Influence of olfactory bulbectomy on mouse neurobehaviors,hippocampal neurotransmission and depression relevant gene expressions / 中国药理学通报

Aim To investigate the changes of behav-iors,neurotransmitters and depression related gene ex-pressions in hippocampus of C57BL/6 mice after olfac-tory bulbectomy (OB).Methods Forced swimming test (FST),tail suspension test (TST),open filed test (OFT),and elevated plus maze test (EPMT) were used to evaluate the behavioral changes 18 days after surgery. LC-MS/MS method was employed to measure the hippocampal neurotransmitters.Quantita-tive PCR approach was established to determine the hippocampal gene expressions associated with depres-sion.Results OB mice were hyperactive in OFT (P<0.05 ) accompanied with increased time spent in open arm of EPMT (P<0.01).Meanwhile,the sur-gery led to the reduction of the ratio of 5-HIAA/5-HT (P <0.01 ) but the increase of Glu/GABA (P <0.01 ) in hippocampus.Moreover,OB elevated the gene expressions of BDNF,Trkb,GDNF,CD1 1 b and TNF-αbut down-regulated that of TPH2 in hippocam-pus (P<0.05 ).Conclusion Behavioral alternation of OB mice was a result of comprehensive effect of the changes of neurotransmission and depression related genes,which call us special attention in using OB as an animal model for research and development of anti-depressants.

Effects of FXR gene deletion on neurobehaviors and neurotransmission systems in female mice / 中国药理学通报

Aim To investigate how Farnesoid X re-ceptor (FXR),a nuclear hormone receptor,acts on neurological behaviors such as emotion,social behav-ior,memory and so on.Methods FXR’s function in central nervous system was evaluated by conducting a battery of behavioral tests including elevated plus maze test (EPMT),forced-swimming test (FST),social in-teraction test (SIT ), and passive avoidance test (PAT),and the contents of neurotransmitters were de-termined by the LC-MS /MS method in FXR knockout (KO)female mice and their wild-type controls.Re-sults FXR KO mice showed significantly increased immobility time in FST (P <0.01 ),and it showed in-creased tendency to enter the open arms in EMPT (P<0.01 ).The number of probing the open arms by FXR KO mice was more than that of the controls.Mo-reover,in SIT,FXR KO mice had remarkably in-creased sniffing interactions with the stranger mouse in the same cage (P <0.01 ).But in PAT,the latency for FXR KO mice to enter the dark chamber on the test day and the number of FXR KO mice to enter the dark chamber didn’t differ from those wild-type mice.In hippocampus,the contents of GABA,Glu,and NE were decreased prominently in FXR KO mice (P <0.05,P <0.05 and P <0.01 ,respectively)as well as the ratio of GABA to Glu (P <0.05).But in pre-frontal cortex,none of the neurotransmitters examined showed any difference between FXR KO mice and their controls.Conclusion FXR may be involved in main-tenance of the homeostasis of neural transmission in the central nervous system,thereby influences the emotion-al and social behavior in animals.

Expression of human estrogen receptor alpha and beta in Escherichia coli / 药学学报

Estrogen participates in many life activities through combination with estrogen receptor alpha (ERalpha) or estrogen receptor beta (ERbeta) in the body. In order to establish an in vitro estrogen-like compound screening model, the coding region of human ERalpha and ERbeta was separately constructed into pET32-ERalpha and pET43-ERbeta prokaryotic expression vector and water-soluble recombinant ERalpha and ERbeta proteins were expressed in Escherichia coli strain BL21. Western blotting revealed that both recombinant proteins have estrogen receptor binding sites. The proteins were purified using S-Tag affinity Purification Kit and digested with enterokinase to get the ERalpha and ERbeta proteins. About 0.90 mg of ERalpha and 0.65 mg of ERbeta were obtained at the concentration of 0.181 and 0.131 mg x mL(-1), respectively.

Determination and biosynthesis of multiple salvianolic acids in hairy roots of Salvia miltiorrhiza / 药学学报

Danshen (Salvia miltiorrhiza Bunge) hairy roots were obtained by infecting Danshen leaves with Agrobacterium rhizogenes 9402. Besides rosmarinic acid (RA) and salvianolic acid B (SAB), the hairy root could also produce salvianolic acid K (SAK), salvianolic acid L, ethyl salvianolic acid B (ESAB), methyl salvianolic acid B (MSAB), and a compound with a molecular weight of 538 (compound 538) identified by using LC-MS. Effects of methyl jasmonate (MeJA) and yeast elicitor (YE) on the accumulation of these compounds had been investigated. MeJA increased the accumulation of SAB, RA, SAK, and compound 538 from 4.21%, 2.48%, 0.29%, and 0.01% of dry weight to 7.11%, 3.38%, 0.68%, and 0.04%, respectively. YE stimulated the biosynthesis of RA from 2.83% to 5.71%, but depressed the synthesis of SAB, SAK and compound 538. It was indicated in all the results that these Danshen hairy roots could be used as alternative resources to produce salvianolic acids. Analysis of the content variation of these compounds after elicitation suggested that SAK and compound 538 might be the intermediates in the biosynthesis from RA to SAB in Danshen hairy roots.

Regulation of Vitreoscilla hemoglobin on biosynthesis of astragaloside IV / 药学学报

In the present study, the regulation of Vitreoscilla hemoglobin (VHb) on astragaloside IV biosynthesis was investigated. An intermediate expression vector consisting of the CaMV35S promoter fused to the vgb and nopaline synthase terminator was transferred into Astragalus membranaceus via Agrobacterium rhizogenes. The transgenic hairy roots were confirmed by PCR amplification and Southern blot hybridization. The expression of vgb in transgenic hairy roots was confirmed by RT-PCR. After 15 days cultivation, the dry weight and growth rate of transgenic hairy roots were higher than that of the non-transgenic hairy root. ELSD-HPLC analysis showed that astragaloside IV content of transgenic hairy roots was 5 to 6 times of non-transgenic hairy root control and 10 to 12 times of Radix Astragali from Shanxi Province. These results suggested that the expression of vgb promoted the growth of transgenic hairy roots, and increased the content of astragaloside IV.

Evaluation on hepatotoxicity caused by Dioscorea bulbifera based on analysis of bile acids / 药学学报

Metabolic profile of bile acids was used to evaluate hepatotoxicity of mice caused by ethanol extraction of Dioscorea bulbifera L. (ethanol extraction, ET) and diosbulbin B (DB), separately. Ultra-performance liquid chromatography coupled with quadrupole mass spectrometry (UPLC-MS) was applied to determine the contents of all kinds of endogenous bile acids including free bile acids, taurine conjugates and glycine conjugates. Obvious liver injuries could be observed in mice after administrated with ET and DB. Based on the analysis using principle components analysis (PCA), toxic groups could be distinguished from their control groups, which suggested that the variance of the contents of bile acids could evaluate hepatotoxicity caused by ET and DB. Meanwhile, ET and DB toxic groups were classified in the same trends comparing to control groups in the loading plot, and difference between the two toxic groups could also be observed. DB proved to be one of the toxic components in Dioscorea bulbifera L. Bile acids of tauroursodeoxycholic acid (TUDCA), taurochenodeoxycholic acid (TCDCA), taurocholic acid (TCA), taurodeoxycholic acid (TDCA), cholic acid (CA) and others proved to be important corresponds to ET and DB induced liver injury according to analysis of partial least square-discriminant analysis (PLS-DA) and the statistical analysis showed that there were significant differences between the control groups and toxic groups (P < 0.01). Furthermore, good correlation could be revealed between the foregoing bile acids and ALT, AST. It indicated that taurine conjugated bile acids as TUDCA, TCDCA, TCA and TDCA along with CA could be considered as sensitive biomarkers of ET and DB induced liver injury. This work can provide the base for the further research on the evaluation and mechanism of hepatotoxicity caused by Dioscorea bulbifera L.

Inhibitory effect on activated renin-angiotensin system by astragaloside IV in rats with pressure-overload induced cardiac hypertrophy / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the effect of astragaloside IV (As IV) on the activation of rennin-angiotensin system in rats with pressure-overload induced cardiac hypertrophy.</p><p><b>METHOD</b>Left ventricle hypertrophy was induced by abdominal aorta banding between bilateral renal aortas for 12 weeks. Rats were given astragaloside IV 1.0 mg x kg(-1) and 3.3 mg x kg(-1) for 12 weeks, respectively. After treatment, the left ventricular mass index (LVMI)was calculated by morphometry methods. Plasma and cardiac tissue angiotensin II, and plasma aldosterone were measured by ELISA method. Gene expressions of ACE, AT1 and AT2 in cardiac tissue were detected by real time PCR. Protein expressions of AT1 and AT2 in cardiac tissue were detected by Western blot.</p><p><b>RESULT</b>Compared with model rats, LVMI was decreased by astragaloside IV treatment. Biochemical results indicated that the contents of angiotensin II in plasma and cardiac tissue as well as aldosterone in plasma were all increased in abdominal aorta banding rats comparing with sham-operated rats, then, decreased by astragaloside IV treatment. Gene expressions of cardiac ACE was downregulated by astragaloside IV, however, gene and protein expressions of cardiac AT2 were upregulated by astragaloside IV. Both elevated gene and protein expressions of AT1 were not attenuated by astragaloside IV.</p><p><b>CONCLUSION</b>Excessive activated rennin-angiotensin system in rats with pressure-overload induced cardiac hypertrophy is inhibited by astragaloside IV treatment.</p>

Influence of Blood Replenishing Angelica Decoction on Enhanced Endothelial Permeability Induced by Histamine / 中草药

Blood Replenishing Angelica Decoction [Danggui Buxue Tang (DGBXT)]is a wellknown TCM prescription composed of Radix Angelicae Sinensis and Radix Astragali with the actions ofinvigorating "Qi" and enriching blood. Its action to curtail endothelial permeability induced by histaminewas studied. Endothelial cells isolated from the aorta of neonatal calf were cultured on polycarbonate mi-croporus filter membrane to develop a confluent endothelial monolayer. After purfused with either plainHank's balanced salt solution or that containing 5 g/L albumin, the monolayer was treated with 10-4 mol/L histamine for 30 min either with or without preincubation for 60 min with 10-4 g/mL of DGBXT. Fluidfiltration coefficient (Kf), filtration volume (Jv) and osmotic reflective coefficient (σ) of protein were thenmeasured. The findings showed that DGBXT could curtail the lowering of Kf and Jv and elevation of σ in-duced by histamine, indicating that DGBXT could inhibit the action of histamine on endothelial permeabili-ty, but its mechanism of action needs further study.

Determination of Epicatechin Content in Uncaria macrophylla by HPLC / 中草药

Epicatechin content in Uncaria macrophylla Wall. was determined by RP-HPLC. As aresult, the epicatechin contents were 0.38％ in the hook and stem, and 0. 820％ in the leaf. Thus it seemedto be more worthwhile to produce epicatechin from the leaf rather than from the hook and stem.

Studies on the structural features and anticoagulant activity of the polysaccharide from Aeodes orbitosa / 中国海洋药物

Objective To study the structural features and anticoagulant activity of the polysaccharide isolated from Aeodes orbitosa.Methods Sugar composition analysis,methylation analysis and IR were used to characterize the structural features.The anticoagulant activity of the polysaccharide was evaluated by cutting tail and capillary methods.Results The polysaccharide was composed of 3,6-anhydrous-galactose,6-methyl-galactose,2-methyl-galactose,galactose,xylose and glucose in the molar ratios of 6.4∶0.9∶5.8∶1.6∶84.1∶1.2.The content of Sulfate group was 37.5%. The main linkage model of the polysaccharide were 1,3 and 1,4 linkages,branch point located at O-2 and 6 of galactose residues.Sulfate group located at 2 and 6 position of 1,4 linked galactose residue and 2 position of 1,3 linked galactose residue.The polysaccharide showed significant activities to extend bleeding time and coagulation time in mice.Conclusion The polysaccharide from Aeodes orbitosa was a sulfate galactan with strong anticoagulant activity.

Studies on liver-protection and promoting bile secretion of gentiopicroside / 中草药

Object To study the liver-protection and bile promoting secretion of gentiopicroside administrated ig in rat. Methods Gentiopicroside was administrated ig to the mice with liver injury induced by CCl 4 and D-GlaN. ALT, AST levels in serum and GSH-Px activity in liver were examined. The bile flow rate and the concentration of the main components in the bile of rats after ig administration of gentiopicroside were estimated. Results Gentiopicroside administrated ig decreased the serum ALT and AST levels, increased the liver GSH-Px activity in the mice treated with CCl 4, promoted the secretion of bile and increased the concentration of bilirubin in the bile. Conclusion Gentiopicroside administrated ig exhibited significant liver-protection and promoting bile secretion.

Study on Chemical Quality Standard of Radix Astragali / 中成药

Objective: To establish the evaluation of the chemial quality standard of Radix Astragali according to their contents of saponin, flavonoid and polyscaccharide. Methods: HPLC ELS was used to determine astragaloside Ⅳ content. RP HPLC was used to determine 10 hydroxy 3,9 dimethoxypterocarpan content. Sulfuric acid phenol method was used to determine total polysaccharide content. Results: Astragaloside Ⅳ, total flavonoid, 10 hydroxy 3,9 dimethoxypterocarpan and total polysaccharide content of Astragalus mongmolicus Fisch. were 0.710? 0.030mg/g, 45?4.243mg/g, 19.955?0.813?g/g, 73.310?0.962mg/g, respectively; Astragaloside Ⅳ, total flavonoid, 10 hydroxy 3,9 dimethoxypeterocarpan and total polysaccharide content of Astragalus membranacens Fisch. were 1.186?0.024mg/g, 46?1.414mg/g,84.640?3.069?g/g, 48.800?1.245mg/g, respectively. Conclusion: The method was practical and provided a new idea to study the chemical quality standard of traditional Chinese drug.

Preparation of Blood-Deficient Model and Effects of Tangkuei Blood-Supplementing Decoction on Hemopoiesis / 中成药

Objective: To prepare a blood deficient model and study the effect of Tangkuei Blood Supplementing Decoction on hemopoiesis in this model. Methods: This model was made in mice by i.p. an accumulate doses of 60mg/kg acetylphenylhydrazine (APH) and 160mg/kg cyclophosphamidum (CY). The RBCs, WBCs, reticulocytes and bone marrow nucleated cells (BMNC) were counted, the micro structure of bone marrow was observed. The swimming time, the body temperature and the plasma cAMP, cGMP levels were measured. The effects of Tangkuei Blood Supplementing Decoction by p.o. administration on promoting the hemopioetic function were observed with the model mice. Results: Tangkuei Blood Supplementing Decoction could remarkably increase RBC, WBC, BMNC, improve the proportion of reticulolytes in peripheral blood and the micro struture of bone marrow, prolony the swimming time, raise the body temperature and the specific value of cAMP/cGMP. Conclusion: The model exhibits the main features of blood deficiency, and can be used as one of models of blood deficiency. Tangkuei Blood Supplementing Decoction can obviously improve the dual deficiention of qi and blood of model mice by supplementing qi and blood.

Extractive B of Evodia to Treat Protopathic Freund's Adjurant Arthritis(AA) / 中成药

Objective: To investigate extractive B of evodia to treat protopathic Freund's adjurant arthritis(AA) of Wistar mouse. Methods: Oncotic degree of the other side lower limb recurring disease; Disease incidence of the former limb, ear and tail; Thymus index and spleed index. Results: It markedly mitigate swelling of AA and regulate index of AA of spleen and thymus. Conclusion: Extractive B of evodia is efficiency treatment on protopathic Freund's adjurant arthritis(AA)