2.Regulation of Vitreoscilla hemoglobin on biosynthesis of astragaloside IV.
Ziyan WANG ; Zhibi HU ; Zhengtao WANG
Acta Pharmaceutica Sinica 2011;46(3):355-60
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.
3.Extractive B of Evodia to Treat Protopathic Freund's Adjurant Arthritis(AA)
Ling GAI ; Yun GAI ; Chunqing SONG ; Zhibi HU
Chinese Traditional Patent Medicine 1992;0(11):-
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)
4.Determination and biosynthesis of multiple salvianolic acids in hairy roots of Salvia miltiorrhiza.
Shujuan ZHAO ; Jinjia ZHANG ; Li YANG ; Zhengtao WANG ; Zhibi HU
Acta Pharmaceutica Sinica 2011;46(11):1352-6
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.
5.Determination of Epicatechin Content in Uncaria macrophylla by HPLC
Jun YANG ; Chunqing SONG ; Ming CHEN ; Haiyan WANG ; Zhibi HU
Chinese Traditional and Herbal Drugs 2000;31(9):662-663
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.
6.Influence of Blood Replenishing Angelica Decoction on Enhanced Endothelial Permeability Induced by Histamine
Zhibi HU ; Yi FAN ; Zhifen HAN ; Dazheng WU ; Chunqing SONG
Chinese Traditional and Herbal Drugs 2000;31(10):756-758
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.
7.Expression of human estrogen receptor alpha and beta in Escherichia coli.
Lu LUO ; Shujuan ZHAO ; Zhengtao WANG ; Zhibi HU
Acta Pharmaceutica Sinica 2012;47(10):1399-402
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.
8.Studies on the structural features and anticoagulant activity of the polysaccharide from Aeodes orbitosa
Shunchun WANG ; Songshan SHI ; Yingqi WENG ; Zhengtao WANG ; Zhibi HU
Chinese Journal of Marine Drugs 1994;0(04):-
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.