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OBJECTIVE：To study the protective effects of butein on oxidative stress injury of PC12 cell and its effects on mitochondrial function. METHODS：Rats PC12 cells were divided into normal control group，model group，solvent control group（1 ‰ dimethyl sulfoxide），butein high，medium and low concentration groups（2，1，0.5 μmol/L）. The latter 4 groups were given relevant reagent/medicine for intervention；24 h later，other groups were given 100 mU/mL glucose oxidase to induce oxidant stress model except for normal control group. After 4 h culture，cell survival rate，apoptosis rate，the levels or activities of ROS，MDA，SOD，CAT，GSH-Px，ATP，IL-1β and TNF-α as well as the change of MMP were detected. RESULTS：Compared with normal control group，cell survival rate，the levels or activities of SOD，CAT，GSH-Px and ATP were all decreased significantly，and apoptotic rate，the content of ROS，the levels of MDA，IL-1β and TNF-α were all increased significantly（P＜0.05 or P＜0.01），while the MMP was decreased significantly. Compared with model group，above indexes of solvent control group had no significant change （P＞0.05），cell survival rates，the levels or activities of SOD （except for medium and low concentration groups），CAT，GSH-Px（except for medium and low concentration groups），ATP（except for low concentration group）were increased significantly in butein high，medium and low concentration groups，while apoptotic rates，the content of ROS，the levels of MDA，IL-1 β and TNF-α were decreased significantly（P＜0.05 or P＜0.01），while the MMP were increased significantly. CONCLUSIONS：Butein can increase the antioxidant enzyme activity， stabilize mitochondrial function， inhibit oxidative stress and inflammationthus， increase energy generation inhibiting neuronal cell apoptosis ultimately exerting a neuroprotective effect.
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Aim To study the effect of butein on apop-tosis of PC12 cells induced by methylglyoxal (MG) and its mechanism. Methods Being pretreated with different concentrations of butein, PC12 cells were damaged by 1.5 mmol·L-1MG. Cell viability and cell toxicity were evaluated by MTT and LDH assay. Cell apoptosis and death were analyzed by PI and Ho-echst 33342. The antioxidant gene and proapoptotic gene expressions were determined by RT-PCR. The protein expression of p53 was detected by Western blot. Results Being pretreated with 2.5~10 μmol· L-1butein for 1 h significantly increased the cell via-bility,decreased LDH release,and protected from cell nuclei shrinkage, condensation and cleavage by MG. Meanwhile, butein increased the gene expression of SOD2, decreased the gene expression of proapoptotic genes p53 and caspase-9, and lowered the protein ex-pression of p53. Conclusion Butein can protect ap-optosis of PC12 cells from MG in a dose-dependent manner,which is linked with antioxidation and inhibi-ting p53 and caspase-9 gene expression.
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Objective:To investigate the effect of mH2A and Butein on mitogen-activation protein kinase (MAPK) signaling pathway through targeting glucose regulated protein of 78 (GRP78) in regulating the biological behavior of melanoma.Methods:Immunohistochemistry was used to detect the expressions of mH2A and GRP78 in melanoma and adjacent normal tissues.The relationship between mH2A and GRP78,and the dinicopathological data was analyzed.The relationship between GRP78 and mH2A protein was detected by immunoprecipitation assay.The protein expressions of mH2A and GRP78 was detected by Western blot.The invasion ability of melanoma after sliencing mH2A was detected by Transwell invasion assay.The migration ability of melanoma after sliencing mH2A was detected by wound healing assays.The protein expressions of MEK and ERK1 after sliencing mH2A was detected by Westem blot.Results:The expressions of mH2A and GRP78 in melanoma tissues was significantly lower than that in adjacent normal tissues (P＜0.05).Butein enhanced the expression of mH2A (P＜0.05).The mH2A regulated the GRP78 protein (P＜0.05).Silencing mH2A promoted the invasion and migration of melanoma A375 cells.The MEK and ERK1 protein expressions were up-regulated after silencing Butein.Conclusion:Butein promotes the role of mH2A in regulating GRP78 and modulating the biological behavior of melanoma cells through MAPK signaling pathway.
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Objective: To investigate the flavonoids from Gleditsiae Spina (thorns of Gleditsia sinensis) and their antitumor activities. Methods: The chemical constituents in the EtOAc fraction from Gleditsiae Spina were isolated and purified by the chromatography on silica gel, Sephadex LH-20 columns, and semi-preparative HPLC. Their structures were identified by various spectroscopic analyses, and the cytotoxicity of compounds 7-16 were evaluated in vitro against MCF-7 cell lines by SRB method. Results: Sixteen compounds were isolated from the extracts of Gleditsiae Spina and identified as tricin (1), liquiritigenin (2), 7,4'-dihydroxy-5,3'-dimethoxyflavanonol (3), garbanzol (4), 7,3',5'-trihydroxyflavanone (5), 7,4'-dihydroxyflavonol (6), dihydrokaempferol (7), butein (8), (2S)-5,7,3',5'- tetrahydroxyflavanone (9), 7,3',5'-trihydroxy-5-methoxyflavanonol (10), quercetin (11), fustin (12), fisetin (13), leucorobinetinidin (14), thevetiaflavon (15), and isovitexin (16). Compound 8 showed the inhibitory effect against MCF-7 cells with IC50 value of 28.53 μmol/L. Conclusion: Compounds 1-6, 8-10, 14 and 15 are isolated from the plants of Gleditsia L. for the first time, compound 8 shows the significant cytotoxic activity against MCF-7 cells.
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Objective: To study the chemical constituents in the dry mature seeds of Myristica fragrans (Myristicae Semen). Methods: Compounds were isolated and purified from the ethyl acetate layer of 75% alcohol extract by means of the silica gel column chromatography, Sephadex LH-20 column chromatography, and preparative HPLC isolation methods. Compound structures were identified by analyzing and comparing the spectral data with those for them in the reference. Results: Eighteen compounds were obtained from the ethyl acetate layer of 75% ethanol extract in Myristicae Semen. On the basis of spectral data and combined with the references, these compounds had been identifited as: vanilic acid (1), butein (2), (2R)-3-(3', 4', 5'-trimethoxyphenyl)-1, 2- propanediol (3), sulphuretin (4), 3-methoxy-4, 5-methylenedioxy-cinnamic acid (5), 7, 3', 4'-trihydroxyflavanone (6), 7-hydroxy- 4-benzopyrone (7), verrucosin (8), (+)-erythro-(7S, 8R)-Δ8'-7-hydroxy-3, 4, 3', 5'-tetramethoxy-8-O-4'-neolignan (9), (-)-erythro-(7R, 8S)-Δ8'-7- acetoxy-3, 4, 3', 5'-tetramethoxy-8-O-4'-neolignan (10), nectandrin B (11), (-)-(7S, 7'R, 8S, 8'R)-4, 4'-dihydroxy-3, 5, 3'-trimethoxy-7, 7'-epoxylignan (12), fragransin B3 (13), fragransin B1 (14), (-)-enantiomer (15), (-)-erythro-(7R, 8S)-Δ8'-7-hydroxy-3, 4, 5, 3', 5'-pentamethoxy-8-O-4'-neolignan (16), (+)-erythro-(7S, 8R)-Δ8'-7, 4-dihydroxy-3, 5, 3', 5'-tetramethoxy-8-O-4'-neolignan (17), and (+)-5-methoxydeydrodiisoeugenol (18). Conclusion: Compounds 2 and 4-7 are isolated from the plants of Myristica Gronov. for the first time. Compound 1 is isolated from this plant for the first time.
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OBJECTIVE: To purify butin and butein from the Vernonia anthelmintica Willd, and preliminary study their effects on proliferation and melanogenesis of A375 human melanoma cells. METHODS: Selica gel and Sephadex LH-20 column chromatography methods were used to separate butin and butein; their effects on proliferation and melanogenesis of A375 human melanoma cells were studied by MTT method, enzyme method and NaOH method. RESULTS: Butin and butein were isolated and identified. In the concentration range of 0.50-10.00 μg · mL-1, butin and butein enhanced the proliferation of A375 human melanoma cells, and the effect of butein was stronger than butin(P < 0.05) ; in the concentration range of 0.10-1.00 μg · mL-1, both butin and butein increased the activity of tyrosinase; in the concentration range of 0.50-5.00 μg · mL-1, butein stimulated melanogenesis, but butin inhibited melanogenesis. CONCLUSION: Butin and butein may be the active components of Vernonia anthelmintica Willd; butein promotes the proliferation and stimulates the melanogenesis of A375 human melanoma cells, however, butin mildly inhibites the melanogenesis.