Protective effect and mechanism of baicalein on hypoxia-induced cortical neuron injury in rats / 中国药房

OBJECTIVE To explore the protective effect and possible mechanism of baicalein on hypoxia-induced cortical neuron injury in rats. METHODS The cortical neurons of rats （RN-C cells） were studied and cultured under hypoxic conditions （5%CO2， 94% N2， 1%O2） for 24 hours； the effects of different concentrations of baicalein （0.01， 0.1， 1， 10， 100 μmol/L） on the survival rate of hypoxic RN-C cells were investigated； the effects of baicalein （0.1 μmol/L） on the activities of lactate dehydrogenase （LDH） and superoxide dismutase （SOD）， the content of malondialdehyde （MDA）， migration rate， apoptotic rate， cell cycle and the expressions of cleaved caspase-3， B-cell lymphoma-2 （Bcl-2） and Bcl-2 X protein （Bax） were all detected. RESULTS Compared with control group， the survival rate of cells in the hypoxia group was significantly reduced （P＜0.01）； 0.01， 0.1 and 1 μmol/L baicalein could reverse survival rate of hypoxia-induced cortical neurons （P＜0.05 or P＜0.01）. Scratch experiments showed that baicalein significantly increased the migration rate of hypoxic RN-C cells （P＜0.01）. Compared with control group， the activity of LDH in the supernatant and the content of MDA in the cells， apoptotic rate and the proportion of cells in G1 phase， were significantly increased in the hypoxia group， while SOD activity and the proportion of cells in G2+S phase was decreased significantly （P＜0.01）. The protein expressions of cleaved caspase-3 were increased significantly， while the ratio of Bcl-2/Bax in cells was significantly reduced （P＜0.05 or P＜0.01）. Compared with hypoxia group， the above indexes were all reversed significantly in baicalein group （P＜0.01）. CONCLUSIONS Baicalein can promote the proliferation and migration of cortical neurons， improve hypoxia-induced cell apoptosis and cell cycle distribution， decrease the activity of LDH in supernatant and the level of cellular lipid peroxidation， and improve antioxidant levels. Its mechanism may be related to regulating the caspase- 3/Bax/Bcl-2 pathway.

Investigation on improvement effects and mechanism of 7-hydroxyethyl chrysin on PC12 cell injury induced by hypobaric hypoxia / 中国药房

OBJECTIVE To investigate the improvement effects and possible mechanism of 7-hydroxyethyl chrysin （7-HEC） on PC12 cell injury induced by hypobaric hypoxia. METHODS The rat adrenal pheochromocytoma cell line PC12 was cultured under low-pressure hypoxia （5%CO2， 94%N2， 1%O2， 54 004 Pa） to investigate the different concentrations of 7-HEC （100， 10， 1， 0.1， 0.01 μmol/L） on the survival rate of hypoxic cells； the effects of 7-HEC（1 μmol/L） on the contents of lactate dehydrogenase （LDH） and malondialdehyde （MDA）， superoxide dismutase （SOD） activity， apoptotic rate， cell cycle， and the expressions of cleaved caspase-3， Bcl-2 and Bax were detected. RESULTS Compared with control group， the survival rate of cells in hypobaric hypoxia group was decreased significantly （P＜0.01）； 10， 1， 0.1 μmol/L 7-HEC could reverse the decrease of cell survival rate caused by hypobaric hypoxia （P＜0.05 or P＜0.01）. Compared with control group， LDH content in supernatant， MDA content in cells， apoptotic rate， the proportion of cells at G1 stage and the protein expression of cleaved caspase-3 were increased significantly in hypobaric hypoxia group， while SOD activity in cells， the proportion of cells at S stage and G2 stage and Bcl-2/Bax ratio were decreased significantly （P＜0.05 or P＜0.01）. Compared with hypobaric hypoxia group， the contents of LDH and MDA， apoptotic rate， the proportion of cells at G1 stage and the expression of cleaved caspase-3 in 7-HEC group were decreased significantly， while SOD activity， the proportion of cells at G2 stage and Bcl-2/Bax ratio were increased significantly （P＜ 0.01）. CONCLUSIONS 7-HEC can significantly increase the survival rate of hypobaric hypoxia cells， reduce the LDH content in supernatant， improve cell cycle arrest， and reduce the rate of apoptosis. Its improvement effects on hypobaric hypoxia cell injury may be related to the inhibition of caspase-3/Bax/Bcl-2 pathway activation.

Research progress on the preparation and application of flavonoid nanocrystals / 浙江大学学报·医学版

Flavonoids have been reported to possess significant pharmacological activities,such as antioxidant, anti-inflammatory and anticancer effects. However, the low solubility and low bioavailability limits their clinical application. Nanocrystal technology can solve the delivery problems of flavonoids by reducing particle size, increasing the solubility of insoluble drugs and improving their bioavailability. This article summaries nanosuspension preparation methods and the stabilizers for flavonoid nanocrystals, and reviews the drug delivery routes including oral, Injection and transdermal of flavonoid nanocrystals, to provide information for further research on nanocrystal delivery system of flavonoids.

Protective effects of active compounds from on heart and brain of mice at simulated high altitude / 浙江大学学报·医学版

To investigate the active compounds from on the heart and brain of mice at simulated high altitude.Fifty healthy male adult BALB/c mice were randomly divided into normal control group, hypoxic model group, acetazolamide group, petroleum ether extract of (PESI) group and octacosan group with 10 mice in each group. Acetazolamide group, PESI group and octacosan group were treated with acetazolamide PESI (200 mg/kg) or octacosan by single tail vein injection, respectively. Except normal control group, the mice were exposed to a simulated high altitude of for in an animal decompression chamber. After the mice were sacrificed by cervical dislocation, the heart and brain were histologically observed by HE staining; superoxide dismutase (SOD) activity, total anti-oxidant capacity (T-AOC) and the content of malondialdehyde (MDA) in plasma, heart and brain tissues were detected by WST-1 method, ABTS method and TBA method, respectively; lactic acid and lactate dehydrogenase (LDH) activity in plasma, heart and brain tissues were detected by colorimetric method and microwell plate method, respectively; ATP content and ATPase activity in heart and brain tissues were detected by colorimetric method. PESI and octacosane significantly attenuated the pathological damages of heart and brain tissue at simulated high altitude; increased SOD activity, T-AOC and LDH activity, and decreased the contents of MDA and lactic acid in plasma, heart and brain tissues; increased the content of ATP in heart and brain tissues; increased the activities of Na-K ATPase, Mg ATPase, Ca ATPase and Ca-Mg ATPase in myocardial tissue; and increased the activities of Mg ATPase, Ca-Mg ATPase in brain tissue. PESI and octacosan exert anti-hypoxic activity by improving the antioxidant capacity, reducing the free radical levels, promoting the anaerobic fermentation, and alleviating the energy deficiency and metabolic disorders caused by hypoxia in mice.

Protective effects of 7-hydroxyethyl chrysin on rats with exercise-induced fatigue in hypobaric hypoxia environment / 浙江大学学报·医学版

: To investigate the protective effect of 7-hydroxyethyl chrysin (7-HEC) on rats with exercise-induced fatigue in hypobaric hypoxic condition.Forty healthy male Wistar rats were randomly divided into four groups with 10 rats in each group: control group, model group, chrysin group and 7-HEC group. The rats in control group were raised at local altitude but other three groups were raised in a simulating altitude of for hypobaric hypoxia treatment. The chrysin group and 7-HEC group were given chrysin or 7-HEC by gavage for respectively; while the control group and model group were given the same amount of sterilized water. The weight-bearing swimming tests were performed 3 d later, and the weight-bearing swimming time was documented. After rats were sacrificed, the liver and skeletal muscle tissue samples were taken for pathological examination and determination of lactate, malondialdehyde (MDA), total superoxide dismutase (T-SOD) and glycogen levels. Blood urea nitrogen was also determined. Compared with the model group, weight-bearing swimming times were significantly prolonged in 7-HEC group [ vs. (4.04±1.30) min, <0.01]; pathological changes in liver and skeletal muscle tissue were attenuated; generation rate of blood urea nitrogen vs. 0.60) mmol·L·min, <0.05], lactate [liver: (0.14±0.05) vs. (0.10±0.03) mg·g·min, skeletal muscle: vs. (0.18±] and MDA [liver: (0.48) vs. (0.78±0.28) nmol·mg·min, skeletal muscle: (0.87±0.19) vs. (0.63±0.11) nmol·mg·min] were significantly reduced (all < 0.05); glycogen content [liver: (15.16±2.69) vs. skeletal muscle: (1.46±0.49) vs.0.48) mg/g] and T-SOD [liver: (1.87±0.01) vs. (2.68±0.12) U/mL, skeletal muscle: 0.42) vs. 0.96) U/mL] were significantly improved (all <0.05). 7-HEC has significant protective effect on the rats with exercise-induced fatigue in hypobaric hypoxia condition.

Establishment of a hypobaric hypoxia-induced cell injury model in PC12 cells / 浙江大学学报·医学版

To construct a hypobaric hypoxia-induced cell injury model. Rat pheochromocytoma PC12 cells were randomly divided into control group, normobaric hypoxia group and hypobaric hypoxia group. The cells in control group were cultured at normal condition, while cells in other two groups were cultured in normobaric hypoxia and hypobaric hypoxia conditions, respectively. CCK-8 method was used to detect cell viability to determine the optimal modeling conditions like the oxygen concentration, atmospheric pressure and low-pressure hypoxia time. The contents of lactate dehydrogenase (LDH), superoxide dismutase (SOD) and malondialdehyde (MDA) were detected by microplate method. The apoptosis ratio and cell cycle were analyzed by flow cytometry. The hypobaric hypoxia-induced cell injury model can be established by culturing for 24 h at 1% oxygen concentration and 41 kPa atmospheric pressure. Compared with the control group and normobaric hypoxia group, the activity of LDH and the content of MDA in hypobaric hypoxia group were significantly increased, the activity of SOD was decreased, the percentage of apoptosis was increased (all <0.05), and the cell cycle was arrested in G0/G1 phase. A stable and reliable cell injury model induced by hypobaric hypoxia has been established with PC12 cells, which provides a suitable cell model for the experimental study on nerve injury induced by hypoxia at high altitude.

Progress in cerebrospinal fluid proteome technology and its clinical application / 生物工程学报

Cerebrospinal fluid surrounds and supports the central nervous system, including the ventricles and subarachnoid spaces. Cerebrospinal fluid should be an important source of biomarkers for central nervous system diseases because it is in direct contact with the central nervous system. Many studies are reported on cerebrospinal fluid proteomics, highlighting many recent progresses. Here, we review recent advances in proteomics technology and clinical application of cerebrospinal fluid.

Anti-hypoxia activity and its protective effects of Lishukang capsule on rat brain tissue at simulated high altitude hypoxia / 药学实践杂志

Objective To investigate anti-hypoxia activity and protective effects of Lishukang capsule on rat brain tissue at simulated high altitude hypoxia.Methods The anti-hypoxic activity of Lishukang capsule was evaluated with normobaric hypoxia test and acute hypobaric hypoxia test in mice.In addition,rats were exposed to large hypobaric hypoxia chamber stim-ulating 8 000 m altitude.The pathological changes of rat brain tissue before and after hypoxia were observed.The oxidative stress indicators and metabolism parameters in brain were measured.Results The low,medium and high Lishukang doses can effectively prolong the survival time of mice(P<0.01)in the dose dependent manner.The medium and high Lishukang doses were significantly better than those of Rhodiola rosea capsules(P<0.05 or P<0.01).The low,medium,high Lishukang dose groups reduced the mortality of acute hypobaric hypoxia mice(P<0.01)with dose dependent effects.The mice mortality in medium and high dose groups was lower than that of Rhodiola rosea group(P< 0.01).Compared with normal control group,the hypoxic model rats exhibited pathological injury in the brain tissue after exposure to hypobaric hypoxia stimulating 8 000 m altitude.The contents of MDA,H2O2,NO,LD and LDH activity increased significantly(P<0.05 or P<0.01), while the activities of SOD,CAT,GPX were significantly decreased(P<0.05 or P<0.01).After pretreatment with Lishu-kang capsule,the pathological damage of rat brain tissue was alleviated and the content of MDA,NO in the brain tissue was re-duced(P<0.05 or P<0.01).The levels of H2O2,LD content and LDH activity in medium and high dose groups were signifi-cantly decreased(P<0.05 or P<0.01).The activities of SOD,CAT and GPX in high dose group were significantly increased(P<0.05).Conclusion Lishukang capsule has good anti-hypoxia activity.It provides protective effect for the injuries induced by hypobaric hypoxia in rats.The mechanism may related to the improvement of antioxidant capability,reduction of free radical damage and amelioration of energy metabolism.

Study on myocardium and brain damage in rats by simulating high altitude / 药学实践杂志

Objective To investigate the effect of hypoxia with cold on the heart and brain damage in rats by simulating 6 000 m high altitude at different exposure time,established a rat model of acute mountain sickness for the related mechanism studies.Methods 32 healthy male Wistar rats were randomly divided into normal control group,hypoxia with cold 1 d,3 d and 5 d group,8 rats in each group.The normal control group was kept in the plain environment(1 500 m)without any treat-ment.The other three groups were placed in large hypobaric hypoxia chamber to simulate 6 000 m altitude with different ex-posed times.HE staining was used to observe the pathological changes of heart and brain tissue.The changes of biochemical indexes were measured to evaluate the damage of heart and brain tissue at different hypoxia times.Results HE staining showed that hypoxia with cold induced rat heart and brain damage with different degrees.The myocardial tissue damage was in-creased with exposure time.The most serious brain damage happened in day 3.Compared with the normal control group,the content of MDA and LD in the myocardial tissue of hypoxia rats were significantly increased(P<0.05 or P<0.01)with pro-longed time,while the contents of GSH,T-SOD and the activity of Na+K+-ATPase were reduced(P<0.05 or P<0.01). The content of MDA in brain tissue was significantly increased at day 1 and day 3(P<0.05 or P<0.01).LD content was sig-nificantly increased(P<0.05)with time.The content of GSH,the activity of T-SOD and Na+K+-ATPase were significantly reduced in day 3(P<0.05).Conclusion Simulating an altitude of 6 000 m caused obvious damage on the heart and brain tis-sues of rats.The degree of damage was related to the exposure time to hypoxia with cold.The decrease of body′s antioxidant capacity,the increase of free radicals and energy metabolism disorders are important factors leading to heart and brain injury.

Pharmacodynamics of the effective anti-hypoxia constituents in Saussurea involu-crat a / 药学实践杂志

Objective To study pharmacodynamics of the effective anti-hypoxia components in the petroleum ether ex-tract of Saussurea Involucrate(PESI)and octacosane.Methods PESI and octacosane were first evaluated by normobaric hy-poxia model,acute decompression model and followed by chemical induced hypoxic models with potassium cyanide,sodium ni-trite and isoprenaline hydrochloride poisoning.Results PESI and octacosane can effectively prolong the survival time of hypo-baric hypoxic mice(P<0.01)and reduce the mortality of acute hypobaric hypoxia mice(P<0.01)in a dose-dependent man-ner.Anti-hypoxic potency of PESI and octacosane obtained by chemical induced hypoxic model indicated that they significantly increase survival time(P<0.05)of hypoxia mice than acetazolamide.Conclusion PESI and octacosane have good anti-hypoxia activity.

Effects of the active constituents from Saussurea involucrata on metabolism of mice at simulated high altitude / 药学实践杂志

Objective To investigate the anti-hypoxia effects of octacosane and the petroleum ether extract from Saus-surea Involucrate(PESI)on the water,sugar,lipid and protein metabolism of mice at simulated high altitude.Methods The healthy adult male BALB/C mice were randomly divided into normal control group,hypoxic model group,acetazolamide group, the petroleum ether of Saussurea involucrata group and octacosane group.Drugs were administered i.v 20 mins before the mice were exposed to a simulated high altitude of 6 000 m for 8 hours in an animal decompression chamber.The mice were sacrificed at the end of 8 hours.Organ water content,organ indexes and metabolism indicators of sugar,protein and lipid were deter-mined.Results The edema of heart,brain and lung was reduced notably(P<0.05,P<0.01)in the mice received PESI at 200 mg/kg and octacosane at 100 mg/kg.In the treated groups,the increase of blood sugar,muscle glycogen,TG(triglycer-ide),TC(total cholesterol)were all significantly inhibited,the decrease of liver glycogen,the protein content of heart and brain was also remarkably blocked(P<0.05,P<0.01).Conclusion PESI and octacosane effectively regulate the metabolism of hypoxic mice and reserve the body′s energy for survival by lowering the basic metabolism.

Effect of resveratrol on peak bone mass in growing rats / 浙江大学学报·医学版

Objective: To investigate the effect of resveratrol on peak bone mineral density and bone mass in growing rats. Methods: Thirty-six female healthy Wistar rats were randomly divided into control group, icariin group and resveratrol group with 12 rats in each group. Icariin (25 mg·kg-1·d-1), resveratrol (8.4 mg·kg-1·d-1) or equal volume of distilled water were given by gavage to icariin group, resveratrol group and control group, respectively. The rats were sacrificed after 12 weeks. The organ indexes were calculated and pathology sections were observed; the bone mineral density (BMD), bone biomechanics, serum bone metabolism index, and results of micro-CT scan were analyzed. Results: During the experiment, the body weight of rats showed an increasing trend and there was no significant difference among three groups (P0.05). There were no significant differences in organ index of vital organs and pathological changes among the groups (all P0.05). Compared with the control group, the whole body BMD, and the BMDs of femur and vertebrae in icariin and resveratrol groups were significantly increased after 12 weeks (all PPPPPPPConclusion: Resveratrol can inhibit bone resorption and enhance bone formation, so as to improve the peak bone mass and bone density, enhance bone strength and improve the microstructure of bone tissue in young rats.

Primary cilium acts as an oxygen sensor in PC12 cells / 浙江大学学报·医学版

Objective: To investigate the function of primary cilium as an oxygen sensor in PC12 cells. Methods: The PC12 cells were transfected with IFT88 siRNA. The nuclear translocation of hypoxia inducible factor-1α (HIF-1α), nuclear factor erythroid-2 related factor 2 (Nrf2), and ciliogenesis were observed by immunofluorescence staining; and the mRNA expressions of HIF-1α, Nrf2, vascular endothelial growth factor (VEGF) and superoxide dismutase (SOD) were detected by real-time RT-PCR. Results: The ciliogenesis was inhibited in PC12 cells transfected with IFT88 siRNA. In hypoxia group and scramble control group, nuclear translocations of HIF-1α and Nrf2 were observed and mRNA expressions of HIF-1α, Nrf2, VEGF were increased, and those of SOD were decreased. While in PC12 cells transfected with IFT88 siRNA, nuclear translocations of HIF-1α and Nrf2 were not observed, and mRNA expressions of HIF-1α, Nrf2, VEGF were inhibited, and mRNA expression of SOD was increased. Conclusion: Primary cilia may act as an oxygen sensor to transfer the information related to hypoxia and oxidative stress into cells, activating intracellular defense mechanism against the hypoxic injuries.

Effect of gastrodin on arterial blood gas and brain injury of rats under simulated high altitude hypoxia environment / 解放军医学杂志

Objective To study the effect of gastrodin on arterial blood gas and brain injury of rats under simulated high altitude hypoxia environment. Methods A total of 60 adult healthy male Wistar rats were randomly divided into normal (N) group, hypoxia model (M) group, rhodiola crenulata (RC) group, low dose of gastrodin (GAS-L) group, medium dose of gastrodin (GAS-M) group and high dose of gastrodin (GAS-H) group (10 for each group). The intragastric administration on rats was continued for 7 days timely in each day. Under simulated 8000m altitude using low pressure oxygen cabin, the arterial blood gas of each group were tested, pathological changes of brain tissues were observed and related indexes of brain were detected after 12h hypoxia. Results Comparing with group N, the blood oxygen partial pressure (PO2), value of blood oxygen saturation (SO2), oxygenation index (PO2/FIO2), Na+ concentration (Na+), actual bicarbonate radical (HCO3–) significantly decreased (P<0.01), lactic acid (Lac), hemoglobin concentration (Hb) significantly increased (P<0.01) and pathological damage was inflicted in group M; and contents of malondialdehyde (MDA), hydrogen peroxide (H2O2) in brain tissue significantly increased (P<0.01), content of glutathione(GSH) and activity of glutathione peroxidase (GSH-Px) in brain tissue significantly decreased (P<0.01) in group M. Compared with group M, PO2, SO2 and PO2/FIO2 significantly increased (P<0.01, P<0.05) in group GAS-L; Na+ and HCO3– significantly increased (P<0.01, P<0.05) in three dose groups of GAS; Lac significantly decreased (P<0.01, P<0.05) in group GAS-L and GAS-H. Hb significantly increased (P<0.01) in group GAS-H, a rising trend appeared in group GAS-L but with no statistical significance. Damages of brain tissue were alleviated in group RC and three dose groups of GAS comparing with group M. Compared with group M, MDA significantly decreased (P<0.01) in three dose groups of GAS; there was a decreasing trend of H2O2 but with no statistical significance in three dose groups of GAS; GSH and GSH-Px significantly increased (P<0.01, P<0.05) in three dose groups of GAS. However, three groups of GAS has no dose dependent. Conclusion There was an protective effect of gastrodin on arterial blood gas and brain injury of rats under simulated high altitude hypoxia environment.

Content Determination of Icariin in Kangguzhi Zengsheng Tablet by HPLC / 中国药房

OBJECTIVE:To establish a method for the content determination of icariin in Kangguzhi zengsheng tablet,and pro-vide a basis for improving the quality standard. METHODS:HPLC was performed on the column of Hypersil ODS2 C18 with mo-bile phase of acetonitrile- water(27∶73,V/V)at a flow rate of 1.0 ml/min,detection wavelength was 270 nm,column temperature was 25℃,and injection volume was 10μl. RESULTS:The linear range of icariin was 3-60μg/ml(r=0.999 9);limit of quantita-tion was 11.2 μg/ml,limit of detection 4.3 μg/ml;RSDs of precision,stability and reproducibility tests were lower than 2.0%;re-covery was 99.00%-100.67%(RSD=0.63%,n=6). CONCLUSIONS:The method is simple,accurate and reproducible,and can be used for the content determination of icariin in Kangguzhi zengsheng tablet.

Research progress in mitochondria-targeted antioxidants / 药学实践杂志

Mitochondria are the main places of cellular respiration as well as the citric acid cycle and oxidative phospho‐rylation .It plays an important role in controlling the life and death of cells .Mitochondrial dysfunction leads to a series of human diseases such as ischemia‐reperfusion injury ,sepsis and diabetes .Mitochondrial become an attractive target for drug transporters strategy and therapeutic targets for neurodegeneration .Although the molecular mechanisms responsible for mitochondria media‐ted disease processes are not fully elucidated yet ,the oxidative stress appears to be critical .Accordingly ,strategies are being de‐veloped for the targeted delivery of antioxidants to mitochondria .The prospect of development of mitochondrial targeted drugs with anti‐oxidative stress protection is tempting .Mitochondrial targeting antioxidants were the antioxidant drugs which took mi‐tochondria as the target site .In this review ,weintroduced the conception and classification of mitochondrial targeted antioxidants and the research progress of disease treatment by mitochondrial targeted antioxidants .

Mechanism s of damga e and treatments of cerebral ischemia and reperfusion / 药学实践杂志

To summarize the advances in mechanisms of damage and treatments of cerebral ischemia and reperfusion and fore -casts future research directions .The existing achievements in literatures were summarized .Cerebral ischemia and reperfusion damage were related to inflammatory response , intracellular calcium overload , free radicals injury , release of excitatory amino acids and other factors.The treatments included reducing ischemia time , blocking glutamate receptors , free radical scavenging , inhibition of apoptosis , reducing inflammation , and promoting neuronal growth , etc.Multi-target treatment would be future directions in treatment of cerebral ischemia.

Effect of icariin and genistein on bone protection / 中国药理学通报

Aim To compare the pharmacological ac-tivity of icariin( ICA) and genistien ( GEN) against os-teoporosis after oral administration with them to growing rats and ovariectomized rats. Methods 25 mg·kg-1 icariin and 10 mg · kg-1 genistein ( equal in molar concentration) were administered to one-month-old fe-male SD rats every day for three months. Treatments at the same dosage were administered to the 6-month-old ovariectomized SD rats every day for three months. Their effects were compared on bone mineral density and biomechanical properties of femurs and vertebrae, serum levels of osteocalcin and tartaric acid phospha-tase 5b ( TRACP 5b) and histomorphometry. Results The results showed that, in young rats, icariin treat-ment significantly increased bone mineral density, the maximum mechanical loads of femurs and vertebrae as well as the bone qualities ( serum markers and microar-chitecture ) , whereas genistein treatment had little effects compared with the non-treatment control. How-ever, genistein treatment was more efficacious than icariin in preventing bone loss and deterioration of bone microarchitecture in ovariectomized rats. Conclusion Our data suggest that, since icariin has a higher os-teogenic activity but lower estrogenic activity, it has been found to be more efficacious than genistein in peak bone mass accrual only in young rats. In the ovariectimized rats, however, as the main force to pre-vent bone loss is the estrogenic activity, genistein has been found to be more efficacious than icariin in reduc-ing bone loss.

Determination of Salvianolic Acid B in Ansheng Yizhi Capsules by HPLC / 中国药师

Objective:To establish an HPLC method for the content determination of salvianolic acid B in Ansheng Yizhi cap-sules. Methods:A Hypersil ODS2 C18 column(150 mm × 4. 6 mm,5 μm) was used and methanol-water-formic acid (40∶60∶1) was used as the mobile phase. The detection wavelength was at 286 nm. The flow rate was 1. 0 ml·min-1 and the sample size was 10 μl. Results:The calibration curve of salvianolic acid B was linear within the range 7. 75-77. 51 μg·ml-1(r=0. 999 6). The average re-covery was 98. 17%(RSD=1. 79%, n=6). Conclusion:The method is simple, accurate and repeatable, which can be used in the quality control of Ansheng Yizhi capsules.

Protective effect of genistein on hypoxic injuries of osteoblasts cultivated in vitro / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the effect of genistein on osteoblast proliferation, cellular cycle, apoptosis and differentiation of osteoblasts cultivated under hypoxia conditions.</p><p><b>METHOD</b>Rat osteoblasts were isolated from calvarias by enzyme digestion and a hypoxic model was established by in a triple-gas incubator. Rat osteoblasts were grouped into the normoxic control group, the hypoxia control group and the hypoxia administration group which was subdivided into Ge-6 group, Ge-5 group and Ge-4 group, to which genistein was administered at doses of 1 x 10(-6), 1 x 10(-5), 1 x 10(-4) mol x L(-1). The cell survival rate, lactic dehydrogenase leakage rate, apoptosis and differentiation of osteoblasts were observed for each group at 3 h after hypoxia, and the gene expression of HIF-1alpha, Bcl-2, Caspase-3 was detected by Real time RT-PCR. Forty-eight hours after hypoxia, osteogenic differentiation markers including alkaline phosphatase activity and nodules were detected.</p><p><b>RESULT</b>Compared with the hypoxia control group, the hypoxia administration group displays a significant increase in the survival rate and a decreased in LDH leakage rate, apoptosis rate and percentage of S + G2 phases. Besides, the mRNA level of HIF-1alpha and Bcl-2 were enhanced, the mRNA level of Caspase-3 was inhibited.</p><p><b>CONCLUSION</b>Genistein has an effect on protecting osteoblasts from hypoxia.</p>