Effect of oxidative stress in SH-SY5Y cell injury induced by deguelin / 中国药理学通报

Aim To study the correlation between oxidative stress and neurotoxicity induced by deguelin, providing the mechanism basis for the structural modification and drug combination about deguelin. Methods SH-SY5Y cells were exposed to deguelin ranging from 1. 56-100 jimol • L"1 for 24 to 72 h, whose survival rate was determined by CCK-8 assay. The content of LDH, MDA levels, GPx and antioidant enzyme activities of SOD were determined using the assay kits ac-cording the manufacturer's protocol. And the content of ROS was determined by flow cytometry. The expressions of MEK, EGF, ERK and CREB were determined by immunoblotting. Results Deguelin obviously inhibited the growth of SH-SY5Y cells in a concentra-tion-and time-dependent manner ( P < 0. 05 ). After the cells were injured by deguelin, the content of ROS, LDH and MDA in SH-SY5Y cells increased obviously (P <0. 05), while the vitality of GPx and SOD decreased obviously ( P < 0. 05) in a dose-dependent manner. Furthermore, the protein expression levels of MEK, EGF, RAS and CREB in MAPK/ERK signaling pathway decreased obviously when treated with deguelin in a dose-dependent manner. (P < 0. 05). Conclusions The trauma of SH-SY5Y cells induced by deguelin is closely related to oxidative stress, in which MAPK/ERK signaling pathway may play a mediating role.

Effect of deguelin on proliferation of non-small cell lung cancer cell in vitro and in vivo / 中国药理学通报

Aim To investigate the effect of deguelin on the proliferation of non-small cell lung cancer cell line A549 and nude mice. Methods CCK-8 assay was used to detect the inhibition of deguelin on proliferation of SH-SY5Y cells; Hoechst stains and Annex-inV-FITC/PI double stained method were employed to observe the apoptotic morphology and apoptotic rate; flow cytometry was applied to determine the effect of deguelin on cell cycle of A549; tumor xenograft experiment and HE staining were conducted to investigate the effect of deguelin on growth of transplanted tumor in nude mice. Results Deguelin inhibited cell proliferation of A549 dose- A nd time-dependently; Hoechst stains and AnnexinV-FITC/PI double stained further confirmed that deguelin could induce the apoptosis of A549 cells, while deguelin blocked A549 cell cycle in G2/M phase in concentration-dependent manner. The nude mice xenograft model and HE staining experiments showed that deguelin significantly inhibited the growth of xenografts in A549 nude npce (P < 0. 01). Conclusions Deguelin has a significant inhibitory effect on non-small cell lung cancer cell line A549, pointing to a basis for the study of the antitumor activity of deguelin.

The protective effect of l-carnosine on neurotoxicity of deguelin-induced SH-SY5Y cells / 中国药理学通报

Aim: To study the protective effect of L-carnosine on deguelin-induced neurotoxicity. Methods: SH-SY5Y cells were treated with 1, 10, 20, 40, 60, 80, 100 mmol · L" L-carnosine for 24 h; cell viability was detected by CCK-8 assay. SH-SY5Y cells were respectively treated with 30 mmol · L-1 L-carnosine, 20 μmol · L-1 deguelin, 20 μmol · L-1 deguelin with 30 mmol · L-1 L-carnosine for 24 h; AO/EB method was employed for observing the morphology and apoptotic morphology of treated cells. SH-SY5Y cells were respectively treated with 8 μmol · L-1 deguelin, 8 μmol · L-1 deguelin with 3 mmol · L-1 L-carnosine, 8 μmol · L-1 deguelin with 30 mmol · L-1 L-carnosine, 30 mmol · L-1 L-carnosine, 20 μmol · L-1 deguelin, 20 μmol · L-1 deguelin with 3 mmol · L-1 L-carnosine, 20 μmol · L-1 deguelin with 30 mmol · L-1 L-carnosine, 50 μmol · L-1 deguelin, 50 μmol · L-1 deguelin with 3 mmol · L-1 L-carnosine, 50 μmol · L-1 deguelin with 30 mmol · L-1 L-carnosine for 24 h. Cell proliferation was detected by CCK-8 assay; apoptotic rate of treated cells was determined by flow cytometry; the reactive oxygen species (ROS) of treated cells was detected by DCFH-DA staining flow cytometry. Results: After 30 mmol · L-1 L-carnosine was co-treated with 20 μmol · L-1 and 50 μmol · L-1 of deguelin, the cell inhibitory rate decreased by 9. 07% and 6. 1%, the number of early apoptotic cells decreased, and the early apoptotic rate decreased by 9. 35%. The total apoptotic rate decreased by 10. 7%, and the ROS level was lower than that of the deguelin alone treatment group. The difference was statistically significant (P < 0. 05). Conclusions: L-carnosine can effectively reduce the neurotoxicity of deguelin-induced SH-SY5Y cells, which may be related to the reduction of oxidative stress levels, thereby inhibiting apoptosis and protecting cells.

Molecular mcchanism of deguelin-induced apoptosis and cell cycle arrest in lung cancer cells in vitro / 中国药理学通报

Aim To explore whether deguelin can block cell cycle and cell migration, inhibit the proliferation of non-small cell lung cancer cells.Methods H1299 cells were treated with 1.562 5, 3.125, 6.25, 12.5, 25, 50 μmol·L-1 deguelin for different time(24, 48, 72 h); cell viability was detected by CCK-8 assay, and cell migration ability was tested by scratch assay.H1299 cells were treated with 1.5, 3, 6 μmol·L-1 deguelin for 24 h.Flow cytometry with PI single staining and Annexin V-FITC/PI double staining experiment were used to evaluate cell cycle and apoptosis.qPCR was used to detect the regulatory effects of deguelin and its carbamate derivative on the Cyclin D-CDK4/6 complex at the gene level.Results Deguelin inhibited cell growth and the IC50 value of deguelin was(5.47±0.97),(4.01±0.45),(2.86±0.19)μmol·L-1 when treated with 24, 48, 72 h respectively.Deguelin also inhibited the healing ability of H1299 cells and the migration of H1299 cells significantly(P<0.05).Deguelin could block H1299 cell cycle in G1 phase.Flow cytometry combined with Annexin V-FITC/PI double staining showed that deguelin could induce apoptosis of H1299 cells.qPCR experiments showed that deguelin could down-regulate the expression of CDK4, CDK6 and Cyclin D1 genes significantly(P<0.05).Conclusions Deguelin may regulate cell cycle by down-regulating CDK4, CDK6 and Cyclin D1 genes in the cell cycle regulation system, and reduce the migration ability of tumor cells to induce apoptosis.

The apoptotic inducing effect of deguelin on SH-SY5Y cells / 中国药理学通报

Aim To study the apoptotic inducing effects of deguelin on SH-SY5Y cells.Methods SH-SY5Y cells were treated with 0,0.625,1.25,2.5,5,10 and 20 μmol·L-1 deguelin for different time(24,48,72 h);cell viability was detected by CCK-8 assay.SH-SY5Y cells were treated with 0,8,20,50 μmol·L-1 deguelin for 24 h;light microscope and AO/EB double stained method were employed for observing the morphology and apoptotic morphology of treated cells.Apoptotic rate of treated cells was determined by flow cytometry.Cells were stained by DCFH-DA,and the whole reactive oxygen species(ROS)was determined by flow cytometry.Spectrophotometry was employed to determine the activation degree of caspase-3.Results Deguelin inhibited cell growth in a time-and dose-dependent manner,and the IC50 value of deguelin was(26.07±2.18),(18.33±0.94),(12.5±1.49)μmol·L-1 when treated with 24,48,72 h respectively.After treated with 8,20,50 μmol·L-1 deguelin for 24 h,cell apoptotic rate,ROS and activation rate of caspase-3 increased markedly(P<0.05),all of which performed a dose related effect.Conclusion Deguelin can inhibit SH-SY5Y cell proliferation and induce cell apoptosis,and the mechanism may be concerned with the elevated ROS and activated caspase-3.

Modulation of signal transduction pathways by natural compounds in cancer / 中国天然药物

Cancer is generally regarded as the result of abnormal growth of cells. According to World Health Organization, cancer is the leading cause of mortality worldwide. Mother nature provides a large source of bioactive compounds with excellent therapeutic efficacy. Numerous phytochemicals from nature have been investigated for anticancer properties. In this review article, we discuss several natural compounds, which have shown anti-cancer activity. Natural compounds induce cell cycle arrest, activate intrinsic and extrinsic apoptosis pathways, generate Reactive Oxygen Species (ROS), and down-regulate activated signaling pathways, resulting in inhibition of cell proliferation, progression and metastasis of cancer. Several preclinical studies have suggested that natural compounds can also increase the sensitivity of resistant cancers to available chemotherapy agents. Furthermore, combining FDA approved anti-cancer drugs with natural compounds results in improved efficacy. On the basis of these exciting outcomes of natural compounds against several cancer types, several agents have already advanced to clinical trials. In conclusion, preclinical results and clinical outcomes against cancer suggest promising anticancer efficacy of agents from natural sources.

Inhibitory Effects of Deguelin on the Viability of Neuro-2 A Cells / 中国药师

Objective:To investigate the effects of deguelin on the viability and apoptosis of Neuro-2A ( N2A) cells. Methods:The cell viability of N2A cells with the density of 5 × 104 ·ml-1 on the time points of 6, 12, 24, 48 and 72 hours after 1 × 10 -8 mol ·L-1 deguelin treatment using CCK-8 kits was determined, and that on the time point of 48 hours after 0, 1 × 10 -11 , 1 × 10 -10 , 1 × 10 -9 , 1 × 10 -8 , 1 × 10 -7 , 1 × 10 -6 or 5 × 10 -4 mol·L-1 deguelin treatment was also detected. The activity of caspase 3 was deter-mined in N2A cells treated by 2 × 10 -8 mol·L-1 deguelin for 24 hours. Results:N2A cells with the density of 5 × 104 ·ml-1 reached the peak level in the growth curve 48 hours after the incubation in DMEM medium with 10% fetal bovine serum. The cell viability of N2A cells was decreased after the treatment of 1 × 10 -8 mol·L-1 deguelin for 24-72 hours in a time-dependant manner or after the treatment of 1 × 10 -8 ~1 × 10 -6 mol·L-1 deguelin for 48 hours in a dose-dependant manner (P<0. 05) with IC50 of 1. 6 × 10 -8 mol ·L-1 . The activity of caspase 3 was increased after the treatment of 2 × 10 -8 mol·L-1 deguelin for 24 hours, which was 5. 6-fold of that in the control group. Conclusion: Cell viability of N2A cells is inhibited by deguelin in a time- and dose-dependent manner, which may be related to the activity increase of caspase 3.

Effect of deguelin on mitochondrial permeability transition pore of human breast cancer cell line MDA-MB-231 / 第二军医大学学报

Objective: To investigate the effect of deguelin on mitochondrial permeability transition pore of human breast cancer cell line MDA-MB-231. Methods: The inhibitory effect of deguelin on cell proliferation was determined by MTT assay; cell apoptosis rate was analyzed by flow cytometry with AnnexinV FITC/PI double staining; MDA-MB-231 cells were stained by Rhodaminel23 to detect the changes of mitochondrial transmembrane potential by FCM; alteration of protein of Cyt c outside of mitochondria was detected by Western blotting analysis; caspase-3 activity was assessed by colorimetric assay; and MDA-MB-231 cells were stained by Fluo-3/AM to detect changes of intracellular Ca 2+ concentration by FCM. The expression of Bcl-2 and Bax was examined by RT-PCR and Western blotting analysis. Results: Deguelin significantly inhibited the growth of MDA-MB-231 cells in a time- and dose-dependent manner ( P < 0.05). After treatment with deguelin, mitochondrial transmembrane potential was decreased, the expression of Cyt c outside of mitochondria was increased, and caspase-3 activity was significantly increased compared with negative control group(P<0.01). FCM analysis showed that the apoptotic rate of MDA-MB-231 cells and intracellular Ca2+ concentration increased gradually with the increase of deguelin concentration. RT-PCR and Western blotting analysis showed that the expression of Bcl-2 mRNA and protein was down-regulated and that of Bax was up-regulated after deguelin treatment. Conclusion: Deguelin can inhibit proliferation and induce apoptosis of MDA-MB-231 cells, and the induction of apoptosis might be related to increased intracellular Ca2+ concentration and changes of mitochondrial permeability transition pore induced by altered Bcl-2, Bax expression.

Anti-cancer Effects of Deguelin on Human Leukemia K562 and K562/ADM Cells In Vitro / 华中科技大学学报（医学）（英德文版）

In order to investigate the anti-cancer effects of deguelin and on K562 and K562/ADM cells in vitro and the underlying molecular mechanism and compare the cytotoxicity of deguelin on K562, K562/ADM cells and human peripheral blood mononuclear cells (PBMCs). The effects of deguelin on cell proliferation were assessed by MTT assay. Apoptosis were detected by AnnexinV/PI double-labeled cytometry. The effects of deguelin on the cell cycle were studied by a propidium iodide method. Our study showed that deguelin inhibited the proliferation of K562 cell and K562/ADM cell in a time- and dose-dependent manner and had minimal effects on normal human peripheral blood mononuclear cells. The ratio of IC50 value of deguelin of 24 h on K562/ADM cells to K562 cells was only 1.27, which was significantly lower than the ratio of IC50 value of ADM (higher than 20). Deguelin could induce apoptosis of K562 cells and K562/ADM cells. K562 cells were arrested at G2/M phase while K562/ADM cells were arrested at G0/G1 phase. Our results suggested that deguelin was a novel anti-leukemia agents with high efficacy and low toxicity and it is also a promising agent for reversing drug resistance.

Effect on proliferation and apoptosis in RPMI-8226 human myeloma cells and regulation on steroid receptor coactivator-3 by deguelin / 中草药

Objective To investigate the effects of deguelin on the proliferation inhibition and apoptosis induction in RPMI-8226 cells in vitro,as well as the regulation of steroid receptor coactivator-3(SRC-3)to explore the relationship between them.Methods The effect of deguelin on the growth of RPMI-8226 cells was studied by MTT assay.Apoptosis was detected through Hoechst 33258 staining and Annexin-V FITC/PI double-labeled cytometry.RT-PCR Technology was applied to assessment of the mRNA expression of SRC-3,whereas,SRC-3 protein expression and localization were determined by using immunohistochemistry method.Results Deguelin presented striking proliferation inhibition potency on RPMI-8226 cells in vitro and apoptosis induction activity in a time-and dose-dependent manner.The IC50 value for 24 h was(54.55?0.40)nmol/L,(17.04?0.73)% RPMI-8226 cells went apoptotic when treated with 25 nmol/L deguelin for 24 h,with the dose of deguelin increasing to 100 nmol/L,(63.57?1.36)% cells were apoptotic.Over-expression of SRC-3 was found in RPMI-8226 cells,whereas the mRNA and protein expression levels of SRC-3 were significantly downregulated in RPMI-8226 cells induced by deguelin in a dose-dependent manner.The disposition of SRC-3 was situated mainly at the nuclear,occasionally in the cytoplasm.Conclusion Deguelin exhibited potent proliferation inhibition to the human myeloma cell line RPMI-8226 cells,furthermore,deguelin might induce RPMI-8226 cells apoptosis in a dose-dependent manner,which might correspond to the regulation of the expression of SRC-3.