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OBJECTIVE: To establish a method for the simultaneous determination of shikonin, acetylshikonin and β, β-dimethylacrylshikonin in Arnebia euchroma. METHODS: RP-HPLC method was adopted. The determination was performed on Kromasil 100-5 C18 column with mobile phase consisted of acetonitrile-0. 1％ formic acid solution (80: 20, V/V) at the flow rate of 1. 0 mL/min. The detection wavelength was set at 516 nm, column temperature was 25 ℃, and sample size was 10 μL. RESULTS: The linear ranges of shikonin, acetylshikonin and β, β-dimethylacrylshikonin were 0. 404-10. 100 μg/mL(r=0. 999 8), 5. 350-107. 000 μg/mL(r=0. 999 6), 2. 035-40. 700 μg/mL(r=0. 999 8), respectively. The limit of quantitation was 0. 40, 2. 91, 1. 34 μg/mL, and the limit of detection was 0. 12, 0. 87, 0. 40 μg/mL. RSDs of precision, stability and reproducibility tests were all lower than 2. 0％ (n=6). The recovery rate were 99. 12％-104. 18％ (RSD=1. 85％, n=6), 96. 51％-100. 21％ (RSD=1. 43％, n=6), 98. 11％-102. 51％ (RSD=1. 42％, n=6), respectively. CONCLUSIONS: The method is simple, precise, stable and reproducible. It can be used for simultaneous determination of shikonin, acetylshikonin and β, β-dimethylacrylshikonin in A. euchroma.
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OBJECTIVE:To study the decoction rate changes of 4 active ingredients in the volatile oil of Curcuma phaeocaulis before and after combined with Sparganium stoloniferum,and provide reference for showing the compatibility mechanism of C. pha-eocaulis and S. stoloniferum. METHODS:HPLC was used to simultaneously determine the contents of 4 active ingredients(curdi-one,curcumol,germacrone and β-elemene) in the volatile oil of C. phaeocaulis after C. phaeocaulis single decoction and com-bined decoction with S. stoloniferum. And the fried rates of each ingredient were compared. RESULTS:Compared with C. phaeo-caulis single decoction,there was significant difference in the fried amounts of curdione,curcumol,germacrone in the volatile oil after combined decoction(P0.05). CONCLUSIONS:The com-bined decoction of C. phaeocaulis and S. stoloniferum can effectively improve the dissolution of curdione,curcumol,germacrone in the volatile oil. It may be the reason for the enhancement of efficacy after compatibility.
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OBJECTIVE:To study the preventive and therapeutic effect and its mechanism of medicine pair decoction liquid of Curcuma phaeocaulis-Sparganium stoloniferum on rats with uterine myoma. METHODS:Rats were randomly divided into normal group,model group,positive group(mifepristone,2.25 mg/kg)and medicine pair decoction liquid group(C. phaeocaulis-S. stolon-iferum decoction liquid,6.0 g/kg),10 in each group. Except for the normal group,rats in other groups were injected Estradiol ben-zoate injection (0.5 mg/kg) intramuscularly every Monday,Wednesday and Friday,for 12 weeks. From the 13th week,rats in modeling group were added Progesterone injection(5 mg/kg)intramuscularly as well as relevant medicines intragastrically,once a day,until the 16th week. After administration,uterine coefficient of rats was detected. HE staining was used to observe the patho-logical changes of uterus and determine the thickness of smooth muscle;immunohistochemical staining was adopted to detect the transforming growth factor β3 (TGF-β3),matrix metalloproteinase 11 (MMP-11) protein expressions in uterus tissue of rats. RE-SULTS:Compared with normal group,uterine coefficient was increased in model group,pathological changes were obvious in uterus,thickness of smooth muscle was increased,TGF-β3 and MMP-11 protein expressions were enhanced(P<0.05 or P<0.01). Compared with model group,above-mentioned changes were improved significantly in positive group and medicine pair decoction liquid groups(P<0.05 or P<0.01). CONCLUSIONS:Medicine pair decoction liquid of C. phaeocaulis-S. stoloniferum shows cer-tain preventive and therapeutic effect on rats with uterine myoma. The mechanism may be associated with downregulating the TGF-β3,MMP-11 protein expressions in uterus tissue.
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Objective To observe the effect of external use of Curcuma oil on the hyperplasia of mammary glands in rats and explore the mechanism. Methods Sixty female Wistar rats were randomly divided into following groups:normal con-trol group, model control group, Sanjierubi plaster group, low-, medium- and high-dose curcuma oil emulsion groups ( n=10 each).The models of hyperplasia of mammary glands were established by intramuscular injection of estradiol benzoate and proges-terone into the medial part of the rat hind limb.Different doses of medicines were given for 4 consecutive weeks.Enzyme-linked im-munosorbent assay, HE staining and immunohistochemistry were used to investigate the action mechanism of curcuma oil emulsion against mammary gland hyperplasia. Results Curcuma oil emulsion had preventive and therapeutic effects on the hyperplasia of mammary glands.The diameter of breasts was significantly reduced, the body weight restored, serum estradiol, follicle-stimulating hormone and prolactin levels profoundly decreased, progesterone, testosterone and luteinizing hormone levels markedly increased and the number and diameter of lobular acini obviously reduced in high-dose curcuma oil emulsion group when compared with those in model control group (P<0.05 or P<0.01). Conclusion Curcuma oil emulsion can remarkably improve the disturb-ance of serum hormones and inhibit the occurrence of hyperplasia of mammary glands.
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Objective To optimize the extraction process of anti-hepatic fibrosis water-soluble component from Carthamus tinctorius L. via the orthogonal method. Methods The influences of type and content of solvent,extraction time,and extraction frequency on processing were investigated by assessing the content of hydroxysafflor yellow A,yield of extraction and anti-hepatic fibrosis potency via the high performance liquid chromatography and enzyme-linked immunosorbent assay by L9(3)4 orthogonal test. Results The optimum extracting condition for the anti-hepatic fibrosis water-soluble component from carthamus tinctorius L. was as follows: extracting with 10 times the volume of 10% ethanol for twice, 0. 5 h in each. Conclusion The process is scientific and feasible, which serves as a guideline for the production of the anti-hepatic fibrosis water-soluble components.
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Aim To investigate the effects of oleanolic acid on proliferation of SGC-7901/CDDP cells in vitro and its potential mechanisms.Methods Model of SGC-7901 cells resistant to cisplatin(SGC-7901/CDDP)was established in vitro.MTT assay was used to evaluate the viability of SGC-7901/CDDP cells in vitro.Real-time PCR was used to analyse the expression of Bcl-2 and Bax gene.Results MTT assay showed that oleanolic acid dramatically inhibited the proliferation of SGC-7901/CDDP cells in vitro,more than 62% SGC-7901/CDDP cells were inhibited when the cells were treated with 100 ?mol?L-1 oleanolic acid for 72 h;Real-time PCR showed that Bax(pro-apoptotic gene)was upregulated and Bcl-2(anti-apoptotic)was downregulated after being treated with oleanolic acid.Conclusions These results suggest that oleanolic acid inhibits proliferation of SGC-7901/CDDP cells in vitro,and its potential mechanisms may be through the upregulation of Bax and the downregulation of Bcl-2.