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Aim To preliminarily investigate the effect of brusatol(BRU), the monomer components fromChinese medicines on H1299 cells and its mechanisms.Methods CCK-8 and EdU staining experiment were used to detect the effect of BRU on cell prolifera-tion.Clone formation experiment was performed to measure the effect of drugs on cell clone formation.Hoechst33258 staining experiment and flow cytometry were employed to observe the cell apoptosis.Western blot assay was used to detect the protein expression levels of Bcl-xL, Bax, Bcl-2, cleaved-caspase-3, caspase-3, Gadd45α, PI3K, p-PI3K, Akt, p-Akt and NF-κB-p65.Results CCK-8 assay revealed that the inhibitory effect of H1299 cells gradually increased with the rising of BRU concentration and action time.Compared with control group, the EdU incorporation rate of the BRU treatment group decreased significantly.Treated with different concentrations of BRU for 24 h, the clone formation rate was significantly reduced in a concentration-dependent manner.Hoechst33258 experiment and flow cytometry showed that BRU could induce apoptosis in H1299 cell nucleus and increase its apoptotic rate.Western blot results revealed that BRU could significantly up-regulate the protein levels of Bax, cleaved-caspase-3, Gadd45α, and significantly down-regulate the expression of Bcl-xL, Bcl-2, caspase-3.In addition, BRU could significantly decrease the expression level of p-PI3K, p-Akt, NF-κB-p65 in cell nucleus.Conclusions BRU can inhibit the proliferation and induce apoptosis of H1299 cells in a concentration and time-dependent manner.The mechanism may be related to the inhibition of PI3K/Akt signaling pathway and the nuclear shuttle of NF-κB-p65.
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Aim To explore the effect of homoharringtonine (HHT) on the prohferation of liver cancer cell PLCS and its possible mechanism. Methods CCK-8 and EdU were used to detect the effect of HHT on the proliferation of PLCS cells; flow cytometry was employed to assess the effect of HHT on cell cycle of PLCS; Western blot was applied to measure the expression levels of cycle-related proteins cyclinA, CDK 2, p 2 1, p53 and A T M. Results Treated with HHT (0, 5, 10, 20, 40, 80 • L
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Objective: To establish the Huangqi-Danshen Decoction (HDD) fingerprint and multicomponent quantitative method, and provide a scientific basis for the quality standard of HDD. Methods: The HPLC fingerprint was performed on ODS Hypersil DIM column with acetonitrile-0.1% formic acid as mobile phase at gradient elution. The column temperature was 25 ℃; The flow rate was 0.8 mL/min, the volume was 20 μL, and the detection wavelength was 254 nm. At the same time, LC-MS was used to establish content determination method for 11 constitutes: astragaloside, astragaloside III, astragaloside II, calycosin 7-O-beta-D- glucoside, calycosin, ononin, salvianolic acid B, protocatechuic aldehyde, caffeic acid, rosmarinic acid, and lithospermic acid; The samples were identified by PCA analysis and OLPS-DA analysis. Results: There were 12 common peaks in the fingerprints of 10 batches of HDD. By comparing with the chemical reference, seven peaks were confirmed, they were: caffeic acid (peak 1), calycosin 7-O-beta-D-glucoside (peak 3), rosmarinic acid (peak 6), lithospermic acid (peak 8), calycosin (peak 9), salvianolic acid B (peak 10), and ononin (peak 11); Ten batches of samples have a high similarity (no less than 0.995). Astragaloside, astragaloside III, astragaloside II, calycosin 7-O-beta-D-glucoside, calycosin, ononin, salvianolic acid B, protocatechuic aldehyde, caffeic acid, rosmarinic acid, and lithospermic acid had good linearity with r2 ≥ 0.995 and recovery was at range of 89.3%-110.9%. Through principal component analysis, 10 batches of HDD can be divided into two categories. It can be seen that there are differences in the quality between different origins and different batches of medicinal materials. Finally, OPLS-DA was used to screen out two substances that cause quality differences: salvianolic acid B and ononin. Conclusion: According to the establishment of the fingerprint of HDD and the quantification by LC-MS, it can provide a reliable reference for its quality control.
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<p><b>OBJECTIVE</b>To establish two strains of mouse developing dampness-heat syndrome models infected by Dengue virus and to compare the difference in infection, so as to choose a suitable mouse strain for modeling.</p><p><b>METHODS</b>According to the modeling methods of the seasonal febrile disease of the dampness-heat syndrome in Chinese medicine, BALB/C and C57BL/6 mice were respectively treated with complex factors as the high glucose and high fat forage + high-temperature chamber + Dengue virus. At the same time a normal control group, the virus infection group (modeled by Dengue virus infection), and the dampness-heat group (modeling by pure dampness heat circumstance) were set up. Changes of the body temperature, platelet counts, virus in the separate serum, pathological changes of the liver, and serological indicators were observed to compare the modeling difference.</p><p><b>RESULTS</b>After modeling low-grade fever appeared in mice in the high-temperature chamber. Compared with the normal control group, the platelet count decreased in mice of the BALB/C model group. AST increased in both BALB/C and C57BL/6 mice and the virus infection group. TC and TG increased in BALB/C model group and the dampness-heat group, with statistical significance (P<0.05). Various degrees of pathological changes were shown in the liver tissue of each group, with the most severe one in the BALB/C model group. The serum virus titers were detected with Real-time PCR after modeling. The virus load was 2.9 x 10(4) - 5.5 x 10(4) copies/mL. No significant difference was found among these groups.</p><p><b>CONCLUSIONS</b>The mouse model of dampness-heat syndrome infected by Dengue virus was primarily established. When compared the infection between BALB/C mice and C57BL/6 mice, BALB/C mice were more suitable for modeling.</p>
