Background: Matrix metalloproteinases (MMPs) are linked with several complications such as metastasis of cancer progression, oxidative stress, and hepatic fibrosis. Brown seaweeds are being extensively studied for their bioactive molecule content against cancer progression. In this context, Sargassu...
Background: Matrix metalloproteinases (MMPs) are linked with several complications such as metastasis of cancer progression, oxidative stress, and hepatic fibrosis. Brown seaweeds are being extensively studied for their bioactive molecule content against cancer progression. In this context, Sargassum horneri was reported to possess various bioactivities including antiviral, antimicrobial, and anti-inflammatory partly due to its phenolic compound content. Methods: In this study, potential of S. horneri was evaluated through anti-MMP effect in HT1080 fibrosarcoma cells. S. horneri crude extract was fractionated with organic solvents, namely, water ($H_2O$), n-buthanol (n-BuOH), 85% aqueous methanol (85% aq. MeOH), and n-hexane. The non-toxicity of fraction samples (Sargassum horneri solvent-partitioned extracts (SHEs)) was confirmed by cell-viability assay. SHEs were tested for their ability to inhibit MMP enzymatic activity through gelatin digestion evaluation and cell migration assay. Expressions of MMP-2 and MMP-9 and tissue inhibitors of MMP (TIMPs) were evaluated by reverse transcription and Western blotting. Results: All fractions inhibited the enzymatic activities of MMP-2 and MMP-9 according to gelatin zymography. Except $H_2O$ fraction, fractions hindered the cell migration significantly. All tested fractions suppressed both mRNA and protein levels of MMP-2, MMP-9, TIMP-1, and TIMP-2. Conclusion: Overall, current results suggested that S. horneri has potential to be a good source for anti-MMP agents, and further investigations are underway for better understanding of the action mechanism and isolation and elucidation of the bioactive molecules.
Background: Matrix metalloproteinases (MMPs) are linked with several complications such as metastasis of cancer progression, oxidative stress, and hepatic fibrosis. Brown seaweeds are being extensively studied for their bioactive molecule content against cancer progression. In this context, Sargassum horneri was reported to possess various bioactivities including antiviral, antimicrobial, and anti-inflammatory partly due to its phenolic compound content. Methods: In this study, potential of S. horneri was evaluated through anti-MMP effect in HT1080 fibrosarcoma cells. S. horneri crude extract was fractionated with organic solvents, namely, water ($H_2O$), n-buthanol (n-BuOH), 85% aqueous methanol (85% aq. MeOH), and n-hexane. The non-toxicity of fraction samples (Sargassum horneri solvent-partitioned extracts (SHEs)) was confirmed by cell-viability assay. SHEs were tested for their ability to inhibit MMP enzymatic activity through gelatin digestion evaluation and cell migration assay. Expressions of MMP-2 and MMP-9 and tissue inhibitors of MMP (TIMPs) were evaluated by reverse transcription and Western blotting. Results: All fractions inhibited the enzymatic activities of MMP-2 and MMP-9 according to gelatin zymography. Except $H_2O$ fraction, fractions hindered the cell migration significantly. All tested fractions suppressed both mRNA and protein levels of MMP-2, MMP-9, TIMP-1, and TIMP-2. Conclusion: Overall, current results suggested that S. horneri has potential to be a good source for anti-MMP agents, and further investigations are underway for better understanding of the action mechanism and isolation and elucidation of the bioactive molecules.
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제안 방법
However, treatment with SHE was observed to produce mixed results regarding the effect on the TIMP levels following the PMA stimuli. Expected results were to inhibit MMP expression while enhancing TIMP expression in order to regulate the extracellular matrix degradation. In these terms, only 85% aq.
FK and SGL carried out the experiments. JAK, and CSK conceived the idea and designed the experiments.
The target cDNA was amplified using the following primers: forward 5′-TGA-AGG-TCG-GTG-TGA-ACG-GA-3′ and reverse 5′-CAT-GTA-GCC-ATG-AGG-TCC-ACC-AC-3′ for MMP-2; forward 5′-CAC-TGT-CCA-CCC-CTC-AG A-GC-3′ and reverse 5′-CAC-TTG-TCG-GCG-ATA-AG G-3′ for MMP-9; forward 5′-AAT-TCC-GAC-CTC-GTCATC-AG-3′ and reverse 5′-TGC-AGT-TTT-CCA-GCAATG-AG-3′ for TIMP-1; forward 5′-TGA-TCC-ACA-CA C-GTT-GGT-CT-3′ and reverse 5′-TTT-GAG-TTG-CT T-GCA-GGA-TG-3′ for TIMP-2; and forward 5′-GCC-A CC-CAG-AAG-ACT-GTG-GAT-3′ and reverse 5′-TGGTCC-AGG-GTT-TCT-TAC-TCC-3′ for β-actin.
대상 데이터
JAK, and CSK conceived the idea and designed the experiments. FK, JHO and JAK participated to write the manuscript and interpreted the data. JHO performed the necessary revisions and updates on the manuscript.
데이터처리
Differences between the calculated means of the each individual group were determined by one-way ANOVA coupled with Duncan’s multiple range tests.
이론/모형
The cells were treated with different concentrations of SHEs (5 and 50 μg/ml) and incubated for 48 h. The cytotoxicity was determined by MTT assay. Values are mean ± SD (n = 3).
Total protein contents of the cells were normalized using Bradford protein determination method.
성능/효과
2, the MMP-2 activity was depicted as a percentage to that of activation of MMP-2 from proMMP-2. Data showed that the n-BuOH and H2O SHEs notably increased the MMP-9 activity while showing decreased MMP-2 activity. This can be linked to the regulatory dynamic between MMP-2 and MMP-9.
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