Part I: Studies on the Superoxide Dismutase (SOD) from the Plants Cardiosperm halicacabum Linn, Ophioglossum petiolatum Hook, Gynostemma pentaphyllum (Thumb.) Markio, Markino, Citrus and Beans Species

• Main Authors: Lee, Lung-ta, 李隆達
• Other Authors: Hsuch-ching Chiang
• Format: Others
• Language: zh-TW
• Published: 1997
• Online Access: http://ndltd.ncl.edu.tw/handle/50664607586452564468

博士 === 國立臺灣大學 === 藥學系 === 85 === Part I: Studies on the Superoxide Dismutase (SOD) from the Plants Cardiosperm halicacabum Linn, Ophioglossum petiolatum Hook, Gynostemma pentaphyllum (Thumb.) Markino, Citrus and Beans Species.A. Cardiospermum halicacabum LinnCardiospermum halicacabum Linn (Sapindaceae) exhibited anti-inflammatory activity. In this study, superoxide dismutases (SOD) were detected from the aerial parts of the fresh plant using polyacrylamide gel electrophoresis, and two major Cu.Zn-SODs were identified by their sensitivity to cyanide and hydrogen peroxide. The molecular weight of these SODs were estimated to be 78 kDa and 29 KDa.By nitro blue tetrazolium (NBT) method, the SOD activity in stems, leaves, fruit capsulae and seeds from the aerial parts of the plant were 475, 221, 535 and 471 mUnits/mg protein respectively. However, the root of the plant did not contain the enzyme.B. Ophioglossum petiolatum HookOphioglossum petiolatum Hook (Ophioglossaceae) has been traditionally used as antipyretic, antidote, antiplatelet, analgesic and anti- inflammatory medicine and the extract of the plant exhibits xanthine oxidase inhibition.In this study, five isozymes of superoxide dismutases(SODs) were characterized from the extract of fresh plant collected in the month of December and April using polyacrylamide gel electrophoresis. Cu.Zn-SODs were identified by their sensitivity to cyanide and hydrogen peroxide for these five isozymes. The molecular weight of these SODs were estimated to be 84, 71, 47, 42 and 34 KDa.By nitro blue tetrazolium (NBT) assay, the total Cu.Zn-SOD activities were 92.4±15 mUnits/mg protein from the plant collected in December and 158.47±24 mUnits/mg protein in April.C. Gynostemma pentaphyllum (Thunb.) MakinoGynostemma pentaphyllum (Thunb.) Makino (Cucurbitaceae) has been used in folk medicine as anti- inflammation for rheumatoid arthritis, chronic bronchitis, liver disease and prostatism.Three Mn-SOD, one Fe-SOD and three Cu.Zn- SOD were detected from the aerial parts of the fresh plant using polyacrylamide gel electrophoresis. The molecular weight of these SODs were estimated to be 115, 100 and 70 KDa for three Mn-SOD, 78, 64 and 57 KDa for three Cu.Zn-SOD and 38 KDa for one Fe-SOD.By nitro blue tetrazolium (NBT) assay, the total SOD activity in the aerial parts of the plant was 248, 3±12.2 mUnit/mg protein.D. Citrus species (Rutaceae)It has been found several isozymes of superoxide dismutases from the peels of the fresh fruits from Citrus species (Rutaceae), such as several variety of Citrus grandis Osbeck, Citrus paradisi, the variety of Citrus tangerina and Fortunella margarita. By nitro blue tetrazolium (NBT) assay, the total SOD activities in the peels of the Citrus species were 22.9±4.3 (Yow-tzyy), 178.0±5.5 (Bair-yow), 93.3±12.2 (Shi-shy-yow), 407.8±20.6 (Pwu-taur- yow), 809.5±37.6 (Tyu, orange), 66.3±7.1 (Huu-tour-gan) and 180.2±21.2 (Jin-tyu) mUnit/mg protein.E. Beans species (Leguminosae)Abundant SODs were detected in the seeds of the Mau-dow and Miin-dow (Leguminosae). By nitro blue tetrazolium (NBT) assay, the total SOD activities in the seeds of the Mau- dow and Miin-dow were 207.0±34.5 and 292.2±23.0 mUnit/mg protein, respectively.Part II: Studies on the Xanthine Oxidase Inhibitors and Antitumor ActivityThe flavonoids are present in the higher plants and their consumption in the normal diet amounts to 1g or more per day. These compounds possess a wide range of pharmacodynamic properties including xanthine oxidase inhibition and anticarcinogenesis.In this study, UV spectrophotometric assay and fluorometric pterin assay of the activity of xanthine oxidase were employed for glycosides and flavonoids. The results showed that glycosides did not display xanthine oxidase inhibition, whereas flavonoids showed the effects of xanthine oxidase inhibition by both assay.In addition, several flavonoids, such as luteolin, quercetin, capillarisin and apigenin showed the inhibitory effects of xanthine oxidase in various cell lines. These compounds not only inhibit xanthine oxidase in NIH3T3 cells but also inhibit the activity of xanthine oxidase induced by PMA (phorbol 12-myristate 13-acetate).In the antitumor study, it has been found that quercetin, luteolin and genistein inhibit the expression of the total kinase activities of various tumor cell lines as well as growth inhibition of the tumor cells.On the other hand, luteolin, genistein and apigenin are able to induce apoptosis of various tumor cells and inhibit metalloproteinases in various tumor cells.