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*

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

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

A new steroid with a long cross-conjugation structure, 15a-hydroxy-(22E, 24R)-ergosta-3, 5, 8 (14), 22-tetraen-7-one (1), was isolated from the marine-derived fungus Aspergillus aculeatus. Its structure was established by the extensive spectroscopic analyses, and its cytotoxicities against P388, HL-60, and PC-3 cell lines were measured in vitro.
Animals ; Aspergillus ; chemistry ; Cell Line, Tumor ; drug effects ; Cholestenones ; chemistry ; isolation & purification ; pharmacology ; HL-60 Cells ; drug effects ; Humans ; Inhibitory Concentration 50 ; Molecular Structure ; Seawater ; microbiology

A new flavonone, named as (2R, 3S)-pinobanksin-3-cinnamate(1), together with six known compounds, pinocem-brin (2), pinobanksin (3), 3-O-acetylpinobanksin (4), galangin (5), kumatakenin(6), and 3-methylkaempferol (7), were isolated from a 95% ethanol extract of seeds of Alpinia katsumadai through a combination of various chromatographic techniques, including silica gel and Sephadex LH-20. The structure of compound 1 was elucidated by spectroscopic data analysis. Compound 1 exhibits a potent neuroprotective effect against the corticosterone-damaged PC12 cells, which may be underlying the effect by scavenging intracellular ROS.
Alpinia ; chemistry ; Animals ; Cell Death ; drug effects ; Cholestenones ; chemistry ; isolation & purification ; pharmacology ; Cinnamates ; chemistry ; isolation & purification ; pharmacology ; DNA Fragmentation ; drug effects ; Neuroprotective Agents ; chemistry ; isolation & purification ; pharmacology ; Oxidative Stress ; drug effects ; PC12 Cells ; Rats ; Reactive Oxygen Species ; metabolism ; Seeds ; chemistry

Ergosta-4,6,8(14),22-tetraen-3-one (ergone) is one of main components in many medicinal fungi. Ergone has been reported to possess the activities of diuresis, cytotoxicity, antitumor, immunosuppression, as well as treatment of chronic kidney disease. According to reported literatures, an overview of spectroscopy characteristics, content determination, pharmacological activity and pharmacokinetics, etc. for ergone is presented in this review. Furthermore, the present review can provide a certain reference value for the further study and development of ergone.
Animals ; Cholestenones ; chemistry ; pharmacokinetics ; pharmacology ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; pharmacology ; Humans

OBJECTIVETo study whether alisol B could inhibit complement 3a (C3a) induced renal tubular epithelial-mesenchymal transition (EMT).

METHODSThe in vitro cultured human renal tubular epithelial HK-2 cells were intervened with 5 ng/mL transforming growth factor-beta (TGF-beta), 0.1 micromol C3a, and 0.1 micromol C3a + 10 micromol alisol B, respectively. The mRNA and protein expressions of alpha-SMA, E-cadherin, and C3 were detected using RT-PCR, Western blot, and immunofluorescence, respectively.

RESULTSThe mRNA and protein expressions of C3 in HK-2 cells were up-regulated after intervention of C3a (P < 0.01), the mRNA and protein expressions of alpha-SMA in HK-2 cells were obviously enhanced (P < 0.01), the mRNA and protein expressions of E-cadherin obviously decreased (P < 0.01). When compared with the group intervened by exogenous C3a, after intervention of alisol B, the mRNA and protein expressions of alpha-SMA in HK-2 cells were obviously reduced (P < 0.01), the mRNA and protein expressions of E-cadherin obviously increased (P < 0.05).

CONCLUSIONSExogenous C3a could induce renal tubular EMT. Alisol B was capable of suppressing C3a induced EMT.

Actins ; metabolism ; Cadherins ; metabolism ; Cell Differentiation ; drug effects ; Cell Line ; Cholestenones ; pharmacology ; Complement C3a ; metabolism ; Epithelial Cells ; drug effects ; metabolism ; Epithelial-Mesenchymal Transition ; drug effects ; Humans ; Kidney Tubules ; cytology ; metabolism

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 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 relationship between tissue quantitative distribution and pharmacokinetics of 3H-achyranthes bidentata ecdysterone and the channel-tropism of herbal drugs in mice.

METHOD3H-achyranthes bidentata ecdysterone was used as a tracer agent and injected into mice by the caudal vein. In 36 hours, the contents of the tracer agent of samples involving 9 different tracing phases and organ or tissue were determined in order to observe the dynamic quantitative distribution and excretion and pharmacokinetics of 3H-achyranthes bidentata ecdysterone and to understand the channel-tropism of herbal drugs achyranthes bidentata.

RESULT3H-achyranthes bidentata ecdysterone of same organs in different tracing phases and the contents of 3H-achyranthes bidentata ecdysterone in same tracing phases of different organs were significantly different (P<0.01). 3H-achyranthes bidentata ecdysterone was mainly distributed, in the liver, kidney, adrenal gland, small intestine and lung. The concentration-time profiles of achyranthes bidentata ecdysterone in rats injected into mice by the caudal vein were shown to fit a two-compartment open model with half-lives of (778.65 +/- 12.36) min, the elimination of achyranthes bidentata ecdysterone from plasma was found to be in accord with linear kinetics.

CONCLUSIONThe above mentioned selective distribution of 3H-achyranthes bidentata ecdysterone basically coincides with the meridian affinity and zang fu selection of the traditional Chinese medicine drug Achyranthes bidentata. This study will provide a scientific basis for the channel-tropism of A. bidentata.

Achyranthes ; chemistry ; Animals ; Drugs, Chinese Herbal ; metabolism ; pharmacokinetics ; Ecdysterone ; metabolism ; pharmacokinetics ; Isotope Labeling ; Male ; Meridians ; Mice ; Organ Specificity ; Tissue Distribution ; Tritium ; chemistry