Improvement effect of diterpene ginkgolides on acute tMCAO rats and chronic tMCAO rats / 药学学报

To study the cognitive effects of diterpene ginkgolides (DG), transient middle cerebral artery occlusion (tMCAO)-induced rats were established. tMCAO-rats induced by suture method were divided into sham operation group, solvent control group, NBP group, DG group. The animal experiments in the present study were performed in accordance with the Ethical Guidelines of the Laboratory Animal Welfare Ethical Committee of Peking Union Medical College (00000646, 00000635). The effects of DG on tMCAO rats were evaluated by neurological severity score, cerebral infarction volume measurement, step-down and Morris water maze test. In the acute tMCAO rat model, 100 mg·kg-1 DG improved the neural score and infarction volume. In the chronic tMCAO rat model, DG 100 mg·kg-1 significantly improved the survival rate of tMCAO-induced rats. The Morris water maze results showed 100 mg·kg-1 DG decreased the latency of tMCAO-induced rats to find the platform, while the effect was weaker than the NBP. However, DG 30 mg·kg-1 did not show a significant effect. In conclusion, DG exerted a therapeutic effect on transient middle cerebral artery occlusion.

Advance in biosynthesis and metabolic regulation of ginkgolides / 中国中药杂志

Ginkgolides,the unique terpenoids in Ginkgo biloba,have a significant effect on the prevention and treatment of cardiovascular and cerebrovascular diseases. Metabolic regulation and synthetic biology strategies are efficient methods to obtain high-quality ginkgolides. The present study reviewed the cloning and functions of genes related to the biosynthetic pathway of ginkgolides,as well as relevant studies of omics,genetic transformation,and metabolic regulation in recent years,and predicted the research trends and prospects,aiming to provide a reference for discovering the key genes related to the biosynthetic pathway and the biosynthesis of ginkgolides.

Comparative study on the effects of total ginkgo flavonol glycosides and total ginkgolides on neurofunctional recovery in mice with subacute stroke / 药学学报

As a Ginkgo biloba extract preparation, shuxuening injection has a unique advantage in the prevention and treatment of acute and subacute stroke, but its main active ingredient is still unclear. Using a subacute model of stroke in mice constructed earlier, we further explored the contribution and mechanism of the two main components of total ginkgo flavonol glycosides and total ginkgolides in facilitating the neurofunctional recovery in stroke-induced mice. The pharmacodynamics was mainly evaluated by neurobehavioral changes, cerebral infarction volume, blood-brain barrier permeability and brain edema. The pathway and targets were predicted by transcriptome and network pharmacology. Finally, the mechanism was verified at the mRNA and protein levels. The results showed that the beneficial effect of total ginkgolides was greater than that of total ginkgo flavonol glycosides in both the pharmacodynamics and the regulatory mechanism of granulocyte adhesion and diapedesis involving granulocyte colony-stimulating factor (G-CSF), macrophage-1 antigen (MAC-1) and E-selectin. These findings suggest that shuxuening injection may improve the prognosis for mice with subacute stroke by down-regulating G-CSF-mediated granulocyte adhesion and diapedesis pathway mainly through the total ginkgolide components. This finding is expected to provide reference for optimizing prescription and searching for natural drugs for targeting the treatment of ischemic stroke prognosis. The animal experiments in this study followed the regulations of Animal Ethics Committee of Tianjin University of Traditional Chinese Medicine.

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.

The efficacy and safety of diterpene ginkgolides meglumine injection in stroke:a Meta-analysis / 药学实践杂志

Objective Diterpene ginkgolides meglumine injection (DGMI) is widely used in patients with stroke, but its efficacy and safety are not consistent.We performed a Meta-analysis to comprehensively evaluate the efficacy and safety of DG-MI in acute ischemic stroke and recovered stroke.Methods The wanfang, VIP, CNKI and PubMed were searched, the randomized controlled trials (RCTs) were enrolled.Data collection and quality evaluation of the included RCTs were performed according to Cochrane systematic evaluation method.Meta-analysis was performed by using Stata software.Results 9RCTs involving 1 129subjects were included with 706subjects in DGMI treatment group and 423subjects in control group. (1) For acute ischemic stroke, DGMI group had superior effective rate compared to conventional therapy group (RR=1.19, 95％CI:1.09, 1.31, P＜0.000 1), improvement of neurologic impairments (SMD=3.23, 95％CI:2.87, 3.60, P＜0.000 1) and improvement of living quality (SMD=3.23, 95％CI:2.87, 3.60, P＜0.000 1). (2) For recovered stroke, DGMI group had better effective rate than Shuxuening injection group (RR=1.17, 95％CI:1.05, 1.30, P＜0.05) and improvement of neurologic impairments (SMD=-0.69, 95％CI:-0.88, -0.49, P＜0.000 1).There was no significant difference in adverse events between DGMI and control groups (P＞0.05).Conclusion DGMI had superior efficacy over control group for both acute ischemic stroke and recovered stroke.There was no significant difference in adverse events between these two groups.However, we still need better quality RCTs to confirm these results.

Effects of ginkgo diterpene lactone meglumine on learning and memory in old mice and its mecha-nisms / 中华行为医学与脑科学杂志

Objective To study the effects of diterpene ginkgolides meglumine injection ( DGMI) on memory impairment, activation of microglia and astrocytes and inflammatory cytokines in aged mice. Methods Twenty aged mice (22 months old) were randomly divided into two groups:aged mouse group(n=10) and DGMI group(n=10). Another 10 mice (2 months old) were selected as young mouse control group. The mice in DGMI group were received 5 mg/kg DGMI per day by tail vain injection for 4 weeks. The mice in the other two groups were received the same amount normal saline for 4 weeks. The Morris water maze was used to evaluate the function of spacial learning and memory after administration of drugs. The ex-pression of CD11b,GFAP,IL-1β,IL-6,TNF-α and NFκB in mice brain hippocampus were detected by West-ern blot. Results (1) The escape latency time of aged mouse group was significantly longer than that of young mouse control group from the 2nd day to the 7th day(P<0. 01). The times of platform crossing,time and distance in target quadrant of aged mouse group were significantly shorter than those of young mouse group (all P<0. 01). Compared with aged mouse group,DGMI significantly reduced the escape latency time of DGMI group (P<0. 01). DGMI increased the times of platform crossing,time and distance in target quad-rant of aged mouse group (P<0. 01). (2) The expressions of CD11b,GFAP in young mouse control group, aged mouse group and DGMI group were as follows respectively:CD11b:(1. 036±0. 023),(1. 757±0. 046), (1. 214±0. 024);GFAP:(1. 022±0. 071),(1. 344±0. 021),(1. 086±0. 073). DGMI reduced the expres-sion of CD11b and GFAP in hippocampus compared with aged mouse group ( t=5. 556,P<0. 01;t=5. 484, P<0. 01). (3) The expressions of IL-1β,IL-6,TNF-α and NFκB in young mouse control group,aged mouse group and DGMI group were as follows respectively:IL-1β:( 1. 003 ± 0. 057),( 2. 062± 0. 105),( 1. 182± 0. 084);IL-6:(1. 018±0. 024),(1. 583± 0. 052),( 1. 152± 0. 031); TNF-α:( 1. 021± 0. 054),(1. 449± 0. 053),(1. 211±0. 036);p-NFκB:(1. 052±0. 034),(1. 782± 0. 113),( 1. 158± 0. 066). DGMI reduced the expression of p-NFκB(t=6. 547,P<0. 01) and pro-inflammatory cytokines including IL-1β(t=8. 513,P<0. 01),IL-6(t=3. 421,P<0. 01) and TNF-α( t=5. 562,P<0. 01) in hippocampus compared with aged mouse group. Conclusion DGMI can improve the ability of learning and memory in aged mice. The mecha-nism may be related with inhibiting activity of microgliosis,astrocytosis,NFκB and neuroinflammaton.

Study on pharmacokinetics and pharmacodynamics of ginkgolides in rat model with cerebral ischemia injury / 中草药

Objective To study the metabolic process of ginkgolides in rats with cerebral ischemic injury based on pharmacokinetic- pharmacodynamics (PK-PD) binding model. Methods The middle cerebral artery occlusion (MCAO) model was established by the suture method. After reperfusion, rats were randomly assigned to nasal administration, ig administration, and iv administration group.The orbital blood was taken at different time points of 0.25, 0.33, 0.5, 0.75, 1.0, 1.25, 1.5, 2.0, 4.0, 6.0, 9.0, and 12.0 h after the administration of the ginkgolides solution. The drug-time curve of ginkgolide B in plasma were drawn according to the concentration measured by LC-MS. The time-effect curve of superoxide dismutase (SOD) and malondialdehyde (MDA) were drawn based on the value measured by the kit. The pharmacokinetic parameters were calculated by DAS 2.0 software to fit the PK-PD binding model. Results The t1/2 of ginkgolide B of the rats in the administration group was smaller than that in the MCAO model group. The area under the curve (AUC) of nasal administration was significantly higher than intragastric administration and intravenous administration. Conclusion Ginkgolide B has a good protective and mitigating effect on ischemic stroke. The pharmacokinetics of nasal administration is better than iv and ig administration, which can provide reference for the development of nasal administration of ginkgolide B.

Study on fragmentation pathways of three ginkgolides by mass spectrometry / 中国中药杂志

The fragmentation pathways of the three ginkgolides (ginkgolides A, ginkgolides B, ginkgolides C) have been studied with high resolution and high mass accuracy using quadrupole time-of-flight mass spectrometry in negative ion mode in this paper. The results indicate that the three ginkgolides have similar fragmentation pathways, including four kinds of common cleavage pathways and one common characteristic ion. In high quality regions, the typical fragmentation pathways of the three ginkgolides are lactone ring opening with continuous loss of CO, CO₂,and loss of H₂O. In low quality regions, the common characteristic fragment ion of the three ginkgolides at 72.993 6 is formed by C rings cleavage. Also, the common fragment ions of ginkgolides A and ginkgolides B at 141.018 8, 125.023 8, 113.024 0, 97.029 1 are formed by A rings cleavage. The study of fragmentation pathways could be adopted for the structural identification of the ginkgolides and their metabolites.

Diterpene ginkgolides meglumine injection inhibits oxidative stress induced by oxygen-glucose deprivation by activating Akt/Nrf2 pathway in SH-SY5Y cells / 中国药理学与毒理学杂志

OBJECTIVE To investigate the protective effects and mechanism of diterpene ginkgolides meglumine injection (DGMI) against oxidative stress induced by oxygen-glucose deprivation (OGD) in SH-SY5Y cells. METHODS SH-SY5Y cells were divided into five groups: normal control, model control (OGD group) and drug(25 mg · L- 1) administration groups including DGMI group, extract of ginkgo biloba leaves injection group (EGBLI) and lactones ginkgo biloba injection group (LGBI). The cells suffered from oxygen-glucose deprivation (OGD) for 4 h, followed by reoxygenation with drugs for 6 h. Then, cell viabilities were detect using CCK-8 assays, reactive oxygen species (ROS) levels using fluorescence probe DCFH-DA and superoxide dismutase (SOD) activities using WST-1 test. Western blotting was used to detected protein levels of hemeoxygenase-1(HO-1), NAD(P)H, quinone oxidore?ductase l (Nqo1), protein kinase B (Akt), phosphorylated Akt (p-Akt), nuclear factor-E2-related factor2 (Nrf2) and phosphorylated Nrf2 (p-Nrf2). The cells were induced by OGD for 4 h, followed by reoxygen?ation and DGMI for 1 h, combined with different concentrations of PI3K inhibitor (LY294002) (at the final concentration of 12.5, 25 and 50 μmol · L-1) before the protein levels of AKT, p-AKT, Nrf2 and p-Nrf2 were detected by Western blotting. RESULTS SH-SY5Y cells induced by OGD for 4 h resulted in an increase in ROS(P<0.01), but a decrease in cell viabilities(P<0.01), SOD activities(P<0.01), and antioxidant protein levels ( Akt, p-Akt, Nrf2, p-Nrf2, HO-1 and Nqo1) (P<0.01). Compared with OGD group, treatment with reoxygenation and drugs (DGMI,EGBLI and LGBI respectively) for 6 h resulted in a decrease in ROS (P<0.01), but an increase in cell viabilities, SOD activities and antioxidant protein levels of p-Nrf2, HO-1, Nqo1 and p-Akt(P<0.05,P<0.01). DGMI group showed the best efficiently. Moreover, after OGD for 4 h, compared with DGMI group, combining reoxygenation and DGMI with LY294002 for 1 h resulted in a concentration-dependent inhibition of the protein levels of p-AKT and p-Nrf2(P<0.01). CONCLUSION DGMI 25 mg · L-1 can inhibit oxidative stress in SH-SY5Y cells induced by OGD by increasing the activity and expression of Nrf2 through PI3K/Akt pathway, which may be one of the mechanisms by which DGMI protects neurons from stroke.

Pharmacokinetics of the prototype and hydrolyzed carboxylic forms of ginkgolides A, B, and K administered as a ginkgo diterpene lactones meglumine injection in beagle dogs / 中国天然药物

Ginkgo diterpene lactones meglumine injection (GDLI) is a commercially available product used for neuroprotection. However, the pharmacokinetic properties of the prototypes and hydrolyzed carboxylic forms of the primary components in GDLI, i.e., ginkgolide A (GA), ginkgolide B (GB), and ginkgolide K (GK), have never been fully evaluated in beagle dogs. In this work, a simple, sensitive, and reliable method based on ultra-fast liquid chromatography-tandem mass spectrometry (UFLC-MS/MS) was developed, and the prototypes and total amounts of GA, GB, and GK were determined in beagle dog plasma. The plasma concentrations of the hydrolyzed carboxylic forms were calculated by subtracting the prototype concentrations from the total lactone concentrations. For the first time, the pharmacokinetics of GA, GB, and GK were fully assessed in three forms, i.e., the prototypes, the hydrolyzed carboxylic forms, and the total amounts, after intravenous administration of GDLI in beagle dogs. It was shown that ginkgolides primarily existed in the hydrolyzed form in plasma, and the ratio of hydrolysates to prototype forms of GA and GB decreased gradually to a homeostatic ratio. All of the three forms of the three ginkgolides showed linear exposure of AUC to the dosages. GA, GB, and GK showed a constant half-life approximately 2.7, 3.4, and 1.2 h, respectively, which were consistent for the forms at three dose levels (0.3, 1.0, and 3.0 mg·kg) and after a consecutive injection of GDLI for 7 days (1.0 mg·kg).

Study on blood-brain distribution of lactones in rats after oral administration of Ginkgo biloba extract by blood-brain synchronization microdialysis / 中草药

Objective: To study the blood and brain drug concentration and drug distribution coefficient of lactones in rats after oral administration of Ginkgo biloba extract (GBE). Methods: Blood-brain synchronization microdialysis was used in combination with HPLC-MS/MS inthis study. Probes were buried into rat brain and jugular vein to collect the blood and brain micro dialysis liquid in rats after oral GBE in different periods. The lactones in the dialysate were analyzed by HPLC-MS/MS, and the blood and brain drug concentration was depicted according to the results. Then the drug distribution coefficients of each component in blood and brain (AUCbrain/AUCblood) were calculated. Results: After GBE suspension (600 mg/kg) oral gavaging in the rats, the blood samples and brain samples were obtained and detected by HPLC/MS/MS. Ginkgolides A, B, and C were detected in both blood and in brain, but bilobalide could be detected just in blood. The drug distribution coefficients in the blood and brain (AUCbrain/AUCblood) were as follows: Ginkgolide A was 2.911%, ginkgolide B was 3.126%, and ginkgolide C was 0.337%. Conclusion: Simultaneous multiple microdialysis technique can be used to detect multiple components in different tissues simultaneously. It also has other characteristics, such as continuous sampling on living animals with small sample volume and little tissue damage, which can save animals. In all, simultaneous multiple microdialysis could be a good method for exploring the effect components in Chinese medicine and their distribution at effect sides.

Protection of Diterpene Ginkgolides Meglumine Injection on acute injury of ischemic stroke in rats / 药物评价研究

Objective To investigate the protective effect and therapeutic window of DGMI on ischemic stroke in rats,and to explore the related mechanism.Method The rats were subjected to middle cerebral artery occlusion (MCAO) for 90 min followed by 72 h of reperfusion.DGMI (i.p.,1.25,2.5,5.0,and 10.0 mg/kg,Bid) was administered at 1 h after the onset of ischemia.Neurological score was evaluated after 24 and 72 h of reperfusion rcspectively.In fact volume,cerebral water content,oxidative stress markers,and IL-1β were evaluated after 72 h of reperfusion.The rats were treated with DGMI 5.0 mg/kg 0.5 h before reperfusion or 1 h,2 h,3 h,and 6 h after reperfusion to determined therapeutic window.Result Treatment with DGMI (2.5,5.0 mg/kg) significantly ameliorated neurological deficit,infarct volume and cerebral water content after cerebral ischemia reperfusion.DGMI also reduced the content of malonaldehyde (MDA),IL-1β,down-regulated the activities of creatine kinase (CK),lacticdehydrogenase (LDH),and up-regulated the activities of superoxide dISmutase (SOD).Treatment with DGMI 5.0 mg/kg exhibited protective effects when administered at all time points except for 6 h after reperfusion.Conclusion DGMI plays a certain protective role in ischemic stroke of rats,and the effect may be related to the improvement on the antioxidant capacity of brain tissue and the inhibition of overproduction of inflammatory cytokine.Moreover,the therapeutic window of DGMI isless than 6 h after reperfusion.

Multicenter evaluation on pharmacoeconomics of Ginkgolide Injection in treatment of ischemic stroke / 药物评价研究

Objective To study the clinical efficacy,safety,and economic efficiency of ginkgolide injection and conventional therapy of patients with cerebral arterial thrombosis in multi center,and to evaluate the economic value of drugs.Methods A prospective cohort study was conducted in this study,patients with ischemic stroke were collected from August 2013 to December 2014.Patients (354 cases) in treatment group were treated with Ginkgolide Injection and routine treatment,and patients (180 cases) in control group could be treated with other drugs for activating blood circulation to dissipate blood stasis on the basis of routine treatment.The patients were telephone followed-up visited 3,6,and 12 months after discharge for pharmacodynamic indexes:evaluation of activities of daily living (ADL) score,self-care rate,cure rate,recurrence rate,and all-cause mortality;economic indicators:the patient work recovery rate,cost effectiveness ratio (CER),and the incidence,and severity of adverse events,to assess the differences in the long-term benefits of different treatment regimens.Results Follow-up in 3,6,and 12 months showed that ADL score,cure rate,self-care rate,and work recovery rate of the treatment group were better than those of control group,and the difference was statistically significant.Follow-up in 12 months showed that,recurrence rate and mortality rate in the treatment group was better than that in the control group,with statistical difference.Follow-up in 6 and 12 months showed that CER of treatment group was smaller than that of the control group.The incidence of adverse reactions was low in the two groups.Conclusion Long term evaluation showed that patients treated with Ginkgolide Injection had better clinical outcomes and better CER than those without it,which proved the effectiveness and economic efficacy of Ginkgolide Inj ection in the treatment of stroke.

Preparation of self-microemulsifying drug delivery system and similarity analysis on dissolution behavior of ginkgolides components / 中草药

Objective: Optimization of ginkgolides components (GC) self-microemulsifying drug delivery system (SMDDS) (GC-SMDDS) and similarity analysis on each drug release. Methods: Using equilibrium solubility to screen the oil phase, emulsifier, and co-emulsifier; Taking appearance, the proportion of microemulsion particle size, Zeta potential, surfactants and co-surfactants, and surfactant mixing ratio of the oil phase as study factors, pseudo-ternary phase diagrams were used to screen GC-SMDDS process. SMEDDS of drug loading, particle size distribution, Zeta potential, and stability were evaluated. With the aid of the similarity factor and the curve linear regression slope analysis, the similarity between the composition of the component and the rate and extent of drug release was analyzed. Results: Optimal prescription of polyoxyethylene and polyethylene glycol 200 mass ratio of 4:1, ethoxylates and polyethylene glycol quality and bitterness total mass of 200 capric triglycerides ratio of 9:1, drug content of 100 mg/g. Particle size under 40 nm, ginkgolides 48 h internal components from microemulsion to room temperature, high temperature, and low temperature stability is good. The release quantity achieves the synchronous drug release with the similarity of 96.9%. Conclusion: The SMDDS not only can improve the dissolution of difficult soluble drugs, but also independently regulate the drug release behavior of each component so as to make the drug release maintain good consistency.

Determination on Human Plasma Protein Binding Rate of Ginkgolide A, B and K in Diterpene Ginkgolides Meglumine Injection / 世界科学技术-中医药现代化

This paper was aimed to study the human plasma protein binding rate of diterpene ginkgolides meglumine injection.The equilibrium dialysis was used to determine the human plasma protein binding rate of ginkgolide A (GA),ginkgolide B (GB) and ginkgolide K (GK) in diterpene ginkgolides meglumine injection.The LC-MS/MS method was used for the content determination of ginkgolides.And then,the plasma protein binding rate was calculated.The results showed that there was no interference from other ingredients for the determination of ginkgolides.The calibration curve of the analytes was in good linearity in certain range of contents.The precision and stability of the analytes met the methodology requirements.After 8 h incubation,the human plasma protein binding rate of GA,GB and GK achieved balance.The human plasma protein binding rate of GA (0.34,1.70 and 8.51μg·mL-1) was 84.03%-88.11%; the human plasma protein binding rate of GB (0.62,3.09 and 15.5μg·mL-1) was 41.21%-53.56%; the human plasma protein binding rate of GK (0.04,0.20 and 1.01μg·mL-1) was 45.24%-59.59%.It was concluded that the method was simple,rapid and sensitive,which met the analysis requirement for biological samples.GA had a high plasma protein binding rate; GB and GK had medium plasma protein binding rate.

Diterpene ginkgolides meglumine injection attenuates oxygen-glucose deprivation-induced apoptosis of nerve cells via inhibition of calpain signaling pathway / 中国药科大学学报

@#To investigate the anti-apoptotic effect of diterpene ginkgolides meglumine injection(DGMI)on SH-SY5Y cells induced by oxygen-glucose deprivation/reoxygenation(OGD/R), and to explore its mechanisms. After 4 h of OGD, the SH-SY5Y cells were treated with 25 mg/L DGMI for 1 h. The release of lactic dehydrogenase(LDH)was measured by cytotoxicity detection kitplus. Cell apoptosis was detected by caspase-3/7 assays. Cell death was detected by ELISA. The concentration of ［Ca2+］i in cytoplasm was measured by Fluo-3 AM and the levels of calpain and cleaved capaease-12 were evaluated by western blot. As we expected, DGMI significantly decreased the release of LDH, the concentration of ［Ca2+］i, the protein levels of calpain and cleaved caspase-12. Furthermore, DGMI injection also attenuated the activities of caspase-3/7 and the contents of cytoplasmic histone-associated- DNA-fragments. These data demonstrated that the DGMI injection showed good anti-apoptotic effect in SH-SY5Y cells induced by OGD/R. The mechanisms may be associated with the inhibition of Ca2+/calpain/caspase-12/caspase-3 signaling pathway.

Mechanism of effect of Ginkgolide B on lipopolysaccharide induced microvascular endothelial cells / 中华胰腺病杂志

Objective To investigate the effect of Ginkgolide B (BN52021) on lipopolysaccharide (LPS) induced pancreas microvascular endothelialv (MS1) cells, and to explore its molecular mechanism.Methods The optimal concentration and best time point of LPS inhibing MS1 cell survival and the optimal concentration of BN52021 increasing survival of LPS induced MS1 cells were determined by MTT.The mRNA and protein expression of adenylate cyclase (AC), phospholipase A2 (PLA2), phospholipase Cβ (PLCβ),protein tyrosine kinase (PTK) and G protein coupled receptor kinase (GRK) in platelet activating factor receptor(PAFR) signal pathway in MS1 cells were determined by real-time PCR and Western blot.Results It was showed that 10 μg/ml LPS for 24 h was the optimal concentration and best time point to induce the decrease of MS1 cells.50 mmol/L of BN52021 was the optimal concentration of increacing survival of LPS induced MS1 cells.After LPS induction, AC, GRK, PLA2, PLCβ, PTK mRNA expressions of MS1 cells were 4.02 ±0.14, 2.63 ± 0.03, 3.31 ± 0.12, 2.09 ± 0.08, 1.85 ± 0.07, which were significantly higher than those in control group (P ＜ 0.01).After BN52021 treatment, AC, GRK, PLA2, PLCβ mRNA expressions of LPS induced MS1 cells were 2.35 ±0.13, 1.17 ±0.14, 1.87 ±0.11, 1.65 ±0.10, which were significantly lower than those in LPS induction group (P ＜ 0.01).The expression of PTK mRNA was 1.83 ± 0.13, which was not significantly different from that in LPS induction group.Western blot showed that the levels of protein expression were consistent with those of mRNA expression.Conclusions BN52021 can down-regulate the up-regulated genes expression of AC, GRK, PLA2 and PLCβ in the PAFR signal pathway in LPS induced MS1 cells.

Pharmacokinetics and Brain Distribution of Ginkgolides after Administration of Sai-Luo-Tong / 世界科学技术-中医药现代化

Sai-Luo-Tong (SLT) is a compound preparation composed of ginseng, ginkgo and saffron for the treatment of vascular dementia. In order to identify its material foundation and provide evidence for therapeutic regimen, the plasma concentration, pharmacokinetics and brain distribution of ginkgolides were investigated after intragastric ad-ministration of SLT. An LC-MS/MS method was developed for the determination of 4 ginkgolides in rat plasma and brain simultaneously. Statistical analysis of obtained data demonstrated that the method had achieved the desired lin-earity, precision, accuracy and sensitivity. The results showed that after administration of SLT at the dose of 60 mg·kg-1, 4 ginkgolides were all absorbed into systemic circulation with AUC value in the order of bilobalide B (BB) >ginkgolide A (GA) > ginkgolide B (GB) > ginkgolide C (GC). All ginkgolides exhibited short half lives less than 2.8 h among which BB showed the shortest t1/2 of 1.61 h. The determination of brain distribution at different time after dos-ing revealed ginkgolides entered into brain promptly dominated by GA and BB. The concentrations of 4 ginkgolides in brain were much lower than these in plasma and declined along with time rapidly. It was concluded that ginkgolides can be absorbed in blood and penetrated into brain rapidly. GA, BB and GB might be main components which effect both periphery and brain collectively by means of their specific mechanism to achieve the therapeutic efficacy on vascular dementia of SLT.

Study on Drug Compatibility between Ginkgolides Meglumine Injection and Neutral Borosilicate Glass Ampoule / 世界科学技术-中医药现代化

This article was aimed to determine the feasibility of neutral borosilicate glass ampoule as the inner packing material of Ginkgolides meglumine injection by studying the drug compatibility between Ginkgolides me-glumine injection and neutral borosilicate glass ampoule. On the basis of the Pharmaceutical Packaging Materials and Drug Compatibility Test Guidance Rules, the study was carried on the compatibility between Ginkgolides me-glumine injection and neutral borosilicate glass ampoule for packaging materials. The results showed that no mi-gration or adsorption quality of change between the Ginkgolides meglumine injection and neutral borosilicate glass ampoule. It had good compatibility between neutral borosilicate glass ampoule and Ginkgolides meglumine injec-tion. It was concluded that neutral borosilicate glass can be used as the packing materials of Ginkgolides meglumine injection.

Phase III clinical trial of Diterpene Ginkgolides Meglumine Injection for syndrome of stagnant phlegm blocking collaterals in convalescence of atherosclerotic thrombotic cerebral infarction / 中草药

Objective: To evaluate the efficacy and safety of Diterpene Ginkgolides Meglumine Injection (DGMI) in the treatment of recovered stroke with syndrome of stagnant phlegm blocking collaterals, especially the clinical efficacy in improving the function of language and movements. Methods: The clinical trial was carried out by the methods of stratification and r andomization (416 cases of patients with atherosclerotic thrombotic cerebral infarction (ATCI) were r andomly divided into experimental and control groups by the ratio of 3:1), blindness, and positive parallel control of Shuxuening Injection (SI), and multi-center clinical study. DGMI (25 mg, 5 mL/amp) or SI (5 mL/amp) was diluted into 250 mL physiological saline, iv drip, once daily for 14 d. The dropping speed must be controlled as 10-15 drops/min for the first infusion. Results: There was statistical difference (P=0.000 1) for the variations of inducing rate of the defect extent of nervous functions before and after the treatment in the experimental and control groups. The experimental group was superior to the control group. There was no statistical difference for the variation of scales of the patient living ability in the two groups (P>0.05). For the comprehensive efficacy of cerebral infarction, the total effective rates were 85.39% and 73.27% in the experimental and control groups, respectively, with statistical difference (P=0.0001). For the efficacy of syndrome of traditional Chinese medicine (TCM), the total effective rates were 62.99% and 40.59% in the experimental and control groups, respectively, with the statistical difference (P=0.0001). For the single indicator of nervous functions, such as arm movements, h and movements, extremity movements, walking, as well as the syndromes of TCM, such as upper limb disable, lower limb disable, deviated tongue, aphasia, abate or deficiency of feeling, dizziness, more and turbid phlegm, abnormal tongue and pulse manifestation, the experimental group was superior to the control group with statistified difference. Conclusion: It is safe and effective for DGMI in the treatment of the syndrome of stagnant phlegm blocking collaterals in convalescence of ATCI. Meanwhile, it is also manifested that DGMI has the certain superiorities in the fields, such as improving the total score of nervous function deficiency for the patients with stroke, the nervous function, the total score of TCM syndromes, including upper limb disable, deviated tongue, dizziness, more and turbid phlegm, white coat of tongue, abnormal pulse, etc.