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Aim To investigate the pharmacological mechanism of the couplet medicines " Cangzhu-Yiy-iren" in treating adenoid hypertrophy (AH) of children based on network pharmacology. Methods To screen the active ingredient and relevant targets of the couplet medicines "Cangzhu-Yiyiren", a visual network map of " Drug-Component-Target " was constructed; related targets of AH were retrieved and standardized, and A PPI network to treat AH of children by " Cangzhu-Yiyiren" was constructed. Enrichment analysis was performed for the core targets, and a " targets -pathways" network was constructed. The expression of target proteins from spleen tissues of different groups was determined by Western blot to verify that atractylone regulated the expression of inflammatory factors by HIF-1 α-SUMOylation. Results A total of 71 drug-related targets and 337 disease-related targets for AH in children were obtained, and there were 30 " Drug-Disease " intersection targets. The main active components of the couplet medicines "Cangzhu-Yiyiren" were stigmaster-ol, atractylone and so on. The biological processes mainly involved in were tube morphogenesis, response to hormone, the main cellular components involved in were membrane raft, transcription regulator complex, and the molecular function of related targets were mainly enriched in the transcription factor binding, protein domain specific binding, etc. The enrichment analysis indicated that it was associated with apoptosis-multiple species, VEGF signaling pathway, and HIF-1 signaling pathway ,etc. The results of animal experiments showed that SUMO-1,HIF-1α,VEGF and VEGF-R protein expression were all down-regulated compared with the model group ( P < 0. 05 ). Conclusions The treatment of pediatric AH which takes the " Activating Spleen Treatment of Nasa" as the guiding ideology, is realized through multi-components, multi-target, multi-pathways, and mainly from the anti-inflammatory, immune regulation, antioxidant and other aspects to play its role in the treatment of children with AH.
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Objective To investigate the effect of atractylone on the viability and apoptosis of hepatoma HepG2 cells and its mechanism of action. Methods Hepatoma HepG2 cells were selected and divided into low-, middle-, and high-dose atractylone groups (5, 10, and 20 μmol/L), and the cells in the control group were added with an equal volume of DMSO. MTT colorimetry was used to measure the viability of HepG2 cells after treatment with different concentrations of atractylone; flow cytometry was used to measure the apoptosis rate and mitochondrial membrane potential of HepG2 cells; the DCFH-DA fluorescent probe labeling method was used to measure the level of reactive oxygen species (ROS) in HepG2 cells; Transwell assay was used to evaluate the effect of atractylone on the migration ability of HepG2 cells; Western blot was used to measure the protein expression levels of Bcl-2, Bax, and cleaved caspase-3. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t -test was used for comparison between two groups. Results After 24 and 48 hours of treatment with atractylone, compared with the control group, the low-, middle-, and high-dose atractylone groups had a tendency of reduction in cell viability (all P < 0.05), with a half inhibitory concentration of 26.19 μmol/L in atractylone treatment of HepG2 cells for 72 hours. The low-, middle-, and high-dose atractylone groups had a significantly higher apoptosis rate than the control group (14.34%/29.32%/50.12% vs 0.32%, all P < 0.05). Compared with the control group, the low-, middle-, and high-dose atractylone groups had a significant increase in the fluorescence intensity of ROS in HepG2 cells (all P < 0.05). After 48 hours of treatment with atractylone, compared with the control group, the low-, middle-, and high-dose atractylone groups had a significant reduction in the number of migrated cells (132.67±18.36/57.00±9.26/31.00±2.45 vs 258.11±38.54, P < 0.05). Compared with the control group, the low-, middle-, and high-dose atractylone groups had a significant reduction in the expression of the anti-apoptotic factor Bcl-2 and significant increases in the expression of the apoptotic factors Bax and cleaved caspase-3 (all P < 0.05). Conclusion Atractylone can induce the apoptosis and inhibit the migration of HepG2 cells, which provides an experimental basis for further development and utilization of atractylone.
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To optimize the processing technology of Atractylodis Macrocephalae Rhizoma fried with honey rice chaff in Jian Chang Bang. With the contents of atractylenolide I, atractylenolide II, atractylenolide III, atractylone, and alcohol extract as indexes, which were determined by HPLC and alcohol extract method in Pharmacopoeia 2010 edition, L9 (34) orthogonal test was used to determine the best processing technology. The optimum processing technology of Atractylodis Macrocephalae Rhizoma fried with honey rice chaff was as following: the amount of honey rice chaff 50%, frying temperature 200℃, and frying time 5 min. The optimum processing technology is stable and feasible by verification.
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AIM: To research the processed products of Rhizoma Atractylodis Macrocephalae at the level of sesquiterpene lactones from it. METHODS: Sesquiterpene lactones were separated by the silica gel column and layer preparation, and the active substances were determined by HPLC. RESULTS: Atractylone、atractylolide Ⅰ, atractylolide Ⅲ, and biepiasterolid were separated from Rhizoma Atractylodis Macrocephalae. Atractylone was (0.535 8)%, atractylolide Ⅰ was (0.044 0)% and actractylolide Ⅲ was (0.081 4)% in Atractylodis stir-fried with wheat bran determined by HPLC. CONCLUSION: The method has good accuracy and repeatability and it can be used for the quality control of Rhizoma Atractylodis Macrocephalae.
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OBJECTIVE: To develop a method for determination of atractylenolide Ⅰ and atractylone in Chang’ankang granule by HPLC. METHODS: The determination was performed on Yilite ODS column (250 mm?4.6 mm,5 ?m). The mobile phase contained of acetonitrile-water (gradient elution) with flow rate of 1 mL?min-1 and detection wavelength of 220 nm. RESULTS: The linear range of atractylenolide Ⅰwere 0.1～2.0 ?g(r=0.999 9) with mean recovery of 98.84%(RSD=0.76%,n=6).The linear range of atractylone were 0.5～10.0 ?g with mean recovery of 98.43%(RSD=1.14%,n=6). CONCLUSION: This method is simple, convenience and accurate for quality control of Chang’ankang granule.