TGAP could be directly bound with FVa. In addition, the binding affinity of FVa to FII was decreased by addition of TGAP in dose-dependant manner ($IC_{50}$ value = 77.9 nM). These results illustrated that TGAP might interact with a heavy chain of FVa ($FVa_H$) bound to FII in prothrombin complex. The present study elucidated that non-cytotoxic T. granosa anticoagulant protein (TGAP) bound to FVa can prolong blood coagulation time by inhibiting conversion of FII to FIIa in blood coagulation cascade. In addition, TGAP did not significantly (P < 0.05) show fibrinolytic activity and cytotoxicity on venous endothelial cell line (ECV 304).

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.6
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• pp.1686-1693
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• 2004

Ekong-san(EKS) was expected to have inhibitory effects on angiogenesis, considering the fact that its constituents such as Ginseng Radix, Glycyrrhizae Radix and Citri Pericarpium were reported to inhibit angiogenesis. Moreover, recently several metabolites transformed by the human intestinal microflora were reported to enhance effectiveness compared to their crude drugs. Based on these data, this study was designed to confirm whether the EKS metabolites (EKS-M) can significantly exert the anti-angiogenic and anti-metastatic activites. Hence, with EKS and EKS-M, viability assay, proliferation assay, in vitro tube formation assay, gelatin zymogram assay, in vitro invasion assay were carried out. EKS showed less toxicity in ECV304 and HT1080 cells than EKS-M. EKS-M inhibited the proliferation of HT1080 cells by 30% at 200㎍/㎖ and 42% at 400 ㎍/㎖ respectively. Also, EKS-M degraded the tube network at 200㎍/㎖. EKS and EKS-M inhibited the expression of MMP-9 at 200 and 400㎍/㎖ in HT1080 cells. EKS reduced the invasive activity of HT1080 cells through matrigel coated transfilter atthe concentration of 200㎍/㎖ more effectively than EKS-M. These data suggest that EKS and EKS-M has anti-angiogenic and anti-metastatic activities.

• The Journal of Internal Korean Medicine
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• v.27 no.3
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• pp.589-602
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• 2006

Hwangryunagyotang is supposed to have significant effects on some sorts of cardiovascular diseases like atherosclerosis. For this study. ACAT inhibitor was put in LDLR -/- mice to derive free cholesterol from it. This was to examine the effectiveness of Hwangryunnagyotang on its protecting and recovering function with endothelial cells damaged by free cholesterol through experimental. The results reported below. Hwangryunagyotang suppressed the crystallization of reactive oxygen species in macrophages and the numbers of free cholesterol crystal plate structured and reduced fragmentation of nucleus in ECV 304 cell strain by ACAT inhibitor significantly. Hwangryunagyotang also suppressed the necrosis of tissue in LDLR -/- mice' (treated with ACAT inhibitor) inflammatory portion which is adjacent to aortic root, proximal aorta and carotid artery by immunohistochemistry and fluorescence microscopy. On the whole, Hwangryunagyotang suppressed the necrosis of endothelial cells and especially it's effcet for the necrosis of para-myocardial tissues by free cholesterol. With this result, I suggest Hwangryunagyotang might have protective and recovery effects on atherosclerosis, so we need to carry on this study henceforth clinically and experimentally as well.

• Journal of Korean Traditional Oncology
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• v.11 no.1
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• pp.41-54
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• 2006

Ekongsan (EKS) was expected to have inhibitory effects on angiogenesis, considering the fact that its constituents such as Ginseng Radix, Glycyrrhizae Radix and Citri Pericarpium were reported to inhibit angiogenesis. Moreover, recently several metabolites transformed by the human intestinal microflora were reported to enhance effectiveness compared to their crude drugs. Based on these data, this study was designed to confirm whether the EKS metabolites (EKS-M) can significantly exert the anti-angiogenic and anti-metastatic activites. Hence, with EKS and EKS-M, viability assay, proliferation assay, in vitro tube formation assay, gelatin zymogram assay, in vitro invasion assay were carried out. EKS showed less toxicity in ECV304 and HT1080 cells than EKS-M. EKS-M inhibited the proliferation of HT1080 cells by 30% at 200 ${\mu}g/m{\ell}$ and 42% at 400 ${\mu}g/m{\ell}$ respectively. Also, EKS-M degraded the tube network at 200 ${\mu}g/m{\ell}$. EKS and EKS-M inhibited the expression of MMP-9 at 200 and 400 ${\mu}g/m{\ell}$in HT1080 cells. EKS reduced the invasive activity of HT1080 cells through matrigel coated transfilter at the concentration of 200 ${\mu}g/m{\ell}$ more effectively than EKS-M. These data suggest that EKS and EKS-M has anti-angiogenic and anti-metastatic activities.

• Journal of Microbiology
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• v.43 no.4
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• pp.337-344
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• 2005

Although pneumococcus is one of the most frequently encountered opportunistic pathogen in the world, the mechanisms responsible for its infectiveness have not yet been fully understood. In this paper, we have attempted to characterize the effects of pneumococcal transformation on the pathogenesis of the organism. We constructed three transformation-deficient pneumococcal strains, which were designated as Nos. 1d, 2d, and 22d. The construction of these altered strains was achieved via the insertion of the inactivated gene, comE, to strains 1, 2 and 22. We then conducted a comparison between the virulence of the transformation-deficient strains and that of the wild-type strains, via an evaluation of the ability of each strain to adhere to endothelial cells, and also assessed psaA mRNA expression, and the survival of hosts after bacterial challenge. Compared to what was observed with the wild-type strains, our results indicated that the ability of all of the transformation-deficient strains to adhere to the ECV304 cells had been significantly reduced (p < 0.05), the expression of psaA mRNA was reduced significantly (p < 0.05) in strains 2d and 22d, and the median survival time of mice infected with strains Id and 2d was increased significantly after intraperitoneal bacterial challenge (p < 0.05). The results of our study also clearly indicated that transformation exerts significant effects on the virulence characteristics of S. pneumoniae, although the degree to which this effect is noted appears to depend primarily on the genetic background of the bacteria.