The cytotoxicological responses to insect growth regulator (IGR), using tebufenozide as ecdysteroid mimic, were investigated in Drosophila Kc cells. Treatment of Kc cells with tebufenozide showed significant growth inhibition and striking morphological changes including aggregation and elongation of the cells. In order to understand the cellular mechanism underlying the response of Drosophila cells to tebufenozide, immunofluorescence microscopy was performed. We found that treatment of Kc cells with tebufenozide enhanced the reorganization of f-actin and stimulated the expression of hsp27. These data suggest a possible association of filamentous actin (f-actin) and hsp27 in the cytotoxicological mechanisms of growth regulators in Drosophila cells.
Actins* ; Drosophila* ; Ecdysteroids ; Insects* ; Microscopy, Fluorescence ; Strikes, Employee

The herbs of Lamium takesimense Nakai (Lamiaceae) is used to treat spasmodic and inflammatory disease. The four polar compounds, ecdysterone, isoacteoside, rutin and lamiuside C, were isolated and identified from the BuOH fraction of the L. takesimense MeOH extract. HPLC quantification was performed on a Capcell Pak C18 column (5 µm, 4.6 mm × 250 mm) with a gradient elution of H₂O and 0.05% acetic acid in MeOH. The HPLC method was validated in terms of linearity, sensitivity, stability, precision, and accuracy. The quantitative level in plant material was determined as the following order: lamiuside C (4, 3.75 mg/g dry weight) > ecdysterone (1, 1.93 mg/g) > isoacteoside (2, 1.32 mg/g) > rutin (3, 0.97 mg/g).
Acetic Acid ; Chromatography, High Pressure Liquid ; Ecdysone ; Ecdysterone ; Glycosides ; Lamiaceae ; Methods ; Phenol ; Plants ; Rutin

AIMTo investigate the chemical constituents of Cyanotis arachnoidea.

METHODSBy using chromatographic methods for separation and combination with spectral analysis, their chemical structures were determined.

RESULTSSix compounds were identified as ajugasterone C-20, 22-acetonide (1), 20-hydroxyecdysone-20, 22-acetonide (2), 22-oxo-ajugasterone C (3), 22-oxo-20-hydroxyecdysone (4), beta-sitosterol (5), daucosterol (6).

CONCLUSIONCompound 3 is a new compound, 4 was a new natural compound.

Commelinaceae ; chemistry ; Ecdysone ; analogs & derivatives ; chemistry ; isolation & purification ; Ecdysterone ; analogs & derivatives ; chemistry ; isolation & purification ; Molecular Structure ; Plants, Medicinal ; chemistry ; Sitosterols ; chemistry ; isolation & purification

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

The effects of the cultivation media, plant growth regulators and inoculum size on the cell growth and 20-hydroxyecdysone production in suspension cultures of Vitex glabrata R. Br. were investigated. The cell growth and 20-hydroxyecdysone formation reach the highest when cells are cultured in the Gamborg's B5 medium supplemented with 2.0 mg/L BAP (6-benzylaminopurine) and 1.0 mg/L 2,4-D. The maximum 20-hydroxyecdysone productivity, of about 1.l mg/L/day, was observed in the culture with 20% PCV (packed cell volume) of inoculum size. These data also show that the increment of the inoculum size to 20% PCV could increase the productivity in 7-folds.
Cell Culture Techniques ; methods ; Culture Media ; Ecdysterone ; biosynthesis ; Vitex ; cytology ; metabolism

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

AIMTo isolate C-25 epimers of inokosterone from Achyranthes bidentata Blume. and identify their structures.

METHODSTo separate C-25 epimers of inokosterone by using various kinds of chromatography methods and identify their structures on basis of spectral analysis and chemical method.

RESULTSThree compounds were isolated and their structures were established as 25S-inokosterone (1), 25R-inokosterone (2) and ecdysterone (3).

CONCLUSIONCompounds 1 and 2 are new C-25 configuration isomers from Achyranthes bidentata Blume., their absolute configurations are elucidated at the first time, and their 13CNMR data are reported for the first time.

Achyranthes ; chemistry ; Cholestenes ; chemistry ; isolation & purification ; Ecdysterone ; chemistry ; isolation & purification ; Molecular Structure ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Stereoisomerism

T-bet is a critical transcription factor that regulates differentiation of Th1 cells from CD4+ precursor cells. Since T-bet directly binds to the promoter of the IFN-gamma gene and activates its transcription, T-bet deficiency impairs IFN-gamma production in Th1 cells. Interestingly, T-bet-deficient Th cells also display substantially augmented the production of IL-2, a T cell growth factor. Exogenous expression of T-bet in T-bet deficient Th cells rescued the IFN-gamma production and suppressed IL-2 expression. IFN-gamma and IL-2 reciprocally regulate Th cell proliferation following TCR stimulation. Therefore, we examined the effect of T-bet on Th cell proliferation and found that T-bet deficiency significantly enhanced Th cell proliferation under non-skewing, Th1-skewing, and Th2-skewing conditions. By using IFN-gamma-null mice to eliminate the anti-proliferative effect of IFN-gamma, T-bet deficiency still enhanced Th cell proliferation under both Th1- and Th2-skewing conditions. Since the anti-proliferative activity of T-bet may be influenced by IL-2 suppression in Th cells, we examined whether T-bet modulates IL-2-independent cell proliferation in a non-T cell population. We demonstrated that T-bet expression induced by ecdysone treatment in human embryonic kidney (HEK) cells increased IFN-gamma promoter activity in a dose dependent manner, and sustained T-bet expression considerably decreased cell proliferation in HEK cells. Although the molecular mechanisms underlying anti-proliferative activity of T-bet remain to be elucidated, T-bet may directly suppress cell proliferation in an IFN-gamma- or an IL-2-independent manner.
Animals ; Cell Proliferation ; Ecdysone ; Humans ; Interleukin-2 ; Kidney ; Mice ; Th1 Cells ; Transcription Factors

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

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