OBJECTIVETo investigate the effect of ecdysterone (EDS) on the proliferation of human bone marrow mesenchymal stem cells (hMSCs) in vitro.

METHODShMSCs were isolated from human bone marrow cell suspension by density gradient centrifugation. The expression of integrins CD44, CD105, CD34 and CD29 were examined by immunocytochemical method. EDS at 10, 25, 50 or 100 microg/ml were added in hMSC culture system, using the routine culture medium for hMSCs as control. The cell viability were analyzed by MTT assay and the cell cycle changes were examined by flow cytometry.

RESULTSThe optical density (OD) differed significant between the EDS treatment groups and the control group (P<0.01), and 25 microg/ml EDS group showed the highest OD value (P<0.01) without significant differences among 10, 50 and 100 microg/ml EDS groups (P>0.05). Flow cytometry showed that treatment of the cells with 25 microg/ml EDS significantly increased the cell percentages in S and G(2)M phases and the proliferation index (PI) of the cells as compared with the control group.

CONCLUSIONWithin a given concentration range, EDS can promote the proliferation of hMSCs in vitro, and this effect can be the most obvious at the concentration of 25 microg/ml. The effect of EDS in promoting the proliferation of hMSCs does not positively correlate to EDS concentration administered.

Adult ; Cell Proliferation ; drug effects ; Cells, Cultured ; Ecdysterone ; pharmacology ; Humans ; Male ; Mesenchymal Stromal Cells ; cytology

OBJECTIVETo prepare ecdysterone cream for promoting wound healing and conduct the dose-effect analysis to determine the optimal concentration.

METHODSThe cream substrate containing 4 concentrations (0.625%, 1.25%, 2.5% and 5%) of ecdysterone was prepared. Full-thickness skin defect was induced in 9 New Zealand rabbits at 5 sites on the dorsal skin, and the wounds were treated with blank cream substrate and ecdysterone cream at the 4 concentrations, respectively. On days 4, 8 and 12 after the injury, the healing area and the healing rate for each wound were determined, and in one rabbit, the tissues around the wounds were sampled for pathological examination.

RESULTSThe ecdysterone cream significantly promoted wound healing as shown by increased percentage of the healing area (P<0.01), and the optimal concentration was 2.5%. Pathologically, the wounds treated with 2.5% ecdysterone cream exhibited more obvious granulation tissue formation and proliferation of the epithelial cells, endothelial cells and fibroblasts than those treated with the cream of the other concentrations.

CONCLUSIONThe ecdysterone cream can obviously promote wound healing in rabbits at the optimal concentration of 2.5%, which may offer a clinical alternative for promoting wound healing.

Administration, Topical ; Animals ; Dose-Response Relationship, Drug ; Ecdysterone ; administration & dosage ; pharmacology ; Female ; Male ; Ointments ; Wound Healing ; drug effects

OBJECTIVETo examine the protective effect of ecdysterone (ECR) against beta-amyloid peptide fragment(25-35) (Abeta(25-35))-induced PC12 cells cytotoxicity, and to further explore its mechanism.

METHODSExperimental PC12 cells were divided into the Abeta group (treated by Abeta(25-35) 100 micromol/L), the blank group (untreated), the positive control group (treated by Vit E 100 micromol/L after induction) and the ECR treated groups (treated by ECR with different concentrations of 1, 50 and 100 micromol/L). The damaged and survival condition of PC12 cells in various groups was monitored by lactate dehydrogenase (LDH) release and MTT assay. The content of malondialdehyde (MDA) was measured by fluorometric assay to indicate the lipid peroxidation. And the antioxidant enzymes activities in PC12 cells, including superoxide dismutases (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px), were detected respectively.

RESULTSAfter PC12 cells were treated with Abeta(25-35) (100 micromol/L) for 24 hrs, they revealed a great decrease in MTT absorbance and activity of antioxidant enzymes, including SOD, CAT and GSH-Px as well as a significant increase of LDH activity and MDA content in PC12 cells (P < 0.01). When the cells was pretreated with 1-100 micromol/L ECR for 24 hrs before Abeta(25-35) treatment, the above-mentioned cytotoxic effect of Abeta(25-35) could be significantly attenuated dose-dependently, for ECR 50 micromol/L, P < 0.05 and for ECR 100 micromol/L, P < 0.01. Moreover, ECR also showed significant inhibition on the Abeta(25-35) induced decrease of SOD and GSH-Px activity, but not on that of CAT.

CONCLUSIONECR could protect PC12 cells from cytotoxicity of Abeta(25-35), and the protective mechanism might be related to the increase of SOD and GSH-Px activities and the decrease of MDA resulting from the ECR-pretreatment.

Amyloid beta-Peptides ; toxicity ; Animals ; Catalase ; analysis ; Ecdysterone ; pharmacology ; Glutathione Peroxidase ; analysis ; L-Lactate Dehydrogenase ; analysis ; Malondialdehyde ; analysis ; PC12 Cells ; Peptide Fragments ; toxicity ; Rats

OBJECTIVETo investigate the effect of ecdysterone on the proliferation of human umbilical cord mesenchymal stem cells (hUCMSCs) in vitro.

METHODShUCMSCs isolated by enzyme digestion from human umbilical cord tissues were cultured and identified for the surface antigens using fluorescence-activated cell sorting (FACS). The cells were treated with ecdysterone at the concentrations of 0, 25, 50, 100, 150, and 200 µg/ml, and the changes in the cell proliferation were detected using MTT assay.

RESULTSThe third-passage hUCMSCs were positive for CD29 and CD105 and negative for CD34 and CD45 as shown by flow cytometry. Treatment with ecdysterone resulted in significantly increased cell proliferation as compared to the control cells (P<0.05), but no significant differences were found in cells treated with 100, 150, and 200 µg/ml ecdysterone (P>0.05). The growth curves of the cells also demonstrated the definite effect of ecdysterone in promoting the proliferation of hUCMSCs.

CONCLUSIONEcdysterone can promote the proliferation of hUCMSCs in vitro with the optimal concentration of 100 µg/ml, suggesting its potential value in the enrichment of mesenchymal stem cells.

Cell Proliferation ; drug effects ; Cells, Cultured ; Ecdysterone ; pharmacology ; Flow Cytometry ; Humans ; Mesenchymal Stromal Cells ; cytology ; drug effects ; Umbilical Cord ; cytology ; drug effects

OBJECTIVETo study the effects of ecdysterone (ECR) on the expression of nuclear factor (NF)-kappaB in H2O2 induced oxidative damage of human lens epithelial cells (HLECs).

METHODSThe cultured HLECs were divided into 5 groups, i.e., the control group, the H2O2 group, the beta-estradiol (E2) group, the ECR group, and the pyrrolidine dithiocarbamate group (PDTC) group. The expression rate of NF-kappaB p65 in the HLECs were detected by flow cytometer (FCM).

RESULTSThe expression of NF-kappaB p65 occurred in normal HLECs (9. 53%). The expression rate of NF-kappaB p65 in the H2O2 group obviously increased (39.87%, P < 0.01). The expression rate of NF-kappaB p65 in the PDTC group obviously decreased (5.90%, P < 0.01). The expression rates of NF-kappaB p65 in the ECR group (13.99%) and the E2 group (25.18%) ranged between the control group and the H2O2 group, but still lower than that of the H2O2 group (P < 0.01).

CONCLUSIONSThe activation of NF-kappaB in the HLECs could be induced by H2O2 ECR with the estrogenic activity could effectively inhibit the activation of NF-kappaB.

Cells, Cultured ; Ecdysterone ; pharmacology ; Epithelial Cells ; drug effects ; metabolism ; Humans ; Hydrogen Peroxide ; adverse effects ; Lens, Crystalline ; cytology ; Oxidative Stress ; drug effects ; Transcription Factor RelA ; metabolism

Objective: The hippocampus is thought to be a vulnerable target of microwave exposure. The aim of the present study was to investigate whether 20-hydroxyecdysone (20E) acted as a fate regulator of adult rat hippocampal neural stem cells (NSCs). Furthermore, we investigated if 20E attenuated high power microwave (HMP) radiation-induced learning and memory deficits. Methods: Sixty male Sprague-Dawley rats were randomly divided into three groups: normal controls, radiation treated, and radiation+20E treated. Rats in the radiation and radiation+20E treatment groups were exposed to HPM radiation from a microwave emission system. The learning and memory abilities of the rats were assessed using the Morris water maze test. Primary adult rat hippocampal NSCs were isolated in vitro and cultured to evaluate their proliferation and differentiation. In addition, hematoxylin & eosin staining, western blotting, and immunofluorescence were used to detect changes in the rat brain and the proliferation and differentiation of the adult rat hippocampal NSCs after HPM radiation exposure. Results: The results showed that 20E induced neuronal differentiation of adult hippocampal NSCs from HPM radiation-exposed rats via the Wnt3a/β-catenin signaling pathway in vitro. Furthermore, 20E facilitated neurogenesis in the subgranular zone of the rat brain following HPM radiation exposure. Administration of 20E attenuated learning and memory deficits in HPM radiation-exposed rats and frizzled-related protein (FRZB) reduced the 20E-induced nuclear translocation of β-catenin, while FRZB treatment also reversed 20E-induced neuronal differentiation of NSCs in vitro. Conclusion These results suggested that 20E was a fate regulator of adult rat hippocampal NSCs, where it played a role in attenuating HPM radiation-induced learning and memory deficits.
Animals ; Cell Proliferation ; Ecdysterone/pharmacology* ; Hippocampus/metabolism* ; Male ; Memory Disorders ; Microwaves ; Neural Stem Cells/physiology* ; Rats ; Rats, Sprague-Dawley ; beta Catenin/metabolism*

AIMTo observe the behavior in learning and memory and the expression of c-fos gene from the brain of rats induced by beta-AP25-35, and the intervention of ecdysterone, in order to explore the protective mechanism of ecdysterone on the dysfunction of learning and memory of the rat induced by beta-AP25-35.

METHODSMicroinjection of beta-AP25-35 into hippocampus induced learning and memory dysfunction of rats. The learning and memory of rats were observed by Morris Water Maze. The expression of c-fos gene in the brain was detected by immunohistochemistry.

RESULTSThe results of Morris Water Maze showed that after rats were microinjected beta-AP25-35 into hippocampus, the rats in model group took longer latency and searching distance compared with the ones in control group (P < 0.01), and the rats in treated group (ECR 4 mg x kg(-1), ECR 8 mg x kg(-1) and nimodipine 7.2 mg x kg(-1)) took shorter latency and searching distance, especially the ECR 8 mg kg(-1) group (P < 0.01). At the same time, after the 5 days training, there was a higher expression of c-fos in hippocampus and cortex from the rats in control group than that in model group (P < 0.01), but in the treated group, there was a relatively higher expression of c-fos, especially the ECR 8 mg x kg(-1) group (P < 0.01).

CONCLUSIONMicroinjection of beta-AP25-35 into the rat hippocampus resulted in dysfunction of learning and memory. Ecdysterone was shown to improve the learning and memory of the rats and increase the expression of c-fos. Increasing the expression of c-fos is probably one of the most molecular mechanism of its protection.

Amyloid beta-Peptides ; antagonists & inhibitors ; toxicity ; Animals ; Ecdysterone ; pharmacology ; Gene Expression ; Genes, fos ; Hippocampus ; metabolism ; Male ; Maze Learning ; drug effects ; Microinjections ; Peptide Fragments ; antagonists & inhibitors ; toxicity ; Proto-Oncogene Proteins c-fos ; metabolism ; Rats ; Rats, Wistar