Effects and mechanism of Danshen decoction on physiological characteristics and function of platelet in hyperli-pidemia model rats / 中国药房

OBJECTIVE To investigate the effects and mechanism of Danshen decoction influencing platelet physiological characteristics and function in hyperlipidemia model rats based on platelet membrane glycoprotein 4 （CD36）/phosphatidylinositol 3- kinase （PI3K）/protein kinase B （Akt） signaling pathway. METHODS The hyperlipidemia model rats were induced by feeding high- fat diet， and then randomly divided into model group， simvastatin group （0.004 g/kg）， Danshen decoction high-dose and low-dose groups （3.6， 0.9 g/kg）， and blank group （fed with basic feed）， with 10 rats in each group. Rats in each administration group were intragastrically administered with corresponding drugs every day， and the other groups were intragastrically administered with equal volume of normal saline for 4 weeks. After the last administration， the contents of blood lipid biochemical indexes [total cholesterol （TC）， triglyceride （TG）， high-density lipoprotein cholesterol （HDL-C）， low-density lipoprotein cholesterol （LDL-C）]， whole blood viscosity， plasma viscosity and fibrinogen （FIB）content in plasma， platelet-related parameters [platelet count （PLT）， platelet distribution width （PDW） and mean plateletvolume （MPV）] were detected. The levels of plateletphysiological characteristics and function-related factors [von Willebrand factor （vWF）， fibronectin （Fn）， phospholipase A2（PLA2）， thromboxane B2 （TXB2）， 6-keto-prostaglandin F1α （6-keto-PGF1α）， thromboxane A2 （TXA2）， prostaglandin I2 （PGI2）， cyclic adenosine monophosphate （cAMP）， cyclic guanosine monophosphate （cGMP）， β-thromboglobulin （β-TG）]， platelet aggregation rate （maximum aggregation rate， 60 s aggregation rate， 180 s aggregation rate） and fibrinolytic system-related factors [tissue plasminogen activator （t-PA）， plasminogen activator inhibitor 1 （PAI-1）] and the expressions of CD36/PI3K/Akt signaling pathway-related proteins [CD36， focal adhesion kinase （FAK）， phosphatidylinositol 4-phosphate 5-kinase （PIP5K）， PI3K， phosphorylated Akt （p-Akt）， p-Akt1/2/3] in platelet were all determined. RESULTS Compared with blank group， the contents of TC， TG and LDL-C in plasma， whole blood viscosity， plasma viscosity， the plasma levels of FIB， PLT， MPV， vWF， Fn， PLA2， TXB2， TXA2， cGMP and β-TG， maximum platelet aggregation rate， 60 s aggregation rate， the expressions of PAI-1 in plasma， protein expressions of CD36， FAK， PIP5K， PI3K， p-Akt and p-Akt1/2/3 were significantly increased in the model group （P＜0.05）. The content of HDL-C and the levels of 6-keto-PGF1α， PGI2 and t-PA were significantly decreased （P＜0.05）. After the intervention of Danshen decoction， most of the above indexes were significantly reversed （P＜0.05）. CONCLUSIONS Danshen decoction can improve the physiological characteristics and function of platelets in hyperlipidemia model rats， and its mechanism may be related to the down-regulation of CD36/PI3K/Akt signaling pathway activity and the reduction of platelet activation.

Effects of Naringin on neuronal apoptosis in mice with memory consolidation disorder / 国际中医中药杂志

Objective:To observe the effect of Naringin on neuronal apoptosis in mice with memory consolidation disorderinduced by sodium nitrite.Methods:Fifty mice were randomly divided into blank group, model group, standardized protocol group, high-dose Naringin group and low-dose Naringin group, with 10 mice in each group. The standardized protocol group was given Donepezil 1 mg/kg, the Naringin high and low dose groups were gavaged with Naringin solution 100 and 50 mg/(kg·d), blank group and model group were gavaged with equal volume of distilled water once a day for 21 days. The model was established on the 22nd day. The blank group was intraperitoneally injected with normal saline, and the other groups were intraperitoneally injected with 100 mg/(kg·d) sodium nitrite solution for 7 days. The cognitive ability of mice in each group was evaluated by platform jumping test, and the hippocampal synaptic structure was observed by electron microscope. The contents of acetylcholine (ACh), SOD, MDA and NO in hippocampus and the activity of choline acetyltransferase (ChAT) and acetylcholinesterase (AChE) was detected by ELISA. The expression of N-methyl-D-aspartate receptor (NMDAR), glutamine receptor 2 (GluR), calcium/calmodulin dependent protease Ⅱ (CaMK Ⅱ), Caspase-3, Bcl-2 and Bad proteins in hippocampus of model mice were detected by Western blot.Results:The number and morphology of hippocampal neurons were normal, nucleus, mitochondria, rough endoplasmic reticulum and synaptic membrane of hippocampal neurons in high-dose Naringin group were clear. Compared with the model group, the latency of mice in the high-dose Naringin group was prolonged and the number of errors was reduced ( P<0.01). The levels of MDA and NO in hippocampus of mice in the high-dose Naringin group significantly decreased ( P<0.01), and the activity of SOD significantly increased ( P<0.01). The content of ACh (23.682 ± 2.835 μg/mg prot vs. 14.939 ± 2.901 μg/mg prot), ChAT (163.302 ± 21.278 U/g vs. 89.612 ± 11.497 U/g) increased, AChE (0.367 ± 0.015 U/mg prot vs. 0.471 ± 0.014 U/mg prot) activity decreased ( P<0.01); The expression of Bad (0.441 ± 0.010 vs. 0.633 ± 0.010), Caspase-3 (0.425 ± 0.036 vs. 0.537 ± 0.024) significantly decreased, and the expression of Bcl-2 (0.890 ± 0.014 vs. 0.727 ± 0.009) significantly increased ( P<0.01); The expression of CAMKⅡ (1.043 ± 0.037 vs. 1.475 ± 0.043) significantly decreased ( P<0.01), and the expression of NMDAR1 (0.407 ± 0.037 vs. 0.345 ± 0.012), GluR2 (1.125 ± 0.033 vs. 0.664 ± 0.023) significantly increased ( P<0.01). Conclusion:Naringin could play the role of protecing the neuron and improving the cognition of mice with memory consolidation disorder by regulating the balance of ACh and glutamate system and reducing neuronal apoptosis and antioxidant stress.

Effects of Buyang Huanwu Decoction on Hemorheology and Platelet Related Biological Indexes in Hyperlipidemia Model Rats / 中国药房

OBJECTIVE：To investigate the effe cts of Buyang huanwu decoction on hemorheology and platelet related biological indexes of hyperlipidemia model rats. METHODS ：Male Wistar rats were randomly divided into blank control group （10 rats）and model group （40 rats）. Blank control group was given normal diet ，and model group was given high-lipid diet for 6 weeks at least to induce hypelipidemia model. After modeling ，rats were randomly divided into model control group ，positive control group （simvastatin，0.004 g/kg），Buyang huanwu decoction low-dose and medium-dose groups （3.5，14.0 g/kg，by crude drug），with 10 rats in each group. Blank control group and model control group were given normal saline intragastrically ， administration groups were given relevant drug intragastrically ，0.01 mL/g，once a day ，for consecutive 4 weeks. Thirty min after last medication ，hemorheological indexes （whole blood viscosity ，plasma viscosity ），platelet adhesion related indexes [adhesion rate，von Willebrand factor （vWF），fibronectin（FN）]，platelet release related indexes [ β-thromboglobulin（β-TG），platelet factor 4 （PF4）] and platelet fibrinolytic system related indexes [tissue plasminogen activator （t-PA），plasminogen activator inhibitor （PAI-1）]，platelet parameters （PLT，PDW，MPV，PCT，PLCR），4 kinds of coagulation parameters （APTT，TT，PT，FIB）were detected. RESULTS ：Compared with blank control group ，the whole blood viscosity （low，medium and high shear rate ），plasma viscosity，platelet adhesion rate ，the contents or levels of vWF ，FN，β-TG，PAI-1，PLT，MPV，PCT，PLCR and FIB in model control group were increased significantly （P＜0.05 or P＜0.01），and t-PA content was significantly decreased （P＜0.05）. Compared with model control group ，the whole blood viscosity （except for whole blood viscosity of high shear rate in Buyang huanwu decoction high-dose group ），plasma viscosity ，platelet adhesion rate ，the contents or levels of vWF ，FN，β-TG，PAI-1， PLT，PDW（except for Buyang huanwu decoction low-dose and high-dose groups ），MPV，PCT，PLCR（except for Byang huanwu decoction low-dose group ）and FIB were decreased significantly （P＜0.05 or P＜0.01），while t-PA content （except for positive control group ） was increased significantly （P＜0.05）. CONCLUSIONS ：Buyang huanwu decoction can significantly improve the pathological state in hyperlipidemia model rats by reducing blood viscosity and FN content ，improving platelet adhesion，enhancing fibrinolytic activity ，improving platelet aggregation ，inhibiting hypercoagulability and hyperplatelet release.

Experimental Study on the Protective Effect of Shenqi Zhilong Decoction Combined with Tripterygium Polygly- coside Tablets on Membranous Nephropathy Model Rats / 中国药房

OBJECTIVE：To investigate the protective effect an d possible m echanism of Shenqi zhilong decoction （SZD） combined with Tripterygium polyglycoside tablets （TPT）on membranous nephropathy （MN）model rats. METHODS ：MN rat model was established by subcutaneous and caudal vein injecting cationic bovine serum albumin + incomplete Freund adjuvant emulsion for 6 weeks. At the 3rd week of modeling ，model rats were randomly divided into model control group ，SZD low-dose and high-dose groups （4，8 g/kg，by total crude drugs ），TPT group （9 mg/kg），low-dose and high-dose of SZD+TPT groups （same dose as single group ），with 10 rats in each group according to 24 h UTP and weight. Another 10 rats withoutmodeling were taken as blank control group. Blank control group was given equal amount of water intragastrically administration groups were given relevant medicine intragastrically，once a day ，for consecutive 4 weeks. The 24 h UTP of rats were detected one day before the last administration；1 h after the la st administration ，blood routine 中indexes（WBC，RBC，PLT），liver func tion indexes （TP，ALB，AST，ALT），blood lipid indexes （TC，TG，HDL-C，LDL-C）， the contents of glucose （GLU），urea nitrogen （BUN）and serum creatinine （Scr）were detected in each group. Uranyl acetate-lead citrate staining was used to observe ultrastructural changes of renal tissue. Immunohistochemical method was used to detect the protein expression of TGF-β1 and HPA- 1 in renal tissue. RESULTS ：Compared with blank control group ，in the model control group，the glomerular podocytes were widely fused or disappeared ，microvilli were formed ，basement membrane was heavily thickened，a large number of electron dense substance was deposited under the epithelium ，TGF-β1 and HPA- 1 positive cells were significantly increased ；24 h UTP ，PLT，the contents of TC ，TG and HDL-C ，the percentage of TGF-β1 and HPA- 1 positive cells were increased significantly ，while RBC ，the contents of TP and ALB were decreased significantly （P＜0.05 or P＜0.01）. Compared with model control group ，above ultrastructural changes of administration groups were improved to different extents ，and TGF-β1 and HPA- 1 positive cells were decreased. The 24 h UTP ，the percentage of TGF-β1 positive cells （except for SZD low-dose group），WBC（except for SZD alone groups and combination groups ），PLT and TC content （except for TPT group ），TG content （except for SZD low-dose alone and its combination group ），the percentage of HPA- 1 positive cells were decreased significantly ； the percentage of TGF-β1 positive cells in SZD high-dose+TPT group as well as the percentage of HPA- 1 positive cells in SZD+TPT groups were significantly lower than TPT group. RBC and GLU content （except for SZD low-dose group and TPT group ），TP and ALB content （except for SZD low-dose group ）were increased significantly ，while the content of GLU in SZD high-dose+TPT group was significantly higher than TPT tablets group （P＜0.05 or P＜0.01）. CONCLUSIONS ：SZD combined with TPT can relieve myelosuppression caused by TPT ，reduce proteinuria of MN model rats and improve pathological damage of renal tissue in rats. Its mechanism is related to the down-regulation of protein expression of TGF-β1 and HPA- 1，and the reduction of TC and TG content in the blood.

Effects of Buyang Huanwu Decoction on Lipid Metabolism and Hepatic Tissue in Hyperlipemia Model Rats / 中国药房

OBJECTIVE：To investigate the effects of Buyang huanwu decoction on lipid metabolism and hepatic tissue in hyperlipemia model rats. METHODS ：Male SD rats were randomly divided into blank control group ，model group ，positive control group （simvastatin 0.004 g/kg），Buyang huanwu decoction low-dose and high-dose groups （3.5，14.0 g/kg，by crude drugs），with 10 rats in each group. Except that blank control group was given water intragastrically ，hyperlipidemia mode of other groups was established by intragastric administration of fat emulsion according to the body weight. From the 21st day after modeling，the rats in each group were given water or fat emulsion in the morning and water or corresponding drugs in the afternoon for consecutive 20 days. After last medication ，the contents of TG ，TC，LDL-C，ALT，AST，LCAT，PLTP and HL in plasma were determined in each group ；the contents of LPL ，MTP and HMG-CoA redutase in liver tissue were also determined. The pathological changes of liver tissue were observed in each group. RESULTS ：Compared with blank control group ，the contents of TG ，TC，LDL-C，ALT and HMG-CoA reductase were increased significantly in model group （P＜0.05 or P＜0.01），while the contents of HL ，LPL and MTP were decreased significantly （P＜0.01）. Compared with model group ，the contents of TG ，TC， LDL-C and HMG-CoA reductase in administration groups as well as the contents of ALT ，AST in Buyang huanwu decoction low-dose and high-dose groups were decreased significantly （P＜0.05 or P＜0.01）；the contents of HL and MTP in administration groups as well as the content of LPL in positive control group were increased significantly （P＜0.05 or P＜0.01）. Pathological sections showed that the hepatocytes of model group were arranged irregularly ，and there was obvious cell steatosis. The hepatocytes in positive control group and Buyang Huanwu decoction high-dose group had almost no abnormal change compared with blank control gr oup，and there was a small amount of slight cell steatosis in Buyang Huanwu decoction low-dose group. CONCLUSIONS ： Buyang huanwu decoction has a 础。E-mail：zhang520-888@163.com significant lipid-lowering effect on hyperlipemia model rats ，which may be related to the regulation of lipid absorptiontransport and metabolism by regulating the expression of lipid 126.com metabolism related enzymes and transporter.

Antitumor components screening of Stellera chamaejasme L. under the case of discrete distribution of active data / 药学学报

This is to report the screening, extracting and validating antitumor components and compounds from Stellera chamaejasme L. under the case of discrete distribution of active data. In this work, different components from Stellera chamaejasme L. were collected by HPD macroporous resin and polyamide resin column, and their antitumor activity on A549 were tested by MTT assay. Activity results indicate that activity of components at 30-39 min is more potent than that of Stellera chamaejasme L. extract, and the activity of components at 33.97 min is equivalent to positive drug, cis-platinum at 100 microg x mL(-1), but with totally different mode of action. Under the case of discrete activity, the weight analysis is capable of screening active components and compounds from natural products.

Structure elucidation of degradation products of Z-ligustilide by UPLC-QTOF-MS and NMR spectroscopy / 药学学报

Z-Ligustilide, a major phthalide isolated from a widely used traditional Chinese medicine Ligusticum chuanxiong, possesses various pharmacological activities including neuroprotective, anti-inflammatory, antiproliferative and vasorelaxing effects. However, it is unstable and inclined to degrade in natural conditions, which limits its study and application greatly. In this study, degradation behavior of Z-ligustilide and its degradation products stored at room temperature under direct sunlight were investigated and structure elucidated by HPLC-UV, UPLC-QTOF-MS and NMR. Z-ligustilide degradation and total five degradation products were generated and detected. Two degradation products were unequivocally identified as senkyunolide I and senkyunolide H by comparison with reference compounds. Another two degradation products were further isolated by semi-preparative HPLC and structure elucidated as (E)-6, 7-trans-dihydroxyligustilide and (Z)-6, 7-epoxyligustilide by 1H and 13C NMR, respectively. The degradation pathways of Z-ligustilide were finally proposed. Oxidation, hydrolysis and isomerization are the major degradation reactions.

Study on Chronic Alcoholic Liver Disease Model of Rats / 中国中医药信息杂志

Objective To establish alcoholic liver disease (ALD) model of rats. Method Rats were given wine by ig for 13 weeks. ALD were assessed by the level of serum glutamic-pyruvic transaminase (ALT), glutamic oxalacetic transaminase (AST), malonaldehyde (MDA) and pathological changes of liver. Result Over-dose wine can increase the level of serum ALT, AST, and MDA, decrease the level of serum albumin (ALB), and induce hepatic steatosis. Conclusion Alcoholic liver disease model of rats can be established by long-time and over-dose wine. The model can be applied in the research of ALD.

Experimental study on effect of Rupixiao Grains on mammary hyperplasia rats / 中华中医药杂志

Objective: To study the curative effects of Rupixiao Grains on mammary hyperplasia rats and provide the bisis for the wider use. Methods: The model of mammary hyperplasia were established with estrogen and progestogen, the changes of nipple appearance and pathological tissue of mammary gland on morphologic change were observed. The E2, P and PRL of serum were determined. Results: The treatment groups of high and middle dose of Rupixiao Grains can siginifi cantly reduce the diameter and height of nipple in rat mammary gland hyperplasia models(P

Studies on chemical constituents of Gastrodia elata / 中草药

Object To study the low polarity constituents of Gastrodia elata Blume Methods The constituents were isolated by column with prepared TLC, and identified by NMR, MS spectra Results Seven compounds were isolated and identified They were p hydroxybenzyl ethyl ether (Ⅰ), ? sitosterol (Ⅱ), palmitic acid (Ⅲ), 2, 2′ methylenebis (6 tert butyl 4 methylphenl) (Ⅳ), p hydroxy benzyl methyl ether (Ⅴ), dimethyl phthalate (Ⅵ), p hydroxybenz aldehyde (Ⅶ) Conclusion Two compounds were isolated from G elata firstly, and compound Ⅳ as tetra benzol dimer is also isolated from the plants of Gastrodia R. Br. and Orchidaceae for the first time

Chemical constituents from Gastrodia elata(Ⅲ) / 中草药

Objective To study the high polar constituents of Gastrodia elata.Methods The constituents were isolated by macroporous resin column chromatography combined with preparative HPLC and identified by spectral analyses and physico-chemical properties comparisons.Results Ten compounds were isolated from H2O and 20% EtOH fractions eluted from resin column and their structures were identified as citric acid monoethyl ester(Ⅰ),uridine(Ⅱ),adenine(Ⅲ),dactylose A(Ⅳ),uracil(Ⅴ),tyrosine(Ⅵ),N2-(p-hydroxybenzyl) guanosine(Ⅶ),1-isoferuloyl-?-D-glucopyranoside(Ⅷ),4-(?-D-glucopyranosyloxy) benzaldehyde(Ⅸ),and p-hydroxyl benzoic acid(Ⅹ).Conclusion Compound Ⅶ is a new compound named gastronucleoside.Other nine compounds are isolated from G.elata for the first time.

Chemical constituents in active fraction from Euonymus alatus with anti-myocardial ischemic effect / 中草药

Objective To study the chemical constituents of the EtOAc soluble fraction obtained from the stems and branches of Euonymus alatus with anti-myocardial ischemic effect. Methods Chemical constituents were isolated and purified by solvent extraction together with various chromatographic techniques, and the structures were identified by physicochemical properties and spectral analyses. ResultsFive compounds, 1-[3-?-D-glucopyranosyloxy)-4, 5-dihydroxyphenyl]-ethanone (Ⅰ), phenethyl alcohol 8-O-?-D-glucopyranosyl-(1→2)-?-D-glucopyranoside (Ⅱ), eugenyl-O-?-D-glucopyranoside (Ⅲ), hyperin (Ⅳ), and hesperidin (Ⅴ) were isolated. Conclusion Compound Ⅰ is determined as a new compound named guijianyuside. Compounds Ⅱ and Ⅲ are isolated from this plant for the first time.

Protective effects of active fraction and constituents from Flos Daturae on Chinese hamster ovary cells injuried by dimethyl sulfoxide / 中草药

Objective To observe the protective effects of various extracts from Flos Daturae (FD), including active fraction (AF-FD), withanolides constituents (WC-FD), and flavonoids constituents (FC-FD) on Chinese hamster ovary (CHO) cells against cytotoxicity induced by DMSO. Methods The survival rate of CHO cells was examined by MTT assay and LDH leakage assays. Results Using MTT assay, coincubation of CHO cells with 3% DMSO for 24 h resulted in a significant reduction of survival rate of CHO cells. AF-FD was tested in a range of 10—80 ?g/mL to improve the survival rate of CHO cells in a dose-dependent manner. FC-FD (2.5—20 ?g/mL), but not WC-FD (30—120 ?g/mL), could significantly relieve the injury induced by 3% DMSO in CHO cells. In the measurement of LDH leakage, coincubation of CHO cells with 4.5% DMSO for 24 h obviously increased LDH release. However, all the compounds tested, including AF-FD (10—80 ?g/mL), WC-FD (30—120 ?g/mL), and FC-FD (2.5—20 ?g/mL) had no effect on LDH leakage induced by 4.5% DMSO. Conclusion The findings suggest that the FC-FD may protect CHO cells from DMSO cytotoxicity assessed by MTT assay, which may be associated with improving mitochondrial function, but not protecting the membrane injury of CHO cells.

HPLC Fingerprint of Gastrodia elata / 中草药

Objective To establish HPLC fingerprint for evaluating and controlling the quality of Gastrodia elata. Methods HPLC Analysis and similarity calculation were used for establishing fingerprint chromatogram and classifying 18 different original samples; LC-MS and pure compound comparisons were employed for the fingerprint peaks identification. Results HPLC Fingerprint chromatogram was cons- tructed with 27 fingerprint peaks, 17 of which were common fingerpring peaks and 12 main compounds were identified. According to the similarity to standardize herbal medicine, 18 samples could be classified three groups, Anhui-Henan, Shanxi-Sichuan, and Yunnan habitats. The two critical points were 0.80 and 0.58 for correlation coefficient and 0.88 and 0.73 for cosine values. Conclusion The selected fingerprint peaks are diagnostic for G. elata and the constituted HPLC fingerprint chromatogram could be used for identifying different habitats and served as a powerful tool for further quality control of G. elata.