Mechanism of Ginkgo biloba L.leaf in the treatment of ischemic stroke based on network pharmacology,bioinformatics and molecular docking

Ginkgo biloba L.leaf(GBL)has been reported to protect against ischemic stroke(IS),one of the leading causes of death and longterm disability worldwide,while there is a lack of systematic study on the exact mechanism.Here,network pharmacology and bioinformatics were used to predict the active components,important targets,and potential mechanisms of GBL in the treatment of IS.Active compounds of GBL were screened based on drug-like index and oral bioavailability,key target genes were screened based on network pharmacology and gene chip,downstream pathways for the regulation of key target genes were predicted based on gene set enrichment analysis,and the interaction between key targets and active compounds was verified based on molecular docking.The results showed that GBL played a protective role in cerebral ischemia with mainly 14 active compounds,such as isoquercitrin,luteolin-4’-glucoside,beta-sitosterol,campesterol,diosmetin,ginkgolide B,ginkgolide C,ginkgolide J,ginkgolide M,isogoycyrol,laricitrin,luteolin,sesamin,and stigmasterol.Further studies revealed that GBL played important role in immunomodulation and inflammation inhibition after cerebral ischemia by acting on its peripheral targets ARG1 and MMP9 to regulate Toll-like receptor,Chemokine and Notch signaling pathway.Meanwhile,GBL played important role in reducing neuroinflammation and blood-brain barrier damage after cerebral ischemia by acting on its central targets,CCL2,PTGS2,IL6,IL1B and MMP9 to regulate the Cytokine-cytokine receptor interaction,Jak-STAT,and Toll-like receptor signaling pathway.Additionally,molecular docking verified that the active compounds mentioned above could bind to ARG1,MMP9,CCL2,PTGS2,IL6,and IL1B.The present study shows the multicomponent,multitarget and multichannel pharmacological effects of GBL on cerebral ischemia and provides a new strategy for the treatment of IS.

The Effect of Ginkgo Biloba Leaf Dropping Pill Combined with Butylphthalide Capsule on Cognitive Dysfunction in Patients after Acute Ischemic Stroke and Its Impact on Serum Cytokines

Objective: To investigate the therapeutic effect of Ginkgo biloba extract dropping pills combined with butylphthalide capsules on cognitive dysfunction in patients after acute ischemic stroke (AIS) and its impact on serum cytokines CRP, IL-6, and Hcy. Methods: This study selected 76 patients with cognitive dysfunction after ischemic stroke who were hospitalized at Zhuji People’s Hospital from January 2023 to January 2024. The patients were divided into two groups. The control group was treated with butylphthalide capsules, while the combination group received Ginkgo biloba extract dropping pills in addition to the treatment given to the control group. The neurological function, cognitive function, activities of daily living, and levels of serum cytokines CRP, IL-6, and Hcy were compared between the two groups after 1 month and 3 months of treatment. Results: The NIHSS scores, MMSE scores, ADL scores, and levels of CRP, IL-6, and Hcy in both groups showed statistically significant differences compared to before treatment (P Conclusion: The combination of Ginkgo biloba extract dropping pills and butylphthalide capsules has a better therapeutic effect on cognitive dysfunction in patients after ischemic stroke. It can improve the neurological function and cognitive function of patients, enhance their ability to perform daily activities, and reduce inflammatory responses.

孑遗植物银杏(Ginkgo biloba L.)伴性光合生理特征与进化生态

银杏类是一类古老的雌雄异株植物,目前仅存单科单属。由于银杏在系统发育等方面的独特地位,吸引了科学家从不同方面进行了广泛的研究。在适宜的条件下,测定了生长在野外环境下的成年银杏的雌雄个体的光合特性。研究结果表明,银杏的雌雄植株对光具有相同的表现趋势,光饱和点,光补偿点,光下呼吸速率等均没有明显差异。但是雌性银杏的净光合速率明显大于雄性银杏的净光合速率。这种差异可能与雌性个体在繁衍后代时需要投入更多的能量有关。研究也表明,在与其它的裸子植物和被子植物比较时,银杏的光合能力并没有明显的弱势,因此光合能力可能并不是银杏在第三纪分布受限的直接原因.

孑遗植物银杏(Ginkgo biloba L.)雌雄株水分生理特征初步研究

[目的]研究银杏雌雄株在水分生理上的差异，探讨其水分利用策略，以及在进化过程中的重要作用。[方法]对银杏雌雄株的树干液流（sap flow）、气孔导度（Gs）、蒸腾速率（Tr）以及水分利用效率（WUE）进行对比研究。[结果]雌雄银杏的液流昼夜进程具有相似的规律，且白天雌雄株液流基本相同，夜间雄株液流大于雌株；不同性别银杏的丹和岱同样呈现为早晚高、午间低的曲线趋势，早晚雌株有比雄株更高的Tr和Gs，但中午11：00-14：00之间雌株的值低于雄株；雌雄银杏的WUE曲线变化趋势相似，但雌株的平均水分利用效率略小于雄株。[结论]与其他伴性植物比较分析得知，不同性别银杏的水分生理有较强的趋同特征。这可能是在长期进化的压力下，雌雄异株植物的一种生存策略。

孑遗植物银杏(Ginkgo biloba L.)雌雄株水分生理特征研究(英文)

[Objective] The experiment aimed to study the difference of water physiology of male and female Ginkgo biloba L. for discussing the strategy of water utilization as well as the important role of this difference during evolution process. [Method] The stem sap flow, stomatal conductance(Gs), transpiration rate(Tr) and water use efficiency (WUE) of male and female Ginkgo biloba L. were comparatively studied. [Result] The day-night processes of flow on male and female Ginkgo biloba L. were similar. The flow on male and female Ginkgo biloba L. in day were almost same while the flow at night on male Ginkgo biloba L. was bigger than that on female Ginkgo biloba L. The Tr and Gs of male and female Ginkgo biloba L. were high in morning and at night but low at noon ,while Tr and Gs of female Ginkgo biloba L. in morning and at night were higher than these of male Ginkgo biloba L. at the same time point. However, these indexes of female plant were lower than these of male plant from 11:00 to 14:00. WUE changing trends of male and female Ginkgo biloba L. were similar, while average water utilization rate of female Ginkgo biloba L. was slightly lower than that of male Ginkgo biloba L. [Conclusion] Compared with other companion plants, water physiology of male and female Ginkgo biloba L. had strong homoplasy. The phenomenon might be a survival strategy of dioecious plants under long term evolutionary pressure.

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.

Physiological and Transcriptomic Changes During Autumn Coloration and Senescence in Ginkgo biloba Leaves

Autumn leaf senescence and coloration is a complex process and a striking natural phenomenon.Here,through biology approach integrating transcriptomic analyses in Ginkgo biloba,we determined that the content of chlorophyll decreased during leaf senescence,while carotenoid components increased until late October in the turning stage(TS)and then decreased in the yellow leaf stage(YS).Simultaneously,chlorophyll biosynthesis genes exhibited significantly lower expression levels while chlorophyll degradation genes showed increased expression from the green leaf stage(GS)to YS.However,carotenoid biosynthesis-related genes showed enhanced expression,especially in TS.An analysis of the expression of genes related to senescence demonstrated that the expression levels of most abscisic acid-and jasmonic acid-related genes,autophagy,WRKY,and NAC genes increased,whereas cytoskeleton-,photosynthesis-,and antioxidation-related genes decreased from GS to YS.Furthermore,G.biloba seedlings exogenously treated with abscisic acid,jasmonic acid,or ultraviolet-B radiation all showed obvious color variation and senescence symptoms.We used these exogenously seedlings to further validate the function of several genes involved in chlorophyll biosynthesis and senescence.Taken together,these results contribute to the elucidation of the molecular mechanisms of leaf coloration and senescence in G.biloba as well as in the identification of candidate genes involved in this process.

Experimental Study on Inhibition of Neuronal Toxical Effect of Levodopa by Ginkgo Biloba Extract on Parkinson Disease in Rats

In order to observe neuronal toxical effect of Levodopa and investigate if using Levodopa together with Ginkgo Bilobar Extract (EGb) would be an workable method to treat Parkinson disease, rat models of Parkinson disease (PD) were made by injecting 6-OHDA stereotaxically to right side of the mesencephic ventral tegmental area (VTA) and substantia nigra pars compacta (SNc). Rotational behavioral observation, TUNEL, immunocytochemistry, Nissl's body staining were performed to measure the difference between group treated by Levodopa (50 mg/kg every day for 3 days, 5 days, 7 days, L-dopa group) and group treated by Levodopa combined with EGb (100 mg/ kg every day, E-D group). The results showed that in the L-dopa group, the numbers of apoptosis of substantial nigra, rings of rotational behavior were more than those in the E-D group (P<0. 05). The numbers of Nissl's cells in L-dopa group were fewer than in E-D group (P<0. 05). The results suggested that Levodopa had neur toxic effect and EGb may decrease the toxicity of levodopa. The combined use of EGb with Levodopa may be a workable method to treat PD and may be better than using Levodopa alone.

Extraction Process and Content Determination of Total Flavonoids in Ginkgo( Ginkgo biloba L. ) Leaves

[Objectives]This study was conducted to study the extraction process and the content determination of flavonoids in ginkgo( Ginkgo biloba L.) leaves.[Methods]Ethanol extraction and methanol extraction of total flavonoids in ginkgo leaves were studied,and the optimal extraction conditions for flavonoids were determined by orthogonal test; and with quercetin as reference substance,total flavonoid content in ginkgo leaves was determined by UV spectrophotometry.[Results]The optimal extraction process was 4 h of Soxhlet extraction with methanol; and the total flavonoid contents had a good linear relation in the range of 0. 006 5-0. 039 mg/ml( R^2= 0. 999 9),the average content was stabilized at 1. 135%,and the average recovery of the method was 102. 0%. [Conclusions]This study selected the optimal extraction process for total flavonoids in ginkgo leaves. The test method is simple with high accuracy and precision,and is suitable for the extraction and determination of total flavonoids in ginkgo leaves.

Dynamic changes of telomeric restriction fragment (TRF) lengths in cells during the developmental process from embryos to seedlings and a comparison with the embryonal calli in Ginkgo biloba L.

Telomeres are the structures that locate at the terminals of linear eukaryotic chromosomes. They can play essential roles in many cellular processes. The terminal location ofArabidopsis-type TTTAGGG tandem repeats were thought to be highly conserved. The terminal location of Ginkgo biloba L. consisting of TTTAGGG tandem repeats, were confirmed by Bal31 exonuclease degradation and Southern blotting. By comparing telomeric restriction fragment （TRF） lengths at different developmental stages from embryos to seedlings, a fluctuant tendency towards variation was found in these samples. The TRF length of embryos was also compared with that of embryonal calli and an upward trend was discovered in callus culture. The results suggest that there should be a telomerase mechanism or/and ALT mechanism for the maintenance of telomere length.

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.

Content Determination of Quercetin in Ferment of Ginkgo biloba L. Leaves by HPLC

[Objectives] This study aimed to determine the content of quercetin in ferment of Ginkgo biloba L.leaves.[Methods]Bacillus licheniformis was selected for solid-state fermentation of G.biloba leaf powder,and the content of quercetin in ferment of G.biloba leaves was determined by reversed-phase high-performance liquid chromatography.First,the flavonoid glycosides were extracted with methanol.Then,the flavonoid glycosides were hydrolyzed with hydrochloric acid to prepare the test solution.The chromatographic conditions were as follows:Platisil ODS C_(18) column(150 mm × 4.6 mm,5 μm);V_(methonal)∶V_(water)(0.4% phosphoric acid solution) =55∶45;flow rate of 1 m L/min;Shimadzu UV detector;detection wavelength of 360 nm.[Results] Quercetin was used as a reference substance.In the range of 0.002 6-0.036 0 g/L,there was a good linear relationship,with correlation coefficient of 0.999 8 and RSD of 1.26%.[Conclusions] This method is simple,easy to operate,accurate,and reproducible.It is suitable for the determination of quercetin content in G.biloba leaves.

Effects of Coupling Treatment of Acid and Aluminum under Soilless Culture on Water Content of Ginkgo biloba L.

In this experiment,two-factor 7-level uniform design scheme was applied.Separate treatment of each factor was performed for single effect test.Then,it analyzed the effects of acid and aluminum stress under soilless culture on the water content of plant during the growth of Ginkgo biloba L.seedlings,so as to provide a theoretical basis for studying the connection between plant water and physiological stress.The results showed that the water content of G.biloba plants declined with the decrease of p H,but the water content of G.biloba plants declined with the increase in the concentration of aluminum treatment,indicating that the stronger the acid and aluminum stress,the lower the water content of the plants,affecting the normal absorption of water content of G.biloba plants,accordingly leading to lack of water.According to the analysis of the degree of influence on the water content,p H is the first factor influencing the growth of G.biloba plants.If p H is lower than 3.5,G.biloba plants will not growth,while p H is higher than 5.0,aluminum has no significant effect on the water content of G.biloba plants.In conclusion,the water content of G.biloba plants can be used as an essential indicator for acid and aluminum stress.

Determination of the Content of Kaempferol from Fermented Ginkgo( Ginkgo biloba L.) Leaves

[Objectives] This study was conducted to determine kaempferol content in ginkgo( Ginkgo biloba L.) leaves subjected to microbial fermentation.[Methods]Bacillus licheniformis was selected for solid-state fermentation of ginkgo leaves,and the content of kaempferol in ginkgo leaves was determined by RPHPLC method. At first,methanol was used to extract flavonoid glycosides,which were then hydrolyzed by hydrochloric acid solution. HPLC was performed with Platisil ODS column C18( 150 mm ×4. 6 mm,5 μm) using mobile phase Vmethanol∶ Vwater( 0. 4% phosphoric acid solution) = 55∶45 at a flow rate of 1 ml/min,and the eluate was detected with a shimadzu HPLC ultraviolet detector at 360 nm. [Results]With kaempferol as the reference substance,the correlation coefficient was0. 999 2 in the range of 0. 001 06-0. 016 96 g/L. The content in the fermented product was less than that in the non-fermented product by 28%. [Conclusions]The method is simple,accurate,and is suitable for determination of kaempferol. This study will provide an experimental basis for the development and utilization of ginkgo.

Cloning and Analysis of Telomere-associated Sequences of Ginkgo biloba L..

Total genomic DNA was extracted from leaves of Ginkgo biloba L. by the method of hot CTAB. After determining quantification of DNA sample by microclorimetric spectrophotography, Arabidopsis-type telomere primer and Sau3AⅠ cassette primer were used to isolate telomere-associated sequences from G. biloba L. by the method of cassette-ligation-mediated polymerase chain reaction (PCR). Using this method, two telomere-associated sequences, TAS1 and TAS2, were isolated. The authors preformed Southern hybridization of EcoRⅠ-treated G. biloba genomic DNA with each clone. The hybridization pattern showed that the clones obtained were derived from G. biloba genomic DNA. There are the Arabidopsis-type TTTAGGG tandem repeats in telomeres of G. biloba.

Historical Changes of Ginkgo Biloba L. Culture

Ginkgo Biloba L.is a rare species endemic to China,strengthening the study of Ginkgo culture is of great significance to eco-economic development.This paper uses the historical research methods to study the Ginkgo and its cultural development process in China.According to the characteristics of the development of Ginkgo culture,the process can be divided into three stages:Shang and Zhou Dynasties to the Northern and Southern Dynasties(which is named theological era),Sui and Tang Dynasties to early Qing Dynasty(which is named the literature era),and the modern China which is named the scientific era.The history of Ginkgo culture is a history of Ginkgo being gradually recognized,and the Ginkgo culture's connotation and extension are gradually deepened and developed.The construction of Ginkgo today's culture should be in the inheritance of historical culture,and combined with the needs of the times comprehensive innovation,take the science and human harmonious development road.

Clinical effect of long term aerobic exercise combined with Ginkgo Biloba Leaves Extract Tablets in the treatment of arteriosclerosis

Objective: To investigate the clinical effect of long-term aerobic exercise combined with Ginkgo biloba leaves on atherosclerosis. Methods: 104 patients with carotid atherosclerosis admitted to the Affiliated Hospital of Xi'an Medical College from January 2016 to January 2018 were randomly divided into exercise training group and control group, 52 cases in each group. The control group was treated with Ginkgo biloba leaves, and the exercise training group was treated with long-term aerobic exercise combined with Ginkgo biloba leaves. Both groups were treated for 6 months. The therapeutic effects of the two groups were compared. Results: After treatment, the levels of TC, TG and LDL-C in the two groups were lower than those before treatment, and the levels of HDL-C were higher than those before treatment (P<0.05);after treatment, the levels of TC, TG and LDL-C in the exercise training group were lower than those in the control group, and the levels of HDL-C were higher than those in the control group, with significant difference between the two groups (P<0.05). After treatment, the CIM T and plaque area of the two groups were smaller than those before treatment, and the blood velocity of cerebral artery was higher than that before treatment (P<0.05);after treatment, the CIM T and plaque area of the exercise training group were smaller than those of the control group, and the blood velocity of left middle cerebral artery, right middle cerebral artery and basilar artery were higher than those of the control group. The difference was statistically significant (P<0.05). Conclusion: Long-term aerobic exercise combined with Ginkgo biloba leaf therapy can effectively reduce the blood lipid concentration in patients with carotid atherosclerosis and improve cerebral blood supply.

银杏叶总黄酮绿色提取工艺及抗氧化活性研究

采用单因素实验考察了低共熔溶剂种类、液料比、提取温度、n(氯化胆碱)∶n(尿素)、低共熔溶剂含水量、提取时间对银杏叶总黄酮提取率的影响,通过响应面法进一步优化了银杏叶总黄酮的提取工艺,并通过测定银杏叶总黄酮对DPPH自由基、ABTS自由基、·OH、·O_(2)^(-)的清除能力,金属离子螯合能力及总还原能力综合评价其抗氧化活性。结果表明,最优提取工艺为:液料比30∶1(mL∶g)、提取温度57℃、n(氯化胆碱)∶n(尿素)为1∶1.6、低共熔溶剂含水量40%、提取时间60 min,在此条件下,银杏叶总黄酮提取率为1.062%,且其抗氧化活性显著优于传统乙醇提取工艺。

银杏叶提取物促进大鼠激素性股骨头坏死中H亚型微血管形成的实验研究

目的观察银杏叶提取物在大鼠激素性股骨头坏死(SONFH)模型中对H亚型微血管的影响。方法采用脂多糖联合甲泼尼龙法复制SONFH模型。将50只大鼠分为空白组、模型组,以及银杏叶提取物高、中、低剂量组,每组10只。银杏叶提取物高、中、低剂量组分别予0.88、0.43、0.21 mL/kg银杏叶提取物溶液尾静脉注射,空白组和模型组分别予等量生理盐水尾静脉注射。4周时Micro-CT断层扫描和HE染色观察股骨头外形、骨小梁结构及囊性变情况;治疗8周后,采用HE染色观察检测股骨头软骨表面光滑度和骨小梁密度,免疫荧光染色观察检测股骨头血小板-内皮细胞黏附分子(CD31)和内皮黏蛋白(Emcn)表达。结果4周后Micro-CT断层扫描可见大鼠股骨头变形,骨小梁稀疏;HE染色结果可见骨小梁稀疏、断裂、股骨头坏死,Micro-CT断层扫描与HE染色结果一致,表明大鼠SONFH模型复制成功。8周后HE染色结果显示,与空白组相比,银杏叶提取物高、中、低剂量组及模型组均有不同程度股骨头坏死,但银杏叶提取物高、中、低剂量组与模型组比较,骨小梁相对密集,股骨头坏死减少。各组大鼠CD31及Emcn阳性率比较,差异均有统计学意义(P<0.05),模型组CD31和Emcn阳性率较空白组及银杏叶提取物高、中、低剂量组均降低(P<0.05),银杏叶提取物高剂量组CD31阳性率与空白组比较,差异无统计学意义(P>0.05),但Emcn阳性率高于空白组(P<0.05),银杏叶提取物高剂量组CD31和Emcn阳性率与中、低剂量组比较,差异均有统计学意义(P<0.05),但中剂量与低剂量比较,差异无统计学意义(P>0.05)。结论银杏叶提取物可促进大鼠SONFH模型中H亚型微血管形成,改善股骨头周围血循,促进血管成骨耦合,延缓疾病进程。