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[Ginkgo biloba extract in peripheral arterial diseases. Meta-analysis of controlled clinical studies].

In the first part the statistical methods of meta-analysis are discussed. Meta-analysis is considered as a statistical tool for quantitatively summarizing the results of clinical trials with comparable aims (treatments) and designs. Meta-analysis can be based on the significance probabilities or effect values. The last procedure is preferable as it gives an estimate (and confidence interval) for the global effect of the treatment of interest, if homogeneity of the effects between the trials can be assumed. Such a homogeneity can be often achieved by a suitable standardization of the effect variables within the trials. In the second part the methods of meta-analysis are applied to controlled clinical trials with Ginkgo biloba extract EGb 761 in patients with peripheral arterial disease. Included were 5 placebo-controlled clinical trials with similar design and inclusion criteria. In all studies treatment effect was quantified by the increase of walking distance (measured in standardized treadmill exercise). The effect value of EGb 761 treatment was expressed by the standardized mean difference in walking distance increase between EGb 761 and placebo, standardized by the standard deviation. It could be shown that this effect value is homogeneous in all trials. The global effect size was estimated as 0.75. This means that the mean increase in walking distance achieved by EGb 761 is 0.75 times of the standard deviation higher than that achieved by placebo. This value is highly significant different from zero. So the meta-analysis revealed a highly significant therapeutic effect of EGb 761 for the treatment of peripheral arterial disease.

Arteries↗

Effect of Ginkgo biloba extract on procarcinogen-bioactivating human CYP1 enzymes: identification of isorhamnetin, kaempferol, and quercetin as potent inhibitors of CYP1B1.

In the present study, we investigated the effect of Ginkgo biloba extracts and some of its individual constituents on the catalytic activity of human cytochrome P450 enzymes CYP1B1, CYP1A1, and CYP1A2. G. biloba extract of known abundance of terpene trilactones and flavonol glycosides inhibited 7-ethoxyresorufin O-dealkylation catalyzed by human recombinant CYP1B1, CYP1A1, and CYP1A2, and human liver microsomes, with apparent Ki values of 2 +/- 0.3, 5 +/- 0.5, 16 +/- 1.4, and 39 +/- 1.2 microg/ml (mean +/- SE), respectively. In each case, the mode of inhibition was of the mixed type. Bilobalide, ginkgolides A, B, C, and J, quercetin 3-O-rutinoside, kaempferol 3-O-rutinoside, and isorhamentin 3-O-rutinoside were not responsible for the inhibition of CYP1 enzymes by G. biloba extract, as determined by experiments with these individual chemicals at the levels present in the extract. In contrast, the aglycones of quercetin, kaempferol, and isorhamentin inhibited CYP1B1, CYP1A1, and CYP1A2. Among the three flavonol aglycones, isorhamentin was the most potent in inhibiting CYP1B1 (apparent Ki = 3 +/- 0.1 nM), whereas quercetin was the least potent in inhibiting CYP1A2 (apparent Ki = 418 +/- 50 nM). The mode of inhibition was competitive, noncompetitive, or mixed, depending on the enzyme and the flavonol. G. biloba extract also reduced benzo[a]pyrene hydroxylation, and the effect was greater with CYP1B1 than with CYP1A1 as the catalyst. Overall, our novel findings indicate that G. biloba extract and the flavonol aglycones isorhamnetin, kaempferol, and quercetin preferentially inhibit the in vitro catalytic activity of human CYP1B1.

Aryl Hydrocarbon Hydroxylases↗

Ginkgo biloba extract: cognitive enhancer or antistress buffer.

Constituents extracted from the leaves of the Ginkgo biloba tree possess beneficial properties that may buffer the aging nervous system from deterioration due to oxidative stress. In the present investigation, a standardized extract of G. biloba (EGb 761) or an equal volume of the vehicle was administered (100 mg/kg/day) to senescent (20-month) C57BL/6 male mice for up to 82 consecutive days. Animals were tested twice in the Morris water maze (MWM) after 28 and 70 days of treatment. No differences were observed in acquisition or retention of performance on the water maze. Elevated-plus maze (EPM) trials were conducted prior to and subsequent to the chronic treatment regimen. Marked baseline differences in plus-maze performance were present in the first experiment. A second experiment used a matched-pairs design to minimize preexisting differences. Results supported the hypothesis that EGb 761 may serve as an antistress buffer, attenuating the increase in anxiety typically observed in animals after cold water exposure. Tissue samples from the hippocampus and cortex were analyzed by Western blot for the transcription factor cyclic-AMP response element binding (CREB) protein. EGb 761 had no significant effect on immunoreactivity to CREB from either the hippocampus or the cerebral cortex.

Aging↗

[Effects of extracts of ginkgo biloba (EGB) on levels of nitric oxide and apoptosis in the retina induced by glutamate in adult rabbits].

OBJECTIVE: To study the effects of extracts of ginkgo biloba (EGB) on the levels of nitric oxide (NO) and apoptosis in the retina induced by glutamate by intravitreal injection in adult rabbit. METHOD: The model of apoptosis in retina induced by glutamate by intravitreal injection was established in adult rabbit and EGB was retrobulbarly injected. The levels of NO were measured by spectrophotometer. Retina DNA fragmentation was determined by agarose gel electrophoresis. RESULT: The levels of NO in retina in experiment groups were significantly increased compared with controls, after treatment with high dosage of EGB, and levels of NO was decreased to normal. They were not decreased by injection of small dosage of EGB. DNA fragmentation of retina apoptosis was detected in experiment groups and small dosage EGB groups. CONCLUSION: The retina apoptosis was induced by glutamate by intravitreal injection in adult rabbit and was inhibited by EGB that may be through blocking the generation of NO free radicals.

Animals↗

[Relationship between endothelin-1 and ischemic brain damage after subarachnoid hemorrhage and protective effect of Ginkgo biloba extract].

OBJECTIVE: To investigate the role of endothelin-1 (ET-1) in development of ischemic brain damage after subarachnoid hemorrhage (SAH), and the protective effect of Ginkgo biloba extract (GBE). METHODS: Wistar rat noncraniotomy models of SAH were divided into SAH group and GBE treated group, the diameter of basilar artery (BA) and dynamic changes of regional cerebral blood flow (rCBF) and ET-1 content of intracranial plasma within 24 hours after SAH of both groups were determined. And pathological examination of CA1 region of hippocampus was performed 3 days later. RESULTS: rCBF decreased and ET-1 content increased obviously and retained in 24 hours after SAH. Spasm of BA occurred half an hour after SAH and neurons of hippocampus CA1 region was damaged severely. GBE could antagonize the above-mentioned pathological changes effectively. CONCLUSION: Increase of ET-1 is an important factor leading to ischemic brain damage after SAH. GBE exerts its protective effect by antagonizing pathological increase of ET-1.

Animals↗

Effect of Ginkgo biloba extract on carrageenan-induced acute local inflammation in gamma irradiated rats.

The effect of low dose whole-body gamma irradiation on inflammation and its possible modulation by Ginkgo biloba extract (GbE) was studied in the carrageenan-induced paw oedema model. Rats were subjected to two doses of gamma radiation (2 Gy or intermittent radiation at 2 Gy increment delivered daily up to cumulative dose of 4 Gy), 4 h before unilateral subplantar injection of carrageenan. The effect of GbE (25 or 50 mg/kg) administered subcutaneously daily for 3 days was also studied. Local oedema (days 1-3), the content of gamma glutamyl transpeptidase (GTT), malondialdehyde (MDA) and glutathione (GSH) in paw (72 h), were determined. In rats subjected to 4 Gy fraction, paw oedema was significantly reduced 1-4 h post-carrageenan injection (-26.2 to -16.2% vs control group). Moreover, at 24, 48 and 72 h after carrageenan, paw oedema was much reduced in the 2 Gy (-33.6, -46.4, -40%) or 4 Gy (-55, -56, -71.8%) irradiated groups compared to carrageenan unirradiated control. In addition, in irradiated rats, the carrageenan oedema was further significantly reduced by the administration of GbE, the effect of the agent being more marked in those irradiated with 2 Gy fraction. Changes in paw oedema were matched by a reduction in GGT and MDA paw tissue levels, while GSH content decreased in inflamed paw tissue 72 h post-treatment. These results indicate that exposure to 4 Gy fraction decreased the carrageenan-induced paw oedema and that the administration of GbE further lessened the severity of this inflammatory response in irradiated rats. The effects observed may be related in part to the inhibition of GGT activity and MDA production, and partly to augmentation of GSH content in the inflamed paw tissue.

Animals↗

Effects of an extract of Ginkgo biloba on the 3',5'-cyclic AMP phosphodiesterase activity of the brain of normal and triethyltin-intoxicated rats.

For clarification of the beneficial effects of the extract of Ginkgo biloba (EGB) on triethyltin (TET) toxicity in rats, the phosphodiesterase (PDE) activities of the cerebral tissue were measured under in vitro and ex vivo conditions. Under in vitro conditions, low concentrations of EGB (0.25-4.0 mg/L) activated the enzyme, whereas after higher concentrations (5-250 mg/L), dose-dependent inhibition of the enzyme activity was observed. In the lower concentration range, the extract also partially restored the high-affinity PDE activity (measured with 0.25 microM cyclic AMP) of the particulate fraction of the brain inhibited by TET in vitro. In contrast, the inhibitory influence of TET on the low-affinity PDE activity (measured with 50 microM cyclic AMP) of the particulate fraction was enhanced by the extract. Although treatment with a single large dose of EGB lowered the particulate PDE activities of the brain of normal rats, no effects of the extract could be detected in animals after repeated daily administrations of EGB during a 4-day period. Curative treatment of the TET-intoxicated rats with EGB during a 7-day period accelerated the recovery of the edematous state of the white matter caused by the intoxication and also normalized the lowered PDE activity of the particulate fraction of the edematous brain tissue. Furthermore, when preventively administered, EGB counteracted both the edema formation and the fall in PDE activity observed with treatment by TET alone. These observations strongly suggest that some beneficial effects of EGB might be due to its modulating influences on cellular cyclic AMP levels via activation of membrane-bound PDE.

3',5'-Cyclic-AMP Phosphodiesterases↗

The effects of Ginkgo biloba extract (LI 1370) supplementation and discontinuation on activities of daily living and mood in free living older volunteers.

The aim of the study was to investigate the effects of continuing treatment with Ginkgo biloba extract (GBE) 120 mg/day on the activities of daily living (ADLs) and mood in healthy older volunteers who had immediately previously participated in a survey of the effects of a 4 month treatment with the drug. Following a prior postal survey investigating the effects of 4 months supplementation with GBE on ADLs and various aspects of mood and sleep, 1570 volunteers continued onto a 6 month follow-up postal survey. Subjects selected their own treatment option for the follow-up survey, which effectively created four groups: a continuation group who received GBE in the initial 4 month study and during the 6 month follow-up (GBE-GBE), a discontinuation group who received GBE in the initial study but not during the follow-up (GBE-NT), a new treatment group who did not receive GBE in the initial 4 month study but who did receive GBE during the 6 month follow-up (NT-GBE), and a no treatment group who received no treatment in either survey (NT-NT). At the end of the 6 month follow-up period each subject completed a line analogue rating scale (LARS) and a self-rating activities of daily living scale (SR-ADL). There were signi fi cant differences in the mean overall LARS and SR-ADL scores between the four treatment combination groups at the end of the follow-up period. A factor analysis of the LARS revealed two factors, 'mood' and 'alertness'. When scores from each of the treatment groups were examined over the whole 10 month period it was evident that the ratings of overall competence in the SR-ADL and both factors of the LARS were diminished on cessation of treatment with GBE, and improved when GBE treatment was initiated. The magnitude of the improvements on all scales was related to the overall duration of GBE supplementation.Signi fi cant differences between the groups of subjects treated with GBE for different periods of time (4-10 months) suggests that the extract has a demonstrable effect in improving mood and the self-assessed performance of the tasks of everyday living.

Activities of Daily Living↗

Effect of an extract of Ginkgo biloba on rat brain energy metabolism in hypoxia.

The purpose of the present investigation was to determine brain energy metabolism under hypoxic conditions as influenced by an extract of Ginkgo biloba (EGB). Male Sprague-Dawley rats treated with EGB were exposed to hypobaric or hypoxic hypoxia, and at various time points during or after hypoxia the levels of high-energy phosphates and some substrates of glycolysis were measured in brain cortical tissue. Rats treated with EGB (100 mg/kg, intraperitoneally) survived hypobaric hypoxia for a much longer period than controls (e.g. controls: 3.9 +/- 1.8 min, EGB-treated: 23.6 +/- 10.5 min). The brain glucose level was elevated by EGB in most experimental series, and the lactate concentration was slightly lower than in control brains. The lowering of lactate/pyruvate ratio was due to the decreased level of lactate and to the enhanced concentration of pyruvate as well. When hypoxia was sufficiently severe the breakdown of high-energy phosphates was less pronounced in EGB-treated animals. After oral application of EGB for 14 days the rats survived hypobaric hypoxia for 25.7 +/- 2.5 min whereas controls survived for 11.5 +/- 5.1 min. However, brain energy metabolism was not significantly influenced by this oral treatment. It is suggested that changes in brain energy metabolism and blood flow may contribute to the protective effect of EGB against hypoxia.

Animals↗

The effects of Ginkgo biloba extract added to haloperidol on peripheral T cell subsets in drug-free schizophrenia: a double-blind, placebo-controlled trial.

OBJECTIVE: To investigate the effects of Ginkgo biloba extract (EGb) administration on T lymphocyte subsets and superoxide dismutase (SOD) levels in schizophrenia. METHODS: One hundred and nine schizophrenic inpatients were randomly assigned to 12 weeks of treatment with 360 mg/day of EGb plus a stable dose of 0.25 mg kg(-1) day(-1) of haloperidol and placebo plus the same dose of haloperidol using a double-blind design. Clinical efficacy was determined using the Brief Psychiatric Rating Scale (BPRS), Scale for Assessment of Positive Symptoms, and Scale for Assessment of Negative Symptoms. T lymphocytes (CD3+), T helper cells (CD4+), T suppressor cells (CD8+), and IL-2-secreting cells were measured using the alkaline phosphatase/antialkaline phosphatase technique; and SOD levels were measured by radioimmunometric assay at baseline and at posttreatment, as compared to 30 sex- and age-matched normal subjects. RESULTS: Patients demonstrated significantly lower CD3+, CD4+, and IL-2-secreting cells, together with CD4/CD8 ratio, and significantly higher blood SOD levels than did healthy controls at baseline. There was a significantly negative relationship between SOD and CD4+ cells in the schizophrenic group at baseline. After a 12-week treatment, CD3+, CD4+, and IL-2-secreting cells, together with CD4/CD8 ratio, showed a significant increase, but a significant decrease in SOD levels in the EGb group. There was only a significant increase in CD4+ cells but no change in SOD levels in the placebo group. There was a significant correlation between the change in CD4+ cells at posttreatment vs pretreatment and a reduction of BPRS total score in the whole patient group. CONCLUSIONS: EGb may improve the decreased peripheral immune functions in schizophrenia. The beneficial effects of EGb on the immune systems and the improvement of schizophrenic symptoms may be medicated through its antioxidant activity.

Adult↗

Induction and recovery of hepatic drug metabolizing enzymes in rats treated with Ginkgo biloba extract.

Herb-drug interactions, especially cytochrome P450 (CYP)-mediated interactions, cause an enhancement or attenuation in efficacy of co-administered drugs. In a previous study, we reported that repeated oral ingestion of Ginkgo biloba extract (GBE) markedly induced hepatic drug metabolizing enzymes in rats. In this study, we focused on the recovery of GBE-induced hepatic drug metabolizing enzymes after the discontinuation of GBE in rats. Feeding of a 0.5% GBE diet to rats for 1 week markedly increased liver weight, content of total CYP, activities of 6 CYP subtypes and glutathione S-transferase (GST). The content and activities of CYP enzymes were recovered to almost basal levels within 1 week after the discontinuation of GBE, while the activity of GST gradually decreased and recovered to the control level after 3 weeks. These results indicated that GBE-induced hepatic drug metabolizing enzymes in rats, especially CYPs, were rapidly recovered by discontinuation of GBE in rats even after excess treatment, and suggested that interactions of GBE with drugs could be avoided by discontinuation of GBE.

Animals↗

Ginkgo biloba extract inhibits tumor necrosis factor-alpha-induced reactive oxygen species generation, transcription factor activation, and cell adhesion molecule expression in human aortic endothelial cells.

OBJECTIVE: This study was conducted to examination whether Ginkgo biloba extract (GBE), a Chinese herb with antioxidant activity, could reduce cytokine-induced monocyte/human aortic endothelial cell (HAEC) interaction, a pivotal early event in atherogenesis. METHODS AND RESULTS: Pretreatment of HAECs with GBE (50 and 100 microg/mL for 18 hours) significantly suppressed cellular binding between the human monocytic cell line U937 and tumor necrosis factor-alpha (TNF-alpha)-stimulated HAECs by using in vitro binding assay (68.7% and 60.1% inhibitions, respectively). Cell enzyme-linked immunosorbent assay and immunoblot analysis showed that GBE (50 microg/mL for 18 hours) significantly attenuated TNF-alpha-induced cell surface and total protein expression of vascular cellular adhesion molecule-1 and intracellular adhesion molecule-1 (63.5% and 69.2%, respectively; P<0.05). However, pretreatment with probucol (5 micromol/L for 18 hours) reduced the expression of vascular cellular adhesion molecule-1 but not intracellular adhesion molecule-1. Preincubation of HAECs with GBE or probucol significantly reduced intracellular reactive oxygen species formation induced by TNF-alpha (76.8% and 68.2% inhibitions, respectively; P<0.05). Electrophoretic mobility shift assay demonstrated that both GBE and probucol inhibited transcription factor nuclear factor-kappaB activation in TNF-alpha-stimulated HAECs (55.2% and 65.6% inhibitions, respectively) but only GBE could inhibit the TNF-alpha-stimulated activator protein 1 activation (45.1% inhibition, P<0.05). CONCLUSIONS: GBE could reduce cytokine-stimulated endothelial adhesiveness by downregulating intracellular reactive oxygen species formation, nuclear factor-kappaB and activator protein 1 activation, and adhesion molecule expression in HAECs, supporting the notion that the natural compound Ginkgo biloba may have potential implications in clinical atherosclerosis disease.

Aorta↗

Stoichiometric and kinetic studies on Ginkgo biloba extract and related antioxidants.

Owing to increasing evidence showing the importance of lipid peroxidation in oxidative stress in vivo, the role and evaluation of antioxidants have received much attention. Ginkgo biloba extract (GBE), well-known as an efficient drug against diseases induced by free radicals, has been suggested to exert its effect by antioxidant action. A method was established to determine the activity of GBE as a hydrogen donor by stoichiometric and kinetic studies, and GBE was compared with several other antioxidants such as alpha-tocopherol, propyl gallate, and two kinds of flavonoids which are found in GBE, quercetin, and kaempferol. It was found that there were 6.62 x 10(19) active hydrogens in 1 g of GBE. Stoichiometric studies showed that one molecule of alpha-tocopherol reacted with one molecule of galvinoxyl radical. For quercetin, kaempferol and propyl gallate, the experimental stoichiometric numbers were 4.0, 1.9, and 3.1, respectively. The rates of reaction of antioxidants with galvinoxyl in ethanol were determined spectrophotometrically, using a stopped-flow technique. The second-order rate constant, k2, obtained at 25 degrees C was 0.13 (g/L)(-1)s(-1) for GBE and 5.9 x 10(3), 2.1 x 10(3), 1.2 x 10(4), and 2.4 x 10(3) M(-1)s(-1) for quercetin, kaempferol, propyl gallate, and alpha-tocopherol, respectively. The second-order rate constant, k2', on the molar basis of active hydroxyl groups in the tested substances obtained at 25 degrees C decreased in the order of propyl gallate > alpha-tocopherol > quercetin > GBE approximately kaempferol. This is the first study on GBE as an antioxidant which reports both stoichiometric and kinetic results.

Antioxidants↗

Sequential uniform designs for fingerprints development of Ginkgo biloba extracts by capillary electrophoresis.

A sequential procedure for the method of development of fingerprints based on a uniform design approach has been described in this paper. The sequential uniform design is used to reach the global optimum for a separation. The procedure is illustrated through developing the fingerprint of a Ginkgo biloba extract by capillary electrophoresis (CE) with diode array detection (DAD). The local overlap index (LOVI) is proposed as a criterion to evaluate the separation quality of two-dimensional hyphenated data in the optimization process. The successful application of the described techniques, including a sequential procedure, the CE-DAD hyphenated method and the LOVI criterion, shows their practicality in the fingerprint development of herbal medicines. However, some critical remarks on their use can also be made.

Buffers↗

Expression of pro-inflammatory and anti-inflammatory cytokines in brain of atherosclerotic rats and effects of Ginkgo biloba extract.

AIM: To study the protein and mRNA expressions of pro-inflammatory and anti-inflammatory cytokines in the brain of rats with atherosclerosis (AS) and the effects of Ginkgo biloba extract (GbE) on expressions of cytokines. METHODS: The experimental model of AS in rats were established by intraperitioneal injection of vitamin D3 with high fat/cholesterol diet. GbE 100 mg/kg was administered to rats by ig. After 8 weeks, the expressions of IL-1beta, TNF-alpha, IL-10, and IL-10R in the brain tissues of AS rats were detected by enzyme-linked immunosorbant assay, immunohistochemistry, Western blotting, and reverse transcriptase polymerase chain reaction. RESULTS: The protein and mRNA expressions of IL-1beta, TNF-alpha, and IL-10 in the brains were markedly higher in AS groups than that in control groups (6.11+/-0.15, 1.55+/-0.14, 0.54+/-0.04 ng/g wet weight vs 0.80+/-0.14, 0.33+/-0.09, and 0.33+/-0.02 ng/g wet weight, respectively). The protein and mRNA expressions of IL-1beta and TNF-alpha in the brains were markedly lower in GbE groups (3.82+/-0.54, 0.95+/-0.08 ng/g wet weight) than that in AS groups, the protein and mRNA expressions of IL-10 and IL-10R in the brains were markedly higher in GbE groups (0.85+/-0.06 ng/g wet weight) than that in AS groups. CONCLUSION: GbE inhibited production of pro-inflammatory cytokines IL-1beta and TNF-alpha, but up-regulated the production of anti-inflammatory cytokines, IL-10 and IL-10R in brain, which might be related with its anti-AS actions.

Animals↗

LC-MS characterization of terpene lactones in plasma of experimental animals treated with Ginkgo biloba extracts Correlation with pharmacological activity.

Liquid chromatography/atmospheric pressure chemical ionization ion trap mass spectrometry (LC/APCI-ITMS) was applied to determine the concentration of terpene lactone in plasma of guinea pigs after chronic administration of Ginkgo biloba extract enriched in ginkgoterpenes in free form (IDN 5380) or complexed with soy phosphlipids (IDN 5381). Oral treatment of the animals with ginkgoterpenes resulted to inhibit the bronchoconstriction (ITP) and the concomitant increase of the levels of thromboxane B2 (TXB2) in the circulation caused by histamine (HIST) and platelet activating factor (PAF) in normal guinea pigs or by ovalbumin (OA) in actively sensitized guinea pigs. To compare the protective activities of G. biloba forms (IDN 5380 and IDN 5381), ED50 and dose ratio (DR) values for both parameters (ITP and TXB2) were evaluated. The phytosomic form (IDN 5381) significantly reduced (two- to four-fold as compared to free form, P < 0.001) the HIST, PAF or OA-induced airway changes and TXB2 release. In addition it has been observed that the absence of ginkgolide C (GC) in plasma samples (in human and animals) was due to its rapid methylation.

Adult↗