ABSTRACT
Aim To study the apoptosis of human hep-atoma cell line ( HepG2 ) induced by different polar parts of Arnebia euchroma ( Royle ) Johnst ( AE ) and to verify its anti-hepatoma effect by a mouse orthotopic liver cancer model so as to explore the anti-cancer effect of AE extract. Methods Firstly, MTT method and Annexin V-FITC/PI double staining method were used to detect the anti-proliferative and pro-apoptotic effects of each polar part of AE on HepG2 cells, and Western blot was used to detect the expression of Bcl-2 apoptosis family proteins incells. Based on the above experimental results, the effective parts with significant pro-apoptotic effect were screened out for anti-in situ liver cancer experiments in mice, and the organ indexes, liver function indexes and tissue sections of mice with orthotopic liver cancer before and after administration were evaluated. Results With the decrease of the polarity of AE extract,the anti-proliferation and pro-apoptotic effects on HepG2 cells were enhanced, and the anti-proliferation and apoptosis-inducing effects of AE petroleum ether fraction ( AEP) were the most significant. When AEP dose was 1.56 (μg • L
ABSTRACT
A method of capillary electrophoresis fingerprint was developed for evaluation of the quality of Arnebia euchroma ( Royle) Johnst. The samples were separated on a 50 μm × 40 cm uncoated capillary separation column at separation voltage of 25 kV with 100 mmol/ L borate buffer ( pH 8. 0) containing 25 mmol/ L SDS and 20% (V/ V) dehydrated alcohol as running buffer. The injection volume of sample was 0. 5 psi ×5 s and the detection wavelength was 214 nm. The results indicated that the samples of Arnebia euchroma (Royle) Johnst were well separated and detected in 35 min. With Shikonin peak as reference peak, 6 characteristic peaks of standard Arnebia euchroma (Royle) Johnst were determined. The quality discriminant analyses were accomplished for different kinds of samples named Arnebia euchroma ( Royle) Johnst that purchased from eight sources by means of characteristic fingerprint peak analysis, similarity evaluation and cluster analysis. This method had good reproducibility, and could be used for the quality control of Arnebia euchroma (Royle) Johnst.
ABSTRACT
Objective: By examining the secondary metabolites content changes of the low temperature stressed Arnebia euchroma suspension cells to explore the effects of unique environmental factors in genuine regional areas on A. euchroma in non-genuine regional areas. Methods: After stressing A. euchroma suspension cells for 24 h at 4 ℃, rosmarinic acid, lithospermum acid, shikonofuran A, shikonofuran E, acetylshikonin, deoxyshikonin, β,β'-dimethylacryl shikonin, isovalerylshikonin, and total shikonin compounds were measured by HPLC on six time points, one of which was before the stress and counted as 0 h, and others were in 12, 24, 48, 72, and 168 h after the stress. Results: The results showed that the different time points after low temperature stress, similar compounds had the same consistent change trends basically. However, the contents had significant differences. In addition to DAS, the maximum value of all the chemical ingredients were in low stress group. Conclusion: Low temperature stress can promote the accumulation of secondary metabolites in A. euchroma suspension cells, and have an important role in revealing the mechanism of the formation of its genuineness.
ABSTRACT
Object In order to promote the artificial cultivation of Arnebia euchroma (Royle) Johnst , and save the wild herb from relieving the lack of resources, the asexual multiplication system of A euchroma was established by tissue culture technique Methods Dedifferentiation in many kinds of explants of A euchroma by inducement succeeded After the embryos were multiplied, there were a lot of germinated transplants in media After callus were multiplied, many adventitious buds had formed in the media, then they were rooting; the transplants formed by inducement and the plants survived after being cultured in soil Results During the dedifferentiation, 2,4 D was essential, the effect of KT was better than BA The shoot sections were dedifferentiated in the most rapid way among the explants The callus from them were in very good quality It is of benefit to the callus multiplication and redifferentiation after reducing the concentration of 2,4 D Both KT and BA can induce adventitious buds, KT was also suitable for the formation of globular shaped embryos In initial period of sphere embryos culture, it was necessary to add the plant growth regulators in media for multiplying sphere embryos and promoting their development Proper temperature difference between day and night (10 ℃-28 ℃) and natural light were helpful for the formation of strong sprouts and survival of transplants The survival rate was 13 2% with 31 plantlets survived among 236 transplants cultured in soil Conclusion The asexual multiplication system of A euchroma established through tissue culture is stable and available
ABSTRACT
Objective To optimize the microemulsion preparation of Fufang Zicao Oil(FZO).Methods The extraction conditions of microemulsion preparation of FZO were optimized by orthogonal experiment;the kinds and contents of surfactants and cosurfactants determined by pseudo-ternary phase diagram were used for the optimization of the prescription of FZO;centrifugation method was used for detecting the stability of FZO.Results The optimum extract conditions were as follows:10-mesh coarse powder of medicinal material,four times amount of solvent(sesame oil),extraction for 25 minutes.With Tween 80 and propylene glycol(7:3)as the surfactant and cosurfactant,and mixing with the oils in the proportion of 20:10,a steady microemulsion of FZO can be obtained after centrifugation by 13000 r? min-1 for 30 min.Conclusion Under the above achieved conditions,the obtained microemulsion of FZO is clear,transparent and steady.
ABSTRACT
Objective Using the contents of naphthaquinine and shikonin as the indices,the influences of ultrasonic extraction and soxhlet extraction on the active constituents in Arnebia euchroma(Royle)Johnst.were studied.Methods An orthogonal design was applied.Naphthaquinine content was determined by spectrophotometry and shikonin content by HPLC.The extraction rate of the two extracting methods was compared to optimize the process condition.Results By using the two extracting methods,particle size had an obvious effect on the extraction rate of naphthaquinine(the bigger particle,the higher extraction rate),but had no effect on the extraction rate of shikonin;the solvent of ethanol showed different effects on the extraction rate of active constituents by using the two extraction methods,the extraction rate being higher by ultrasonic extraction while lower by soxhlet extraction.Conclusion Ultrasonic extraction is efficient,and with energy and time saving in extracting active constituents of Arnebia euchroma(Royle)Johnst.,which is superior to soxhlet extraction.