博士 / 中國醫藥大學 / 中國藥學暨中藥資源學系博士班 / 99 / In this study, the chemical profile of water extract of Crossostephium chinensis (L.) Makino (CCW) and methanol extract of Limonium wrightii (Hance) Ktze.(LWM) was analyzed by LC-MS-MS spectrometer (high-performance liquid chromatography tandem mass). Chromatographic pattersn from HPLC analysis (254 nm) of rutin and scopoletin were detected in CCW. We then compared the results in anti-oxidant experiment with water and methanol extract of Crossostephium chinensis (L.) Makino (CCW, CCM) and Limonium wrightii (Hance) Ktze.(LWW, LWM). We also compared the anti-oxidant capability of different partition layers of CCM and LWM.Eventually, the effects of CCW and LWM on hepatoprotectic, analgesic and anti-inflammation activity were evaluated. Further assessment of the anti-carcinogenic activity and the mechanisms of its chemopreventive action were also performed.
The antioxidative assays included: ABTS ([2,2’-azinobis (3-ethylbenzothiazoline)-6-sulfonic acid] diammonium salt) free radical scavenging assay, TPC (determination of total phenolics contents) , (FRAP) ferric reducing antioxidant power assay, rapid screening of antioxidant by dot-blot DPPH (1,1-diphenyl-2-picrylhydrazyl) staining, DPPH radical-scavenging activities, reducing power measurement, Chelating ability, superoxide radical scavenging, nitric oxide (NO) radical scavenging and lipid peroxidation assay. In the hepatoprotective studies, serum levels of GOT (glutamate oxaloacetate transaminase) , GPT (glutamate pyruvate transaminase), NO production, antioxidant capacity, TNF-α (tumor necrosis factor-α) concentration in serum, the liver SOD (superoxide dismutase), and GPx (glutathione peroxidase), the CAT (catalase) activities, GSH (glutathione) contents, TBARS (thiobarbituric acid reactive substances) formation and the liver tissue examinations were determined. Activities of MMP-9 (matrix metalloproteinases) in SDS-PAGE zymography of serum and liver were also determined. The full time lick induction test and the λ-carrageenan paw edema induction test were also performed. Light microscopy observations of mice paw sections stained with Van Gieson of iNOS (inducible nitric oxide synthase) and COX-2 (cyclo-oxygenase- 2) were performed. The expression of iNOS and COX-2 were inhibited by western blotting. NO concentration, tissue MDA (malondialdehyde) concentration, tissue TNF-α concentration of foot and serum in mice were determined.
The results showed that CCW did contain the active contents rutin and scopoletin, with the retention time 8 min between CCW and rutin, and retention time 13 min between CCW and scopoletin, respectively. And LWM did contain the active content gallic acid, with the retention time 8 min between LWM and gallic acid. In the anti-oxidant capability of this study, water extract of Crossostephium chinensis (L.) Makino and Limonium wrightii (Hance) Ktze.(CCW, LWW) are better than methanol extract of Crossostephium chinensis (L.) Makino and Limonium wrightii (Hance) Ktze.(CCM, LWM). Significant in vitro oxidative resistance phenomena by EtOAC and n-BuOH fractions were observed. Liver functional, inflammation, morphological and mechanism studies GOT, GPT, SOD, GPx, CAT, GSH, NO, antioxidant capacity, TNF-α concentration, TBARS formation, zymography showed reduction of liver damage by administration of CCW. Accordingly, analgesia and foot edema reduction together and MDA, Nitric Oxide concentration and TNF-α concentration were also significantly observed in CCW and LWM.
In conclusion, CCW contained active compounds, rutin and scopoletin whereas LWM contained active compound gallic acid, both have the strong anti-oxidation. Significant in vitro oxidative resistance phenomena by EtOAC and n-BuOH fractions were observed to be better than other partition layer. CCW and LWM both have liver protection, analgesic, anti-inflammative and anti-hepatoma activities.
| Identifer | oai:union.ndltd.org:TW/099CMCH5049006 |
| Date | January 2011 |
| Creators | Tien-Ning Chang, 張恬寧 |
| Contributors | 張永勳 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 201 |
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