• The Journal of Korean Medicine
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• v.28 no.4
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• pp.114-123
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• 2007

Osteoarthritis is characterized by cartilage degradation and chondrocytes death. Chondrocyte death is induced by the apotosis through special mechanisms including the activation of caspase-3. On the basis of this background, this study was designed to examine the cartilage protective and anti-apototic effects of Aralia Cordata in in vtro and in collagenase-induced arthritis rabbit model. To conduct in vitro study, chondrocytes culturedfrom rabbit knee joint were treated by 5 ng/ml IL-1a.For in vivo experiment, collagenase-induced arthritis (CIA) rabbit model was made via intraarticular injection with 0.25 ml of collagenase solution. Aralia cordata pharmacopuncture (ACP) was administrated on bilateral Dokbi acupoint (ST35) of rabbits at a dosage of 150 ${\mu}g/kg$ once a day for 28 days after the initiation of the CIA induction. In the study by using CIA rabbit model in vivo, ACP showed the inhibition of cartilage degradation in histological analysis. Aralia cordata also showed anti-apoptotic effect both in vitro and in vivo study. In chondrocytes treated by IL-1a, Aralia cordata inhibited caspase-3 activity and enhanced the proliferation of IL-1a-induced dedifferentiated chondrocytes. ACP showed the inhibition effect on the caspase-3 expression and activity from CIA rabbit model. This study indicates that ACP inhibits the cartilage destruction and the chondrocyte apotosis through downregulation of caspase-3 activity. These data suggest that ACP has a beneficial effect on preventing articular cartilage destruction in osteoarthrtis.

• Journal of Life Science
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• v.14 no.2
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• pp.362-367
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• 2004

For better understanding of the host infection mechanism of Vibrio, a Vibrio parahaemolyticus collagenase mutant was generated by insertional inactivation of a vppC gene encoding extracellular collagenase. A recombinant DNA containing vppC::nptII was cloned into a suicide plasmid pDMS197, resulted in pVCM03. The recombinant suicide plasmid pVCM03 contained in E. coli $\chi$7213 was transferred to a wild-type V. parahaemolyticus 04 through conjugation. The recombinant vppC::nptII DNA in pVCM03 was exchanged with wild-type allele by homologous recombination resulting vppC mutant, V. parahaemolyticus CM. The mutant was selected and screened on TCBS media containing 10% sucrose and kanamycin. The mutation by allele exchange was confirmed with the comparison of the size of DNAs amplified by PCR. V. parahaemolyticus CM showed at least 4-fold less collagen-degrading activity than those of wild-type, and the mutant exhibited less cytotoxicity than that of wild-type in MTT assay.

• Journal of Korean Medicine Rehabilitation
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• v.26 no.1
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• pp.1-11
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• 2016

Objectives The purpose of this study was to investigate the effect of Hyulboochucke-tang on the collagenase induced intracerebral hemorrhage in white rats. Methods To identify the effect of the Hyulboochucke-tang on intracerebral hemorrhage, intracerebral hemorrhage was induced in the right caudate nuclei of white rats. For normal group (n=12) and comparative group (n=12), saline was dosed, and vaccum evaporated Hyulboochucke-tang extract was dosed to treatment group (n=12), 3 and 10 days after the collagenase injection, the body weight, the brain weight, the size of hematoma, the size of the area of malacia, the number of apoptotic cell and the change in pathological histology were observed. Results 3 days after the injection, the brain weight(g) was considerably decreased in treatment group (n=12) compared to comparative group (n=12). The brain weight after 10 days of the injection was also considerably decreased in treatment group (n=6) against comparative group (n=6). The cross section(mm) of cerebral malacia after 10 days of the injection was considerably decreased in treatment group (n=6) compared to comparative group (n=6). The number of apoptotic cell in normal intracerebral around the area of malacia did not show considerable change between treatment group and comparative group. 12 days after the injection, the multiplication of gitter cells, astrocyte and newly formed capillaries around the area of malacia was distinct. Conclusions On the basis of these results, We sugggest that Hyulboochucke-tang controls swelling caused by hemorrhage and contributes to absorption of hematoma by multiplication of newly formed capillaries and recovery of damaged cerebral tissue by multiplication of gitter cells and astrocyte.

• Food Science of Animal Resources
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• v.34 no.6
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• pp.844-851
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• 2014

This study focused on the anti-oxidative and collagenase- and elastase inhibition effects of low molecular weight peptides (LMP) from commercial Jeju horse leg bone hydrolysates (JHLB) on pancreatin, via enzymatic hydrolysis. Cell viability of dermal fibroblasts exposed to UVB radiation upon treatment with LMP from JHLB was evaluated. Determination of the antioxidant activity of various concentrations of LMP from JHLB were carried out by assessing 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2-azino-bis-3-ethybenzothiazoline-6-sulphonic acid (ABTS) radical scavenging activity, ferric reducing antioxidant power (FRAP), and oxygen radical absorbance capacity (ORAC). The DPPH radical scavenging activity of LMP from JHLB (20 mg/mL) was 92.21% and ABTS radical scavenging activity (15 mg/mL) was 99.50%. FRAP activity (30 mg/mL) was $364.72{\mu}M/TE$ and ORAC activity (1 mg/mL) was $101.85{\mu}M/TE$. The anti-wrinkle potential was assessed by evaluating the elastase- and collagenase inhibition potential of these LMP. We found that 200 mg/mL of LMP from JHLB inhibited elastase activity by 41.32%, and 100 mg/mL of LMP from JHLB inhibited collagenase activity by 91.32%. The cell viability of untreated HS68 human dermal fibroblasts was 45% when exposed to a UVB radiation dose of $100mJ/cm^2$. After 24 h of incubation with $500{\mu}g/mL$ LMP from JHLB, the cell viability increased to 60%. These results indicate that LMP from JHLB has potential utility as an anti-oxidant and anti-wrinkle agent in the food and cosmetic industry. Additional in vivo tests should be carried out to further characterize these potential benefits.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.25 no.2
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• pp.59-75
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• 1999

Skin is continuously exposed to external stimuli including ultraviolet radiation, which is a major cause of skin photoaging. According to recent discoveries, UVA with a lower energy but deep-penetrating properties, compared to UVB, is likely to play a major part in causing skin photoaging. The clinical and histochemical changes of photoaging are well characterized, but the biochemical mechanisms are poorly understood partly due to the lack of suitable experimental systems. In this work, three-dimensional, reconstituted skin culture models were prepared. After certain period of maturation, the equivalent models were shown to be similar in structure and biochemical characteristics to normal skin. Mature dermal and skin equivalent models were exposed to sub-lethal doses of UVA, and the effects of UVA relevant to dermal photoaging were monitored, including the production of elastin, collagen, collagenase(MMP-1), and tissue inhibitor of metalloproteinases-1 (TIMP-1). Interestingly, dermal and skin equivalents reacted differently to acute and chronic exposure to UVA. Elastin production was increased as soon as one week after commencing UVA irradiation by chronic exposure, although a single exposure failed to do so. This early response could be an important advantage of equivalent models in studying elastosis in photoaged skin. Collagenase activity was increased by acute UVA irradiation, but returned to control levels after repeated exposure. On the other hand, collagen biosynthesis, which was increased by a single exposure, decreased slightly during 5 weeks of prolonged UVA exposure. Collagenase has been thought to be responsible for collagen degeneration in dermal photoaging. However, according to the results obtained in this study, elevated collagenase activity is not likely to be responsible for the degeneration of collagen in dermal photoagig, while reduced production of collagen may be the main reason. It can be concluded that reconstituted skin culture models can serve as useful experimental tools for the study of skin photoaging. These culture models are relatively simple to construct, easy to handle, and are reproducible Moreover the changes of dermal photoaging can be observed within 1-4 weeks of exposure to ultraviolet light compared to 4 months to 2 years for human or animal studies. These models will be useful for biochemical and mechanistic studies in a large number of fields including dermatology, toxicology, and pharmacology.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2019.10a
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• pp.63-63
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• 2019

The thinning Green ball apple was extracted using water and ethanol and a phenolic concentration of thinning Green ball apple was $50-200{\mu}g/mL$. The water and ethanol extracts of thinning Green ball apple showed 94.69% and 92.24% 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity and 100.30% and 99.16% 2,2'-Azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging activity at phenolic concentration of $200{\mu}g/mL$, respectively. The water and ethanol extracts of thinning Green ball apple showed antioxidant protection factor of 1.76 antioxidant protection factor and 1.76 antioxidant protection factor, respectively. The water and ethanol extracts showed 101.46% and 99.64% anti-oxidative effect on thiobarbituric acid reactive substances at phenolic concentration of $200{\mu}g/mL$. Hence, the water and ethanol extracts of thinning Green ball apple can be considered a potential anti-oxidant. The water and ethanol extracts showed 33.28% and 32.14% hyaluronidase inhibition, respectively, at phenolic concentration of $150{\mu}g/mL$. The water and ethanol extracts showed 47.33% and 40.92% elastase inhibition and 46.19% and 65.58% collagenase inhibition at phenolic concentration of $200{\mu}g/mL$, respectively. About these experiments, thinning Green ball apple was found to exhibit anti-oxidation activity as well as hyaluronidase, elastase and collagenase inhibitory activities. Therefore, thinning Green ball apple can be considered a potential source for functional food.

• Journal of Periodontal and Implant Science
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• v.28 no.4
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• pp.703-721
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• 1998

In order to observe the effects of Nicotine and NNK on cultured human gingival fibroblast, several factors were examined including mutagenicity, the number of cells attached culture plate surface through MTT test, the abundance of collagen & collagenase in mRNA level and collagenolytic activity in extracellular matrix. The results were as follows; 1. Regardless of the co-existence of S9, Nicotine did not show the mutagenicity by itself and NNK by itself showd the same result; However, dose related mutagenicity was shown in NNK with S9. 2. The number of fibroblasts attached cultured plate surface was measured by MTT procedure. The number of cells in Non-smokers increased at all time periods as compared to those of smoker. 3. Non-smoker's fibroblast treated by NNK or Nicotine was dosedependently dosedependently decreased in the number of cells when compared to untreated control. In higher dose, Nicotine showed the cellular toxicity, but NNK did not. 4. No change in the abundance of mRNA for pro${\alpha}1$ and pro${\alpha}2$ was shown in Nicotine treated group but in gingival fibroblasts following treatment with NNK, the abundance of mRNA for pro${\alpha}1$, but not pro${\alpha}2$ collagen was decreased. 5. The abundance of mRNA for collagenase was decreased when NNK was treated but no change occurred in Nicotine treated group. 6. The effect of NNK and Nicotine in collagenolytic activity showed that, collagenase activity exclusively react to type I collagen, was increased in both group, but gelatinase exclusively react to type IV collagen was not influenced at all. Collagenase activity of smoker's fibroblast was also increased as much as Nicotine and NNK group. The findings suggest that both of Nicotine and NNK lead gingival fibroblast to decrease in the abundance of collagen. And it seems to be that Nicotine and NNK have independent pathway toward the gingival fibroblast.

• Korean Journal of Food Science and Technology
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• v.39 no.6
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• pp.669-675
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• 2007

The present study was conducted to explore the anti-inflammatory action of $2-hydroxyethyl-{\beta}-undecenate$ (HPS) purified from Cumin (Cuminum cymium L.) seed against periodontitis. From the study in which human leukocyte was employed to detect the inhibiting effects of 5-lipokygenase and cyclooxygenase, enzymes generating carriers of infection like $LTB_4$ and PGs, as well as of collagenase and elastase, organ-destroying enzymes, following conclusions could be drawn: HPS was found to inhibit leukotrien $B_4$ biosynthesis by stimulating more than 97% of human polymorphonuclear leukocyte (PMNL) with addition of $5\;{\times}\;10^{-2}\;M$ when $IC_{50}$ was set at $2\;{\times}\;10^{-4}\;M$. Ninety-two percent of enzyme activation turned out to be inhibited when $5\;{\times}\;10^{-2}\;M$ was added in a test to prove inhibiting effects of HPS against activation of PMNL 5-lipoxygenase from homogeneous humans and purified 5-lipoxygenase on the market. Besides, $IC_{50}$ for enzyme activation was valued at $2.5\;{\times}\;10^{-4}\;M$, while the value of $IC_{50}$ for purified 5-lipoxygenase was $2.3\;{\times}\;10^{-4}\;M$. The $IC_{50}$ values of COX-activated leukocyte and purified collagenase were $5.1\;{\times}\;10^{-4}\;M$ and $2.3\;{\times}\;10^{-4}\;M$, respectively. Moreover, the value of $IC_{50}$ for activation of leukocyte collagenase was $2\;{\times}\;10^{-3}\;M$, whereas that for purified collagenase was $5\;{\times}\;10^{-2}\;M$. In case of leukocyte elastase, addition of $5\;{\times}\;10^{-2}\;M$ inhibited its activation by 66%. In case of purified one, however, activation of enzyme was inhibited by 25% with addition of $5\;{\times}\;10^{-2}\;M$. Furthermore, the $IC_{50}$ value for activation of leukocyte elastase was revealed to be $7.5\;{\times}\;10^{-3}\;M$. From the virulence test with human gingiva cell, it was shown that, on the second day of cultivation, 47.83% of the cell had been activated when HPS was added by $5\;{\times}\;10^{-2}\;M$. Even the addition of HPS by $1\;{\times}\;10^{-2}\;M$ featured 68.53% of cell activation, suggesting relatively strong toxicity of the substance against gingiva cell.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.3
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• pp.353-357
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• 2008

A series of studies has been conducted to establish a base infrastructure for an ovarian follicle culture system in the porcine and this study was designed to develop an effective retrieval protocol of preantral follicles. Five different methods using collagenase type I (A) or IV (B, C1, C2 and C3), which employed different treatment durations and/or conditions, were employed and sliced ovarian tissue of prepubertal gilts was provided for the retrieval. A significant increase in total number of follicles retrieved was detected when collagenase IV (methods B or C) was used. In total, more ovarian follicles were retrieved by method B undertaking agitation and method C2 without the agitation than method C1 and C3, while the number of preantral follicles collected was the largest in method B. Neither incubation in 5% $CO_2$ in air atmosphere instead of the agitation nor increased duration of enzymatic treatment up to 120 minutes improved the efficiency of follicle retrieval. There were no differences in the number of follicles retrieved from intact ovaries and from used ovaries for oocyte collection. These results demonstrate the collagenase IV treatment with agitation is effective for retrieving porcine preantral follicles from the ovaries.

• BMB Reports
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• v.48 no.8
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• pp.467-472
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• 2015

Heat shock increases skin temperature during sun exposure and some evidence indicates that it may be involved in skin aging. The antioxidant response mediated by the transcription factor NF-E2-related factor 2 (Nrf2) is a critically important cellular defense mechanism that serves to limit skin aging. We investigated the effects of heat shock on collagenase expression when the antioxidant defense system was downregulated by knockdown of Nrf2. GSH and collagenases were analyzed, and the expression of inducible Nrf2, HO-1, and NQO1 was measured. HS68 cells were transfected with small interfering RNA against Nrf2. Heat shock induced the downregulation of Nrf2 in both the cytosol and nucleus and reduced the expression of HO-1, GSH, and NQO1. In addition, heat-exposed Nrf2-knockdown cells showed significantly increased levels of collagenase protein and decreased levels of procollagen. Our data suggest that Nrf2 plays an important role in protection against heat shock-induced collagen breakdown in skin. [BMB Reports 2015; 48(8): 467-472]