Marie-Thérèse Droy-Lefaix

Antioxidant Action of Ginkgo Biloba Extract (EGB 761)

Extracts from the leaves of Ginkgo biloba trees have been used therapeutically for centuries in traditional Chinese medicine, and in modern Chinese pharmacopoeias both the leaves and fruits are recommended for treating problems of heart and lungs. EGb 761 is a dry, powdered extract prepared from Ginkgo biloba leaves. It is a standardized mixture of several different chemical constituents; its two major classes of compounds are flavonoid glycosides and terpenoids (Drieu 1986; DeFeudis 1991). The flavonoid fraction is mainly composed of three flavonols: quercetin, kaempferol, and isorhamnetin, which are linked to a sugar(DeFeudis 1991). The terpenoid fraction is composed of ginkgolides and bilobalides(Drieu 1988). It also contains some organic acids, which help make it water-soluble(Drieu 1986).

[46] Antioxidant action of Ginkgo biloba extract EGb 761

Publisher Summary Extracts of Ginkgo biloba leaves (EGb 761) are complex titrated, standardized mixtures of active ingredients introduced into medical practice as polyvalent therapeutic agents. This chapter discusses the radical scavenging properties of EGb 761, in several in vitro systems. The interaction of EGb 761 with superoxide and hydroxyl radicals generated by using the water radiolysis method is presented in the chapter. The method of radiolysis is useful because it specifically generates superoxide and hydroxyl radicals in known quantities, thus reducing the risk of possible interferences. To evaluate the role of terpenes contained in EGb 761, a comparative study is described using CP 202, a similar mixture lacking the terpene fraction. The interaction of EGb 761 with peroxyl radicals is assayed by a chemiluminescent method. The interaction of EGb 761 with nitric oxide is assessed by two different spectrophotometric methods. Finally, the effect of EGb 761 on xanthine oxidase activity is described.

A Ginkgo Biloba Extract (EGb 761) Prevents Mitochondrial Aging by Protecting Against Oxidative Stress

The effect of aging on indices of oxidative damage in rat mitochondria and the protective effect of the Ginkgo biloba extract EGb 761 was investigated. Mitochondrial DNA from brain and liver of old rats exhibited oxidative damage that is significantly higher than that from young rats. Mitochondrial glutathione is also more oxidized in old than in young rats. Peroxide formation in mitochondria from old animals was higher than in those from young ones. According to morphological parameters (size and complexity), there are two populations of mitochondria. One is composed of large, highly complex mitochondria, and the other population is smaller and less complex. Brain and liver from old animals had a higher proportion of the large, highly complex mitochondria than seen in organs from young animals. Treatment with the Ginkgo biloba extract EGb 761 partially prevented these morphological changes as well as the indices of oxidative damage observed in brain and liver mitochondria from old animals.

Effects of natural antioxidant ginkgo biloba extract (EGB 761) on myocardial ischemia-reperfusion injury.

Recently, it was reported that Ginkgo biloba extract (EGb 761), which is known to have antioxidant properties, also has antiarrhythmic effects on cardiac reperfusion-induced arrhythmias. In the present study, effects of EGb 761 on cardiac ischemia-reperfusion injury were investigated from the point of view of recovery of mechanical function as well as the endogenous antioxidant status of ascorbate. Isolated rat hearts were perfused using the Langendorff technique, and 40 min of global ischemia were followed by 20 min of reperfusion. EGb 761 improved cardiac mechanical recovery and suppressed the leakage of lactate dehydrogenase (LDH) during reperfusion. Furthermore, EGb 761 diminished the decrease of myocardial ascorbate content after 40 min of ischemia and 20 min of reperfusion. Interestingly, EGb 761 also suppressed the increase of dehydroascorbate. These results indicate that EGb 761 protects against cardiac ischemia-reperfusion injury and suggest that the protective effects of EGb 761 depend on its antioxidant properties.

Superoxide Dismutase (SOD) and the Paf-Antagonist (BN 52021) Reduce Small Intestinal Damage Induced by Ischemia-Reperfusion

Oxygenated free-radicals appear to play a prominent role in mediating damage associated with gastrointestinal diseases. Production of reactive oxygen metabolites in ischemia-reperfusion involves oxidases found in resident phagocytic cells and microvascular and mucosal epithelial cells. Platelet activating factor (PAF), a phospholipid associated with inflammatory disorders, has been shown to both prime and amplify the release of superoxide anion and hydrogen peroxide from polymorphonuclear neutrophils and macrophages stimulated by FMLP or PMA. To further elucidate the involvement of free radicals in intestinal damage and the potential role of PAF in their production, we examined the effect of superoxide dismutase (SOD) and BN 52021 (ginkgolide B) on ischemia-reperfusion induced damage in the small intestine. The study involved 32 Sprague-Dawley rats (100-200 g) divided into four groups. Three of these groups were subjected to occlusion of the mesenteric artery 30 mins followed by 24 h reperfusion. On 2 groups SOD (15,000 U/kg/iv) and BN 52021 (20 mg/kg/po) were administered 45 mins before arterial occlusion. Following the 24 h reperfusion, the rats were sacrificed after overnight fasting. The jejunum and ileon were removed and fixed for morphological examination. Lesions in the small intestine were quantified. The results showed extensive necrosis, hemorrhage, oedema and neutrophil invasion in the jejunal and ileal mucosa. This injury was significantly reduced by SOD (15,000 U/kg/iv) and BN 52021 (20 mg/kg/po) pretreatment. In conclusion, free-oxygenated radicals appear to mediate reperfusion damage in the small intestine and PAF appears to be involved in the genesis of these toxic products. Thus, SOD and BN 52021 may be considered as protectors against ischemic disorders.

EGb 761 protects liver mitochondria against injury induced by in vitro anoxia/reoxygenation

The present study investigated the protective effects of Ginkgo biloba extract (EGb 761) on rat liver mitochondrial damage induced by in vitro anoxia/reoxygenation. Anoxia/reoxygenation was known to impair respiratory activities and mitochondrial oxidative phosphorylation efficiency. ADP/O (2.57 +/- 0.11) decreased after anoxia/reoxygenation (1.75 +/- 0.09, p < .01), as well as state 3 and uncoupled respiration (-20%, p < .01), but state 4 respiration increased (p < .01). EGb 761 (50-200 microg/ml) had no effect on mitochondrial functions before anoxia, but had a specific dose-dependent protective effect after anoxia/reoxygenation. When mitochondria were incubated with 200 microg/ml EGb 761, they showed an increase in ADP/O (2.09 +/- 0.14, p < .05) and a decrease in state 4 respiration (-22%) after anoxia/reoxygenation. In EPR spin-trapping measurement, EGb 761 decreased the EPR signal of superoxide anion produced during reoxygenation. In conclusion, EGb 761 specially protects mitochondrial ATP synthesis against anoxia/reoxygenation injury by scavenging the superoxide anion generated by mitochondria.

OXIDATIVE STRESS IN DIABETIC RETINA

The authors describe the alterations usually associated with diabetic retinopathy. They concern the classical thickening of the basal membrane of retinal capillaries and the associated modification of retinal vessel permeability. These alterations correspond to the blood-retinal barrier disruption. The authors then discuss the participation of oxygenated free radicals in the pathogenesis of diabetic retinopathy. They report several experimental studies establishing such a participation and finally describe their own results obtained on a model of retinas isolated from alloxan-induced diabetic rats. After one month of evolution, the electroretinograms (ERG) recorded on isolated retinas from diabetic rats had an amplitude about 20% lower than the controls, whereas after two months of diabetes, this decrease was about 60%. Under these conditions, the authors tested the protective properties of Ginkgo biloba extract (EGb 761) on their model. They observed that in EGb-treated animals (100 mg/kg/day), the ERG had a significantly (p less than 0.001) greater amplitude than untreated animals after two months of diabetes evolution. In conclusion, the authors discuss the possible utilization of a free radical scavenger, such as EGb 761, in the prevention of the retinal impairment in diabetes.

Role of platelet-activating factor (PAF) and prostaglandins in colonic motor and secretory disturbances induced by Escerichia coli endotoxin in conscious rats

The involvement of prostaglandins (PGs) and platelet-activating factor (PAF) in the effects of Escherichia coli endotoxin on colonic motility, transit time, and fecal dry matter (DM) in rats were evaluated in this study. Myoelectric activity was investigated in a first group of male Wistar rats chronically implanted with intraparietal nichrome electrodes in the proximal colon. A second group of animals was chronically fitted with an intracolonic catheter (+2 cm from the cecocolonic junction); colonic transit time was then calculated as the Mean Retention Time (MRT) of 51Cr chromate sodium (1 microCi 0.1ml) administered through the intracolonic catheter and determined in the faeces collected at 1 hour interval on a conveyor belt. Fecal DM was measured after 24h dessication at 103 degrees C. E. coli endotoxin (50 micrograms/kg ip) increased the frequency of colonic contractions and decreased both colonic MRT and fecal DM. PAF (25 micrograms/kg ip) also decreased colonic MRT and fecal DM, and increased the frequency of colonic contractions. BN 50730 (10 mg/kg ip), a specific PAF receptor antagonist, blocked the effects of PAF and reduced those of endotoxin on colonic motility and transit time, but did not affect either the endotoxin- or the PAF-induced fecal DM decrease. Indomethacin (10 mg/kg ip) and SC 19220 (5 mg/kg ip) reduced the increased frequency of colonic contractions induced by endotoxin and PAF, and antagonized their effects on MRT and fecal DM. These results indicate that 1) E. coli endotoxin increases both colonic motility and transit, and decreases fecal DM, 2) PAF displays similar effects, 3) the action of endotoxin on colonic motility and transit is partly mediated by PAF and PGs while its secretory effects only depend upon PGs generation.

Effects of EGb761 and superoxide dismutase in an experimental model of retinopathy generated by intravitreal production of superoxide anion radical

Abstract · Background: A study was carried out to investigate the effect of two antioxidants –Ginkgo biloba extract (EGb761) and superoxide dismutase (SOD) – in an experimental model of vitreoretinopathy obtained by direct production of oxygen free radicals in the vitreous cavity. · Methods: Twenty-eight pigmented rabbits were used. Vitreoretinopathy was induced by intravitreal injection of 50 µl of a mixture composed of 40 nmol of xanthine and 0.001 IU of xanthine oxidase. Rabbits were randomly distributed into four groups: Group 1 (n=8) did not receive any treatment and served as a positive control. Groups 2 (n=8) and 3 (n=8) received for 1 month EGb761 given orally at a dose of 100 mg/kg/day, respectively 1 day after and 1 week before induction of retinopathy. Group 4 (n=4) was treated by three intramuscular injections of 15 000 IU/kg of SOD, 24 h before induction and 24 and 48 h thereafter. Clinical evaluations and electroretinograms (ERG) were repeatedly performed until the animals were killed at day 28. Histological examinations and immunohistological procedures were performed to ascertain the origin and characteristics of the cellular proliferation and to compare vitreoretinal structures in the four groups. · Results: Intravitreal injection of xanthine–xanthine oxidase produced a strong inflammatory response with vitreous infiltrates and epiretinal membrane formation, inconstantly associated with retinal detachment. ERG showed a decrease of the a-, b- and c-waves beginning within a few hours after injection. Histologic evaluation found an intravitreal and epiretinal infiltration by leukocytes and epithelial-derived cells, dense vitreoretinal membranes and retinal detachments with occasional neovascularization. In the treated groups (groups 2–4), all clinical, electric and histologic data were significantly improved compared to the control group. However, no difference could be found among the three treated groups. · Conclusion: This study demonstrates the strong pathologic effects of free radical production on the retina and the close relationships between free radicals, inflammatory pathways and vitreoretinal proliferative disorders. It also confirms the pharmacological interest of prevention by antioxidants and free radical scavengers.

Myoelectric intestinal disturbances in Escherichia coli endotoxic shock in rats. Involvement of platelet-activating factor

Administration of BN 52021 (50 mg/kgi.v.), a specific antagonist of platelet-activating factor (PAF), significantly reduced the intestinal myoelectric disturbances induced byE. coli endotoxin injection (50 μg/kgi.v.) by 62%. Thus, PAF may be involved in the intestinal motor alterations observed in endotoxic shock. When given in combination with indomethacin (10 mg/kgi.p.), BN 52021 inhibited endotoxic shock intestinal disturbances. Indomethacin alone also reduced PAF induced (25 μg/kgi.p.) disruption of migrating myoelectric complexes. Endotoxins may act on intestinal motilityvia release of endogenous PAF and prostaglandins, the effects of PAF being mediated through the release of prostaglandins.