Protective Effect of Ginkgo Biloba Leaf Extract on Learning and Memory Deficit Induced by Aluminum in Model Rats

Objective： To examine the protective effect of Ginkgo biloba leaf extract （GbE） on learning and memory deficit induced by aluminum chloride （AlCl3）, and explore its mechanisms. Methods： The rat models with learning and memory deficit were induced by administering via gastrogavage and drinking of AlCl3 solution. And the model rats were treated with GbE at the dose of 50, 100, 200 mg/kg every day for 2 months accompanied with drinking of AlCl3 solution, respectively. Their abilities of spatial learning and memory were tested by Morris water maze, and the acetylcholinesterase （ACHE） activity in serum was assayed with chemical method, the AChE expression in hippocampus was observed by immunohistochemistry assay, and then quantitative analysis was done by BI 2000 image analysis system. Results： Learning and memory deficit of rats could be induced by AlCl3 solution （P〈0.01）, and AChE expressions in rats hippocampus were increased （P〈0.01）; GbE ameliorated learning and memory deficit and reduced AChE expression in rats hippocampus in a dose-dependent manner, while GbE significantly increased serum AChE activity at the dose of 200 mg/kg each day （P〈0.05）. Conclusion： GbE can ameliorate learning and memory deficit induced by AlCl3, which may be due to its inhibition of the AChE expression in hippocampus.

Ginkgo biloba leaf extract effects on inducible nitric oxide synthase, Bcl-2, and Bax expression in rat models of spinal cord injury

BACKGROUND：Ginkgo biloba leaf extract exhibits neuroprotective effects in spinal cord injury. However, the mechanisms of action remain unclear. OBJECTIVE： To investigate inducible nitric oxide synthase （iNOS） and Bcl-2/Bax expression in the injured spinal cord, and to explore the neuroprotective mechanisms of ginkgo biloba leaf extract in rats with spinal cord injury. DESIGN, TIME AND SETTING： The randomized, controlled, cell molecular biology experiment was performed at Soochow University, China from March 2007 to March 2008. MATERIALS： A total of 120 healthy, adult Sprague Dawley rats were selected for this study. Rat models of moderate acute thoracic （T9） spinal cord injury were established using the modified Allen method. Shuxuening injection was obtained from Zhenbaodao Pharmaceutical Co., Ltd., China. Methylprednisolone was purchased from North China Pharmaceutical Co., Ltd. METHODS： All rats were equally and randomly divided into four groups. Only the spinal cord was exposed in the sham operation group rats. In the trauma group, rats were not treated with drugs following spinal cord injury. Rats in the hormone group were intraperitoneally injected with 30 mg/kg methylprednisolone following spinal cord injury. Rats in the ginkgo biloba leaf extract group were intraperitoneally infused with a 1.0 mL/kg Shuxuening injection per day. MAIN OUTCOME MEASURES： At 1 hour, as well as 1, 3, 5, 7, and 14 days after spinal cord injury, iNOS- and Bcl-2/Bax-positive cells were quantified with immunohistochemistry. Pathological changes were detected using hematoxylineosin staining under an optical microscope. RESULTS： Spinal cord injury in the ginkgo biloba leaf extract and hormone groups was milder compared with the trauma group. Demyelination was significantly ameliorated and the necrotic cavity was obviously reduced in the injured spinal cord of rats in the ginkgo biloba leaf extract and hormone groups at each time point. iNOS expression was increased in the injured spinal cord, and reached a peak at 5 days. The number of iNOS-positive cells was lower in the ginkgo biloba leaf extract and hormone groups compared with the trauma group （P 〈 0.05-0.01）. The number of iNOS-positive cells was lower in the ginkgo biloba leaf extract group compared with the hormone group at 7 and 14 days after spinal cord injury （P 〈 0.05）. Bcl-2 expression reached a peak at 3 days, and Bax expression reached a peak at 5 days following rat spinal cord injury. Bcl-2 expression was increased, but Bax expression was decreased in the ginkgo biloba leaf extract and hormone groups compared with the trauma group （P 〈 0.05-0.01）. Bcl-2 expression was greater, but Bax expression was reduced in the ginkgo biloba leaf extract group compared with the hormone group at 7 and 14 days after spinal cord injury （P 〈 0.05）. CONCLUSION： Ginkgo biloba leaf extract exhibits neuroprotective effects by upregulating Bcl-2 expression, downregulating Bax expression, and significantly inhibiting high expressions of iNOS in the injured spinal cord. The neuroprotective effects of ginkgo biloba leaf extract are greater compared with methylprednisolone at 1 week after spinal cord injury.

Effects of Ginkgo Leaf Extract on Function of Dendritic Cells and Th1/Th2 Cytokines in Patients with Unstable Angina Pectoris

Objective： To investigate the effects of Ginkgo leaf extract （GLE） on function of dendritic cells （DO） and Th1/Th2 cytokines in patients with unstable angina pectoris（UAP）. Methods： Fifty-four patients with UAP were equally assigned into two groups, the treated group and the control group, both treated with conventional Western medicine, but with GLE given additionally to the treated group. Blood of all patients was taken before and 4 weeks after treatment to prepare the peripheral mononuclear cells, then which were incubated in the completed medium containing granulocyte-macrophage colony stimulatory factor （GMCSF） and interleukin-4 （IL-4） to induce mature DO. The expression of co-stimulating factor CD86 （B7-2） on the surface of DC was detected by flow cytometry, and the stimulating capacity of DC was determined by mixed lymphocyte reaction （MLR）. The blood levels of cytokines, interferon-γ （IFN-γ）, and IL-4, were analyzed by ELISA, and blood C-reactive protein （CRP） level by turbidimetry. Moreover, the direct effect of Ginkgolide B on CD86 expression on DO were also tested in vitro. Results： After treatment, CD86 expression on DO, the stimulating capacity of DO as well as levels of IFN-γ and ORP were lowered in both groups （P〈0.05 or P〈0.01）, but the changes were much more significant in the treated group than those in the control group. Ginkgolide B showed a direct inhibitory effect on the CD86 expression on DO. Conclusion： The inhibition of GLE on DO and thereby the suppression on inflammatory reaction may be one of the mechanisms of GLE in treating patients with UAP.

Ginkgo Leaf Extract and Armillariella Mellea Powders Oral(银杏蜜环口服溶液) attenuates inflammation of microglia in habenular nucleus through CX3CL1-CX3CR1 axis in post stroke depression

OBJECTIVE The Ginkgo Leaf Extract and Armillariella Mellea Powders Oral(Yinxingmihuan Koufu Rongye,YXMH),a representative drug for"Treating both Brain and Heart",showed considerable clinical effects in isch⁃emic cardiovascular and cerebral vascular diseases.Recently,it is reported that YXMH has the potential for treating myocardial and cerebral ischemia related mental disorders,such as post stroke depression(PSD)and chronic heart disease(CHD)associated anxiety disorder.However,its mechanism has not been clearly elucidated.Meanwhile,increasing evidence revealed that there are close functional links between depression and habenular nucleus.The present study investigates the underlying mechanism of YXMH on attenuating the inflammation of microglia in habenular nucleus through CX3CL1-CX3CR1 axis in in a rat model of PSD.METHODS Rats were randomly devided into sham group,model group,Ginaton group(18 mg·kg^-1),Armillariella Mellea group(600 mg·kg-1),Fluoxetine group(10 mg·kg^-1),YXMH high-dose group(618 mg·kg^-1)and YXMH low-dose group(309 mg·kg^-1).The PSD model was induced by transarterial microembolization combined with sleep deprivation(2-Chloro-D-phenylalanine,PCPA,IH,200 mg·kg^-1,for 3 times,before the behavior test)in SD male rats.Then rats were treated with corresponding medicaments through gavage once a day until 3 weeks later,followed by body mass measurement,neurological deficit score evaluation,gripping strength and thermal withdrawl latency measurement,as well as depression related behavioral indicators,the open field test(OFT)and sucrose preference test.The pathological morphological changes of habenular nucleus was observed by HE staining,the expression of IBA-1 was measured and analyzed by immunohistochemistry staining,and alterations of proteins and genes related to the CX3CL1-CX3CR1 axis were analyzed using Western blotting(CX3CL1,CX3CR1)and real-time polymerase chain reaction(PCR)(CX3CL1,CX3CR1).RESULTS Compared with the sham group,rats in the model group manifested as decreased body mass,deficient neurological behavior and gripping strength,reduced loco⁃motor activity and sugar water consumption,as well as elevated thermal withdrawl latency(P<0.05,P<0.01).Mean⁃while,the pathological morphology of the habenular nucleus on the ischemic hemisphere showed significant neuronal degeneration,microglial proliferation,inflammatory cells and glia cells infiltration,together with up-regualted expression of IBA-1,CX3CL1,CX3CR1 protein and CX3CL1,CX3CR1 mRNA.YXMH attenuated inflammation of microglia in habenular nucleus through improving pathological morphology,inhibiting IBA-1 activation,down-regulating the expres⁃sion of CX3CL1 and CX3CR1 proteins and genes,and thus improved the behavior performance of ischemic injury and depression.CONCLUSION YXMH ameliorates neurological deficit and depressive behavior in rat model of PSD induced by transarterial microembolization combined with sleep deprivation,and the mechanism is probably related to attenu⁃ating inflammation of microglia in habenular nucleus through CX3CL1-CX3CR1 axis.

Simultaneous quantification of flavonol glycosides, terpene lactones, polyphenols and carboxylic acids in Ginkgo biloba leaf extract by UPLC-QTOF-MS^E based metabolomic approach

Abstract： In the present study, we established an ultra performance liquid chromatography coupled with time-of-flight mass spectrometry （UPLC-QTOF-MSE） method to simultaneously quantify 33 components in Ginkgo biloba leaf extracts （GBEs）, including 17 flavonol glycosides, five terpene trilactones （TTLs）, four polyphenols and seven carboxylic acids. This optimized method was successfully applied to analyze the explicit compositions of GBE samples collected from different places. Furthermore, the data were processed through unsupervised principal component analysis （PCA） and supervised orthogonal partial least squared discrimination analysis （OPLS-DA） to evaluate the quality and compare the differences between the samples according to the contents of the 33 chemical constituents. Bilobalide, protocatechuic acid, shikimic acid, quinic acid, ginkgolide B, ginkgolide J, kaempferol-3-O-rutinoside, isorhamnetin-3-O-rutinoside, quercetin-3-O-ct-L-rhamnopyranocyl-2＂-（6＇＂-p-coumaroyl）-β-D-glucoside and rutin were recognized as characteristic chemical markers that contributed most to control the quality of GBEs. Based on the fact that GBEs should be standardized with the characteristic components as quality control chemical markers, it is most important to maintain the quality of GBEs stable and reliable, and this method also provided a good strategy to further rectify and standardize the GBEs market.

Progress in Researches on the Pharmaceutical Mechanism and Clinical Application of Ginkgo Biloba Extract on Various Kinds of Diseases

Progress made over the pharmaceutical mechanism and clinical application of Ginkgo Biloba extract （GBE） on various kinds of diseases were reviewed in this paper. The effective elements con- tained in GBE are mainly kinds of Ginkgo flavonoid and Ginkgolide, which have marked protective effects on cardio-cerebral vascular and central nerve systems. In clinical practice, it is applied mostly in treatment of cardio-cerebral vascular diseases. Also it shows apparent effects in the treatment processes of some other diseases as an adjuvant, and therefore, has been gradually accepted by the medical circle in the world, proving to be a medicine of wide prospect in development and application.

Effect of Ginkgo Leaf Extract on Vascular Endothelial Function in Patients with Early Stage Diabetic Nephropathy

Objective:To explore the effect of ginkgo leaf extract(GLE) on vascular endothelial function(VEF) in patients with early stage diabetic nephropathy(DN).Methods:Sixty-four patients were randomized equally by a randomzing digital table into two groups,the treated group and the control group.They were all treated for 8 weeks with conventional therapy for diabetes,but GLE tablets were given to the treated group additionally. Changes in VEF were estimated before and after treatment by ultrasonic examination of t...

Ginkgo Leaf Extract and Dipyridamole Injection as Adjuvant Treatment for Angina Pectoris:A Meta-Analysis of 41 Randomized Controlled Trials

Objective： To provide information about the effectiveness and safety of Ginkgo Leaf Extract and Dipyridamole Injection （GD） as one adjuvant therapy for treating angina pectoris （AP） and to evaluate the relevant randomized controlled trials （RCTs） with meta-analysis. Methods： RCTs concerning AP treated by GD were searched in China Biology Medicine Disc （SinoMed）, PubMed, the China National Knowledge Infrastructure Database （CNKI）, the Chinese Scientific Journals Database （VIP）, Wanfang Database, Embase, and the Cochrane Library, from inception to February, 2017. The Cochrane Risk Assessment Tool was adopted to assess the methodological quality of the RCTs. The Review Manager 5.3 software was utilized to conduct the meta-analysis. Results： A total of 41 RCTs involving 4,462 patients were included in the meta-analysis. The results indicated that the combined use of GD and Western medicine （WM） against AP was associated with a higher total effective rate [risk ratio （RR）=1.25, 95% confidence interval （CI）： 1.21-1.29, P〈0.01], total effective rate of electrocardiogram （RR=1.29, 95% CI： 1.21-1.36, P〈0.01）. Additional, GD combined with WM could decrease the level of plasma viscosity [mean difference （MD）=-0.56, 95% CI： -0,81 to -0.30, P〈0.01], flbrinogen [MD=-1.02, 95% CI： -1.50 to -0.54, P〈0.01], whole blood low shear viscosity [MD=-2.27, 95% CI： -3.04 to -1.49, P〈0.01], and whole blood high shear viscosity （MD=-0.90, 95% CI： 1.37 to -0.44, P〈0.01）. Conclusions： Comparing with receiving WM only, the combine use of GD and WM was associated with a better curative effect for patients with AP. Nevertheless, limited by the methodological quality of included RCTs more large-sample, multi-center RCTs were needed to confirm our findings and provide further evidence for the clinical utility of GD.

Antibacterial mechanism of Ginkgo biloba leaf extract when applied to Shewanella putrefaciens and Saprophytic staphylococcus

The antimicrobial mechanism of Ginkgo biloba leaf extracts(GBLE)when applied to predominant spoilage bacteria(Shewanella putrefaciens and Saprophytic staphylococcus)on refrigerated pomfret and minimal inhibitory concentrations(MICs)were measured by the plate counting method.GBLE at MIC and 2MIC were prepared in tryptic soy broth(TSB)medium and equivalent amounts of sterile distilled water were used in place of GBLE as a control group.The impact of GBLE on the growth of bacteria,the permeability of cell membrane,and cell wall were also investigated by growth curve of bacteria,alkaline phosphates activity(AKP),and electrical conductivity.A scanning electron microscope(SEM)was used to study the effects of GBLE on the cellular structure of S.putrefaciens and S.staphylococcus.The results showed that the MICs of GBLE when applied to S.putrefaciens and S.staphylococcus were 100 mg/mL,the inhibitory rates of MIC and 2MIC concentrations of GBLE when applied to S.putrefaciens were 36.11%and 100%,while 27.78%and 62.22%for S.staphylococcus.Meanwhile,GBLE inhibited the growth of S.putrefaciens and S.staphylococcus until the number of cells at 2MIC values decreased to 0 and 4.29 log CFU/mL,respectively,after 24 h.The electrical conductivity of bacteria increased with GBLE treatment,which was followed by an increased leakage of AKP.The SEM revealed that the structure of bacterial cells was destroyed and the bacteria began to be adhere to each other.The inhibition effect of GBLE when applied to S.putrefaciens and S.staphylococcus was related to the damage of cell membrane and cell wall.It was also revealed that GBLE damages the morphology of bacteria and had stronger effects on the cell membrane of S.putrefaciens than that of S.staphylococcus.

Sensory and chemical assessment of silver pomfret (Pampus argenteus) treated with Ginkgo biloba leaf extract treatment during storage in ice

This study investigated the physical(L*,a*,b*,texture profile analyses,pH),chemical(TVB-N,K value and TBA),microbiological,amino acid content,and flavor effects that Gingko biloba leaf extract(GBLE)had on silver pomfret(Pampus argenteus)stored at 4±1
C in ice for 18 days.Fresh pomfret samples were obtained directly from the local fish market and transported to the laboratory with ice immediately.After being gutted,washed,filleted and trimmed in a water-ice mixture,samples were treated with different concentrations of GBLE(0.0 mg/mL,2.5 mg/mL,5.0 mg/mL,10.0 mg/mL)and packaged in Polyethylene bag,then stored in a refrigerator at 4±1
C with ice.The results show that the shelf-life of untreated(0.0 mg/mL)pomfret samples was 8e9 days compared to 14e15 days for the GBLE1(2.5 mg/mL)treated group.The assessment results showed that different concentrations of GBLE had variable effects on preserving the texture parameters of acceptability limit,inhibit lipid oxidation,protein degradation,and microorganism growth.2.5 mg/mL of GBLE was the best for the preservation of pomfret during storage in ice.Therefore,there is potential use for GBLE as a preservative to extend the shelf-life of pomfret during chilled storage in ice.