Ginkgolide A attenuated apoptosis via inhibition of oxidative stress in mice with traumatic brain injury.

Traumatic brain injury (TBI) is the main cause of death among young adults and the main cause of mortality and disability for all ages groups worldwide. Ginkgolides terpenoid compounds unique to Ginkgo biloba, which have protective effects on cardiovascular and cerebrovascular diseases. The aim of this study is to investigate whether ginkgolide A (GA) can improve TBI in mice and whether it can alleviate cell apoptosis in the brain of TBI mice by reducing oxidative stress. Mice received TBI and GA administration for 7 days. Neurological deficits were monitored and brain tissues were examined for molecular pathological markers. TBI mice had more severer neurobehavioral deficits compared with sham group, which could be improved by administration of GA. GA administration improveed Modified Neurological Severity Scale (mNSS) scores, Grid-Walking test and Rotarod test of TBI mice. The apoptosis increased in TBI mice, and reduced after GA treatment. The biomarkers of oxidative stress 8-OHdG and malondialdehyde (MDA) in the brain of TBI mice increased, while SOD reduced. These changes were reversed after GA administration. These outcomes showed that GA could raise neurobehavioral deficiency of TBI mice. GA treatment could attenuate apoptosis in TBI mice by reducing oxidative stress.

Ginkgolide A downregulates transient receptor potential (melastatin) 2 to protect cisplatin-induced acute kidney injury in rats through the TWEAK/Fn14 pathway: Ginkgolide A improve acute renal injury.

PURPOSE: In order to seek effective drugs for treating cisplatin-induced acute renal injury and explore the corresponding potential mechanism. METHODS: Mouse kidney injury model was established by intraperitoneal injection of 20 mg/kg cisplatin. The temporal expression of TRPM2 and the regulation of Ginkgolide A on its expression were analyzed by western blot. In order to perform the mechanical analysis, we used TRPM2-KO knockout mice. In this study, we evaluated the repair effect of GA on acute kidney injury through renal function factors, inflammatory factors and calcium and potassium content. Pathological injury and cell apoptosis were detected by H&E and TUNEL, respectively. RESULT: Ginkgolide A inhibited inflammatory reaction and excessive oxidative stress, reduced renal function parameters, and improved pathological injury. Meanwhile, we also found that the repair effect of Ginkgolide A on renal injury is related to TRPM2, and Ginkgolide A downregulated TRPM2 expression and inactivated TWEAK/Fn14 pathway in cisplatin-induced renal injury model. We also found that inhibition of TWEAK/Fn14 pathway was more effective in TRPM2-KO mice than TRPM2-WT mice. CONCLUSION: Ginkgolide A was the effective therapeutic drug for cisplatin-induced renal injury through acting on TRPM2, and TWEAK/Fn14 pathway was the downstream pathway of Ginkgolide A in acute renal injury, and Ginkgolide A inhibited TWEAK/Fn14 pathway in cisplatin-induced renal injury model.

Ginkgolide A improves the pleiotropic function and reinforces the neuroprotective effects by mesenchymal stem cell-derived exosomes in 6-OHDA-induced cell model of Parkinson's disease.

Parkinson's disease (PD) is a common disorder attributed to the loss of midbrain dopamine (mDA) neurons and reduced dopamine secretion. Currently, the treatment regimes for PD comprise deep brain stimulations, however, it attenuates the PD progression marginally and does not improve neuronal cell death. We investigated the function of Ginkgolide A (GA) to reinforce Wharton's Jelly-derived mesenchymal stem cells (WJMSCs) for treating the in vitro model of PD. GA enhanced the self-renewal, proliferation, and cell homing function of WJMSCs as assessed by MTT and transwell co-culture assay with a neuroblastoma cell line. GA pre-treated WJMSCs can restore 6-hydroxydopamine (6-OHDA)-induced cell death in a co-culture assay. Furthermore, exosomes isolated from GA pre-treated WJMSCs significantly rescued 6-OHDA-induced cell death as determined by MTT assay, flow cytometry, and TUNEL assay. Western blotting showed that apoptosis-related proteins were decreased following GA-WJMSCs exosomal treatment which further improved mitochondrial dysfunction. We further demonstrated that exosomes isolated from GA-WJMSCs could restore autophagy using immunofluorescence staining and immunoblotting assay. Finally, we used the alpha-synuclein recombinant protein and found that exosomes derived from GA-WJMSCs led to the reduced aggregation of alpha-synuclein compared to that in control. Our results suggested that GA could be a potential candidate for strengthening stem cell and exosome therapy for PD.

Ginkgolide A targets forkhead box O1 to protect against lipopolysaccharide-induced septic cardiomyopathy.

Ginkgolide A (GA), a main terpenoid extracted from Ginkgo biloba, possesses biological activities such as anti-inflammatory, anti-tumor, and liver protection. However, the inhibitory effects of GA on septic cardiomyopathy remain unclear. This study aimed to explore the effects and mechanisms of GA in countering sepsis-induced cardiac dysfunction and injury. In lipopolysaccharide (LPS)-induced mouse model, GA alleviated mitochondrial injury and cardiac dysfunction. GA also significantly reduced the production of inflammatory and apoptotic cells, the release of inflammatory indicators, and the expression of oxidative stress-associated and apoptosis-associated markers, but increased the expression of pivotal antioxidant enzymes in hearts from LPS group. These results were consistent with those of in vitro experiments based on H9C2 cells. Database analysis and molecular docking suggested that FoxO1 was targeted by GA, as shown by stable hydrogen bonds formed between GA with SER-39 and ASN-29 of FoxO1. GA reversed LPS-induced downregulation of nucleus FoxO1 and upregulation of p-FoxO1 in H9C2 cells. FoxO1 knockdown abolished the protective properties of GA in vitro. KLF15, TXN2, NOTCH1, and XBP1, as the downstream genes of FoxO1, also exerted protective effects. We concluded that GA could alleviate LPS-induced septic cardiomyopathy via binding to FoxO1 to attenuate cardiomyocyte inflammation, oxidative stress, and apoptosis.

γ-Aminobutyric acid promotes the inhibition of hair growth induced by chronic restraint stress.

Stress plays a critical role in hair loss, although the underlying mechanisms are largely unknown. γ-aminobutyric acid (GABA) has been reported to be associated with stress; however, whether it affects stress-induced hair growth inhibition is unclear. This study aimed to investigate the potential roles and mechanisms of action of GABA in chronic restraint stress (CRS)-induced hair growth inhibition. We performed RNA-seq analysis and found that differentially expressed genes (DEGs) associated with neuroactive ligand-receptor interaction, including genes related to GABA receptors, significantly changed after mice were treated with CRS. Targeted metabolomics analysis and enzyme-linked immunosorbent assay (ELISA) also showed that GABA levels in back skin tissues and serum significantly elevated in the CRS group. Notably, CRS-induced hair growth inhibition got aggravated by GABA and alleviated through GABAA antagonists, such as picrotoxin and ginkgolide A. RNA sequencing analysis revealed that DEGs related to the cell cycle, DNA replication, purine metabolism, and pyrimidine metabolism pathways were significantly downregulated in dermal papilla (DP) cells after GABA treatment. Moreover, ginkgolide A, a GABAA antagonist extracted from the leaves of Ginkgo biloba, promoted the cell cycle of DP cells. Therefore, the present study demonstrated that the increase in GABA could promote CRS-induced hair growth inhibition by downregulating the cell cycle of DP cells and suggested that ginkgolide A may be a promising therapeutic drug for hair loss.

Ginkgolide A alleviates cardiac remodeling in mice with myocardial infarction via binding to matrix metalloproteinase-9 to attenuate inflammation.

Ginkgolides are terpenoids peculiar to Ginkgo biloba, which have protective properties against cardiac diseases. This study aims to explore whether ginkgolide A (GA) could improve cardiac dysfunction of MI mice, and whether it could alleviate cardiac remodeling via binding to matrix metalloproteinase-9 (MMP9) to attenuate inflammation. Cardiac remodeling in mice induced by left coronary artery ligation were used in the in vivo model, and angiotensin (Ang) II-induced cardiac fibroblasts (NRCFs) and cardiomyocytes (NRCMs) isolated from neonatal rats were used in in vitro fibrosis and hypertrophy models, respectively. Cardiac dysfunction and fibrosis in MI mice were alleviated by GA treatment. Upregulations of collagen I (Col I), collagen III (Col III) and fibronectin in NRCFs, and enhanced levels of atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP) and beta-myosin heavy chain (ß-MHC) in NRCMs were inhibited by GA treatment. A total of 100 potential targets were found in 5 databases (TCMSP, BATMAN-TCM, PharmMapper, ETCM and SWISS Target). According to Protein Data Bank database GA could form hydrogen bonds between LYS65, GLU157, ASN17, ARG109, ARG106 of MMP9 protein, a target of GA. The regulatory role of GA in downregulating Col I, Col III, fibronectin in NRCFs, and enhancing levels of ANP, BNP and ß-MHC in NRCMs were reversed by MMP9 overexpression, so as the downregulation of IL-1ß, IL-6 and TNF-α in Ang II-induced NRCFs and NRCMs. GA could alleviate cardiac dysfunction and remodeling via binding to MMP9 to attenuate inflammation. Therefore, GA is a potential drug for cardiac remodeling therapy.

Ginkgolide A attenuates sepsis-associated kidney damage via upregulating microRNA-25 with NADPH oxidase 4 as the target.

The aim of the present study was to explore the effects of Ginkgolide A (GA) on renal function of mice with sepsis and whether GA could attenuate sepsis-associated inflammation and apoptosis in kidney via upregulating microRNA (miR)-25 with NADPH oxidase 4 (Nox4) as the target. Experiments were carried out on lipopolysaccharide (LPS)-treated mice and kidney tubular (NRK-52E) cells. GA significantly inhibited the increases of creatinine (Cr), blood urea nitrogen (BUN) and cystatin C (CysC) in the serum of LPS-treated mice. The increases of inflammatory factors including tumor necrosis factor (TNF)-α, interleukin (IL)-1ß and IL-6 in the kidneys of LPS-treated mice or NRK-52E cells were inhibited by GA administration. The changes of cleaved-caspase 3, cleaved-caspase 8, Bax, Bcl2 in mouse kidney and NRK-52E cells treated by LPS were reversed by GA administration. The sepsis-induced decrease of miR-25 was enhanced by GA treatment. The LPS-induced increases of inflammatory factors and apoptosis in mouse kidney or NRK-52E cells were attenuated after miR-25 agomiR administration. The bioinformatics analysis and luciferase reporter assays showed that Nox4 was a direct target gene of miR-25. Treatment with miR-25 inhibited Nox4 expression, while Nox4 over-expression reversed the inhibiting effects of miR-25 agomiR on LPS-induced increases of inflammatory factors and apoptosis in NRK-52E cells. These results indicated that GA could improve sepsis-induced renal damage by attenuating renal inflammation and apoptosis via upregulating miR-25 with Nox4 as the target.

Therapeutic promises of ginkgolide A: A literature-based review.

Ginkgolide A is a highly active platelet activating factor antagonist cage molecule which was isolated from the leaves of the Ginkgo biloba L. It is known for its inflammatory and immunological potentials. This review aims to sketch a current scenario on its therapeutic activities on the basis of scientific reports in the databases. A total 30 articles included in this review suggests that ginkgolide A has many important biological activities, including anti-inflammatory, anticancer, anxiolytic-like, anti-atherosclerosis and anti-atherombosis, neuro- and hepatoprotective effects. There is a lack of its toxicological (e.g. toxicity, cytotoxicity, genotoxicity and mutagenitcity) profile. In conclusion, ginkgolide A may be one of the potential therapeutic lead compounds, especially for the treatment of cardiovascular, hepatological, and neurological diseases and disorders. More studies are necessary on this hopeful therapeutic agent.

Identification of the in vivo relevant dissolution media for the three active components in EGb 761 tablet for better correlation with their pharmacokinetics in healthy subjects.

Although EGb 761, the standardized dry extract of Ginkgo biloba leaves, exhibited numerous pharmacological activities and widely used in Asia, European and North America, the quality control of its dosage forms such as tablet mainly relies on monitoring the contents of the active marker components, namely quercetin, kaempferol, isorhamnetin, bilobalide, ginkgolide A, ginkgolide B and ginkgolide C. So far, the in vitro dissolution profiles of EGb761 tablet were barely used to monitor its quality and how these dissolution profiles correlate with their in vivo pharmacokinetics was not known. Thus, the present study was proposed aiming to 1) develop the in vitro-in vivo correlations (IVIVCs) for the marker components in EGb 761 tablet; 2) identify the in vivo relevant dissolution media for the marker components in EGb 761 tablet based on the established IVIVCs. The content analyses of the marker components in EGb 761 tablet was first carried out. Then, the dissolution profiles were further obtained using paddle method of United States Pharmacopeia for bilobalide, ginkgolides A, and ginkgolide B, that have previously reported human plasma pharmacokinetics after EGb 761 tablet oral administrations. About seven different media including 0.1 M hydrochloric acid (HCl), acetate buffer, H2O, fasted state simulated gastric fluid (FaSSGF), fasted state simulated intestinal fluid version 2 (FaSSIF-V2), fed state simulated intestinal fluid version 2 (FeSSIF-V2), and sequential medium (0.1 M HCl for 2 h with pH adjusted to 7 for another 2 h) were tested in the current investigation. The obtained in vitro dissolution profiles of bilobalide, ginkgolides A and ginkgolide B from EGb 761 tablet were first fitted with four dissolution models, namely Weibull, Double Weibull, Hill and Makoid-Banakar, to obtain the best-fit model for each component in each medium. The human plasma concentration versus time profiles of the above three components were then inputted into the Phoenix WinNonlin IVIVC Toolkit to obtain their in vivo absorption profiles using numerical deconvolution. The best-fit dissolution profiles of each marker component in the seven studied media were further used to correlate with its obtained in vivo absorption profile by the linear correlation models to establish the corresponding IVIVCs in each studied medium. Finally, the best in vivo correlated medium for each investigated marker component was selected based on their adjusted correlation coefficients, Akaike Information Criterion (AIC) and Schwarz's Bayesian Criterion (SBC) values. As a result, the dissolution profiles of bilobalide, ginkgolide A, ginkgolide B from EGb 761 tablet in 0.1 M HCl, FaSSGF, FaSSIF-V2 demonstrated the best correlation with their in vivo absorption profiles, respectively. Our current studies for the first time applied the concept of IVIVC to EGb 761 tablet and successfully identified the in vivo relevant dissolution media for its three active marker components to improve its quality control.

Ginkgolide-A attenuates bacterial translocation through activating PXR and improving antimicrobial peptide Reg 3A in experimental cirrhosis.

BACKGROUND AND AIMS: Bacterial translocation (BT) is strongly associated with disease progression and poor outcome in cirrhotic patients. The role of Pregnane X receptor (PXR) in regulating bacterial translocation in cirrhosis is unknown. We previously showed that Ginkgolide-A (GA), a natural PXR ligand, attenuated BT in cirrhotic mice by abrogating inflammation along the gut-liver-axis, and by protecting small intestinal tight junctions (TJ). Here, we aimed to investigate the effect of GA in activating PXR and associated antimicrobial peptides (AMPs) in regulating BT in experimental cirrhosis. METHODS: Male Swiss albino mice were administered CCl4 (0.5 mL/kg body-weight, i.p twice a week) for 12 consecutive weeks. After the 12th week, mice were randomized and administered with GA (100-mg/kg body-weight, oral) every-day for 2 weeks. At termination, blood, gut and liver tissues were collected for molecular studies. RESULTS: GA treatment to cirrhotic mice significantly increased the expression of small intestinal PXR and Regenerating family member 3 alpha (Reg3A), which were otherwise reduced in CCl4 cirrhotic mice. Moreover, compared to naive mice a significantly reduced Lactobacillus, and increased Bacteroides and Enterococcus 16s rRNA levels were observed in the small intestine and liver of cirrhotic mice. Treatment with GA to cirrhotic mice significantly reduced intestinal overgrowth and translocation of Enterococcus and Bacteroides to the liver. Furthermore, GA treatment significantly attenuated intestinal permeability and BT marker soluble-CD14 (sCD14), which were increased in CCl4 cirrhotic mice. CONCLUSION: The study showed for the first time that, GA treatment to cirrhotic rodents attenuates BT, by improving PXR and Reg3A expression.

Effect of GSK-137647A, the first non-carboxylic FFA4 agonist, on the osteogenic and adipogenic differentiation of bone mesenchymal stem cells in db/db mice.

OBJECTIVE: To investigate the effect of GSK-137647A, the first non-carboxylic FFA4 agonist, on osteogenic and adipogenic differentiation of bone mesenchymal stem cells (BMSCs) of db/db mice. METHODS: Bone mesenchymal stem cells were extracted from 8-week-old db/db mice. Cell Counting Kit-8 was used to evaluate the toxicity of GSK-137647A on BMSCs, and the optimal concentration of GSK-137647A was selected to investigate the osteogenic and adipogenic differentiation of BMSCs, and relevant indicators of osteoblasts and adipocytes were detected. KEY FINDINGS: GSK-137647A had no significant toxicity on cell growth and proliferation. Moreover, GSK-137647A showed a significant increase in mineralization of BMSCs differentiated osteoblasts compared to the control group and elevated the alkaline phosphatase (ALP) activity in a time-dependent manner. Meanwhile, the treatment of GSK-137647A decreased the adipogenic differentiation of BMSCs. The expression levels of ALP, runt-related transcription factor 2, bone morphogenetic protein 4, osterix and ß-catenin were significantly increased in GSK-137647A-treated group, while the gene and protein levels of peroxisome proliferator-activated receptor γ and CCAAT/enhancer binding protein α were significantly reduced. CONCLUSIONS: All of these results demonstrated that GSK-137647A suppressed the adipogenic differentiation and promoted osteogenic differentiation of BMSCs, which is partly attributed to the increased expression of ß-catenin in wingless/integrated signalling pathway.

Pregnane X receptor activation by its natural ligand Ginkgolide-A improves tight junction proteins expression and attenuates bacterial translocation in cirrhosis.

BACKGROUND AND AIMS: Pregnane X receptor (PXR) is a ligand-activated transcription factor and nuclear receptor expressed ubiquitously along gut-liver-axis. Inflammatory bowel disorders have been reported to implicate PXR in maintaining tight junction (TJ) integrity and countering inflammation. However, the hepatoprotective role of PXR activation in soothing bacterial translocation in liver cirrhosis has not been explored. Ginkgolide A (GA), a terpene trilactone from Ginkgo Biloba extract, is a natural ligand of rodent and human PXR. This study aims to investigate the effect of GA in activating PXR and improving associated tight junction integrity and reducing bacterial translocation in gut-liver axis of CCl4 induced cirrhosis model. METHODS: Swiss albino mice were administered with CCl4 (0.5 ml/kg body weight, i.p) in corn oil for 12 weeks at an interval of two times a week. Following ascites induction, mice were randomized & administered 100 mg/kg body weight of GA through oral gavage for 2 weeks. At termination, blood, gut and liver tissues were collected for biochemical and molecular studies. RESULTS: When compared to naïve mice, protein expression of hepatic and small intestinal PXR, CYP3A, ZO-1 and occludin were found to be significantly (p < 0.01) decreased in CCl4 induced cirrhotic mice. Treatment with GA to cirrhotic mice significantly (p < 0.05) induced the expression of both hepatic and small intestinal PXR, CYP3A, ZO-1 and Occludin. Furthermore, increased (p < 0.01) hepatic and small intestinal NFκB was observed in CCl4 induced cirrhotic mice that was significantly (p < 0.05) lowered following GA treatment. Over expression of TLR4/MyD88/NFκB axis and its downstream pro-inflammatory mediators TNF-α, IL6 and IFN-γ were observed in CCl4 induced mice, and these indices were abrogated significantly after GA treatment. Furthermore, significantly increased plasma levels of bacterial translocation markers LBP and procalcitonin were found in CCl4 mice, which were reduced significantly (p < 0.05 & p < 0.0001) after GA treatment. CONCLUSION: In conclusion, our data supports the hypothesis that, GA treatment to CCl4 induced cirrhotic mice, activated hepatic and small intestinal PXR and diminished inflammation, thereby improving tight junction integrity and attenuating bacterial translocation.

Clinical pharmacokinetics of Ginkgolides Dropping Pills in healthy subjects / 中草药

Objective: In order to describe the pharmacokinetic profiles of two effective constituents ginkgolide A and ginkgolide B in healthy subjects, and to provide supports for setting out the clinical application of Ginkgolides Dropping Pills. Methods: Ten healthy subjects were enrolled in a randomized and open experimental design. Following a single oral administration of Ginkgolides Dropping Pills, blood samples which were anticoagulated by heparin sodium were collected at predetermined time, and then centrifuged to separate plasma samples. The total concentration of ginkgolide A and ginkgolide B in plasma samples and the lactone concentration of ginkgolide A and ginkgolide B were determined by a verified LC-MS/MS method, the pharmacokinetic parameters were calculated by WinNonlin 6.3 with non-compartment model. Results: After a single oral administration of Ginkgolides Dropping Pills, the tmax of lactone, total concentration of ginkgolide A respectively were (3.05 ± 1.40), (3.40 ± 1.22) h, the Cmax were (84.3 ± 32.8), (92.2 ± 35.0) ng/mL, respectively, and its Cmax ratio was 91.4%. The AUC0-t were (636 ± 183), (753 ± 205) ng∙h/mL, respectively, and its AUC0-t ratio was 84.5%, half-life time (t1/2) were (13.0 ± 10.3), (12.9 ± 8.49) h, respectively. The Tmax of lactone, total concentration of ginkgolide B were (3.15 ± 1.42), (3.35 ± 1.25) h, The Cmax were (74.1 ± 31.5), (148 ± 60.1) ng/mL, respectively, and its Cmax ratio was 50.1%.The AUC0-t were (627 ± 202), (1410 ± 431) ng∙h/mL, respectively, and its AUC0-t ratio was 44.5%, t1/2 were (13.2 ± 5.83), (13.7 ± 5.83) h, respectively. Conclusion: The results demonstrated that ginkgolide A and ginkgolide B were both at a moderate absorption and elimination rate, ginkgolide A mainly existed in human plasma upon lactone, while ginkgolide B presented as hydrolyzed forms with one or two lactone groups hydrolyzed and lactone, and the two forms of ginkgolide B were at equal exposure level after single oral administration of Ginkgolides Dropping Pills.

Mixing Ginkgo biloba Extract with Sesame Extract and Turmeric Oil Increases Bioavailability of Ginkgolide A in Mice Brain.

Ginkgo biloba extract (GBE) is widely used as herbal medicine. Preventive effect of GBE against dementia, including Alzheimer's disease, has been reported. The bioactive compounds in GBE that impart these beneficial effects, flavonoids and terpene lactones, have poor bioavailability. Our previous study found distribution of bioactive compounds of sesame extract in mice brain after mixing it with turmeric oil. Here, we evaluate the distribution of bioactive compounds of GBE by combining it with the mixture of sesame extract and turmeric oil (MST). The content of terpene lactones in mice serum was significantly increased in a dose-dependent manner after administration of GBE. However, the contents of terpene lactones in mice brain were not significantly changed. Concentration of ginkgolide A in mice brain increased significantly when GBE was co-administrated with MST than when GBE was administered alone. These results suggest that MST may be effective in enhancing the bioavailability of ginkgolide A in GBE.

Ginkgolide A Prevents the Amyloid-ß-Induced Depolarization of Cortical Neurons.

Utilizing the N-methyl-d-aspartate (NMDA) receptor antagonist as a strategy, memantine is the only agent available for clinically treating mild to severe Alzheimer's disease (AD). Our aim was to develop novel similar herb-based drugs. Using a screening platform, ginkgolide A (GA), a pure compound extracted from Ginkgo biloba, was found to attenuate amyloid ß (Aß)-induced abnormal depolarization in mouse primary cortical neurons. Using receptor agonists, it was determined that GA inhibits both NMDA receptors and α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors. Furthermore, the Aß-induced increase in c-Jun N-terminal kinase phosphorylation in neurons was prevented by GA. Body weight, glutamate oxaloacetate transaminase, glutamic-pyruvic transaminase, liver histology, and kidney histology were similar when the wild-type/AD animal model mice with and without GA treatment were compared. This pure compound improves the memory of wild-type mice. Our findings indicate that GA has great potential clinically for the treatment of AD because it might target NMDA receptors just like memantine.

Ginkgolide A reduces airway inflammation in a neutrophil-predominant murine model of asthma via suppression of immune response of helper T cell 17 / 中国中西医结合急救杂志

Objective To determine the effects of ginkgolide A (GA) in a neutrophil-predominant murine model of asthma and explore underlying mechanisms. Methods Thirty-five female BALB/c mice were randomly divided into sham operation group (Sham group), asthma group, dexamethasone intervention control group (DEX group), low dose GA intervention group (L-GA group) and high dose GA intervention group (H-GA group), with 7 mice in each group. The asthma model was induced by intraperitoneal injection of 20 μg ovalbumin (OVA) and 75 μL Fluorine complete adjuvant (FCA) on day 0, 14 and 21, and challenged 30 minutes with 5% OVA atomization on days 22-24 consecutively; phosphate buffer (PBS) was sensitized and stimulated in Sham group. The mice in L-GA group and H-GA group were intraperitoneally injected with GA of 40 mg/kg and 80 mg/kg at 1 hour before each challenge, while the mice in DEX group were intraperitoneally injected with dexamethasone of 1 mg/kg. After 24 hours of the last OVA stimulation, the airway resistance was measured at the time of 0, 3, 6, 12, 25, 50 g/L acetylmethacholine aerosol stimulation. The total number of cells and cell classification in bronchoalveolar lavage fluid (BALF) were counted. The transforming growth factor-β1 (TGF-β1) and interleukin-17 (IL-17) in BALF were detected by enzyme linked immunosorbent assay (ELISA). The proportion of helper T cell 17 (Th17) to CD4+ T cell in lung tissue was detected by flow cytometry, and the pathological characteristics of lung tissue were evaluated. Results Compared with the Sham group, the airway hyper responsiveness (AHR), the total cells, the neutrophil counts, the levels of TGF-β1, IL-17 in BALF, and the proportion of Th17 cells in the lung tissue in the asthma group were significantly increased, obvious inflammatory cell infiltration and collagen fiber deposition around airway were observed, and airway inflammation score and mucus score were significantly increased. Compared with the asthma group, low and high doses of GA significantly reduced AHR, and there was a significant difference in airway resistance at the time of 50 g/L acetylmethacholine stimulation (cmH2O·s-1·mL-1: 5.29±0.40, 3.99±0.57 vs. 7.34±0.77, both P < 0.05); the total cells, neutrophil counts, and levels of TGF-β1, IL-17 in BALF, and the proportion of Th17 cells in lung tissue were significantly decreased [total cells count (×104/L): 21.00±1.00, 17.00±1.02 vs. 27.50±2.50; neutrophil count (×104/L): 12.600±0.600, 10.610±0.210 vs. 16.875±1.125; TGF-β1 (ng/L): 371.40±107.80, 289.60±70.76 vs. 551.90±68.34; IL-17 (ng/L): 60.75±11.79, 44.77±7.09 vs. 122.50±38.87; the proportion of Th17 cells: (5.53±0.40)%, (3.76±1.10)% vs. (8.30±1.19)%, all P < 0.05]; inflammatory cell infiltration around the airway and mucus secretion was significantly reduced, airway inflammation score and mucus score were significantly decreased (2.16±0.28, 1.16±0.28 vs. 3.77±0.25; 1.33±0.58, 1.17±0.29 vs. 3.67±0.58, all P < 0.05). The AHR, total cells, neutrophil counts, and IL-17 level in BALF, the proportion of Th17 cells in lung tissue and airway inflammation score decreased more obviously with the increase of GA dosage (all P < 0.05). For index mentioned above, no significant differences were observed between DEX group and asthma group. Conclusion GA treatment was effective in a murine model of neutrophil-predominant asthma via inhibiting response in the immune cells Th17.

Applying risk management to analytical methods for the desorbing process of ginkgo diterpene lactone meglumine injection.

Analysis errors can occur in the desorbing process of ginkgo diterpene lactone meglumine injection (GDMI) by a conventional analysis method, due to several factors, such as easily crystallized samples, solvent volatility, time-consuming sample pre-processing, fixed method, and offline analysis. Based on risk management, near-infrared (NIR) and mid-infrared (MIR) spectroscopy techniques were introduced to solve the above problems with the advantage of timely analysis and non-destructive nature towards samples. The objective of the present study was to identify the feasibility of using NIR or MIR spectroscopy techniques to increase the analysis accuracy of samples from the desorbing process of GDMI. Quantitative models of NIR and MIR were established based on partial least square method and the performances were calculated. Compared to NIR model, MIR model showed greater accuracy and applicability for the analysis of the GDMI desorbing solutions. The relative errors of the concentrations of Ginkgolide A (GA) and Ginkgolide B (GB) were 2.40% and 2.89%, respectively, which were less than 5.00%. The research demonstrated the potential of the MIR spectroscopy technique for the rapid and non-destructive quantitative analysis of the concentrations of GA and GB.

[Effect of compatibility of ginkgolide A, ginkgolide B and ginkgolide K].

To investigate the best active compatibility of ginkgolide A, B and K (GA，GB，GK). The effects of GA, GB, GK alone, combinations of each two of them, and combinations of these three components on platelet-activating factor (PAF)-induced platelet aggregation activity and rat cerebral ischemia reperfusion model (tMCAO) were compared in this study. Different compatibilities of GA, GB and GK could significantly reduce the maximum aggregation rate of PAF-induced platelet aggregation, and the effect was most obvious in combination of the three. Different compatibilities of GA, GB and GK could alleviate the neural function, cerebral infarction volume and cerebral edema in the tMCAO model of rats to different degrees, and the effect of combinations of the three was stronger than those of combinations of two and single use. The combination of all of GA, GB and GK had the strongest effect on nerve injury caused by anti-platelet aggregation in tMCAO rats.

Applying risk management to analytical methods for the desorbing process of ginkgo diterpene lactone meglumine injection / 中国天然药物

Analysis errors can occur in the desorbing process of ginkgo diterpene lactone meglumine injection (GDMI) by a conventional analysis method, due to several factors, such as easily crystallized samples, solvent volatility, time-consuming sample pre-processing, fixed method, and offline analysis. Based on risk management, near-infrared (NIR) and mid-infrared (MIR) spectroscopy techniques were introduced to solve the above problems with the advantage of timely analysis and non-destructive nature towards samples. The objective of the present study was to identify the feasibility of using NIR or MIR spectroscopy techniques to increase the analysis accuracy of samples from the desorbing process of GDMI. Quantitative models of NIR and MIR were established based on partial least square method and the performances were calculated. Compared to NIR model, MIR model showed greater accuracy and applicability for the analysis of the GDMI desorbing solutions. The relative errors of the concentrations of Ginkgolide A (GA) and Ginkgolide B (GB) were 2.40% and 2.89%, respectively, which were less than 5.00%. The research demonstrated the potential of the MIR spectroscopy technique for the rapid and non-destructive quantitative analysis of the concentrations of GA and GB.

Relative bioavailability of terpene lactones in Beagle dogs after ig administration of domestic and imported Ginkgo Leaf Tablets / 中草药

Objective To compare the pharmacokinetic parameters and bioavailability of terpene lactones in Beagle dogs between domestic and imported Ginkgo Leaf Tablets. Methods Beagle dogs were ig administrated demestic and imported Ginkgo Leaf Tablets, and then the plasma of Beagle dogs were detected. LC-MS was used to determine the contents of terpene lactones (including ginkgolide A, ginkgolide B, and ginkgo lactone) in plasma of Beagle dogs. Plasma concentration-time curves were drawn and analyzed by DAS software to obtain pharmacokinetics parameter. Results The area under curve (AUC0-t) of GA, GB, and BB in Beagle dogs after ig administration domestic Ginkgo Leaf Tablets was 51.64, 19.86, and 72.90 ng∙h/mL, while it was 69.98, 24.35, and 169.60 ng∙h/mL after ig administration imported G. biloba leaf extract tablets, respectively. According to the contents of three components in two preparations, the relative bioavailability of GA, GB, and BB of domestic Ginkgo Leaf Tablets respectively was 37.77%, 33.70%, and 95.98%. Conclusion The oral bioavailability of the terpene lactones in imported Ginkgo Leaf Tablets was significantly higher than that of domestic tablets.