Mechanisms of Dangua Recipe in Improving Glycolipid Metabolic Disorders Based on Transcriptomics / 中国结合医学杂志

OBJECTIVE@#To explore the mechanisms of Dangua Recipe (DGR) in improving glycolipid metabolism based on transcriptomics.@*METHODS@#Sprague-Dawley rats with normal glucose level were divided into 3 groups according to a random number table, including a conventional diet group (Group A), a DGR group (Group B, high-calorie diet + 20.5 g DGR), and a high-calorie fodder model group (Group C). After 12 weeks of intervention, the liver tissue of rats was taken. Gene sequence and transcriptional analysis were performed to identify the key genes related to glycolipid metabolism reflecting DGR efficacy, and then gene or protein validation of liver tissue were performed. Nicotinamide phosphoribosyl transferase (Nampt) and phosphoenolpyruvate carboxykinase (PEPCK) proteins in liver tissues were detected by enzyme linked immunosorbent assay, fatty acid synthase (FASN) protein was detected by Western blot, and fatty acid binding protein 5 (FABP5)-mRNA was detected by quantitative real-time polymerase chain reaction. Furthermore, the functional verification was performed on the diabetic model rats by Nampt blocker (GEN-617) injected in vivo. Hemoglobin A@*RESULTS@#Totally, 257 differential-dominant genes of Group A vs. Group C and 392 differential-dominant genes of Group B vs. Group C were found. Moreover, 11 Gene Ontology molecular function terms and 7 Kyoto Encyclopedia of Genes and Genomes enrichment pathways owned by both Group A vs. Group C and Group C vs. Group B were confirmed. The liver tissue target validation showed that Nampt, FASN, PEPCK protein and FABP5-mRNA had the same changes consistent with transcriptome. The in vivo functional tests showed that GEN-617 increased body weight, HbA@*CONCLUSION@#Nampt activation was one of the mechanisms about DGR regulating glycolipid metabolism.

Polysaccharides from Bupleurum Induce Immune Reversal in Late Sepsis.

BACKGROUND: Bupleurum chinense, a well-known Traditional Chinese Medicine, has been used for thousands of years in China. In this study, we would suggest that Bupleurum polysaccharides (BPS) could improve the prognosis of sepsis through its impact on redistribution of BMCs, which triggers immune reversal in late sepsis. METHODS: BALB/c mice were divided into five groups: sham burn group, burn plus P aeruginosa group, burn plus P aeruginosa with BPS (40 mg/kg, 100 mg/kg, and 250 mg/kg) treatment group, and they were sacrificed at post-burn day (PBD) 0, 3, 5, and 7. BMCs, liver cells, and dendritic cells (DCs) were harvested. Flow cytometry was used to determine the change of phenotypes of DCs and isolate these cells. Cytometric beads array was utilized to analyze the level of inflammatory factors. Cell therapy of BMCs, liver cells, and DCs was administrated to explore the protective role of regional organ immunity. RESULTS: BPS could decrease the lethality of burn sepsis in a dose-dependent fashion and increase both the percentage of CD11cCD45RB DCs in bone marrow (BM) and liver and the number of BMCs and liver cells significantly. Cell therapy of BMCs, liver cells, and CD11cCD45RB DCs at PBD7 could protect septic mice from sepsis. CONCLUSION: BPS has shown its potential in promoting the prognosis of post-burn sepsis through its effect on immune redistribution of BMCs, especially via differentiation of CD11cCD45RB DC cells in BM and nonimmune organs to induce immune reversal in late sepsis.

Dan-gua fang improves glycolipid metabolic disorders by promoting hepatic adenosine 5'-monophosphate activated protein kinase expression in diabetic Goto-Kakizaki rats / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the effect of Dan-gua Fang on adenosine 5'-monophosphate (AMP) activated protein kinase (AMPK) α expression in liver and subsequent improvement of glucose and lipid metabolism.</p><p><b>METHODS</b>Forty 13-week-old diabetic Goto-Kakizaki (GK) rats were randomly divided into model, Dan-gua Fang, metformin and simvastatin groups (n=10 for each), and fed high-fat diet ad libitum. Ten Wistar rats were used as normal group and fed normal diet. After 24 weeks, liver expression of AMPKα mRNA was assessed by real-time PCR. AMPKα and phospho-AMPKα protein expression in liver was evaluated by Western blot. Liver histomorphology was carried out after hematoxylin-eosin staining, and blood glucose (BG), glycosylated hemoglobin A1c (HbA1c), food intake and body weight recorded.</p><p><b>RESULTS</b>Similar AMPKα mRNA levels were found in the Dan-gua Fang group and normal group, slightly higher than the values obtained for the remaining groups (P<0.05). AMPKα protein expression in the Dan-gua Fang group animals was similar to other diabetic rats, whereas phospho-AMPKα (Thr-172) protein levels were markedly higher than in the metformin group and simvastatin group (P<0.05), respectively. However, phosphor-AMPKα/AMPKα ratios were similar in all groups. Dan-gua Fang reduced fasting blood glucose with similar strength to metformin, and was superior in reducing cholesterol, triglycerides, high-density lipoprotein cholesterol as well as improving low-density lipoprotein cholesterol in comparison with simvastatin and metformin. Dan-gua Fang decreases plasma alanine aminotransferase (ALT) significantly.</p><p><b>CONCLUSION</b>Dan-gua Fang, while treating phlegm-stasis, could decrease BG and lipid in type 2 diabetic GK rats fed with high-fat diet, and effectively protect liver histomorphology and function. This may be partly explained by increased AMPK expression in liver. Therefore, Dan-gua Fang might be an ideal drug for comprehensive intervention for glucose and lipid metabolism disorders in type 2 diabetes mellitus.</p>

Paradox of using intensive lowering of blood glucose in diabetics and strategies to overcome it and decrease cardiovascular risks / 中国结合医学杂志

Hyperglycemia significantly increases the risk of cardiovascular disease (CVD) in diabetics. However, it has been shown by a series of large scale international studies that intensive lowering of blood glucose levels not only has very limited benefits against cardiovascular problems in patients, but may even be harmful to patients at a high risk for CVD and/or poor long-term control of blood glucose levels. Therefore, Western medicine is faced with a paradox. One way to solve this may be administration of Chinese herbal medicines that not only regulate blood glucose, blood fat levels and blood pressure, but also act on multiple targets. These medicines can eliminate cytotoxicity of high glucose through anti-inflammatory and anti-oxidant methods, regulation of cytokines and multiple signaling molecules, and maintenance of cell vitality and the cell cycle, etc. This allows hyperglycemic conditions to exist in a healthy manner, which is called "harmless hyperglycemia" Furthermore, these cardiovascular benefits go beyond lowering blood glucose levels. The mechanisms of action not only avoid cardiovascular injury caused by intensive lowering of blood glucose levels, but also decrease the cardiovascular dangers posed by hyperglycemia.

Effect of dangua recipe on glycolipid metabolism and VCAM-1 and its mRNA expression level in Apo E(-/-) mice with diabetes mellitus / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the effect of Dangua Recipe (DGR) on glycolipid metabolism, vascular cell adhesion molecule-1 (VCAM-1) and its mRNA expression level of transgenic Apo E(-/-) mouse with spontaneous atherosclerosis, thus revealing its partial mechanism for curing diabetes mellitus (DM) with angiopathy.</p><p><b>METHODS</b>Diabetic model was prepared by peritoneally injecting streptozotocin (STZ) to Apo E(-/-) mouse. Totally 32 modeled mice were stratified by body weight, and then divided into 4 groups referring to blood glucose levels from low to high by random digit table, i.e., the model group (MOD, fed with sterile water, at the daily dose of 15 mL/kg), the DGR group (fed with DGR at the daily dose of 15 mL/kg), the combination group (COM, fed with DGR at the daily dose of 15 mL/kg and pioglitazone at the daily dose of 4.3 mg/kg), and the pioglitazone group (PIO, at the daily dose of 4.3 mg/kg), 8 in each group. Another 8 normal glucose C57 mouse of the same age and strain were recruited as the control group. All interventions lasted for 12 weeks by gastrogavage. The fasting blood glucose (FBG), body weight, food intake, water intake, skin temperature, the length of tail, and the degree of fatty liver were monitored. The hemoglobin A1c (HbA1c), total cholesterol (TC), and LDL-C were determined. Endothelin-1 (ET-1) was determined by radioimmunoassay. Nitrogen monoxidum (NO) was determined by nitrate reductase. The kidney tissue VCAM-1 level was analyzed with ELISA. The expression of VCAM-1 mRNA in the kidney tissue was detected with real time quantitative PCR.</p><p><b>RESULTS</b>Compared with the control group, the body weight and food intake decreased, water intake increased in all the other model groups (P < 0.05). Besides, the curve of blood glucose was higher in all the other model groups than in the control group (P < 0.01). Compared with the model group, the body weight increased; levels of HbAlc, TC, LDL-C, ET-1, and VCAM-1 were significantly lower; and skin temperature was higher in the DGR group (P < 0.05, P < 0.01). Compared with the PIO group, body weight, the increment of body weight, FBG, TC, and LDL-C were lower (P < 0.05, P < 0.01); food intake and water intake increased more and the tail length was longer in the DRG group (P < 0.01). There was no statistical difference in the level of NO among groups. The degree of fatty liver in the model group was significantly severer than that in the control group (P < 0.05). It was obviously alleviated in the DGR group (P < 0.05) when compared with the model group and the PIO group (P < 0.05, P < 0.01). But it was severer in the PIO group than in the model group (P < 0.01). The degree of fatty liver in the combination group ranged between that of the DGR group and the PIO group (P < 0.05). The level of VCAM-1 mRNA expression was significantly lower in the DGR group than in the model group, the PIO group, and the combination group (P < 0.05).</p><p><b>CONCLUSIONS</b>DGR had effect in lowering blood glucose and blood lipids, and fighting against fatty liver of transgenic Apo E(-/-) mouse with spontaneous atherosclerosis. DGR played an effective role in preventing and treating DM with angiopathy by comprehensively regulating glycolipid metabolism and promoting the vascular function.</p>

Research of Dangua Recipe on intervening the glycolipid metabolism and oxidative stress in diabetic rats with atherosclerosis / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the effects of Dangua Recipe (DGR) on glycolipid metabolism, serum reactive oxygen species (ROS) level, nuclear factor kappa B (NF-kappaB) positive expression and its mRNA expression level in the thoracic aorta of diabetic rats with atherosclerosis, thus revealing its partial mechanisms for intervening chronic diabetic complications.</p><p><b>METHODS</b>Recruited 40 Goto-Kakisaki (GK) Wistar rats were fed with high fat forage containing metabolic inhibition Propylthiouracil, and peritoneally injected with endothelial NOS inhibitor N-nitro-L-arginine methyl ester to establish a high fat diabetes model with atherosclerosis. The modeled GK rats were stratified by body weight, and then, by blood glucose level from high to low, randomly divided into the DGR group (at the daily dose of 8 mL/kg), the metformin group (MET, at the daily dose of 150 mg/kg), the simvastatin group (SIM, at the daily dose of 2 mg/kg), and the model group (MOD, fed with pure water, at the daily dose of 8 mL/kg) according to the random number table, 10 in each group. Another 10 Wistar rats of the same ages and comparable body weight level were recruited as the normal control group. All the interventions lasted for 24 weeks by gastrogavage. The fasting blood glucose (FBG) and body weight were monitored. The HbA1c, TC, LDL-C, HDL-C, TG, serum ROS were determined. The aortic NF-kappaB level was analyzed with immunohistochemical assay. The expression of NF-kappaB (P65) mRNA in the aorta was detected with Real-time PCR.</p><p><b>RESULTS</b>The body weight in the normal control group was eventually heavier than others (P < 0.01). There was no difference among the four groups of GK modeled rats (P > 0.05). The FBG in the four GK modeled groups were higher than that in the normal control group (P < 0.01, P < 0.05). There was no statistical difference in the blood glucose level at the first visit and at the baseline among the GK modeled groups (P > 0.05). The last FBG level was obviously lower in the MET and DGR groups than in the MOD group (P < 0.01) and the SIM group (P < 0.05). Twenty-four weeks after intervention, the level of FBG, HbA1c, TC, LDL-C, HDL-C, and NF-kappaB positive expression rate of the thoracic aorta of the four groups of GK modeled rats, and NF-kappaB mRNA expression in the thoracic aorta in the MOD group, the MET group, and the DGR group were significantly higher than those in the normal control group (P < 0.01, P < 0.05). The TG level, serum ROS in the MET, DGR, and SIM groups, and the NF-kappaB mRNA expression level in the thoracic aorta in the SIM group were significantly lower than those in the normal control group (P < 0.01, P < 0.05). The levels of FBG, TC, LDL-C, serum ROS, NF-kappaB mRNA expression level in the thoracic aorta in three drug intervention groups, and NF-kappaB positive expression rate in the DGR and MET groups, and the levels of HbA1c, TG in the DGR group were significantly lower than those in the MOD group (P < 0.01, P < 0.05). The level of FBG in the MET and DGR groups were lower than that in the SIM group (P < 0.05). The level of NF-kappaB mRNA expression in the thoracic aorta of the SIM and DGR groups, and the levels of TC and LDL-C in the DGR group were significantly lower than those in the MET group (P < 0.01).</p><p><b>CONCLUSION</b>DGR played a role in preventing and treating chronic diabetic complications by comprehensively regulating blood glucose and serum lipids, as well as down-regulating oxidative stress.</p>

Toxicity features of high glucose on endothelial cell cycle and protection by Dan Gua-Fang in ECV-304 in high glucose medium / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the toxicity features of high glucose on the endothelial cell cycle and the influence of Dan Gua-Fang, a Chinese herbal compound prescription, on the reproductive cycle of vascular endothelial cells cultivated under a high glucose condition; to reveal the partial mechanisms of Dan Gua-Fang in the prevention and treatment of endothelial injury caused by hyperglycemia in diabetes mellitus (DM); and offer a reference for dealing with the vascular complications of DM patients with long-term high blood glucose.</p><p><b>METHODS</b>Based on the previous 3-(4,5)-dimethylthiahiazo (z-y1)-3-5-diphenytetrazoliumromide (MTT) experiment, under different medium concentrations of glucose and Dangua liquor, the endothelial cells of vein-304 (ECV-304) were divided into 6 groups as follows: standard culture group (Group A, 5.56 mmol/L glucose); 1/300 herb-standard group (Group B); high glucose culture group (Group C, 16.67 mmol/L glucose); 1/150 herb-high glucose group (Group D); 1/300 herb-high glucose group (Group E); and 1/600 herb-high glucose group (Group F). The cell cycle was assayed using flow cytometry after cells were cultivated for 36, 72 and 108 h, respectively.</p><p><b>RESULTS</b>(1) The percentage of cells in the G0/G1 phase was significantly increased in Group C compared with that in Group A (P<0.05), while the percentage of S-phase (S%) cells in Group C was significantly reduced compared with Group A (P<0.05); the latter difference was dynamically related to the length of growing time of the endothelial cells in a high glucose environment. (2) The S% cells in Group A was decreased by 30.25% (from 40.23% to 28.06%) from 36 h to 72 h, and 12.33% (from 28.06% to 24.60%) from 72 h to 108 h; while in Group C, the corresponding decreases were 23.05% and 21.87%, respectively. The difference of S% cells between the two groups reached statistical significance at 108 h (P<0.05). (3) The percentage difference of cells in the G2/M phase between Group C and Group A was statistically significant at 72 h (P<0.01). (4) 1/300 Dan Gua-Fang completely reversed the harmful effect caused by 16.67 mmol/L high glucose on the cell cycle; moreover it did not disturb the cell cycle when the cell was cultivated in a glucose concentration of 5.56 mmol/L.</p><p><b>CONCLUSIONS</b>High glucose produces an independent impact on the cell cycle. Persistent blocking of the cell cycle and its arrest at the G0/G1 phase are toxic effects of high glucose on the endothelial cell cycle. The corresponding variation of the arrest appears in the S phase. 1/300 Dan Gua-Fang completely eliminates the blockage of high glucose on the endothelial cell cycle.</p>

Astragalus polysaccharides regulate T cell-mediated immunity via CD11c(high)CD45RB(low) DCs in vitro.

ETHNOPHARMACOLOGICAL RELEVANCE: Astragalus polysaccharides (APS) isolated from one of the Chinese herbs, Astragalus mongholicus, are known to have a variety of immunomodulatory activities. However, it is not yet clear whether APS can induce the activation and differentiation of dendritic cells (DCs) and subsequently activate T cells. AIM OF THE STUDY: This study was carried out to investigate the effect of APS on the differentiation of splenic DCs and its influence on T cell-mediated immunity through interleukin (IL)-12-producing CD11c(high)CD45RB(low) DCs in vitro. METHODOLOGY: MACS microbeads were used to isolate splenic DCs, CD11c(high)CD45RB(low) DCs, CD11c(low)CD45RB(high) DCs and CD4(+) T cells. Phenotypes were analyzed by flow cytometry, and cytokine levels were determined with cytometric bead array or ELISA. RESULT: The percentage of CD11c(high)CD45RB(low) DCs was significantly increased after treatment with APS compared to their counterparts. The cytokine secretion pattern of CD11c(high)CD45RB(low) DCs and CD11c(low)CD45RB(high) DCs was detected, and it was found that unlike the stable IL-10 secretion pattern of CD11c(low)CD45RB(high) DCs induced by APS, CD11c(high)CD45RB(low) DCs showed a dose-dependent relationship between IL-12 production and APS stimulation. In order to verify whether the activation of CD4(+) T was associated with the differentiation of splenic DCs mediated by APS to CD11c(high)CD45RB(low) DCs, anti-IL-12 receptor (IL-12R) as well as anti-IL-10R monoclonal antibody was used to inhibit the effect of CD11c(high)CD45RB(low) DCs and CD11c(low)CD45RB(high) DCs in CD4(+) T mixed lymphocyte reaction culture. After treatment with anti-IL-12R or anti-IL-10 monoclonal antibody in CD4(+) T+CD11c(high)CD45RB(low) DCs or CD11c(low)CD45RB(high) DCs mixed lymphocyte reaction, the inductions of these DCs on T cells were inhibited dramatically. CONCLUSION: APS might induce the differentiation of splenic DCs to CD11c(high)CD45RB(low) DCs followed by shifting of Th2 to Th1 with enhancement of T lymphocyte immune function in vitro. Also, the effect of APS on T-cell differentiation to Th1 was not associated with the inhibition of IL-10 production in CD11c(low)CD45RB(high) DCs.

Anticolchicine cytotoxicity enhanced by Dan Gua-Fang, a Chinese herb prescription in ECV304 in mediums / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the effect of anticolchicine cytotoxicity of Dan Gua-Fang, a Chinesea Chinese), a Chinese herbal compound prescription on endothelial cells of vein (ECV304) cultivated in mediums of different glucose concentrations as well as the proliferation of those cells in the same conditions, in order to reveal the value of Dan Gua-Fang in preventing and treating endothelial damage caused by hyperglycemia in diabetes mellitus.</p><p><b>METHODS</b>The research was designed as three stages. The growing state and morphological changes were observed when ECV304 were cultivated in the culture mediums, which have different glucose concentrations with or without Dan Gua-Fang and at the same time with or without colchicine.</p><p><b>RESULTS</b>(1) Dan Gua-Fang at all concentrations reduced the floating cell population of ECV304 cultivated in hyperglycemia mediums. (2) Dan Gua-Fang at all concentrations and hyperglycemia both had a function of promoting "pseudopod-like" structure formation in cultivated ECV304, but the function was not superimposed in mediums containing both hyperglycemia and Dan Gua-Fang. (3) Colchicine reduced and even vanished the "pseudopod-like" structure of the endotheliocyte apparently cultivated in mediums of hyperglycemia or with Dan Gua-Fang. The "pseudopod-like" structure of the endotheliocyte emerged quickly in Dan Gua-Fang groups after colchicine was removed, but it was not the case in hyperglycemia only without Dan Gua-Fang groups. (4) Dan Gua-Fang reduced the mortality of cells cultivated in mediums containing colchicine. The cell revived to its normal state fast after colchicine was removed.</p><p><b>CONCLUSION</b>Dan Gua-Fang has the functions of promoting the formation of cytoskeleton and fighting against colchicine cytotoxicity.</p>

Inflammatory response and immune regulation of high mobility group box-1 protein in treatment of sepsis.

Sepsis is an infection induced systemic inflammatory response syndrome and is a major cause of morbidity as well as mortality in intensive care units. A growing body of evidence suggests that the activation of a proinflammatory cascade is responsible for the development of immune dysfunction, susceptibility to severe sepsis and septic shock. The present theories of sepsis as a dysregulated inflammatory response and immune function, as manifested by excessive release of inflammatory mediators such as high mobility group box 1 protein (HMGB1), are supported by increasing studies employing animal models and clinical observations of sepsis. HMGB1, originally described as a DNA-binding protein and released passively by necrotic cells and actively by macrophages/monocytes, has been discovered to be one of essential cytokines that mediates the response to infection, injury and inflammation. A growing number of studies still focus on the inflammation-regulatory function and its contribution to infectious and inflammatory disorders, recent data suggest that HMGB1 formation can also markedly influence the host cell-mediated immunity, including T lymphocytes and macrophages. Here we review emerging evidence that support extracellular HMGB1 as a late mediator of septic complications, and discuss the therapeutic potential of several HMGB1-targeting agents in experimental sepsis. In addition, with the development of traditional Chinese medicine in recent years, it has been proven that traditional Chinese herbal materials and their extracts have remarkable effective in treating severe sepsis. In this review, we therefore provide some new concepts of HMGB1-targeted Chinese herbal therapies in sepsis.

Glucose endothelial cytotoxicity and protection by Dan Gua-Fang, a Chinese herb prescription in huVEC in hyperglycemia medium.

BACKGROUND: Low success rate of blood glucose in diabetes is an international problem. The endothelia cytotoxicity of hyperglycemia has been widely accepted. However, it has not been seen in reports of the value of concentration of high glucose beginning to produce cytotoxicity and the relationship between hyperglycemia and cytotoxicity as well as how to effectively prevent and control hyperglycemia cytotoxicity. Dan Gua prescription is an effective Chinese herb prescription for diabetic vascular complications. METHODS: Dan Gua prescription was contained in Dan Gua liquor utilized in experiments. (1) The cytotoxicity experiment of Dan Gua was carried out with M199 medium whose glucose (Glu) was 5.55 mmol/l to seek for a suitable experimental concentration of Dan Gua. (2) The human vessel endotheliocyte was cultivated for 72 h with mediums containing glucose in different concentrations (Group G1 to Group G11, Glu: 5.5 to 99.9 mmol/l, respectively), and assayed an optical density (OD) value using the 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide method. (3) Experiment 2 was repeated. However, the medium of each group (Groups Y1 to Y11) contained Dan Gua liquor whose concentration was 1/300. RESULTS: There was a negative correlation between means of cell OD values and glucose concentrations (r=-.927, R(2)=.844), and it presented a notable linear correlation (y=0.681-0.002x). Based on the OD value of 5.5-mmol/l glucose concentration (group G1), when glucose concentration reached 22.2 mmol/l (G4), the difference in OD values has a statistical significance. OD values in Y1-Y11 were not less than that of G1. CONCLUSION: There is a notable linear correlation between the endothelial cytotoxicities of Glu and its concentrations. The spinodal point concentration of statistical significance of hyperglycemia cytotoxicity is 22.2 mmol/l; 1/300 Dan Gua can reverse the endothelia cytotoxicity in different concentrations of hyperglycemia.

Research progress on determination of lignans from Schiandra chinensis and its preparations / 中国中药杂志

The latest research progress on quantitative determination methods of main active components-lignans from Schisandra chinensis and its preparations has been summarized, such as spectrophotometry, thin-layer chromatography scanning, high performance liquid chromatograpy, gas chromatography-mass spectrometry and capillary electrochromatography. The characteristics and application areas of every analytical method have also been stated. It offers reference on quality control of crude drug and its preparations of S. chinensis.

Progress in research on constituents and pharmacological activities of sarcotestas of Ginkgo biloba / 中国中药杂志

The latest progress in research on constituents and pharmacological activities of sarcotestas of Ginkgo biloba has been studied. The main constituents in sarcotestas of G. biloba include flavones, ginkgolides, alkylphenols, polysaccharides and amino acids, etc. They show the following activities, such as bacteriostatic, bactericidal and pesticidal activities, antitumor and mutagenic, carcinogenic effects, antianaphylaxis and allergenic activity, effects on immunologic function, scavenging free radical, antisenile action, etc. The problems at present and the reseach direction for the future on sarcotestas of G. biloba have been put forward.

Determination of ginkgolic acids in Ginkgo biloba extract and its preparations by high performance liquid chromatography / 药学学报

<p><b>AIM</b>To establish a high performance liquid chromatographic method for determination of ginkgolic acids in Ginkgo biloba extract and its preparations.</p><p><b>METHODS</b>Ginkgo biloba extract and its preparations were extracted with petroleum ether in Soxhlet apparatus, and then concentrated under vacuum. The ginkgolic acids were determined directly by HPLC, and identified by LC/DAD/ESI/MS. The chromatographic column was Inertsil ODS-2; the mobile phase was methanol-3% aqueous acetic acid(92:8); the flow rate was 1.0 mL.min-1; the column temperature was 40 degrees C; the detection wavelength was 310 nm.</p><p><b>RESULTS</b>There were six kinds of ginkgolic acid (C13:0, C15:1, C17:2, C15:0, C17:1 and an unknown compound C17:3 tentatively) in the Ginkgo biloba extract. The relative percentage content of ginkgolic acids C13:0, C15:1 and C17:1 was above 94%. The content of ginkgolic acids in Ginkgo biloba extract containing high content ginkgolic acids was 1.12%, and RSD was 2.4% (n = 5). The content of ginkgolic acids in one kind of EGb preparations (tablet) was 49.2 micrograms.g-1, and RSD was 4.3% (n = 5). The average recovery was 98.2%, RSD was 2.6% (n = 5).</p><p><b>CONCLUSION</b>The method is accurate, fast, simple, and can be used for determination of ginkgolic acids in Ginkgo biloba extract and its preparations.</p>

Determination of ginkgolic acids by high performance liquid chromatography / 药学学报

<p><b>AIM</b>To establish a simple pre-treated method and high performance liquid chromatographic method for separation and determination of ginkgolic acids in Ginkgo biloba leaves.</p><p><b>METHODS</b>The ginkgolic acids in the German standard sample were identified by LC/DAD/ESI/MS. The methods for pre-treatment and high performance liquid chromatography determination of ginkgolic acids were studied. Ginkgo biloba leaves were extracted with n-hexane in Soxhlet apparatus, then concentrated under vacuum. The ginkgolic acids can be determined directly by HPLC after one-pre-purified-step by silica gel column chromatography. The eluant was petroleum ether-diethyl ether-formic acid (89:11:1). The chromatographic column was Inertsil ODS-2; the mobile phase was methanol-3% acetic acid (92:8); the flow rate was 1.0 mL.min-1; the column temperature was 40 degrees C; the detection wavelength was at 310 nm.</p><p><b>RESULTS</b>There were five kinds of ginkgolic acid (C13:0, C15:1, C17:2, C15:0 and C17:1) in ginkgo biloba leaves. The relative percentage content of ginkgolic acids C15:1 and C17:1 was about 85%. Ginkgolic acid C17:2 had not been reported in China. The HPLC indicates that there was nearly no impurities except ginkgolic acids after treated by column chromatography. The results showed that the content of ginkgolic acids in the leaves of Ginkgo biloba collected in April, May and June was 1.48%, 1.19% and 1.11% respectively. The average recovery of Ginkgo biloba leaves collected in June was 97.0%, RSD was 1.7% (n = 6).</p><p><b>CONCLUSION</b>The method is accurate, simple and reliable, and can be used for determination of ginkgolic acids in Ginkgo biloba leaves.</p>