• Title/Summary/Keyword: TIMP-1

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Combined Treatment of Activin A and Heparin Binding-EGF (HB-EGF) Enhances In Vitro Production of Bovine Embryos

  • Kim, Se-Woong;Jung, Yeon-Gil;Park, Jong-Im;Roh, Sangho
    • Journal of Embryo Transfer
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    • v.29 no.2
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    • pp.127-132
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    • 2014
  • This study was carried out to investigate the effects of tissue inhibitor of matalloproteinase-1 (TIMP-1), Activin A and Heparin binding epidermal growth factor (HB-EGF) on in vitro production of bovine embryos. In experiment 1, presumptive zygotes were cultured in the medium supplemented with TIMP-1 ($0.5{\mu}g/ml$), Activin A (100 ng/ml), or HB-EGF (100 ng/ml) at $39^{\circ}C$ in a humidified atmosphere of 5% (v/v) $CO_2$, 5% (v/v) $O_2$ and 90% (v/v) $N_2$. In experiment 2, TIMP-1 + HB-EGF or Activin A + HB-EGF combinations were supplemented in the culture medium. The developmental rate to blastocysts, hatching rate and total cell numbers of the blastocysts were evaluated in both experiments. The embryos cultured in medium without growth factor supplementation was used as control group. In experiment 1, the embryos cultured in medium supplemented with TIMP-1 and Activin A showed significantly higher developmental rate to blastocysts than those cultured with HB-EGF and control (36.9%, 34.1%, 21.2% and 23.1%, respectively) (P<0.0001). However, the hatching rate of blastocyst was significantly higher in embryos with HB-EGF than those with TIMP-1, Actvin A and Control groups (84.4%, 58.8%, 51.4% and 49.3%, respectively) (P<0.001). Total cell number per blastocyst was also significantly higher in embryos with HB-EGF group ($174.3{\pm}2.5$) than those with TIMP-1, Activin A (149.7 and 150.0, respectively) (P<0.05) and Control (119.0) (P<0.001). In experiment 2, embryos cultured with combined treatment of Activin A and HB-EGF resulted in significantly higher rates of blastocysts formation (48.0%), hatching rate (89.7%) and total cell number in blastocyst ($182.3{\pm}2.1$) than those with TIMP-1 and HB-EGF combination group (32.0%, P<0.001; 76.6%, P<0.05; $165.7{\pm}4.2$, P<0.001, respectively). Our data demonstrate that in vitro production of bovine embryos could be improved by combined supplementation of Activin A and HB-EGF in culture medium.

Anti-aging Effect of Amino Acid Complex on the Skin (Amino Acid Complex의 항노화 작용)

  • Kim, Ki-Ho;Kim, Ki-Soo;Kim, Young-Heui;Ko, Kang-Il;Park, Sun-Hee;Kim, Jin-Guk;Chio, Hyun-Jin;Ko, Su-Yeon;Bae, Yoon-Joo;Kim, Yong-Min
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.32 no.2 s.57
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    • pp.81-88
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    • 2006
  • Amino acid complex was made with the composition of amino acids quite similar to that of stratum corneum on the skin. In order to evaluate the efficacy of amino acid complex on the skin as an active cosmetic ingredient, we measured cytotoxicity. TIMP-1 expression, moisturizing effect and the amount of horny substance. In cytotoxicity assay, amino acid complex did not show any cytotoxicity. In moisturizing effect test using corneometer, it showed very good moisturizing effect. In TIMP-1 mRNA assay using RT-PCR, moo acid complex showed the increase of TIMP-1 expression, suggesting that amino acid complex have anti-wrinkle effect. Therefore, amino acid complex may be useful as an active ingredient for anti-aging.

The Influence of Diabetes on of $PGE_2$, MMP-14 and TIMP Expressions in Human Chronic Periodontitis (만성 치주염 환자에서 2형 당뇨병이 $PGE_2$, MMP-14 및 TIMP의 발현양상에 미치는 영향)

  • Kim, Seon-Woo;Park, Jin-Woo;Suh, Jo-Young;Lee, Jae-Mok
    • Journal of Periodontal and Implant Science
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    • v.37 no.4
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    • pp.755-766
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    • 2007
  • Purpose: The purposes of this study were to compare and quantify the expression of $PGE_2$, MMP-14 and TIMP-1 in the gingival tissues of patients with type 2 diabetes mellitus and healthy adults of chronic periodontitis with alveolar bone resorption. Material and methods: Gingival tissue samples were obtained during periodontal surgery or tooth extraction. According to the patient's systemic condition & clinical criteria of gingiva, each gingival sample was devided into three groups. Group 1 (n=8) is clinically healthy gingiva without bleeding and no evidence of bone resorption or periodontal pockets, obtained from systemically healthy 8 patients. Group 2 (n=8) is inflammed gingiva from patients of chronic periodontitis with alveolar bone resorption. Group 3(n=8) is inflammed gingiva from patients of chronic periodontitis with alveolar bone resorption associated with type 2 diabetes. Tissue samples were prepared and analyzed by Western blotting. The quantification of $PGE_2$ MMP-14 and TIMP-1 were performed using a densitometer and statistically analyzed by one-way ANOVA followed by Tukey test. Results: The expressions of MMP-14 and TIMP-1 were showed increasing tendency in group 2 & 3 compared to group 1. The expressions of $PGE_2$, MMP-14 were showed increasing tendency in group 3 compared to group 1 and group 2. According to MMP-14 levels were increasing, $PGE_2$ showed increasing tendency in group 3, and although $PGE_2$, MMP-14 levels were increasing, TIMP-1 levels were similar expressed comparing to group 2. Conclusion: In conclusion, this study demonstrated that the expression levels of MMP-14 and TIMP-1 had increasing tendency in inflammed tissue. It can be assumed that $PGE_2$ and MMP-14 may be partly involved in alveolar bone resorptive process and the progression of periodontal inflammation associated to type 2 DM.

Expression Analysis of Matrix Metalloproteinases and Tissue Inhibitor of Matrix Metalloproteinases from In Vitro Maturation Oocytes Complexes in Porcine (돼지 체외성숙난자에서 MMPs와 TIMPs의 발현 분석)

  • Kim, Sang-Hwan;Kang, Hyun-Ah;Kim, Dae-Seung;Lee, Myeong-Seop;Seo, Kang-Suk;Min, Kwan-Sik;Yoon, Jong-Taek
    • Reproductive and Developmental Biology
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    • v.34 no.1
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    • pp.55-62
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    • 2010
  • Matrix metalloproteinases (MMP) play important roles in extracellular matrix (ECM) remodeling during ovarian follicular development, oocytes development and ovulation. In an attempt to investigate the effect of MMP activation in development cumulus-oocytes complexes, we examined the localization and expression of MMP, and monitored MMP expression profile. Cumulus-oocytes complexes were collected and matured in vitro for 24 hr, 36 hr and 48 hr. A mRNA expression of MMP-2, MMP-9, TIMP-2 and TIMP-3 was detected in all culture medium regardless of CC, DC and CDCs. Activity of MMP-2 in the DC progressively was increased from 24 hr to 48 hr. But MMP-9 was not detected in all culture medium. The localization of MMP-2 was also measured by immunohistochemistry analysis. The MMP-2 and TIMP-2 was detected in cumulus cell and oocyte zone pellucida. Expression of MMP-2 protein in the COCs was progressively increased from 24 hr to 48 hr. However, MMP-9 protein was progressively decreased from 24 hr to 48 hr. And TIMP-2 protein was most highly expressed in the CDCs 36 hr. Expression of TIMP-3 protein in the CDCs was progressively increased from 24 hr to 48 hr. In conclusion, these results suggest that MMP-2 plays a role in maintaining normal maturation and development by controlling the ECM inhibitor concentration on cumulus cell and oocytes.

Suppression of Human Breast Cancer Cell Metastasis by Coptisine in Vitro

  • Li, Jing;Qiu, Dong-Min;Chen, Shao-Hua;Cao, Su-Ping;Xia, Xue-Lan
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.14
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    • pp.5747-5751
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    • 2014
  • Background: Coptisine, an isoquinoline alkaloid extracted from Coptidis rhizoma, has many biological activities such as antidiabetic, antimicrobial and antiviral actions. However, whether coptisine exerts anti-cancer metastasis effects remains unknown. Materials and Methods: Effects of coptisine on highly metastatic human breast cancer cell MDA-MB-231 proliferation were evaluated by trypan blue assay and on cell adhesion, migration and invasion by gelatin adhesion, wound-healing and matrigel invasion chamber assays, respectively. Expression of two matrix metalloproteinases (MMPs), MMP-9, MMP-2 and their specific inhibitors tissue inhibitor of metalloproteinase 1 (TIMP-1) and tissue inhibitor of metalloproteinase 2 (TIMP-2) were analyzed by RT-PCR. Results: Coptisine obviously inhibited adhesion to an ECM-coated substrate, wound healing migration, and invasion through the matrigel in MDA-MB-231 breast cancer cells. RT-PCR revealed that coptisine reduced the expression of the ECM degradation-associated gene MMP-9 at the mRNA level, and the expression of TIMP-1 was upregulated in MDA-MB-231 cells, while the expression of MMP-2 and its specific inhibitor TIMP-2 was not affected. Conclusions: Taken together, our data showed that coptisine suppressed adhesion, migration and invasion of MDA-MB-231 breast cancer cells in vitro, the down-regulation of MMP-9 in combination with the increase of TIMP-1 possibly contributing to the anti-metastatic function. Coptisine might be a potential drug candidate for breast cancer therapy.

Associations of matrix metalloproteinase (MMP)-8, MMP-9, and their inhibitor, tissue inhibitor of metalloproteinase-1, with obesity-related biomarkers in apparently healthy adolescent boys

  • Shin, Youn Ho;Kim, Ki Eun;Lee, Yong-Jae;Nam, Jae-Hwan;Hong, Young Mi;Shin, Hye-Jung
    • Clinical and Experimental Pediatrics
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    • v.57 no.12
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    • pp.526-532
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    • 2014
  • Purpose: Matrix metalloproteinases (MMPs) have been implicated in atherosclerosis, and therefore, are considered risk factors for metabolic dysfunction in adults. However, there is little data on circulating levels of MMPs and tissue inhibitors of MMPs (TIMPs) with regard to obesity-related biomarkers in the general adolescent population. In the present study, we determined the associations of MMP-8, MMP-9, and TIMP-1 levels and MMP-8/TIMP-1 and MMP-9/TIMP-1 ratios with obesity-related biomarkers in apparently healthy adolescent boys. Methods: We measured MMP and TIMP concentrations in plasma samples using the enzyme-linked immunosorbent assay and analyzed their associations with obesity-related biomarkers, such as liver enzymes and lipid profiles, in a sample of 91 Korean boys aged 13-14 years who participated in a general health check-up. Results: The mean age of the boys was $13.8{\pm}0.3years$; 72 boys were normal weight and 19 were overweight/obese. The Pearson correlation coefficients revealed a significant correlation between MMP-8 and aspartate aminotransferase (r=0.217, P=0.039) and alanine aminotransferase (r=0.250, P=0.017) and between TIMP-1 and aspartate aminotransferase (r=0.267, P=0.011). In a multivariate linear regression analysis, serum alanine aminotransferase was positively associated with the MMP-8 level. There were no significant differences in the MMP-8, MMP-9, and TIMP-1 levels or MMP-8/TIMP-1 and MMP-9/TIMP-1 ratios between control and overweight/obese subjects. Conclusion: We found a significant association between the MMP-8 level and alanine aminotransferase in the apparently healthy adolescent boys. These findings indicate that there may be a pathophysiological mechanism underlying the relationship between MMP-8 and liver enzymes in young adolescents.

The comparison of inflammatory mediator expression in gingival tissues from human chronic periodontitis patients with and without type 2 diabetes mellitus (단순 만성 치주염 환자 및 2형 당뇨병환자의 만성 치주염 치은조직에서 염증성 매개인자의 발현 양상 비교)

  • Joo, Sang-Don;Lee, Jae-Mok
    • Journal of Periodontal and Implant Science
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    • v.37 no.sup2
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    • pp.353-369
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    • 2007
  • Gingival tissue samples were obtained during periodontal surgery or tooth extraction. According to the patient's systemic condition & clinical criteria of gingiva, each gingival sample was divided into three groups. Group 1 (n=8) is clinically healthy gingiva without bleeding and no evidence of bone resorption or periodontal pockets, obtained from systemically healthy 8 patients. Group 2 (n=8) is inflamed gingiva from patients with chronic periodontitis. Group 3 (n=8) is inflamed gingiva from patients with chronic periodontitis associated with type 2 diabetes. Tissue samples were prepared and analyzed by Western blotting. The quantification of $IL-1{\beta}$, MMP-13 and TIMP-1 were performed using a densitometer and statistically analyzed by one-way ANOVA followed by Tukey test. 1. The expressions of MMP-13 and TIMP-1 showed increasing tendency in group 2 & 3 compared to group 1. 2. The expressions of $IL-1{\beta}$ & MMP-13 were showed increasing tendency in group 3 compared to group 2. 3. As $IL-1{\beta}$ levels were increasing, MMP-13 showed increasing tendency in group 3, and although $IL-1{\beta}$ , MMP-13 levels were increasing, TIMP-1 levels were similar expressed comparing to group 2. In conclusion, this study demonstrated that the expression levels of MMP-13 and TIMP-1 had increasing tendency in inflamed tissue. It can be assumed that $IL-1{\beta}$ and MMP-13 may be partly involved in the progression of periodontal inflammation associated to type 2 DM.

Expression of Matrix Metalloproteinase-2 and Tissue Inhibitor of Metalloproteinase-2 in Radiation Exposed Small Intestinal Mucosa of the Rat (방사선조사를 받은 흰쥐 소장 점막의 손상과 재생과정 중 금속단백효소 및 억제자의 발현)

  • Kwag, Hyon-Joo;Lee, Kyoung-Ja;Rhee, Chung-Sik
    • Radiation Oncology Journal
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    • v.21 no.1
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    • pp.66-74
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    • 2003
  • Purpose : The matrix metalloprotelnases (MMPs) are a family of enzymes whose main function is the degradation of the extracellular matrix. Several studies have revealed that MMPs and TIMPS are related to the wound heating process and in photoaging caused by ultraviolet Irradiation. However, the expressions of MMP and TIMP after irradiation have not, to the best of our knowledge, been studied. This study investigates the expressions of MMP-2 and TIMP-2 in rat Intestinal mucosa following irradiation. Materials and Methods : The entire abdomen of Sprague-Dawley rats was irradiated using a single dose method. The rats were sacrificed on day 1, 2, 3, 5, 7 and 14 following irradiation. Histopathological observations were made using hematoxilin & eosin staining. The expressions of MMP-2 and TIMP-2 were examined using immunohistochemistry, Irnrnunoblotting and ELISA. Results : Radiation induced damage associated with atrophic villi, and infiltration of inflammatory cell was observed from the first postirradiation day, and severe tissue damage was observed on the second and the third postirradiation days. An increase in mitosis and the number of regenerating crypts, as evidence of regeneration, were most noticeable on the fifth postirradiation day. From the immunohistochemlstry, the MMP-2 expression was observed from the first postirradiation day, but was most conspicuous on the third and the fifth postirradiation days. The TIMP-2 expression was most conspicuous on the fifth postirradiation day. From the irnrnunoblotting, the MMP-2 expression was strongly positive on the third postirradlatlon day, and that of TIMP-2 showed a strong positive response on the fifth postirradiation day. In ELISA tests, the expressions of MMP-2 and TIMP-2 were increased in the postirradiation groups compared to those of the normal controls, and showed a maximum increase on the fifth postirradiatlon day. These results were statistically significant. Conclusion : The expressions of MMP-2 and TIMP-2 were increased in the intestinal mucosa of the rats following irradiation, and these results correlated with the histopathological findings, such as tissue damage and regeneration. Therefore, this study suggests that MMP-2 and TIMP-2 play roles in the mechanisms of radiation-induced damage and regeneration of intestinal mucosa of rats.

THE mRNA EXPRESSION OF MMP-1, TIMP-1, $TGF-{\beta}_1$ IN GINGIVAL KERATOCYTES FROM GINGIVAL HYPERPLASIA INDUCED BY CYCLOSPORINE A (정상 치은 조직에서 배양된 각화세포에서 Cyclosporine A에 의한 MMP-1, TIMP-1, $TGF-{\beta}_1$의 발현에 관한 연구)

  • Kang, Hag-Soo;Lee, Jae-Sun;Bing, Jung-Ho;Park, Chang-Joo;Im, Jae-Jung;Hwang, Kyung-Gyun;Shim, Kwang-Sup
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.34 no.4
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    • pp.405-411
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    • 2008
  • Purpose : Cyclosporine A (CsA) is a versatile immunosuppresive agent used to prevent graft rejection syndrome and treat autoimmune disease. One of the major side effects associated with CsA is the abnormal gingival hyperplasia. The purpose of this study was to investigate the relationship between the mRNA expression of the MMP-1, TIMP-1, and $TGF-{\beta}_1$ and the concentration of CsA in cultured human gingival keratinocytes. Materials & Methods : Gingival keratocytes were obtained from gingival tissues of 4 healthy donors. The cultured gingival keratocytes were incubated with increasing concentrations of CsA (0-2000 ng/ml) for 24 hours and the expression of MMP-1, TIMP-1, and $TGF-{\beta}_1$ were determined by reverse transcription-polymerase chain reaction (RT-PCR). Results : The expressions of MMP-1 and $TGF-{\beta}_1$ were not significantly different according to the concentrations of CsA. The expression of TIMP-1 was significantly increased at the CsA concentration of 500 ng/ml. Conclusion : We concluded that the gingival hyperplasia induced by CsA was more related with TIMP-1 than MMP-1 or $TGF-{\beta}_1$ on gingival collagen metabolism in patients treated with CsA.