Pharmacodynamic of cilostazol for anti-altitude hypoxia / 中南大学学报(医学版)

OBJECTIVES@#The plateau environment is characterized by low oxygen partial pressure, leading to the reduction of oxygen carrying capacity in alveoli and the reduction of available oxygen in tissues, and thus causing tissue damage. Cilostazol is a phosphodiesterase III inhibitor that has been reported to increase the oxygen release of hemoglobin (Hb) in tissues. This study aims to explore the anti-hypoxic activity of cilostazol and its anti-hypoxic effect.@*METHODS@#A total of 40 male BALB/C mice were randomly divided into a low-dose cilostazol (6.5 mg/kg) group, a medium-dose (13 mg/kg) group, a high-dose (26 mg/kg) group, and a control group. The atmospheric airtight hypoxia experiment was used to investigate the anti-hypoxic activity of cilostazol and to screen the optimal dosage. Twenty-four male Wistar rats were randomly divided into a normoxia control group, a hypoxia model group, an acetazolamide (22.33 mg/kg) group, and a cilostazol (9 mg/kg) group. After 3 days of hypoxia in the 4 010 m high altitude, blood from the abdominal aorta was collected to determine blood gas indicators, the levels of IL-6 and TNF-α in plasma were determined by enzyme-linked immunosorbent assay, and the levels of malondialdehyde (MDA), superoxide dismutase (SOD), and glutataione (GSH) were measured. The degree of pathological damage for rat tissues was observed with HE staining.@*RESULTS@#Compared with the control group, the survival time of mice in the low, medium, and high dose group of cilostazol was significantly prolonged, and the survival time of mice in the medium dose group was the longest, with an extension rate at 29.34%, so the medium dose was the best dose. Compared with the hypoxia model group, the P50 (oxygen partial pressure at Hb oxygen saturation of 50%) value of rats in the cilostazol group was significantly increased by 1.03%; Hb and Hct were significantly reduced by 8.46% and 8.43%, and the levels of IL-6 and TNF-α in plasma were reduced by 50.65% and 30.77%. The MDA contents in heart, brain, lung, liver, and kidney tissues were reduced by 37.12%, 29.55%, 25.00%, 39.34%, and 21.47%, respectively. The SOD activities were increased by 94.93%, 9.14%, 9.42%, 13.29%, and 20.80%, respectively. The GSH contents were increased by 95.24%, 28.62%, 28.57%, 20.80%, and 44.00%, respectively. The results of HE staining showed that compared with the hypoxia model group, cilostazol significantly improved the damage of heart, lung, and kidney tissues in rats after hypoxia.@*CONCLUSIONS@#Cilostazol can significantly improve the oxidative stress and inflammatory reaction caused by rapid altitude hypoxia, and it has a significant protective effect on tissue damage caused by hypoxia, suggesting that it has obvious anti-hypoxic activity.

Research progress on mechanism in adaptation of hemoglobin to plateau hypoxia / 浙江大学学报·医学版

Low oxygen partial pressure is the main cause of acute mountain sickness.Hemoglobin plays a crucial physiological role in the binding, utilization, transportation and release of oxygen in the body. To increase the capacity of oxygen binding of hemoglobin or the capacity of oxygen supply in tissues can help alleviate altitude sickness. However, increasing hemoglobin content has certain limitations. Using techniques from molecular biology, researchers are looking for endogenous or exogenous substances that can regulate the conformation of hemoglobin to increase oxygen uptake in the alveoli, or the availability of alveolar oxygen in the tissues. At present, the research on allosteric modulators to improve the affinity of hemoglobin has made some progress, and research on applying this mechanism to plateau hypoxia is also underway. This article reviews the relationship between hemoglobin and hypoxia, the structure of hemoglobin and the role of various allosteric modulators in hypoxia, which would provide information for finding new substances regulating the conformation of hemoglobin.

Qualitative and Quantitative Analysis of Bushen Jiannao Grains by Ultra Performance Convergence Chromatography / 分析化学

An ultra performance convergence chromatographic ( UPC2 ) method was established for the attribution analysis of the main peaks as well as the quantitative determination of echinacoside andβ-ecdyserone in Bushen Jiannao Grains. The samples were extracted with ethanol and separated on Waters ACQUITY UPC2TM BEH column (100 mm × 3. 00 mm, 1. 7 μm), with a gradient supercritical CO2-0. 05%phosphoric acid-methanol solvent system at 40 ℃. The flow rate was 0. 8 mL/min, the detection wavelength was set at 248 nm and the injection volume was 1 μL. Results showed that all the main peaks in the fingerprint were clearly attributed. The peak named 12 wasβ-ecdyserone with the content of 380μg/g and the peak named 15 was echinacoside with the content of 9. 562 mg/g. The method was simple, eco-friendly, accurate and reliable compared with HPLC and UPLC.

Sulfated modification and anticoagulant activity in vitro of sulfated glucan isolated from the aqueous extract of Hedysarum polybotrys / 药学学报

SHG was sulfated by chlorosulfonic acid-pyridine method, and six samples which we got were prepared in different reaction conditions. There is a characteristic absorption peak near 260 nm in UV spectra and there are two characteristic absorption peaks near 1240 cm(-1) and 810 cm(-1) in the FT-IR. Degree of sulfation (DS) was calculated by elemental analysis and turbidimetry. Under the same conditions the absorption peaks become strong with the DS increase. The anticoagulant activity of SHG and sulfated modification samples was evaluated by the classic coagulant assays of prothrombin time (PT), activated partial thrombin time (APTT) live enzymes, and plasma thrombin time (TT). Results show that sulfated SHG has a good anticoagulant activity in vitro, and DS increased activity within a certain range.

Study on molecular characteristics of four components contained in Hedysari Radix polysaccharide by gel permeation chromatography-multiangle laser light scattering technology (GPC-MALLS) / 中国中药杂志

<p><b>OBJECTIVE</b>To determine such molecular characteristic parameters as absolute molecular weight, molecular weight distribution, root-mean-square turning radius (Rg) and polydispersity index (Mw/Mn) of four components contained in Hedysari Radix polysaccharide 3 (HPS-3) and map weight-average molecular weight (Mw) with root-mean-square turning radius (Rg), in order to calculate conformations of the four components at solution state.</p><p><b>METHOD</b>The gel permeation chromatography-multi angle laser light scatting (GPC-MALLS) was adopted, with 0.1 mol x L(-1) NaNO3 contained 0.02% NaN3 as the mobilephase, Ultrahydrogel 1000 connected in series with Ultrahydrogel500.</p><p><b>RESULT</b>Among the four components of HPS-3, HPS-3-C showed the highest weight average molecular weight of 1.986 x 10(5) g x mol(-1), followed by HPS-3-B 1.113 x 10(5) g x mol(-1) and HPS-3-D 8.457 x 10(4) g x mol(-1) HPS-3-A showed the lowest weight average molecular weight of 1. 223 x 10(4) g x mol(-1) but the highest square radius of gyration, that is 55.5 nm. HPS-3-D had the widest range of molecular weight distribution in four components, with the polydispersity index (Mw/Mn) of 2.543. In the mobile phase, HPS-3-A was globular structure, HPS-3-C was random coil, HPS-3-B and HPS-3-D were both highly branched structure.</p><p><b>CONCLUSION</b>The results provided necessary basis for further studies on molecular characteristics of the four components contained in HPS-3 and their relationship with bioactivity.</p>

Phamacokinetic study of bergapten in rats plasma by LC-MS/MS / 中国中药杂志

<p><b>OBJECTIVE</b>To determine bergapten's concentration in plasma and observe its pharmacokinetics in rats using a combined LC-MS/MS analytical method.</p><p><b>METHOD</b>Blood samples were separated on a Hypersil ODS column (4.6 mm x 250 mm, 5 mm) at a temperature of 30 degrees C, and mobile phase consisted of water and methanol (22.5: 77.5) at a flow rate of 0.8 mL x min(-1).</p><p><b>RESULT</b>The methodological study showed a good linear relationship of 8.12-162.4 g x L(-1) (r = 0.9999). The inner and inter-days relative standard deviations were both less than 10% , indicating legitimate precise and accuracy to the requirement of biological sample analysis.</p><p><b>CONCLUSION</b>The method is suitable for in vivo quantitative analysis for bergapten due to its accuracy, sensitivity and specificity. The pharmacokinetic process in rats forms a two-compartment model with first-order absorption.</p>

Quantification of six bioactive compounds in Zhenqi Fuzheng preparation by high-performance liquid chromatography coupled with diode array detector and evaporative light scattering detector / 药物分析学报

A simple and accurate high-performance liquid chromatography（HPLC）coupled with diode array detector（DAD）and evaporative light scattering detector（ELSD）was established for the determination of six bioactive compounds in Zhenqi Fuzheng preparation（ZFP）.The monitoring wavelengths were 254,275 and 328 nm.Under the optimum conditions,good separation was achieved,and the assay was fully validated in respect of precision,repeatability and accuracy.The proposed method was successfully applied to quantify the six ingredients in 31 batches of ZFP samples and evaluate the variation by hierarchical cluster analysis（HCA）,which demonstrated significant variations on the content of these compounds in the samples from different manufacturers with different preparation procedures.The developed HPLC method can be used as a valid analytical method to evaluate the intrinsic quality of this preparation.

Quantification of six bioactive compounds in Zhenqi Fuzheng preparation by high-performance liquid chromatography coupled with diode array detector and evaporative light scattering detector / 药物分析学报

A simple and accurate high-performance liquid chromatography (HPLC) coupled with diode array detector (DAD) and evaporative light scattering detector (ELSD) was established for the determination of six bioactive compounds in Zhenqi Fuzheng preparation (ZFP).The monitoring wavelengths were 254,275 and 328 nm.Under the optimum conditions,good separation was achieved,and the assay was fully validated in respect of precision,repeatability and accuracy.The proposed method was successfully applied to quantify the six ingredients in 31 batches of ZFP samples and evaluate the variation by hierarchical cluster analysis (HCA),which demonstrated significant variations on the content of these compounds in the samples from different manufacturers with different preparation procedures.The developed HPLC method can be used as a valid analytical method to evaluate the intrinsic quality of this preparation.

Evaluation preparation technology of Xiaochaihu granules using fingerprint-peak pattern matching / 药物分析学报

An approach was proposed to evaluate preparation technology by means of fingerprint-peak matching technology of high performance liquid chromatography with diode array detector (HPLC-DAD).Similarity and hierarchical clustering analysis (HCA) were applied to identify the 15 batches of Xiaochaihu granules from different manufacturers and our laboratory,and peak pattern matching between the composite formulae and Radix Bupleuri Chinensis,which was one of the main ingredients of Xiaochaihu granules,was utilized to evaluate the preparation technology of Xiaochaihu granules via the indexes of the relative deviation of retention time (RT) and UV spectrum feature similarity of their corresponding peaks.Eleven matching peaks were found between Xiaochalhu granules and Radix Bupleuri Chinensis.However,the saikosaponin A and saikosaponin D,which are the important active components in Radix Bupleuri Chinensis,were not found in Xiaochaihu granules from any manufacturers.The peak areas of 11 characteristic peaks of Xiaochaihu granules samples formed a matrix of 11 × 15.The result of HCA showed that Xiaochaihu granules samples were divided into four kinds of category.Xiaochaihu granules samples from the same manufacturer were basically clustered of the same category.The results suggested that the saikosaponin A and saikosaponin D are prone to structural transformation under the condition of decoction and in the presence of the organic acidic components.These active components,existing in raw herb,might transform to a series of non-active secondary saikosaponin due to unfavourable preparation technology.So the conventional decoction-based preparation technology of Xiaochaihu granules might greatly affect its quality and therapeutic effectiveness. This study demonstrates that fingerprint-peak matching technology can not only be used for quality control of this composite formulae,but also provide some guidance for preparation technology of Xiaochaihu granules.

Simultaneous determination of five active components in compound preparation of Dangguishuan by high-performance liquid chromatography / 中国中药杂志

<p><b>OBJECTIVE</b>To develop a HPLC method for determination of matrine, oxymatrine; ferulic acid, L-shikonin and beta, beta-dimethylacrylshikonin in compound preparatioti of Dangguishuan.</p><p><b>METHOD</b>The chromatogtaphic separation was performed on a Kromasil ODS C18 column (4.6 mm x 250 mm, 5 microm) maintaining at 30 degrees C during the whole process. The mobile phase consisted of methanol and 0.1% triethylamine aqueous solution (adjusted with phosphoric acid, pH 3) at a flow rate of 1.0 mL x min(-1). The detection wavelength was set at 220 nm for matririne and oxymatrine, 316 nm for ferulic acid, 516 nm for L-shikonin and beta, beta-dimethylacrylshikonin, respectively.</p><p><b>RESULT</b>All the compounds showed good linearity (r > 0.9996) in the range of the test concentrations, and the average recoveries of the method is betwuen 96.92% and 102.22%, RSD < 3.1%.</p><p><b>CONCLUSION</b>The method is proved to be credible, sensitive, accurate and repeatable. It can be applied to determine of matrine, oxymatrine, ferulic acid, L-shikonin and beta, beta-dimethylacrylshikonin in compound preparation of Dangguishuan simultaneously, and provide a basal method of quality control to this preparation and other relative preparations.</p>

Coumarins from Peucedanum harry-smithii var. subglabrum / 中国中药杂志

The root of Peucedanum harry-smithii var. subglabrum was extracted with methanol, then separated with solvents at different polarity into four fractions: aqueous (H2O), ethyl acetate (AcOEt), chloroform (CHCl3) and petroleum ether (DAB-6). From AcOEt psoralen, bargapten, xanthotoxin, marmesin, umbelliferone, scopoletin, (+/-) peuformosin, Pd-I b, (+/-) selinidin, praeruptorin D were isolated by column chromatography on silica gel, using petroleum ether/ethyl acetate as eluent. The structures of the coumarins were identified by 1H-NMR and 13C-NMR.

Determination of jasminoidin and paeoniflorin in Chizhihuang Capsules by HPLC / 中成药

AIM: To establish a HPLC quality control for Chizhihuang Capsules(Radix Paeoniae Rubra, Fructus Gardeniae, Radix et Rhizoma Rhei, etc.). METHODS: Shim pack C 18 column was used, and jasminoidin and paeoniflorin were determined at the same time and detection wavelength at 230nm. RESULTS: The standard curve of jasminoidin was linear in the range of 0.30～1.50?g. The average recovery and RSD were 99.58 and 1.57% ( n =5), respectively. The standard curve of paeoniflorin was linear in the range of 0.26～ 1.30?g . The average recovery and RSD were 99.11 and 1.29% ( n =5), respectively. CONCLUSIONS: This method is a convenient, reliable and accurate for the quality control of Chizhihuang Capsules.

Determination of Content of Ginsenoside R_(g1) ,R_e, and Paeoniflorin from Shenshao Tablet by HPLC / 中成药

Objective:To determine the content of ginsenoside R g1 ,R e and Paeoniflorin by HPLC in Shenshao tablet.Methods: HPLC Symmetry C 18 column was used and CH 3CN H 3PO 4 (11∶89) as mobile phase. The flow rate was 0.8mL/min, the column temperature at 30 ?C and the detective wavelength 203nm and 230nm. Results:The content of ginsenoside R g1 ,R e, and paeoniflorin can be determined by HPLC. The linear concentration ranges of these compound were 0.960?g～4.800?g, 0.912?g～4.560?g,1.050?g～ 5.250 ?g. The average recoveries were 105.2%, 98.5% and 103.5%, respectively.Conclusion: This method is simple and accurate and can be used to determine the content of Shenshao Tablet.

Determination of polysaccaride content in Radix Hedysari by HPLC / 中成药

AIM: To establish a method for determing polysaccaride content of Radix Hedysari(Hedysarum Polybotrys Hand-Mazz). METHODS: To determine content of Radix Hedysari polysaccaride by HPLC and ELSD. RESULTS: The composes of Radix Hedysari polysaccaride Ⅰ、Ⅱ、Ⅲ、Ⅳ are all Rhamnose、Arabinose、Xylose、Glucose、Galactose,the average content of polysaccharides is 25.34%、24.23%、15.08%、17.24%. CONCLUSION: The contents of Radix Hedysari polysaccaride in Radix Hedysari can be determined by HPLC.

Chromatography fingerprint of Baitiao Radix Codonopsis in Gansu Province by HPLC / 中成药

AIM To establish chromatography fingerprint of Radix Codonopsis. METHODS HPLC was applyed on a Hypersil ODS column with water-methanol gradient elution, flow-rate of 0.8 ml?min -1 . Determination of content of atractylenodie Ⅲ adopted ELSD and PAD with methanol-water(67∶ 33), flow-rate of 1.0 ml?min -1 , column temperature at 30?C , ELSD:carried-gas flow-rate of 2.2 L?min -1 , drift-tuber temperature was 80?C ;PAD:wavelength was 220 nm. RESULTS 25 common peaks were picked up, their similarity among 10 batchs of samples was more than 98.12 % based on marker composition(Atractylenolide Ⅲ). CONCLUSION The chromatography fingerprint of Baitiao Radix Codonopsis could be used as their characteristic chromatography fingerprint of hydrophobic fraction.

Determination of formononetin and calycosin from Zhengqifuzheng Preparation by HPLC / 中成药

AIM: To establish a method of RP-HPLC for determination of calycosin and formononetin in Zhengqifuzheng Preparation(Radix Astragali, Fructus Ligustri Lucidi, etc.). METHODS: Kromasil ODS column(5?m,4.6mm?250mm) was used to determine two isoflavoniods with water-methanol gradient elution, at column temperature of 25℃, flow-rate of 1.0mL?min -1 and detection wavelength at 254nm. RESULTS: The average recoveries of formononetin and calycosin in Zhengqifuzheng Capsules were 96.53% and 98.34% and RSD were 2.25% and 4.01%, respectively, the average recoveries of formonoenetin and calycosin in Zhengqifuzheng Granules were 97.20% and 95.65% and RSD were 2.66% and 2.32%, respectively. CONCLUSION: The method is sample, reliable, accurate, practical, and can provid a new idea to study the quality standard of this preparation.