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OBJECTIVE To investigate the inhibitory effects of Ginkgo biloba extract （GBE） on renal inflammation in diabetic nephropathy （DN） model mice， and its potential mechanism. METHODS KK/Ay mice were fed with high fat and high sugar to induce DN model. They were divided into model group， positive control group [metformin 200 mg/（kg·d）]， GBE low-dose and high-dose groups [100， 200 mg/（kg·d）]， with 6 mice in each group. Six C57BL/6J mice were fed with a regular diet as the control group. Administration groups were given relevant liquid intragastrically， control group and model group were given constant volume of normal saline intragastrically， once a day， for 8 consecutive weeks. The body weight， fasting blood glucose， 24-hour food intake， 24-hour urine output， monocyte chemoattractant protein-1 （MCP-1）， interleukin-12 （IL-12）， IL-10， advanced glycation end products （AGEs）， blood urea nitrogen （BUN） and serum creatinine （Scr） of mice were measured， and the ratio of bilateral kidneys to body weight was also calculated. The pathological injury and fibrotic changes of the renal cortex were observed， and the expressions of macrophage polarization marker proteins [type M1： inducible nitric oxide synthase （iNOS）； type M2： arginase-1 （Arg-1）] and AGEs-the receptor of advanced glycation end products （RAGE）/Ras homolog gene pharm_chenjing@163.com family member A （RhoA）/Rho-associated coiled-coil forming protein kinase （ROCK） signaling pathway-related proteins were determined in renal cortex. RESULTS Compared with the model group， the symptoms such as renal cortical hyperplasia， vacuoles， infiltration of inflammatory cells， and renal cortical fibrosis had been improved in GBE low-dose and high-dose groups； body weight， serum level of IL-10， the expression of Arg-1 in the renal cortex were significantly higher than model group （P＜ 0.01）； fasting blood glucose， 24-hour food intake， 24-hour urine output， serum levels of MCP-1， IL-12， BUN， Scr and AGEs， the ratio of bilateral kidneys to body weight， renal injury score， the proportion of renal interstitial fibrosis， the protein expressions of iNOS， RAGE， RhoA and ROCK1 （except for GBE low-dose group） in renal cortex were significantly lower than model group （P＜0.01）. CONCLUSIONS GBE could improve kidney damage and alleviate inflammatory response in DN model mice， the mechanism of which may be related to inhibiting the AGEs-RAGE/RhoA/ROCK signaling pathway and regulating macrophage polarization.
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Background Fluorine accumulates in the brain tissue after long-term excessive intake and subsequently cause nerve damage and decline of learning and memory ability. Receptor of advanced glycation end-products (RAGE)/p38 mitogen-activated protein kinase (p38MAPK)/nuclear factor kappa-B (NF-κB) signaling pathway is considered to be involved in the associated mechanism. Objective To study the changes of RAGE/ p38MAPK/ NF-κB signaling pathway in rats with subchronic fluorosis, and to explore the protective effects of extract of Ginkgo biloba 761 (EGb761) and RAGE antagonist (FPS-ZM1) on neuromemory ability. Methods Ninety male clean SD rats were divided into 9 groups with 10 rats in each group. The modeling period was 6 months. Control group (C group): free drinking tap water (fluoride content <0.5 mg·L−1), low- and high-dose fluoride groups (LF group, HF group): free drinking tap water with 10 or 50 mg·L−1 fluoride; intervention group of Ginkgo biloba extract (CE, LFE, and HFE groups): on the basis of the C group, LF group, and HF group, 100 mg·kg−1·d−1 EGb761 was given daily via intragastric administration; FPS-ZM1 intervention groups (CF, LFF, and HFF groups): 7 d before the end of modeling, 1 mg·kg−1·d−1 FPS-ZM1 was injected intraperitoneally daily on the basis of the C group, LF group, and HF group. The contents of fluoride in brain and blood of each group were detected. The learning and memory ability was tested by water maze experiment. The histopathologic changes of the hippocampus were detected by Nissl staining. The protein expression levels of RAGE and its ligand high mobility group protein B1 (HMGB1), NF-κB, p38MAPK, phospho-p38MAPK (p-p38MAPK), interleukin-6 (IL-6), and tumour necrosis factor-α (TNF-α) in brain tissue were detected by Western blotting. The mRNA expression levels of RAGE, HMGB1, and p38MAPK were detected by quantitative real-time PCR. Results Compared with the C group, the contents of blood fluoride and brain fluoride in the LF and the HF groups were increased (P<0.05). The results of the water maze experiment showed that, compared with the C group, the escape latency time of the LF group and the HF group was longer and the crossing times were reduced; compared with the HF group, the escape latency time of the HFE group and the HFF group was shortened, and the crossing times were increased (P<0.05). The Nissl staining results showed that the number of Nissl body in the HF group decreased compared with the C group; compared with the HF group, the number of Nissl body in the HFE group and the HFF group increased. The Western blotting results showed that compared with the relative protein expression levels of RAGE, HMGB1, NF-κB, p38MAPK, p-p38MAPK, IL-6, and TNF-α in the C group , the levels of above indicators in the HF group and the levels of RAGE, HMGB1, NF-κB, p-p38MAPK, and IL-6 in the LF group were up-regulated (P<0.05); compared with the HF group, the levels of above indicators in the HFE group and the HFF group were all down-regulated (P<0.05); compared with the relative protein expression levels of RAGE and HMGB1 in the LF group, the levels in the LFE group and the LFF group were all down-regulated (P<0.05). The quantitative real-time PCR results showed that compared with the C group, the mRNA expression levels of RAGE and HMGB1 in the LF group and the HF group were up-regulated; compared with the LF group, the mRNA expression levels of RAGE in the LFE group and the LFF group were down-regulated ; compared with the HF group, the mRNA expression levels of RAGE and HMGB1 in the HFE group and the HFF group were down-regulated (P<0.05). Conclusion The central nervous system injury caused by subchronic fluorosis may be related to the activation of RAGE/p38-MAPK/NF-κB signaling pathway, which can impair the learning and memory ability of rats, while EGb761 and FPS-ZM1 may have certain protective effects on the nerve injury.
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Ginkgo biloba Extract( GBE50) Dispersible Tablets is a new standardized prescription,which is widely used in the treatment of ischemic cardiovascular and cerebrovascular diseases. However,there are still many problems in its clinical application.Rational and safe use of GBE50 Dispersible Tablets is pivotal to the medication safety and clinical prognosis of patients. This consensus has been jointly formulated by clinical experts of traditional Chinese medicine and western medicine in cardiovascular and cerebrovascular diseases and followed the Manual for the Clinical Experts Consensus of Chinese Patent Medicine published by the China Association of Chinese Medicine. The present study identified clinical problems based on clinical investigation,searched the research papers according to PICO clinical problems,carried out evidence evaluation,classification,and recommendation by GRADE system,and reached the expert consensus with nominal group technique. The consensus combines evidence with expert experience. Sufficient evidence of clinical problems corresponds to " recommendations",while insufficient evidence to " suggestions". Safety issues of GBE50 Dispersible Tablets,such as indications,usage and dosage,and medication for special populations,are defined to improve clinical efficacy,promote rational medication,and reduce drug risks. This consensus needs to be revised based on emerging clinical issues and evidencebased updates in practical applications in the future.
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Humans , Cerebrovascular Disorders/drug therapy , Consensus , Drugs, Chinese Herbal/therapeutic use , Medicine, Chinese Traditional , TabletsABSTRACT
The present study aimed to systematically evaluate the efficacy and safety of Ginkgo biloba extract 50(GBE50) in the treatment of ischemic stroke. The databases including CNKI, Wanfang, VIP, SinoMed, PubMed, EMbase, and Cochrane Library were searched for randomized controlled trial(RCT) of GBE50 for the treatment of ischemic stroke reported between database inception and May 2020. The methodological quality of the included RCTs was evaluated via the Cochrane risk of bias tool. The RevMan 5.4 was used for Meta-analysis. Sixteen RCTs were included, involving 1 615 patients with acute ischemic stroke. Most of the included RCTs reported the methods of random sequence generation, but only two performed the concealment of random sequence. All RCTs failed in blinding. Two RCTs reported the information of cases lost to follow-up and drop-outs. Since the number was small, the baselines of groups remained balanced. All RCTs reported key outcomes of ischemic stroke, which made selective reporting bias in a low risk. Meta-analysis results revealed that GBE50 combined with routine therapies could effectively lower the score of the National Institutes of Health stroke scale(NIHSS) and restore cognitive function and daily activity in ischemic stroke patients. Compared with routine therapies, the combination is advantageous in treating patients with ischemic stroke. However, high-quality multicenter RCTs with large sample sizes are still required for verification.
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Humans , Brain Ischemia/drug therapy , Ginkgo biloba , Ischemic Stroke , Multicenter Studies as Topic , Plant Extracts , Stroke/drug therapyABSTRACT
This study aimed to assess the effects of Ginkgo Biloba Extract and Troxerutin on the hippocampus of induced diabetes mellitus in adult albino rats using histological methods.50 adult male albino rats were divided into three groups; Group I (Control); Group II (diabetic): subdivided into Subgroup IIa (T1DM)), Subgroup IIb (T1DM+GBE), Subgroup IIc (T1DM+ troxerutin); Group III: subdivided into Subgroup IIIa (GBE) and Subgroup IIIb (troxerutin). The brain was removed and the cerebral hemisphere was coronally cut at the hippocampal level and used for light microscopic study (H&E staining and PCNA immunostaining). There was a statistically insignificant improvement in animal weights in subgroup IIb and subgroup IIc. Subgroup IIb showed a statistically significant reduction of blood glucose levels while the subgroup IIc showed insignificant reduction of blood glucose levels. Diabetes disturbed the light microscopic structure of the hippocampus. In subgroup IIb and subgroup IIc the hippocampus retained an apparently normal appearance and the stratum pyramidale exhibited the pyramidal cells with rounded vesicular nuclei and acidophilic cytoplasm. Diabetic hippocampal sections revealed negative PCNA immunoreactivity inall layers of DG. In subgroup IIb and subgroup IIc, hippocampal sections showed positive immunoreactivity
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OBJECTIVE：To rapidly evaluate the effectiveness ，safety and economics of Ginkgo biloba extract（EGb）in the treatment of Alzheimer ’s disease （AD）patients，and to provide evidence-based reference for clinical drug selection and decision. METHODS：Retrieved from PubMed ，Embase，Cochrane Library ，Web of Science ，CNKI，CBM，Wanfang database ，health technology assessment （HTA）organization websites and database during the inception to Aug. 10，2020，HTA reports ，systematic reviews/Meta-analysis，and pharmacoeconomic studies of EGb versus placebo in the treatment of AD were collected. After literature screening and data extration ，HTA checklist ，AMSTAR-2 scale and CHEERS scale were used respectively to evaluate the literature quality of the included HTA report ，systematic review/Meta-analysis and pharmacoeconomics studies. The conclusion of the included studies were summarized by using qualitative description. RESULTS ：A total of 9 literatures were included ，involving 8 systematic reviews and 1 economic studies. In terms of effectiveness ，there was no statistical significance in MMSE score of EGb group，ADAS-Cog score of 120 mg EGb group ，compared with placebo group （P＞0.05）. Dementia Quality of Life （DQoL）score of EGb group was significantly higher than that of placebo group. The scores of short cognitive aptitude tests ，neuropsychiatric inventory（NPI），NPI caregiver version score ，ADAS-Cog score of 160 mg EGb group and 240 mg EGb group were significantly lower than those of control group （P＜0.05）. ADL scores of patients were inconsistent ；ADL scores of EGb group were significantly lower than those o f placebo group （P＜0.05），or there was no significant diff erence between 2 groups（P＞0.05）； . subgroup analysis by dose showed that there was no RDY2019-39） significant difference in ADL score between 120 mg EGb group and placebo group （P＞0.05）；ADL score of 240 mg E-mail：renxiaolei83@126.com EGb group were signicantly lower than that of placebo group （P＜0.05）. Subgroup analysis of clinical global impression 010-88325751。E-mail：lyi1267@126.com change （CGIC） score showed that there was no significant difference in CGIC score between EGb group and placebo group after receiving ＜200 mg EGb and 26 weeks of treatment （P＞ 0.05）；CGIC score of EGb group was significantly higher than that of placebo group after receiving ＞200 mg EGb and 24 weeks of treatment （P＜0.05）. In terms of safety ，there was no statistical significance in the incidence of ADR or the incidence of severe ADR between EGb group and placebo group （P＞0.05）. Subgroup analysis by dose showed that the incidence of ADR in 240 mg EGb group was significantly higher than placebo group （P＜0.05）. Economically ，EGb treatment for AD is cost-effective ，which could indirectly save the nursing costs of AD patients. CONCLUSIONS ：The efficacy of EGb in the treatment of AD is uncertain ， and the safety and economy are good.
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@#To explore the effect of Ginkgo biloba extract (GBE) on anticoagulation of 4 new oral anticoagulants (NOACs), dabigatran, apixaban, rivaroxaban and edoxaban in vitro, thrombin time (TT), prothrombin time (PT), activated partial thrombin time (APTT) and the activity of coagulation factor Xa (FXa) of rat plasma were measured at different concentrations of NOACs, GBE or NOACs combined with GBE, respectively. The results showed that TT, PT and APTT were prolonged with the increase of NOACs concentration in the range of 0-500 ng/mL; that except for TT of rivaroxaban, other results showed a good linear correlation with NOACs concentration (r2= 0.78-0.98); and that FXa activity decreased with increased concentration of FXa inhibitors (apixaban, rivaroxaban and edoxaban), with a good linear correlation with concentration of FXa inhibitors in the range of 0-250 ng/mL (r2= 0.85-0.94). GBE had no significant effect on TT, PT and APTT (P>0.05) in the concentration range of 0-500 μg/mL, but FXa activity had a positive linear correlation with GBE concentration (r2= 0.840 4). TT was prolonged with increasing GBE concentration when dabigatran was combined with GBE. When the above FXa inhibitors were combined with GBE, TT shortened and FXa activity increased with rising GBE concentration. There were no significant changes in PT and APTT (P>0.05) when NOACs were combined with GBE. The study results suggest that GBE may synergize with the anticoagulant activity of dabigatran and antagonize the anticoagulant activity of FXa inhibitors, possibly due to its role in increasing FXa activity.
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Ginkgo biloba is a kind of ancient relict woody. It is a common homologous plant of medicine and food. Its leaves and fruits can produce secondary metabolites, such as flavonoids and ginkgolide, which have good medicinal effects. Ginkgo biloba leaves have low toxicity and broad-spectrum pharmacological activity. The research showed that ginkgo flavonoid has the effect of anticancer and cancer prevention. It is a potential chemical for cancer prophylaxis, which can significantly reduce the incidence of many malignant cancers. The progress in research on the structures, antitumor effects, and mechanism of flavonoids of Ginkgo biloba is summarized in this article, which provides a pharmacological basis for the deep processing and comprehensive utilization of Ginkgo biloba.
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Objective: To observe the effect of Ginkgo biloba extract on cerebral cortical ischemia in mice with acute cerebral ischemia and explore its effect on Toll-like receptor 4 (TLR4)/nuclear factor-κB (NF-κB) pathway. Methods: Sixty adult healthy male CD1 mice were randomly divided into sham operated group, model group, positive control group (3 × 104 IU/kg ulinastatin) and Ginkgo biloba extract low, medium and high doses (10, 20, 40 mg/kg) groups. The acute cerebral ischemia model in other five groups were all established except sham operated group. After successful modeling, mice in each group were given corresponding drugs iv tail, sham operation group and model group were given the same amount of saline. The pathological changes and histological grades of the cerebral cortex were observed. The survival neuron density of each group was compared. Malondialdehyde (MDA) and superoxide dismutase (SOD) were measured by colorimetry and xanthine oxidation respectively. The expressions of Toll like receptor 4 (TLR4) and nuclear transcription factor NF-κB p65 (NF-κB p65) mRNA were detected by real-time fluorescent quantitative PCR (qRT-PCR). Western blotting was used to detect TLR4 and NF-κB p65 protein expression. Results: In sham-operated group, the structure of cerebral cortex was normal, and the cells were full and arranged tightly and orderly. In the blank control group, the structure of cerebral cortex was severely damaged, and the cells were severely shrunk and disordered. Pathological changes in the Ulinastatin group and Ginkgo biloba extract groups were alleviated and dose-dependent. Compared with the sham operation group, the other five groups showed higher histological grade and MDA level in cerebral cortex (P < 0.05). Compared with the model group, the histological grade and MDA level of cerebral cortex decreased in ulinastatin group and Ginkgo biloba extract groups (P < 0.05). Compared with ulinastatin group, the histological grade and MDA content of brain cortex decreased in high dose group (P < 0.05). Compared with the sham operated group, the density of surviving neurons and the level of SOD in the other five groups decreased (P < 0.05). Compared with the model group, the density of survival neurons and the level of SOD in cortex increased in ulinastatin group and Ginkgo biloba extract group (P < 0.05). Compared with ulinastatin group, the density of survival neurons and the level of SOD in cortex increased in high dose group (P < 0.05). Compared with the sham operated group, the expression of TLR4, NF-κB p65 mRNA and protein in the other five groups increased (P < 0.05). Compared with the model group, the expression of TLR4, NF-κB p65 mRNA and protein in cortex of mice in ulinastatin group and Ginkgo biloba extract group decreased (P < 0.05). Compared with ulinastatin group, the expression of TLR4, NF-κB p65 mRNA and protein in the brain cortex of mice decreased in the high dose group (P < 0.05). Conclusion: Ginkgo biloba extract can alleviate the pathological changes caused by cerebral ischemia in mice with acute cerebral ischemia, alleviate neurological impairment, and improve oxidative stress indexes of cerebral cortex tissues. It is presumed that it may be achieved by regulating TLR4/NF-κB p65 signaling pathway and inhibiting the expressions of TLR4 and NF-κB p65 proteins.
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To investigate if ginkgo biloba extract (Egb-761) can improve tardive dyskinesia (TD) symptoms through increasing the activity of plasma MnSOD. Methods We enrolled a total of 384 schizophrenia patients including 157 TD patients and 227 non-TD patients, as well as 280 normal subjects. The difference of MnSOD level in plasma among these groups were compared. TD patients were then randomly divided into two groups. The treatment group (n=77) and the placebo group (n=75) were treated with 240 mg of Egb-761 or placebo per day for 12 weeks, respectively. The abnormal involuntary movement scale (AIMS) and the positive and negative symptoms scale (PANSS) were used to evaluate the severity of the symptoms in baseline, the sixth week and the twelfth week after treatment. The level of MnSOD activity in plasma was also detected before and after the treatment. Results The level of MnSOD activity was lower in schizophrenia groups than in healthy control group (P＜0.01). In addition, the level of MnSOD activity was significantly lower in TD group than in non-TD group (P＜0.05). Repeated measures analysis of variance showed that group effect (F=4.00, P=0.05), time effect (F=32.17, P＜0.01) and interactive effect of group and time (F=39.04, P＜0.01) were significant in AIMS total score. The AIMS total score of treatment group was significantly lower than that of placebo group at 6-week and 12-week time points (all P＜0.01). Repeated measures analysis of variance showed that time effect (F=23.04, P＜0.01) and interactive effect of group and time (F=6.41, P＜0.05) were significant in the level of MnSOD activity. In addition, the level of MnSOD at baseline was significantly correlated with the reduction of AIMS total score during the treatment period (r=0.27, P=0.018). Conclusion Treatment of Egb-761 can improve symptoms of TD and activity of MnSOD.
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Objective To observe the clinical effect of treating Alzheimer's disease (AD) using transcranial magnetic stimulation.Methods One hundred and ninety-six patients with Alzheimer's disease were randomly divided into an observation group and a control group,each of 98.The observation group was given transcranial magnetic stimulation of the left and right dorsolateral frontal lobes of the brain and simultaneously given "8-shaped" coil stimulation.The stimulation intensity was 80％ of the motor threshold with a sequence of 2 s of stimulation at 5 Hz and 30 s rest for 30 min in each session.There were two sessions a day for 28 days.The control group was treated with identical pseudo-stimulation.Moreover,both groups were treated with intravenous injections of 20 ml of Ginkgo biloba extract dissolved in 250 ml of sodium chloride,or in the control group a glucose injection,one daily for two weeks.Before and after the treatment,the cognition,behavior and neuropsychological symptoms of both groups were evaluated using the mini mental state examination scale (MMSE),the AD rating scale (ADAS-cog),the activity of daily living (ADL) scale,a neuropsychiatric questionnaire (NPI) and an AD behavioral pathology rating scale (BEHAVE-AD) to compare the clinical effects.Results There were no significant differences in the groups' average scores on any of the evaluations before the treatment.After the treatment,the average MMSE and ADAS-cog scores in both groups had improved significantly,but with significantly greater improvement in the observation group.After the treatment,the average ADL,NPI and BEHAVE-AD scores of the observation group and the average ADL score of the control group were significantly lower than before the treatment.No significant differences were observed in the average NPI and BEHAVE-AD scores of the control group.After the intervention,the average ADL,NPI and BEHAVE-AD scores in the observation group were significantly lower than those of the control group.The total effectiveness rate of the observation group (90.8％) was significantly higher than that of the control group (62.2％).Conclusion Transcranial magnetic stimulation can significantly improve the cognitive,behavioral and neuropsychological status of patients with Alzheimer's disease.
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OBJECTIVES.: Sodium salicylate (SS) is well known for its ototoxic properties that induce functional and morphological changes in the cochlea and brain. Ginkgo biloba extract (GBE) has been widely used for treatment of various neurodegenerative diseases; however, its effects on salicylate-induced ototoxicity remain unclear. Herein, we examined the effects of EGb 761 (EGb), a standard form of GBE, on the plasticity of the N-methyl-D-aspartate receptor subunit 2B (GluN2B) in the inferior colliculus (IC) following SS administration. METHODS.: Seven-week-old Sprague Dawley rats (n=24) were randomly allocated to control, SS, EGb, and EGb+SS groups. The SS group received a single intraperitoneal SS injection (350 mg/kg), the EGb group received EGb orally for 5 consecutive days (40 mg/kg), and the EGb+SS group received EGb for 5 consecutive days, followed by an SS injection. The auditory brainstem responses (ABRs) were assessed at baseline and 2 hours after SS administration. GluN2B expression was examined by Western blot and immunohistochemistry. RESULTS.: There were no significant differences in ABR threshold shifts among the groups. The expression of the GluN2B protein normalized by which of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was significantly lower in the EGb+SS group, as compared to the SS group (P=0.012). Weak and diffused GluN2B immunoreactivity was detected in the IC neural cells of the EGb+SS group, while those of the SS group exhibited strong and diffused GluN2B positivity. CONCLUSION.: EGb may play a role in regulating the GluN2B expression in the IC of salicylate-induced ototoxicity model.
Subject(s)
Blotting, Western , Brain , Cochlea , Evoked Potentials, Auditory, Brain Stem , Ginkgo biloba , Glyceraldehyde 3-Phosphate , Immunohistochemistry , Inferior Colliculi , N-Methylaspartate , Neurodegenerative Diseases , Oxidoreductases , Plastics , Rats, Sprague-Dawley , Sodium SalicylateABSTRACT
OBJECTIVE: To develop on-line high performance liquid chromatography-biochemical detection (HPLC-BCD) METHODS: to screen effective components in Ginkgo biloba extract for treatment of Alzheimer's disease.METHODS: Radical scavengers and AchE inhibitors in G.biloba leave extract were screened by HPLC-UV-DPPH/ABTS and HPLC-UV-AchE METHODS:, respectively. The stability and repeatability of the on-line HPLC-UV-AchE method were investigated using galanthamine as a positive reference. The main active ingredients in the extract were identified by ultra high performance liquid chromatography linear ion trap/orbitrap high resolution mass spectrometry (UPLC-LTQ/orbitrap MS) method. RESULTS: Fourteen antioxidants in G.biloba extract were detected by the HPLC-UV-DPPH/ABTS method, while three AchE inhibitors were found by the HPLC-UV-AchE method. The three AchE inhibitors were tentatively identified by UPLC-LTQ/orbitrap MS as 4-O-beta-glucopyranosyl-cis-coumaric acid ginkgolide A and 3-O-[2-O-(β-D-Glucosyl)-α-L-rhamnosyl]quercetin, respectively. CONCLUSION: The proposed on-line HPLC-BCD method shows high sensitivity, good repeatability and stability. It can be used to screen antioxidants and AchE inhibitors in G. biloba extract. This will provide an effective, quick and convenient analysis tool to quickly find bioactive ingredients of traditional Chinese medicines for treating related diseases.
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Ischemic cerebrovascular disease and cerebral ischemia/reperfusion injury threaten the health of human being. We studied the protective effect of Ginkgo biloba extract 50 (EGb50) on the mitochondrial function in SH-SY5Y cells after hypoxia/reoxygenation (H/R) injury and explored its mechanisms, so as to provide new ideas for studies on the treatment for ischemic cerebrovascular disease. We established the H/R injury model in SH-SY5Y cells after administrating EGb50. Subsequently, the mitochondrial membrane potential and the concentration of intracellular Ca²⁺ were measured by flow cytometer. The levels of optic atrophy1 (Opa1) and dynamin-like protein 1 (Drp1) were evaluated by immunofluorescence and western blot. The results showed that the mitochondrial membrane potential was decreased and the level of intracellular Ca²⁺ was increased after H/R injury. Moreover, the expression of mitochondrial fusion protein Opa1 was decreased, while the expression of mitochondrial fission protein Drp1 was increased. However, EGb50 significantly increased the mitochondrial membrane potential and suppressed the level of intracellular Ca²⁺. In addition, EGb50 increased the expression of Opa1 and decreased the expression of Drp1. The results demonstrated that EGb50 has a neuroprotective effect on SH-SY5Y cells after H/R injury, and could improve the energy metabolism and mitochondrial function. The underlying mechanisms may be associated with the regulation of mitochondrial fusion and fission, which provided data support for the treatment of ischemic cerebrovascular disease with EGb50.
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Humans , Cell Hypoxia , Membrane Potential, Mitochondrial , Mitochondria , Plant Extracts , Reperfusion InjuryABSTRACT
Objective:To determine and investigate the stability of ginkgo biloba extract injection from three different manufactur-ers respectively in 0.9% NaCl infusion and 5% glucose infusion under different conditions (room temperature, high temperature and light). Methods:Ginkgo biloba extract injection was mixed with the two kinds of infusions,and then divided into the normal tempera-ture group,the high temperature group and the light group. The appearance,insoluble particles,pH and content of flavonoids after the relevant treatment were investigated. The appearance and insoluble particles were tested according to the characteristics of the inspec-tion method described in Chinese Pharmacopoeia(2015 edition,volume IV,the general rule),and the content of flavonoids was detec-ted by HPLC-UV. Results:All the mixed solutions were yellow. No significant changes were found in the appearance,pH value,in-soluble particles and contents of quercetin and isorhamnetin in all the mixed solutions in 24 h. The pH value of the mixed solution with 5% glucose infusion was lower than that with 0.9% NaCl infusion,and all the pH values met the standard in Chinese Pharmacopeias. The kaemphenol content in the injection from Shenwei pharmaceutical company was higher than that from the other manufacturers, while the content of kaemphenol in all the injections was within the standard range. Conclusion:The quality of Ginkgo biloba extract injection from the three different manufacturers is stable under different conditions.
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Objective @#To investigate the effect and potential molecular mechanisms of isorhamnetin (ISO) extracted from Ginkgo biloba on the differentiation of osteoclasts.@*Methods@#Osteoclast precursor RAW264.7 cells were induced with RANKL to differentiate into mature osteoclasts. Different concentrations of ISO were added to RAW264.7 cells to determine its effect on osteoclast differentiation. CCK8 was used to evaluate the effect of ISO on cytotoxicity. The impact of ISO on the osteoclast differentiation process was investigated by analyzing tartrate resistance and bone resorption lacuna. Real-time PCR was performed to analyze the levels of differentiation marker genes, including tartrate resistant acid phosphatase (Trap), cathepsin K (Ctsk), and matrix metalloproteinase 9 (MMP-9); differentiation-related transcription factors, including the proto-oncogene protein c-Fos, nuclear factor of activated T-cells cytoplasmic 1(NFATc1); and the levels of downstream NF-κB p65 signaling pathway phosphorylation. Using the above-described method, we verified that ISO exerted an inhibitory effect on osteoclast differentiation and explored related molecular mechanisms. @*Results @#Different concentrations of ISO (1-10 μM) had no cytotoxic effects on RAW264.7 cells, inhibited TRAP activity and decreased the number of bone resorption lacuna during osteoclast differentiation. When applied at a concentration of 10 μM, its inhibitory effect was significant. In addition, ISO significantly reduced the expression levels of Trap, Ctsk, MMP-9, c-Fos, NFATc1 and NF-κB p65 mRNA. @*Conclusion@# ISO extracted from Ginkgo biloba extract exerted an inhibitory effect on osteoclast differentiation, and the mechanism underlying its activity may involve the inhibition of the classical NF-κB pathway.
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In this study, the anti-inflammatory mechanism of Ginkgo biloba extract 50 (GBE50) and its mechanism of action on NLRP3 inflammatory corpuscles were observed by primary microglia cells. LPS/ATP was used to stimulate microglia, and the best time for stimulation and optimal concentration of GBE50 were screened. Pro-inflammatory cytokine IL-1 and TNF-α was determined by ELISA. Western blot was performed to observe the protein expression of NLRP3, ASC, caspase-1 and IL-1 in cultured primary rat microglia. Effect of GBE50 on NLRP3 inflammatory corpuscle aggregation was detected by CO-IP. GBE50 (10 mg·L⁻¹) preconditioning could significantly inhibit the expression of LPS (100 μg·L⁻¹,12 h), ATP (5 mmol·L⁻¹, 30 min) induced primary microglia corpuscle associated protein, inflammatory corpuscle aggregation, and the release of inflammatory factors IL-6 and TNF-α. These results indicated that GBE50 could inhibit the secretion of inflammatory factors after microglia activation, which is related to down regulating the protein expression and activity of NLRP3 inflammatory corpuscle.
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OBJECTIVES: Local administration of 3-nitropropionic acid (3-NP) to the inner ear induces sensorineural hearing loss. Several studies have shown the otoprotective effects of ginkgo biloba extract EGb 761. Moreover, EGb 761 has been reported to activate Sirtuin 1 (SIRT1). The present study was designed to investigate whether EGb 761 prevents 3-NP-induced sensorineural hearing loss and determine its effects on the expression of SIRT1. METHODS: Sprague Dawley rats were divided into four experimental groups: control group receiving vehicle of 3-NP, EGb group receiving EGb 761, 3-NP group receiving 3-NP, and EGb+3-NP group receiving EGb 761 and 3-NP. EGb 761 was given orally for 5 days. The 3-NP solution was injected into the tympanum 3 days after the start of EGb 761 administration. The auditory brainstem response was recorded before and after the injection. At 4 weeks after the administration of 3-NP or vehicle of 3-NP, cochleae were harvested, and hematoxylin and eosin staining and immunohistochemistry for SIRT1 antibody were performed. RESULTS: EGb+3-NP group showed significantly lower threshold shifts than 3-NP group. There was a significant preservation of type II fibrocytes and spiral ganglion cells in EGb+3-NP group than in 3-NP group. In EGb+3-NP group, there was a significantly greater number of SIRT1 immunopositive type II fibrocytes and spiral ganglion cells than in 3-NP group. Calculating the percentage of SIRT1 immunoreactive type II fibrocytes and spiral ganglion cells in viable type II fibrocytes and spiral ganglion cells, respectively, EGb+3-NP group showed significantly higher SIRT1 immunoreactive cells than 3-NP group. CONCLUSION: These results suggest that EGb 761 may prevent hearing loss induced by 3-NP in an acute ototoxic animal model, which appears to be related with SIRT1 expression.
Subject(s)
Cochlea , Ear, Inner , Ear, Middle , Eosine Yellowish-(YS) , Evoked Potentials, Auditory, Brain Stem , Ginkgo biloba , Hearing Loss , Hearing Loss, Sensorineural , Hearing , Hematoxylin , Immunohistochemistry , Models, Animal , Rats, Sprague-Dawley , Sirtuin 1 , Spiral GanglionABSTRACT
OBJECTIVE@#To investigate the protective effects of Ginkgo biloba extract(GBE) on paracetamol(APAP)-induced acute hepatic injury in mice and its mechanism.@*METHODS@#Thirty mice were randomly divided into control group, model group, GBE low, medium and high-dose(50,100,and 200 mg·kg)groups,with 6 mice in each group. All mice except control group were administered with APAP(300 mg/kg)for one time by intraperitoneal injection. The mice in GBE low, medium and high-dose groups were intragastric administered with GBE for 2 d consecutively, then samples were harvested for analysis. The appearance and pathology of liver were observed. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in serum and the levels of superoxide dismutase (SOD), myeloperoxidase(MPO), glutathione (GSH) and malondialdehyde (MDA) in hepatic tissue were measured. Western blot was used to detect the protein expressions of Nrf2 and HO-1.@*RESULTS@#Compared with control group, in model group, the appearance and pathology of liver were bad, the levels of ALT,AST,TNF-α and IL-6 in serum were increased significantly(<0.01),the levels of GSH and SOD were decreased while the levels of MDA and MPO were increased in hepatic tissue(<0.01), the expressions of Nrf2 and HO-1 were increased in hepatic tissue(<0.05). Compared with model group, in GBE groups, the appearance and pathology of liver were improved, the levels of ALT,AST,TNF-α and IL-6 in serum were decreased significantly(<0.01), the levels of GSH and SOD were increased while the levels of MDA and MPO were decreased in hepatic tissue(<0.01), the expression of Nrf2 and HO-1 were increased in hepatic tissue(<0.05). The high-dose of GBE possessed the most obvious treatment effect among them.@*CONCLUSIONS@#GBE may play a protective role in APAP-induced acute hepatic injury through Nrf2/HO-1 pathway.
Subject(s)
Animals , Mice , Acetaminophen , Alanine Transaminase , Aspartate Aminotransferases , Chemical and Drug Induced Liver Injury , Ginkgo biloba , Liver , Malondialdehyde , Oxidative Stress , Plant ExtractsABSTRACT
To determine whether ginkgo biloba extract(GBE)combined with dexamethasone(DEX)plays a role in the treatment of allergic rhinitis-related olfactory dysfunction using an animal model.Six week old BALB/C mice were sensitized and challenged with ovalbumin.30 sensitized mice were divided into three groups:Group 1 was given high-dose GBE and DEX(n=10);Group 2 was given low dose GBE and DEX(n=10);Group 3 was given DEX alone(n=10).We assessed the histology of the olfactory mucosa and serum IL-4,IFN-γ,and caspase 1.A significant higher fraction of mice in group 1 could find the food pellet within300 scompared to group 3(<0.05).Caspase-1 levels improved during the second week compared with the first week in each group.IFN-γlevels were significantly lower during the second week compared with the first week(<0.05,all).IL-4 levels also were significantly lower during the second week compared with the first week in all groups except those receiving DEX alone.IFN-γ/IL-4 levels in each group were significantly lower during the second week compared with the first week(<0.05,all).In this animal model of allergic rhinitis-related olfactory dysfunction,the addition of ginkgo biloba extract to dexamethasone have a better anti-inflammatory effect,which can partly improve the therapeutic effect on olfactory dysfunction caused by allergic rhinitis.