Effect of Ginkgo biloba extract-761 on motor functions in permanent middle cerebral artery occlusion rats

doi:10.1016/j.phymed.2018.05.003

Phytomedicine

Volume 48, 15 September 2018, Pages 94-103

https://doi.org/10.1016/j.phymed.2018.05.003 Get rights and content

Abstract

Background

Ginkgo biloba extract (EGb-761) has been in use to treat variety of ailments including memory loss and emotional disorders usually experienced after ischemic stroke. However, data regarding its protective role in stroke associated motor dysfunction is scarce.

Purpose

The present work was designed to investigate the long-term effects of EGb-761 on the motor dysfunctions associated with permanent middle cerebral artery occlusion (pMCAO) in rats.

Study Design/Methods

Focal ischemic stroke was induced in male Sprague–Dawley rats by pMCAO. These rats were orally administered with EGb-761 (25, 50, 100 mg/kg) and positive control butylphthalide (50 mg/kg) for up to 28 consecutive days. The motor function was evaluated by assessing neurological scores, rotarod performance and gait analysis after 7, 14, 21 and 28 days. After 28 days, the histological examination of in frontal cortex and hippocampus was also carried out.

Results

EGb-761 treatment significantly improved motor function with better outcome in coordination and gait impairment rats. EGb-761 (25, 50, 100 mg/kg) treatment for 28 days significantly decreased the neurological scores. After 28 days of treatment EGb-761 (50 and 100 mg/kg) significantly increased the latency in rotarod test, walk speed, and the body rotation, whereas, decreased the stride time and the left posterior swing length in gait were observed. EGb-761 (50, 100 mg/kg). EGb-761 (50, 100 mg/kg) significantly improved the pathological changes related to pMCAO.

Conclusions

EGb 761 could improve motor function especially gait impairments among pMCAO rat model related to the decreased neuronal damage. Therefore, it might be the potential to be explored further as an effective therapeutic drug to treat post stroke motor dysfunctions.

Graphical abstract

Introduction

Stroke is considered as one of the critical devastating medical condition with high rates of mortality and neurological disorders. Regulatory steps taken by different countries overcome its related mortalities but post stroke complications still remain a major concern (Minino et al., 2010). Majority of stroke survivors suffered from neurological deficit (Sun et al., 2013). and it has been estimated that about 15 million peoples encounter stroke every year. More than 50% disabled stroke survivor present a great economic burden to society and families as they require special attention for routine activities. About one third of stroke survivors experience abnormalities related to motor functions such as gait impairments (Ejupi et al., 2014). Gait impairments and imbalance has been observed in stroke individuals. Functional impairments of upper limbs in walking are the most common reported deficits among post stroke disabilities.

EGb 761 is popular and standardized extract of Ginkgo biloba leaves which belongs to division Gingkophyta and known for its nutritional and medicinal value (Lang et al., 2012). It was reported that Ginkgo biloba extract could cure neurodegenerative disorders such as ischemic stroke, epilepsy, Alzheimer disease (Lautenschlager et al., 2012). Meanwhile, it could improve functional outcome related to anxiety and mood disorders in ischemic stroke. Preliminary clinical and pre-clinical trials demonstrated that EGb761 successfully modulated dementia and emotional disorders after ischemic stroke. However, to our best of knowledge, there is no well-established evidence for the protective of EGb761 against permanent middle carotid artery (pMCAO) induced motor dysfunction is scarce.

The pMCAO is acknowledged model which is widely used in preclinical research for drug screening, drug development and it related mechanism of action (Strom et al., 2013). It is also used to study neural functional recovery in ischemic stroke. So far, the effect of Gingko biloba was mainly evaluated for reducing ischemic brain damage, delayed hippocampal neuronal death and apoptosis, cognitive deficits and emotional disorders after ischemic stroke (Clostre, 1999, Sun et al., 2013 and Tulsulkar et al., 2016). Thus, this study was designed to evaluate the protective effect of Gingko biloba extracts on motor function among pMCAO rats model.

Section snippets

Materials

EGb-761 was provided from Hunan Fang sheng Pharmaceutical Co. Ltd, China. It mainly contains ginkgo flavonoides 25.5%, terpene lactonesb 7.3%. Butylphthalide was provided by Shijiazhuang Pharmaceutical Co. Ltd, China. Monofilament coated with poly L lysine was provided from Beijing Cinontech Co. Ltd, China.

Standardization of Ginkgo biloba extract. The chemical fingerprinting of the extracts was analyzed. The chemicals used for the identification and quantification of compounds in the Ginkgo

Neurological scores

Neurological score was evaluated by using a five-point scale on day 1st, 7th, 14th, 21th and 28th day after pMCAO as mentioned experimental design section.

Rotarod performance test

The impairment of coordinated motor function was analyzed by using computer aided accelerated rotarod (Chinese Academy of Medical Science, Beijing, China) as described previously with few modification (Yousuf et al., 2011). Before surgery, rats were trained for 3 consecutive days for five sessions a day. Each session consisted of 3 min with a

Effects of EGb761 on neurological scores

Gingko biloba extract at 25, 50 and 100 mg/kg and positive control drug butylphthalide (50 mg/kg) significantly reverse the neurological deficit after 7 to 28 days of treatment as compared to model group (p < 0.05; Fig. 3). Whereas, sham and control group did not show any neurological deficits throughout the experiments.

Effects of EGb761 on rotarod test

The rotarod test was conducted following the gait test. Fig. 4 depicts the latency time of rats to perform on rotarod. It demonstrated that control and sham rats were similar

Discussion

Ischemic stroke is characterized by high morbidity, disability and mortality rate. Approximately 90% of patients suffer permanent neurological deficits such as cognitive impairment and mood disorders after stroke (Jorgensen et al., 1999). In particular, post stroke motor impairments are common and are characterized by a slowness or paucity of movement and hemiplegia (Hendricks et al., 2002). The behavioral and pathophysiology of pMCAO induced stroke closely matches clinical symptoms in stroke

Conclusions

In conclusion, EGb-761 effectively improved functional motor deficits which might be related to the decreased neuronal damage in cerebral ischemic rats. Therefore, EGb-761 might be considered as a potential drug for alleviating the post ischemic stroke related complications to improve motor dysfunctions.

Conflict of interest

We wish to confirm that there are no known conflicts of interest associated with this publication and there has been no significant financial support for this work that could have influenced its outcome.

Acknowledgments

The present study was supported by Department of Science & Technology, Hunan province, China (No. 2015DK3010, 2016TP1026), as well as CAMS Initiative for Innovative Medicine, China (2016-I2M-2-006).

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Ginkgo biloba: An updated review on pharmacological, ethnobotanical, and phytochemical studies
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Ginkgo biloba L. (Ginkgoaceae), known as a "living fossil,"has a long history of medicinal use. Both its leaves and nutshave been widely prescribed for the treatment of various ailments, including pulmonary diseases, alcohol abuse, bladder inflammation, heart and lung dysfunction, and skin infections. Currently, there are different forms of G. biloba available in the market, including extracts and isolated compounds such as terpenoids, flavonoids, bioflavonoids, and organic acids. Despite the significant medicinal importance of this plant, there is a lack of updated reviews encompassing its pharmacology and other relevant aspects. Therefore, the objective of this review is to collect and analyze existing literature on the phytochemical, ethnobotanical, and pharmacological aspects of G. biloba.
Electronic databases including PubMed, Embase, Scopus, and Google Scholar were searched for keywords “Ginkgo biloba” or “ginkgo” in combination with “phytochemistry”, “ethnobotany”, “pharmacology”, “nervous system”, and “cardiovascular”, etc. to collect the relevant articles. There was no language restriction.
Retrieved studies showed that the extracts and isolated compounds from G. bilobahave beneficial activities in cerebrovascular, nervous, cardiovascular, endocrine, muscular and skeletal, renal, respiratory, digestive, and immunity systems.
Numerous in vitro and in vivo studieshave been conducted to investigate the therapeutic potential of G. biloba in the treatment and improvement of various diseases. Additionally, there is a substantial body of clinical research on G. biloba preparations and isolated compounds. However, further investigations, particularly human-controlled trials, are recommended to determine the efficacy and safety of G. biloba. These trials would provide more robust evidence regarding the potential benefits and risks associated with the use of G. biloba in clinical settings.
Comparative efficacy and safety of ginkgo biloba related agents in patients with acute ischemic stroke: A Bayesian network meta-analysis of randomized controlled trials
2023, Pharmacological Research - Modern Chinese Medicine
Acute Ischemic stroke (AIS) has been a leading cause of long-term disability and even death, accounting for significant economic costs and leading to heavy care and financial burdens for patients and their families. Ginkgo biloba extract is highly valued for its efficacy and broad implementation into clinical practice. Therefore, a Bayesian network meta-analysis (BNMA) was conducted to determine the differences in efficacy and safety among various ginkgo biloba-related agents and to prescribe the optimal medication for decision-making.
Inclusion and exclusion eligibility criteria were determined in advance. From the inception of the literature until December 2022, a systematic literature search was conducted in seven electronic databases. An assessment of the methodological quality of the included studies was carried out according to the Cochrane Collaborations tool. A previous registration (PROSPERO CRD42022319301) of this review protocol was undertaken.
There were 94 eligible studies included in the NMA involving 10,071 patients, with the experimental control group of 5,063 and the control group of 5,008, involving five ginkgo biloba-related agents included in the NMA. Findings of the studies revealed that supplementation with ginkgo biloba-related agents significantly improved the clinical effective rate, functional independence, neurological function impairments and activities of daily living function compared with the conventional western medicine (WM) group in a relatively safe manner. Comprehensive therapeutic efficacy analysis of neurological function impairments and activities of daily living function showed that GI+WM and GDLM+WM manifest at the first grade among these five therapeutic strategies.
This Bayesian network meta-analysis results showed that the combination of ginkgo biloba-related agents and WM was more effective for AIS patients than WM alone. GI+WM and GDLM+WM stand out for the comprehensive therapeutic efficacy of patients with stroke through comparisons among various therapeutic approaches. Moreover, SXN+WM, GK+WM and GDLM+WM might be among the most effective in improving the functional efficacy outcome, while GI+WM might be among the least effective. GI+WM, GDLM+WM, SXN+WM and GK+WM might be among the most effective in improving neurofunctional deficits, while GNT+WM might be among the least effective. In terms of improving activities of daily living function, five ginkgo biloba-related agents might be among the most effective. However, well-designed researches with standardized functional outcome measures for AIS, further adequately powered, blinding and suitable comparators are still required to contribute to the current clinical practice.
Rotator cuff muscle degeneration in a mouse model of glenohumeral osteoarthritis induced by monoiodoacetic acid
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Osteoarthritis (OA) is a disease of joint degeneration and impaired function. Muscle atrophy, fatty infiltration, and fibrosis are degenerative features of muscle injury and predict poor outcomes in some degenerative and exercise-related injuries. Patients with glenohumeral joint OA usually have rotator cuff muscle degeneration, even though the rotator cuff is intact. However, the mechanism and correlation between OA and degeneration of muscles around joints are still unknown.
Forty-five 12-month-old C57BL/6J mice received a single injection of monoiodoacetic acid into the right glenohumeral joint. The sham group was injected with saline on the same day in the right glenohumeral joint. Three and 6 weeks after the operation, gait analysis was conducted to evaluate the function of the forelimb. Then, the shoulder joint and supraspinatus muscle were collected for histologic staining, reverse transcription quantitative polymerase chain reaction, and biomechanics test. Correlations between fat area fraction in muscle, percentage wet muscle weight change or Osteoarthritis Research Society International score, and gait analysis/muscle mechanics tests were assessed using Pearson's correlation coefficient or Spearman's correlation coefficient.
Compared with the sham group, the monoiodoacetic acid group developed significant glenohumeral joint OA and the supraspinatus muscle developed significant fatty infiltration and muscle atrophy. Shoulder function correlated with glenohumeral joint OA/rotator cuff muscle severity, weight loss, and fatty infiltration.
In mice, glenohumeral joint OA can lead to rotator cuff degeneration and inferior limb function. The small animal model could be a powerful tool to further study the potential mechanisms between glenohumeral OA and rotator cuff muscle degeneration.
Ginkgo biloba extract (EGb 761) mitigates methotrexate-induced testicular insult in rats: Targeting oxidative stress, energy deficit and spermatogenesis
2021, Biomedicine and Pharmacotherapy
Citation Excerpt :
A total of 40 rats were allocated into five equal groups comprising control group in which animals received the vehicles; methotrexate rats group (MTX) in which rats were injected intraperitoneally with methotrexate (MTX; 0.5 mg/kg body weight in DMSO) twice a week for four successive weeks [21]. EGb-761 groups; in which rats are treated orally with EGb-761 (25, 50 or 100 mg/Kg /day) for four weeks [22] in addition to intraperitoneally administration of MTX (0.5 mg/kg /twice a week). Twenty-four hours after surgery, blood samples were taken through the retro-orbital sinus under Ketamine–Xylazine anesthesia (50 mg/kg-5 mg/kg, i.p) [23] and Sera samples were collected and stored at 20 °C until needed.
Methotrexate (MTX) is commonly used as a therapeutic agent in the treatment of malignancies and autoimmune disorders. Risk of subsequent infertility following MTX administration has been reported as a significant side effect due to testicular toxicity. The aim of the study was to evaluate the modulatory effects of Ginkgo biloba (standardized extract, EGb 761) on MTX-induced testicular oxidative stress, energy deficits and spermatogenic status in rats. All groups received intraperitoneal injection of MTX (0.5 mg/kg) twice weekly for four weeks except the control group that received the corresponding vehicles. Other groups received oral EGb761, at doses 25, 50 or 100 mg/kg/day, for four weeks, concurrently with MTX. Blood and semen sampling followed by testis dissection were performed 24 h after last EGb 761 treatment. Semen was examined for sperm progressive motility, percent of normal spermatozoa and sperm cell concentration as well as seminal plasma essential and non-essential amino acids. Serum LH, FSH and testosterone were detected as well as testicular MDA, GSH, GSSG, TNF-α, IL-1β, IL-6, NF-κB and the nuclear, cytoplasmic and mRNA expression levels of Nrf-2 besides the testicular cell energy; AMP, ADP and ATP. Histopathological studies of interstitial fibrosis and the severity of germ cell degeneration were also conducted. MTX induced significant decline in sperm quality along with decreased essential and non-essential amino acids in seminal plasma. MTX reduced serum FSH, LH and testosterone as well as testicular ATP, GSH and Nrf2, while increased levels of testicular ADP, AMP, MDA, GSSG and TNF-α. Results were confirmed by prominent interstitial fibrosis and severe germ cell degeneration in MTX group. Concurrent treatment with EGb 761 alleviated MTX-induced testicular insult evidenced by amelioration of oxidative stress biomarkers, energy functions, seminal sperms abnormalities and spermatogenesis status. The present study suggests a beneficial role of EGb 761 in MTX-induced testicular injury and subsequent distortion of spermatogenesis.
Xiao-Xu-Ming decoction prevented hemorrhagic transformation induced by acute hyperglycemia through inhibiting AGE-RAGE-mediated neuroinflammation
2021, Pharmacological Research
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Some studies have been shown that EGB761 plays neuroprotective effects against various cardiovascular and neurological disorders, such as Alzheimer’s disease, stroke and depression [31]. In stroke, EGB761 can decrease infarct volume, improve motor function and cognitive function [32,33], and promote neurovascular restoration and axonal regeneration [34]. Therefore, EGB761 was used as positive control in this study.
Stroke is one of the leading causes of death worldwide. Hemorrhagic transformation (HT) is a common serious complication of ischemic stroke (IS) and is related to poor prognosis. Hyperglycemia after stroke is associated with the occurrence of HT and seriously affects the clinical treatment of stroke. Our previous experiments demonstrated that the Xiao-Xu-Ming decoction effective components group (XXMD), which is a Chinese medicine formula reconstituted by active ingredients, has multiple pharmacological effects in the treatment of IS. However, the effects of XXMD on HT after IS remain unclear. Thus, we investigated the preventive effects of XXMD on hyperglycemia-induced HT and further explored the underlying mechanism. Acute hyperglycemia combined with the electrocoagulation cerebral ischemia model was used to establish the HT model. XXMD (37.5, 75, 150 mg/kg/d) was given by gavage for 5 days. Network pharmacology was used to predict potential targets and pathways of XXMD in HT occurrence, and further studies confirmed the related targets. The results showed that hyperglycemia aggravated neurological deficits and blood-brain barrier (BBB) disruption, leading to intracerebral hemorrhage. Pretreatment with XXMD improved neurological function and BBB integrity and inhibited HT occurrence. Network pharmacology revealed that AGE-RAGE-mediated neuroinflammation may be associated with hyperglycemia-induced HT. Further studies confirmed that hyperglycemia activated the AGE-RAGE signaling pathway, increased the expression of HMGB1, TLR4 and p-p65, and induced the release of inflammatory factors and neutrophil infiltration, leading to HT. XXMD could inhibit AGE-RAGE-mediated neuroinflammation. These findings indicated that pretreatment with XXMD alleviated hyperglycemia-induced HT, which may be associated with the inhibition of AGE-RAGE-mediated neuroinflammation. Therefore, XXMD may be a potential therapeutic drug for HT.
Shuxuening injection facilitates neurofunctional recovery via down-regulation of G-CSF-mediated granulocyte adhesion and diapedesis pathway in a subacute stroke mouse model
2020, Biomedicine and Pharmacotherapy
Citation Excerpt :
It is reported that Ginkgo biloba extract can treat neurodegenerative diseases [32], such as ischemic stroke [33,34], memory loss [35,36], epilepsy [37,38] and Alzheimer's disease [39–41]. It can also improve cognitive and neurological dysfunction after stroke [42,43]. Shuxuening Injection (SXNI) is a GBE approved by the Sino Food and Drug Administration [44].
Post-stroke neural damage is a serious health concern which does not yet have an effective treatment. We have shown previously that Shuxuening injection (SXNI), a Ginkgo biloba extract-based natural medicine, protects brain after an acute ischemic stroke, but its efficacy for post-stroke recovery is not known. This study was to investigate whether SXNI can improve the prognosis of stroke at a subacute phase. Mice with cerebral ischemia-reperfusion injury (CIRI) were established by middle cerebral artery occlusion (MCAO), and drugs or saline were injected by the tail vein every 12 h after reperfusion. The therapeutic effect of SXNI was evaluated by survival rate, modified neurologic severity scores (mNSS), open-field test, locomotive gait patterns, cerebral infarction volume, brain edema and histopathological changes. Subsequently, a combined method of RNA-seq and Ingenuity® Pathway Analysis (IPA) was performed to identify key targets and pathways of SXNI facilitating the prognosis of stroke in mouse brain. The results of the transcriptome analysis were verified by real time reverse transcription-polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA), western blot (WB) and immunohistochemistry (IHC). The experimental results showed that in the new subacute stroke model, SXNI markedly improves the survival rate, neurological and motor functions and histopathological changes, and significantly reduces cerebral infarction and edema volume. RNA-seq analysis of subacute stroke mice with or without SXNI (3 mL/kg) indicated 963 differentially expressed genes (DEGs) with a fold change ≥ 1.5 and a P-value ≤ 0.01. IPA analysis of DEGs showed that granulocyte adhesion and diapedesis ranked first in the pathway ranking, and the most critical gene regulated by SXNI was G-csf. Simultaneously, RT-PCR, ELISA, WB and IHC results demonstrated that SXNI not only obviously reduced the mRNA expression levels of key genes G-csf, Sele and Mac-1 in this pathway, but also significantly decreased the protein expression levels of G-CSF in serum and E-selectin and MAC-1 in brain tissues. In summary, our research suggested that SXNI can exert a remarkable neurofunctional therapeutic effect on stroke mice via down-regulating G-CSF to inhibit granulocyte adhesion and diapedesis. This study provides experimental evidence that SXNI may fulfill the need for stroke medicine targeting specifically at the recovery stage.

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Original ArticleEffect of Ginkgo biloba extract-761 on motor functions in permanent middle cerebral artery occlusion rats

Abstract

Background

Purpose

Study Design/Methods

Results

Conclusions

Graphical abstract

Introduction

Section snippets

Materials

Neurological scores

Rotarod performance test

Effects of EGb761 on neurological scores

Effects of EGb761 on rotarod test

Discussion

Conclusions

Conflict of interest

Acknowledgments

Arch. Phys. Med. Rehabil.

Neurochem. Int.

Behav. Brain Res.

Arch. Phys. Med. Rehabil.

Lancet Neurol.

Behav. Brain

Lancet

Rat middle cerebral artery occlusion: evaluation of the model and development of a neurologic examination

Stroke

Music therapy for acquired brain injury

Cochrane Database Syst. Rev.

The need for speed in rodent locomotion analyses

Anat. Rec.: Adv. Integr. Anat. Evol. Biol.

Ginkgo biloba extract (EGb 761). State of knowledge in the dawn of the year 2000

Ann. Pharm. F

Preclinical assessment of CNS drug action using eye movements in mice

J. Clin. Invest.

Weaver, Effects of polyethyl eneglycol and magnesium sulfate administration on clinically relevant neurological outcomes after spinal cord injury in the rat

J. Neurosci.

New methods for fall risk prediction

Curr. Opin. Clin. Nutr. Metab. Care

Long-term functional outcome following transient middle cerebral artery occlusion in the rat: correlation between brain damage and behavioral impairment

Behav. Neurosci.

Movement disorders after stroke

Age Ageing

Stroke neurologic and functional recovery the Copenhagen stroke study

Phys. Med. Rehabil. Clin. North Am.

Original Article
Effect of Ginkgo biloba extract-761 on motor functions in permanent middle cerebral artery occlusion rats