The physiological effect of plant antioxidants on bone cells

• Main Authors: Min-Ting Chen, 陳敏婷
• Other Authors: Wen-Tao Huang
• Format: Others
• Language: zh-TW
• Online Access: http://ndltd.ncl.edu.tw/handle/t8zb4g

碩士 === 元培科技大學 === 放射技術研究所 === 100 === The purpose of this study was to explore the in vitro antioxidative activities of five plant antioxidants and their biological effects on the osteoblast-like cells. These antioxidants include ferulic acid, tetramethylpyrazine (TMP), caffeic acid, curcumin, and vitamin C. The study was divided into three parts: the first part was to evaluate the DPPH radical scavenging, ferrous ions chelating, and total reducing abilities of five antioxidants ; the second part was to assess the proliferation of MG-63 or 143b osteoblast-like cells with or without five oxidants treatment under hydrogen peroxide stress; the third part was using clonogenic assay and migration proficiency test to testing five antioxidants on cancer cell growth and metastatic ability.. The results show that : (1) for DPPH scavenging capability, we compare five antioxidants to dibutylhydroxytoluene (BHT) and the DPPH scavenging effects are caffeic acid (94.3 ± 0.9%), vitamin C (66.6 ± 0.7%), curcumin (56.9 ± 0.9%), ferulic acid (45.1 ± 6.9%), TMP (0.82 ± 4%), and BHT (54.7 ± 8.3%), respectively. We also find no significant ferrous ions chelating tendency of five antioxidants. Total reducing capability are determined at 50 μM for vitamin C (1.34 ± 0.02), followed by curcumin (0.70 ± 0.01), caffeic acid (0.6 ± 0.02), ferulic acid (0.52 ± 0.02), TMP (0) and BHT (0.56 ± 0.02) at 50 μM concentration. (2) TMP, ferulic acid, vitamin C in various concentrations (25, 50, 100 μM) show significant protection ability of cell damage induced by hydrogen peroxide on MG-63 osteoblast-like cancer cells. Caffeic acid has a significant protective effect at 25 μM; however, caffeic acid exhibits no protective effects of cell damage at 50 and 100μM concentration. We also find no protective effect of curcumin in various concentrations. For 143b cells, TMP, ferulic acid, caffeic acid in low concentration (25 μM) have a protective effect, five antioxidants at high concentrations (100 μM) are cell damage unprotected. (3) for clonogenic assay and cell migration analysis, curcumin and caffeic acid show significant inhibition of clongenicity on osteoblast-like cells. Low concentration of curcumin (25 μM) and caffeic acid (50 μM) can apparently restrain colony formation, i.e., at such concentration as osteoblast-like cancer cells can’t survive. In addition, curcumin and caffeic acid can effectively inhibit osteoblast-like cancer cells migration, we use 143b cell, after adding 50μM of caffeic acid, the cell migration distance at 3, 6, and 12 hours are 220.3 ± 2.1, 219.3 ± 4.5 and 214.7 ± 5.5 μm, respectively. Comparison with the migration distance at original time (0 hours, 223.3 ± 10.8 μm), the cell migration distance is slightly decreasing for 3, 4, 8.6 μm, respectively. After treating with 50μM of curcumin, the 143b cell migration distance at 3, 6, and 12 hours are 280.3 ± 5.7, 278.7 ± 2.5 and 271.7 ±10 μm, respectively. As comparing with the migration distance at 0 hours (294.7 ± 2.5 μm), the cell migration distance is apparently decreasing for 14.4, 16, 23μm, respectively. Similar experimental results also appear in MG63 osteoblast-like cancer cells. The MG63 cell migration distance at 3, 6, and 12 hours, after treating with 50μM of caffeic acid, are 199.7 ± 14.1, 200.0 ± 11.0 and 207.7 ± 8.0μm, respectively. As we compare with the migration distance at original time (200.3 ± 15.2 μm), the cell migration distance is slightly decreasing for 0.6, 0.3, 7.4 μm, respectively. After adding 50μM of curcumin, the MG63 cell migration distance at 3, 6, and 12 hours are 371.7 ± 18.0, 372.0 ± 23.9 and 370.7 ± 22.0 μm, respectively. The cell migration distance, comparing to the original time, is apparently decreasing for 20.7, 21, 19.7 μm (P <0.05), respectively. Under 50μM concentration, TMP, ferulic acid and vitamin C present no significant inhibitory effect on cancer cells colongenic assay and cell migration test. Based on the above results, TMP, ferulic acid, caffeic acid, vitamin C at low concentrations (25μm) exhibit good antioxidant capacities. Curcumin and caffeic acid at 50μm can effectively inhibit 143b or MG63 cancer cells colongenicity, furthermore, curcumin and caffeic acid also exhibit apparently inhibition of 143b or MG63 cancer cells migration. Keywords: osteoblast-like cancer cell, tetramethylpyrazine, ferulic acid, curcumin, caffeic acid, vitamin C, antioxidants, reactive oxygen species