Research progress on culture technologies,chemical components,and pharmacological activities of Saussurea involucrata cells / 中国中药杂志

Saussurea involucrata,a traditional Chinese medicinal material,is effective in the treatment of rheumatoid arthritis with cold-dampness blockage syndrome,cold pain in lower abdomen,and menstrual irregularities. However,due to the specific habitat,low natural reproduction rate,slow growth,and overexploitation,it is at the high risk of extinction. S. involucrata cells can be obtained through callus culture,suspension culture,and hairy root culture. This study highlighted the influences of reactor type,culture system,precursor,elicitor type, and light wavelength on the suspension culture of S. involucrate cells. The chemical components of S. involucrata cells mainly include phenylpropanoids,flavonoids,lignans,and steroids,among which phenylpropanoids are the most abundant. S. involucrata cells have multiple pharmacological activities of anti-inflammation,analgesia,activating blood and resolving stasis,immunoregulation,increasing bone density,lowering blood lipids,anti-hypoxia,anti-exercise fatigue,anti-radiation,anti-obesity,and anti-oxidation. Moreover,it has the potential of treating aplastic anemia. This study reviews the cell culture technologies,chemical components,and pharmacological activities of S. involucrata cells,laying a basis for the further research,development,and utilization.

Analysis of flavonoids and antitumor activity of transgenic Saussurea involucrate / 中国中药杂志

The aim of this paper was to investigate the flavonoids of callus of transgenic and non-transgenic Saussurea involucrate and its antitumor activity on the esophageal cancer CaEs-17 cells. The species and content of mono-phenols were detected by high performance liquid chromatography. The growth of human esophageal cancer CaEs-17 cells was detected using CCK-8 assay, apoptosis morphology observation and flow cytometry. Expression of related apoptotic genes Bax and Bcl-2 were determined by qPCR. The results showed that the content of total flavonoids in the transgenic callus was 2.72 times that of the non-transgenic callus. The cyanidin-galactoside was detected in the transgenic callus, but not in the non-transgenic callus. The inhibitory effect of the extracts from the transgenic callus on CaEs-17 cells was more significant than that of the non-transgenic callus, and the IC₅₀ value had a decreased of 26.4%. Flow cytometry analysis results showed that the apoptosis induction effect of the extracts from the transgenic callus on CaEs-17 cells was significantly better than that of the non-transgenic callus. Fluorescence quantitative PCR analysis results showed that the extracts from the transgenic callus could up-regulate the expression of proapoptotic gene Bax and down-regulate the expression of apoptotic gene Bcl-2, and the regulation effect of the transgenic callus was more significant. Therefore, compared with the non-transgenic callus, the antitumor activity of the extracts from the callus of transgenic S. involucrate on the esophageal cancer CaEs-17 cells was significantly increased, which was closely related to the accumulation of cyanidin-galactoside and its metabolism-related flavonoid compounds in the transgenic callus.

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.

Study on mechanism of regulating synthesis of β-sitosterol by SiSQS1 and SiSQS2 in Saussurea involucrata cells / 中草药

Objective: To explore the synthetic mechanism of β-sitosterol by comparing the locus mutation, prokaryotic expression, expression level of SiSQS1 and SiSQS2, and the content of β-sitosterol in three color types of cells. Methods: Firstly, we preformed the cloning and bioinformatic analysis of SiSQS1 and SiSQS2 which were key enzymes involved in the biosynthesis of β-sitosterol in Saussurea involucrata cells. Secondly, we compared the differences of prokaryotic expression between SiSQS1 and SiSQS2, then optimized the expression conditions. Finally, we compared the expression levels of SiSQS1 and SiSQS2 by Real-time PCR and the content of β-sitosterol by GC-MS in three color types of cells, and made the correlative analysis on the expression level and the content of β-sitosterol. Results: There was a locus mutation of amino acid residues in 242E/D between SiSQS1 and SiSQS2. The results of prokaryotic expression analysis and conditions optimization showed that both target proteins had been expressed successfully, but the optimal prokaryotic expression system was different. The results of expression level and quantitative analysis showed a positive correlation to the expression levels of SiSQS1 and SiSQS2 and the content of β-sitosterol, the correlation coefficients were 0.92 and 0.89, respectively. Conclusion: A locus mutation of amino acid residues in 242E/D between SiSQS1 and SiSQS2 may influence the expression of SiSQS, and there may exist the functional differences in catalytic activity and the accumulation of β-sitosterol. The study will provide technical support and lay a theoretical foundation for studying the accumulated mechanism of β-sitosterol regulated by SQS in S. involucrata cells.

Effect of extract from callus of Saussurea involucrate on proliferation and differentiation of osteosarcoma cell MG-63 / 中草药

Objective: To investigate the effect of extract from callus of Saussurea involucrate (ESI) on the proliferation and differentiation of osteosarcoma cells MG-63 and explore its anti-osteoporosis and mechanism. Methods: Using human osteosarcoma cells MG-63 for the study, the effect of ESI on the proliferation was detected by MTT and LDH methods; The activity of alkaline phosphatase activity (ALP) and the level of hydroxyproline (Hyp) were investigated by related reagent kit; The effect of ESI on mineralized nodules was observed by Alizarin red staining and quantified by CPC, the expression of OPG/RANKL was measured by RT-PCR and Western blotting, and the proliferation, differentiation, mineralization, and OPG/RANKL expression levels of MG-63 were assayed when medium contains both p38 inhibitor SB203580 and ESI. Results: MTT and LDH assay showed that ESI (< 125 μg/mL) was non-toxic to MG-63, and it promoted the proliferation at the concentration of 31.25 and 62.5 μg/mL; ESI could significantly increase the ALP activity, Hyp content, and amount of mineralized nodules; At the mRNA level, ESI could significantly up-regulate OPG expression and down-regulate RANKL expression; At the protein level, ESI could significantly increase the ratio of OPG/RANKL; SB203580 could reverse the acceleration of ESI on the proliferation, differentiation, mineralization, and OPG/RANKL expression of MG-63. Conclusion: ESI has the facilitating effect on the osteoblast proliferation, differentiation, and mineralization; The mechanism may be associated with the expression of OPG and RANKL, as well as the signal transduction pathway of p38 Mitogen-activated potein kinase (MAPK).

A new polyphenol, 1, 3-di-O-caffeoyl-5-O-(1-methoxyl-2-O-caffeoyl-4-maloyl)-quinic acid, isolated from cultured cells of Saussurea involucrata / 中国天然药物

The present study was designed to isolate the polyphenol constituents of cultured cells of Saussurea involucrata. The polyphenol type constituents were isolated using chromatography methods, and then characterized by spectral analysis. 1,1-Diphenyl-2-picrylhydrazyl radical 2,2-Diphenyl-1-(2,4,6-trinitrophenyl)-hydrazyl (DPPH) and 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) free radical scavenging were assayed using Vitamin C as the positive control. One new polyphenol 18, 1, 3-di-O-caffeoyl-5-O-(1-methoxyl-2-O-caffeoyl-4-maloyl)-quinic acid, together with 17 known compounds, was isolated and characterized. In conclusion, Compound 18 was a new caffeoyl maloyl quinic acid type polyphenol and showed desired vitro anti-oxidant activity. Compounds 1-5, 9, 10, 14, 15, and 17 were isolated from cultured cells of Saussurea involucrata for the first time.

Effects of different physicochemical factors on growth of tissue culture seedlings and accumulation of total flavonoids in Saussurea involucrata / 中草药

Objective To investigate the effects of physicochemical factors on the growth and total flavonoids accumulation of Saussurea involucrata. Methods The dry weight and total flavonoids accumulation in multiple seedings of S. involucrata were measured. Results The optimal cultural condition for the growth and total flavonoid accumulation of S. involucrata was as following: MS culture medium adding 6-BA 0.5 mg/L + NAA 0.05 mg/L, 40 mmol/L of nitrogen source, NH4+-NO3- (15 : 25), 40 g/L of carbon sucrose, pH values 5.0-6.0, illumination intensity of 2 500 Ix, temperature 25°C. Conclusion The optimal culture conditions could significantly improve the growth and total flavonoids accumulation of S. involucrata. And the nitrogen, carbon, and temperature have the most obvious effects on the growth and total flavonoids accumulation of S. involucrata.

Application of macroporous absorption resin in purification for active constituents from Saussurea involucrata / 中草药

Objective To study the technological parameters of the purification process for active cons-tituents from Saussurea involucrata.Methods Using absorption and desorption rates of syringin,chologenic acid,and rutin as the primary screening indexes,four resins were selected,and the conditions of absorption and desorption were optimized for active constituents.Results Resin HPD400 gave good separation performance and was selected to purify the active constituents in S.involucrata.The optimum parameters were established as followings:4 BV(bed volume)sample extract whose pH value was adjusted to pH 2-3 by HCl was passed through the column with a flow rate of 1 BV/h,12 h later,the column was washed with 4 BV water,6 BV 30% ethanol and 4 BV 50% ethanol,respectively.The combined 30% and 50% ethanolic elutes were taken as object products.Conclusion The contents of active constituents were increased to about five times.Macroporous absorption resin HPD400 could be well used in separating and purifying the active constittuents from S.involucrata.

Effect of Saussurea involucrata Injection on Counteracting Adjuvant-induced Arthritis and Its Immunoregulation Function / 中药新药与临床药理

Objective To investigate the effect of Saussurea involucrata Injection(SII)on counteracting adjuvant-induced arthritis and its immunoregulation function,thus to supply the pharmacological evidences for the clinical treatment of arthritis.Methods Adjuvant arthritis was induced by plantar injection of Freundi' s complete adjuvant.MTT method was used to detect T and B lymphocytes proliferation,and sheep red blood cell immunization was used for hemolysin determination.Results(1)Rats right posterior metatarsus which was injected adjuvant was swollen from the second day to the 22nd day(the primary injury),and left posterior metatarsus not receiving adjuvant injection was swollen from the sixth day to the 20 th day(the secondary lesion).The differences of swelling degree were significant between SII group and NS group.(2)Consecutive intramuscular injection of SII 0.2,0.4,0.8 mL? kg-1? d-1 for 22 days suppressed the primary injury and the secondary lesion of adjuvant arthritis in rats,relieved swelling significantly from the sixth day(P

Analysis of Saussurea involucrata Kar. et Kir.from different sources by HPLC-FPS / 中成药

Objective:To study comparatively HPLC FPS of seven kinds of snowlotus, Saussurea involucrata Kar. et Kir. from different sources of Xinjian area. Methods: HPLC fingerprint analytical method of Snowlotus, from Urumqi was developed, and the HPLC FPS of seven samples were established. Results: The methodological evaluation showed that this method had a good repeatability, and the ratio of common peaks area of different samples were different. Conclusion: The method can be used to distinguish Snowlotus from different sources conveniently.

Study on the Pharmacokinetics of Powerful Saussurea lnvolucrata Spirit in Rabbits / 中国药房

It has been shown that powerful Saussurea involucrata spirit has effects of expelling wind and removing dampness, anti-inflammation and alleviating pain, and activating blood flow and diminishing swelling. The pharmacokinetic findings show that this drug has advantages of slow-absorption, long-action and high-bioavailability.