• Korean Journal of Plant Resources
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• v.22 no.4
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• pp.317-322
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• 2009

In the present study, the effect of ethanol extracts of leaf and root of Rosa multiflora on the proliferation of B16 cells, tyrosinase activity and melanin synthesis were investigated. 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide(MTT) assay demonstrated that the cell viability upon treatment with Rosa multiflora extract(0-200 ${\mu}g$/ml) was similar to that of untreated control. Treatment with leaf or root extracts(200 ${\mu}g$/ml) decreased the in vitro tyrosinase activity to about 65% of that in untreated control. Similarly, the intracellular tyrosinase activity of B16 cells decreased in a concentration-dependent manner. Furthermore, the melanin synthesis of B16 cells decreased by the two extracts in a concentration dependent manner. However, the extracts did not change the level of tyrosinase mRNA, as determined by RT-PCR. These results demonstrated that the Rosa multiflora extracts inhibit the tyrosinase dependent melanin biosynthesis, and therefore, are candidates for skin-whitening agents.

• The Korean Journal of Food And Nutrition
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• v.13 no.5
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• pp.411-418
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• 2000

The purpose of this study was to investigate effects of chestnut inner skin tea, brown rice-green tea and Cassia lora tea on the activation of physiological functions (regional cerebral blood flow, mean arterial blood pressure, proliferation of immunocytes in vitro and in vitro, suppression of cancer cell proliferation) in mouse and rat. We used 8 weeks-old balb/c male mice, 300g ICR rats and L1210 cell lines. Regional cerebral blood flow(rCBF) and mean arterial blood pressure(BP) were measured using Leser-Doppler Flowmetry(LDF) and the proliferation of cells was measured using a colorimetric tetrazolium assay(MTT assay). The experimental results are as follows : 1. rCBF was increased by Cassia tora tea, but decreased by chestnut inner skin tea in rats. 2. BP was increased by brown rice-green tea in rats. 3. Proliferation of mouse thymocytes and splenocytes were significantly increased by chestnut inner skin tea in vitro. 4. Proliferation of mouse thymocytes was decreased by Cassia tora tea and brown rice-green tea in vitro. 5. Proliferation of mouse thymocytes was decreased by Cassia tora tea and brown rice-green tea in L1210 transplanted mice. 6. Proliferation of splenocytes was accelerated by chestnut inner skin tea in L1210 transplanted mice. 7. Proliferation of L1210 cells was inhibited by chestnut inner skin tea and Cassia tora tea in L1210 transplanted mice.

• The Journal of Korean Medicine
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• v.26 no.2 s.62
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• pp.217-230
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• 2005

Objectives: Chungpaesagan-tang (CPSGT), which is frequently used for treating patients of cerebrovascular disease, has not been reported by clinical doctors concerning the effect of neuronal aptosis caused by brain ischemia. To study the effect of CPSGT on focal cerebral ischemia in normal and diabetic rats and SHR, focal cerebral ischemia was induced by transient MCAO, and after onset CPSGT was administrated. Methods: Rats (Sprague-Dawley) were divided into four groups: sham-operated group, MCA-occluded group, CPSGT­administrated group after MCA occlusion, and normal group. The MCA was occluded by intraluminal method. CPSGT was administrated orally twice (l and 4 hours) after middle cerebral artery occlusion. All groups were sacrificed at 24 hours after the surgery. The brain tissue Was stained with $2\%$ triphenyl tetrazolium chloride (TTC) or $1\%$ cresyl violet solution, to examine effect of CPSGT on ischemic brain tissue. The blood samples were obtained from the heart.~. Tumor necrosis $factor-\alpha$ level and interleukin-6 level of serum was measured from sera using enzyme-linked immunoabsorbent assay (ELISA). Then changes of immunohistochemical expression of $TNF-\alpha$ in ischemic damaged areas were observed. Results: In NC+MCAO+CP and DM+MCAO+CP, CPSGT significantly (p<0.01) decreased the number of neuron cells compared to the control group. CPSGT markedly reduced (p<0.01) the infarct size of the forebrain in distance from the interaural line on cerebral ischemia in diabetic rats. CPSGT significantly reduced the $TNF-\alpha$ expression in penumbra region of damaged hemisphere in diabetic rats. Conclusions: CPSGT had a protective effect on cerebral ischemia in SD rats, especially in diabetic rats compared with normal SD rats.

• Archives of Plastic Surgery
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• v.36 no.3
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• pp.247-253
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• 2009

Purpose: Hydroxyapatite(HA) has been widely used due to its chemical similarity to bone and good biocompatibility. HA is composed of macropores and micropores. Too much irregularities of the micropores are ineffective against the adhesion and proliferation of osteoblast. Many efforts have been tried to overcome these drawbacks. HA crystal coating on the irregular surface of HA scaffold, crystallized HA, is one of the method to improve cell adhesion. Meanwhile, the collagen has been incorporated with HA to create composite scaffold that chemically resembles the natural extracellular matrix components of bone. The authors proposed to examine the effect of collagen - coated crystallized HA on the adhesion and proliferation of osteoblast. Method: HA powder containing $10{\mu}m$ pore size was manufactured as 1 cm pellet size. For the making crystallized HA, 0.1 M EDTA solution was used to dissolve HA powder and heated $100^{\circ}C$ for 48 hours. Next, the crystallized HA pellets were coated with collagen (0.1, 0.5, and 1%). The osteoblasts were seeded into HA pellets and incubated for the various times (1, 5, and 9 days). After the indicating days, methylthiazol tetrazolium (MTT) assay was performed for cell proliferation and alkaline phosphatase (ALP) activty was measured for bone formation. Result: In SEM study, the surface of crystallized HA pellet was more regular than HA pellet. MTT assay showed that the proliferation of osteoblasts increased in a collagen dose - dependent and time - dependent manner and had a maximum effect at 1% collagen concentration. ALP activity also increased in a collagen dose - dependent manner and had a highest effect at 1% collagen concentration. Conclusion: These data showed that crystallization and collagen coating of HA was effective for osteoblast proliferation and ALP activity. Therefore, our results suggest that crystallized - HA scaffold with collagen coating is may be a good strategy for tissue engineering application for bone formation.

• Journal of Sericultural and Entomological Science
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• v.52 no.1
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• pp.6-9
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• 2014

This study was for the investigation of the stability of purified bee venom (PBV) during the treatment in the pH range from pH2 to pH9 for 24 hours, respectively. Changes of components and physiological functionalities in PBV were by evaluated silver staining, and melittin contents were measured by liquid chromatography. The antimicrobial activity against bacteria by minimum inhibitory concentration (MIC) and effect of the cell regeneration were measured by 3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl tetrazolium bromide (MTT) assay using human dermal fibroblast (HDF) cell. The main proteins such as melittin and phospholipase $A_2$ showed no characteristic changes. The antimicrobial activity and effect of cell regeneration showed no difference from pH2 to pH9. From this study, we suggest that components and physiological functionalities of PBV against treated pH were kept stability at from pH2 to pH9.

• Journal of Chest Surgery
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• v.29 no.4
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• pp.365-372
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• 1996

In an attempt to evaluate the effect of St. Thomas' hospital cardioplegic solution (STH) I and II on the cultured rat myocardial cells, beating rate, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (Irtrazolium MTT) and lactate dehydrogenase activity were investigated, and also light and electron microscopic studies were carried out. After rat myocardial cells were cultured for 72 hours, cells were treated with STH I or with STH H solution for 30 and 120 min. and thereafter myocardial cells were cultured in control medium for 24 hours. The results obtained were as follows : 1. Beating rate was 154 times per min. in control group, 141 times per min. in STH I solution(for 2. 120 min.)-treated group and 145 times in STH ll solution (for 120 min.)-treated group. MTT absorbances by MTT assay were 102% in STH I solution (for 120 min.)-treated group and 93% in STH ll solution (for 120 min.)-treated group compared with control group. 3. The amount of lactate dehydrogenase released into the medium were 123% in STH I solution (for 120 min.)-treated group and 109% in STH H solution (for 120 min.)-treated group compared with control group. 4. In the light microscopy examination, myocardial cells showed no differences between experimental and control groups in their number and shape. 5. In the electron microscopy examination, myocardial cells treated with STH I solution showed fewer destroyed mitochondria compared to STH ll solution-treated group. These results suggest that both 51. Thomas'cardioplegic solution STH 1 and STH H have no cytotoxicity on cultured rat myocardial cells, but STH H solution has more protective effect on myocardial cells compared to STH I solution.

• Journal of Applied Biological Chemistry
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• v.61 no.1
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• pp.9-15
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• 2018

We analyzed the antioxidant and anti-wrinkle activities of berberine, isolated from dried rhizome of Coptis japonica Makino, to determine its cosmetic potential. We performed the 3-[4,5-dimethylthiazol]-2-yl]-2,5-diphenyl-tetrazoliumbromide (MTT) assay to evaluate the toxicity of the berberine. We also measured the ROS and hyaluronic acid production, and expression of MMP-2, MMP-9, TIMP-1, TIMP-2, and tumor necrosis factor-alpha ($TNF-{\alpha}$) to evaluate the antioxidant and anti-wrinkle activities of berberine, respectively. The cytotoxicity of ultraviolet light, in presence of berberine, was measured by the MTT assay using CCD-986sk fibroblasts, and no cytotoxicity was observed at concentrations less than $25{\mu}g/mL$. We also found that berberine decreased ROS production in a concentration-dependent manner and promoted the synthesis of hyaluronic acid. Further, berberine reduced the protein levels and mRNA expression of MMP-2 and MMP-9, which are associated with wrinkle formation, and increased the expression of TIMP-1 and TIMP-2. In addition, the inhibitory effect of berberine on $TNF-{\alpha}$, known as pro-inflammatory cytokine, was inhibited by $TNF-{\alpha}$ gene in a concentration-dependent manner. These results suggest that berberine holds cosmetic value owing to its antioxidant activity, by inhibiting ROS production and anti-wrinkle activity by reducing MMP-2 and MMP-9 and increasing TIMP-1 and TIMP-2 expression.

• The Korean Journal of Physiology and Pharmacology
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• v.24 no.3
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• pp.241-248
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• 2020

Luminespib (AUY922), a heat shock proteins 90 inhibitor, has anti-neoplastic and antitumor effects. However, it is not clear whether AUY922 affects events in vascular diseases. We investigated the effects of AUY922 on the platelet-derived growth factor (PDGF)-BB-stimulated proliferation and migration of vascular smooth muscle cells (VSMC). VSMC viability was detected using the XTT (2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide) reagent. To detect the attenuating effects of AUY922 on PDGF-BB-induced VSMCs migration in vitro, we performed the Boyden chamber and scratch wound healing assays. To identify AUY922-mediated changes in the signaling pathway, the phosphorylation of protein kinase B (Akt) and extracellular signal-regulated kinase (ERK) 1/2 was analyzed by immunoblotting. The inhibitory effects of AUY922 on migration and proliferation ex vivo were tested using an aortic ring assay. AUY922 was not cytotoxic at concentrations up to 5 nM. PDGF-BB-induced VSMC proliferation, migration, and sprout outgrowth were significantly decreased by AUY922 in a dose-dependent manner. AUY922 significantly reduced the PDGF-BB-stimulated phosphorylation of Akt and ERK1/2. Furthermore, PD98059 (a selective ERK1/2 inhibitor) and LY294002 (a selective Akt inhibitor) decreased VSMC migration and proliferation by inhibiting phosphorylation of Akt and ERK1/2. Greater attenuation of PDGF-BB-induced cell viability and migration was observed upon treatment with PD98059 or LY294002 in combination with AUY922. AUY922 showed anti-proliferation and anti-migration effects towards PDGF-BB-induced VSMCs by regulating the phosphorylation of ERK1/2 and Akt. Thus, AUY922 is a candidate for the treatment of atherosclerosis and restenosis.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.42 no.6
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• pp.614-620
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• 2009

We report non-specific immune responses and its disease resistance against Edwardsiella tarda by alga mixture (HE; Hizikia:Ecklonia) in olive flounder for the first time. Five isonitrogenous (44% crude protein) and isocaloric (17.1 MJ $kg^{-1}$) diets were formulated to have 0%, 2%, 4%, 6% and 8% of the alga mixture. One of five experimental diets was fed triplicate groups of fish (30 fish/group) to apparent satiation in a flow through system. After a two week feeding, blood was sampled at 3, 6, 12, 24 h after the last feeding for a kinetic measurement of nitroblue tetrazolium (NBT) activity and healthy fish with similar sizes in each tank were selected and injected with 1 mL of E. tarda suspension ($1.0\times10^7$ CFU/mL) to evaluate the disease resistance of the fish. Dietary supplementation of alga mixtures resulted in significantly higher non-specific immune responses compared with the fish fed the control diet. The cumulative mortality was significantly lower in the fish groups fed alga mixture containing diets than control group in the challenge test with E. tarda. Therefore, the results in this study indicate that dietary supplementation of Hizikia and Ecklonia mixtures enhance the non-specific immune responses and a disease resistance of olive flounder.

• The Korean Journal of Physiology and Pharmacology
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• v.14 no.3
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• pp.177-183
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• 2010

Tetrahydrobiopterin (BH4), an essential cofactor for nitric oxide synthase (NOS) activity, is known to play important roles in modulating both NO and superoxide production during vascular diseases such as atherosclerosis. However, the role of BH4 in functions of vascular smooth muscle cells is not fully known. In this study, we tested the effects of BH4 and dihydrobiopterin (BH2), a BH4 precursor, on migration and proliferation in response to platelet-derived growth factor-BB (PDGF-BB) in rat aortic smooth muscle cells (RASMCs). Cell migration and proliferation were measured using a Boyden chamber and a 5-bromo-2'-deoxyuridine incorporation assay, respectively, and these results were confirmed with an ex vivo aortic sprout assay. Cell viability was examined by 2,3-bis [2-methoxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxanilide assays. BH4 and BH2 decreased PDGF-BBinduced cell migration and proliferation in a dose-dependent manner. The inhibition of cell migration and proliferation by BH4 and BH2 was not affected by pretreatment with $N^G$-nitro-L-arginine methyl ester, a NOS inhibitor. Moreover, the sprout outgrowth formation of aortic rings induced by PDGF-BB was inhibited by BH4 and BH2. Cell viability was not inhibited by BH4 and BH2 treatment. The present results suggest that BH4 and BH2 may inhibit PDGF-stimulated RASMC migration and proliferation via the NOS-independent pathway. Therefore, BH4 and its derivative could be useful for the development of a candidate molecule with an NO-independent anti-atherosclerotic function.