Aging ; Health ; Humans ; Longevity

In order to clarify material basis of effective parts of liangxue tongyu prescription, blood-heat and blood-stasis rat model induced by dry yeast was established. The changes of rectal temperature, blood viscosity and plasma viscosity were used to evaluate the cooling-blood and activating-blood effects of liangxue tongyu prescription and its parts. Compared with the model group, the extract from liangxue tongyu prescription, its volatile oil and n-butanol part could significantly reduce rectal temperature (P<0.01), and also reduce blood viscosity and plasma viscosity to various degrees (P<0.01 or P<0.05). So volatile oil and n-butanol part were primarily identified as effective parts of liangxue tongyu prescription. By using GC-MS with normalization method of area to analyze volatile oil of liangxue tongyu prescription, 70 compounds were identified, accounting for about 92.54%, mainly as β-asarone, paeonol, α-asarone and shyobunone. 42 compounds such as peony glycosides, tannins, and iridoid glycosides were identified by HPLC-MS techniques and standard comparison. The study determined the effective parts of liangxue tongyu prescription and clarified the chemical composition providing the foundation for further studies on material basis of liangxue tongyu prescription.
Acetophenones ; chemistry ; Animals ; Anisoles ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Gas Chromatography-Mass Spectrometry ; Oils, Volatile ; chemistry ; Rats ; Tannins ; chemistry

OBJECTIVETo investigate the mechanism of senescence delay of human diploid fibroblast (2BS) and protecting telomere length by epimedium flavonoids (EF).

METHODSThe drug sera of EF were used to treat the 2BS. The population doublings of 2BS cells were observed, the mRNA expression of p16 gene were determined by fluorescence real-time quantitative RT-PCR, the telomerase activation of 2BS cells were determined by TRAP-Hyb, the total retinoblastoma (Rb) and phosphorated Rb protein content were detected by ELISA, the telomere length of 2BS cells were determined by telomere restriction fragment (TRF) Southern blot assay.

RESULTSEF could significantly extend the population doublings of 2BS cells, the expression of p16 mRNA was decreased and the content of phosphorated Rb protein were increased by EF. The telomere lengthening of 2BS cells were improved by EF, but the telomerase was not activated.

CONCLUSIONIn senescence human fibroblasts 2BS cells, p16 gene mRNA expression increased, content of phosphorated Rb protein decreased and the telomere length of 2BS shortened, EF might delay the aging of cells through inhibiting the p16 gene expression, promoting the production of phosphorated Rb protein and to protect the length of telomere, but not activating the telomerase.

Animals ; Cells, Cultured ; Cellular Senescence ; genetics ; Cyclin-Dependent Kinase Inhibitor p16 ; biosynthesis ; genetics ; Epimedium ; chemistry ; Fibroblasts ; cytology ; Flavonoids ; pharmacology ; Male ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Sprague-Dawley ; Retinoblastoma Protein ; metabolism ; Telomerase ; biosynthesis ; Telomere ; genetics ; metabolism ; Transduction, Genetic

UNLABELLEDTo investigate the regulatory effects of epimedium flavonoids (EF) on adrenocortical regeneration in rats with inhibited hypothalamic-pituitary-adrenal (HPA) axis.

METHODSCell distribution in cell cycle and cell apoptotic rate were measured with PI stain and flow-cytometry; apoptosis cells were showed by in situ terminal-deoxynucleotidyl transferase-mediated deoxy-uridine triphosphate-fluorescene nick end labeling assay (TUNEL), and the genome-wide gene mRNA expression was detected by oligonucleotide microarrays.

RESULTSCompared to the normal control, adrenal cells isolated from the HPA axis inhibited model group were arrested in Go/GI phase, and showed a higher apoptotic rate (P < 0.05). After treated with EF, cells in G0/G1 phase decreased and those in G2/M phase increased (P < 0.01), and the elevated apoptotic rate reduced significantly (P < 0.05). TUNEL assay showed the number of apoptotic cells per section was 4.67 1.53 in the normal control group, 70.67 +/- 9.29 in the model group, and 18.67 +/- 7.64 in the EF-treated group respectively (n=3). Gene expressions in adrenal were mostly restrained in the model group, including 7 cytocycle promoting genes, including V-ras, V-jun, etc., while after treatment with EF, 6 cytocycle promoting genes, 1 anti-apoptotic gene, and genes that closely related with adrenocortical regeneration as IGF-II and FGF7 and their receptors, as well as 7 steroid biosynthesis participated genes were all up-regulated. Conclusion EF can accelerate adrenocortical cell proliferation, inhibit its apoptosis, and promote steroid biosynthesis so as to enhance adrenocortical regeneration in HPA axis inhibited rats, which may contribute to the beneficial effects of EF in protecting adrenocortical function during glucocorticoid withdrawal.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Epimedium ; chemistry ; Flavonoids ; pharmacology ; Gene Expression ; Gene Expression Profiling ; Glucocorticoids ; adverse effects ; Hypothalamo-Hypophyseal System ; drug effects ; Male ; Oligonucleotide Array Sequence Analysis ; Pituitary-Adrenal System ; drug effects ; physiology ; Rats ; Rats, Sprague-Dawley ; Substance Withdrawal Syndrome ; prevention & control

OBJECTIVETo study the regulatory pathways and rules of the gene networks in Shen deficiency syndrome.

METHODSTissues of hypothalamus, pituitary, adrenal, lymphocyte, bone, liver and kidney were taken as samples from 4 months' and 24 months' old SD rats and rats after treatment with Epimedium flavonoids (EF), differences of gene expression profile in Shen deficiency syndrome were studied repeatedly with gene chip rat expression set U230 2.0 array from Affymetrix Co.

RESULTSGene expressions in the aged rats all decreased including neurotransmitter of gamma-aminobutyric acid (gammaGABA), gonadotropin releasing hormone (GnRH), thyrotropin-releasing hormone (TRH), thyroid stimulating hormone (TSH), growth hormone-releasing hormone receptor (GHRH), insulin-like growth factor (IGF) and binding proteins (IGFBP) in hypothalamus, pituitary and adrenal (HPA axis), cell growth-related gene, growth factor related protein, and immune regulatory genes such as interferon gamma (IFN-gamma), interleukin 4 (IL-4) and interleukin 6 (IL-6) in lymphocytes, parathyroid hormone (PTH), calcitonin, procollagen, collagen, connective tissue growth factor in bone, and oxidative phosphorylation genes such as cytochrome P450 and NADH dehydrogenase, glutamate dehydrogenase related with protein metabolism, and glucose-6-phosphatase related with glucose metabolism in liver, most of which were up-regulated after treatment with EF as well as genes related with ageing and cell cycle, such as cyclin B, metabolism related genes and proteins of sodium and chloride channel in kidney.

CONCLUSIONDysfunction of the two regulatory pathways of gene networks as nerve-endocrine-immunity and nerve-endocrine-bone metabolism exists in Shen deficiency syndrome differentiated by effects of drugs, which could be improved by strengthening Shen therapy.

Aging ; genetics ; physiology ; Animals ; Diagnosis, Differential ; Drugs, Chinese Herbal ; pharmacology ; Epimedium ; chemistry ; Flavonoids ; pharmacology ; Gene Expression ; Hypothalamo-Hypophyseal System ; drug effects ; physiology ; Kidney Diseases ; genetics ; physiopathology ; Male ; Medicine, Chinese Traditional ; Neurosecretory Systems ; drug effects ; physiology ; Pituitary-Adrenal System ; drug effects ; physiology ; Rats ; Rats, Sprague-Dawley ; Yang Deficiency ; genetics ; physiopathology

OBJECTIVETo investigate the effect and its mechanism of Epimedium flavanoids (EF) in retarding aging with different systematic viewpoints.

METHODSHypothalamus, pituitary, adrenal and lymphocytes taken from 4-, 10-, 18-, 24-month old rats and from EF treated 24-month old rats were used to measure whole genome mRNA expression by gene array. Serum samples were used for metabonomic assay with high performance liquid chromatography. Using specific gene chip for NF-kappaB signaling pathway to detect the gene expression of the molecule related to that pathway in lymphocytes. Then, a neural network (NN) model was established upon the data obtained to quantitatively evaluate the degree of aging and the efficacy of drug intervention.

RESULTSGene expression of 199 genes showedan age-dependent pattern, most of which were reversed by EF, and the output of NN model showed that EF made the transcriptomics of 24-month old rats to 8-13 months. Seventeen metabolites among the 1,885 peaks detected were identified to have significant age-depending changes, and EF intervention reset the level of metabolites to a younger (18-month) level. The integral level of gene expression for NF-kappaB signaling pathway decreased significantly along with the increasing of age, and was significantly elevated by EF, NN model showed it approached to 10.5-month old.

CONCLUSIONPhenotype of aging at different levels demonstrates a common age-dependent trend; EF can reverse this age-dependent change at different levels in a synchronous manner.

Adrenal Glands ; drug effects ; growth & development ; metabolism ; Aging ; drug effects ; genetics ; metabolism ; Animals ; Chromatography, High Pressure Liquid ; Epimedium ; chemistry ; Flavonoids ; pharmacology ; Gene Expression Profiling ; Gene Expression Regulation, Developmental ; drug effects ; Hypothalamus ; drug effects ; growth & development ; metabolism ; Lymphocytes ; cytology ; drug effects ; metabolism ; Male ; NF-kappa B ; blood ; genetics ; metabolism ; Oligonucleotide Array Sequence Analysis ; methods ; Pituitary Gland ; drug effects ; growth & development ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Time Factors ; Transcription, Genetic ; drug effects

OBJECTIVETo explore similarity and difference of connotation between Shen deficiency syndrome (SDS) and Shen-yang deficiency syndrome (SYDS) on the molecular level.

METHODSThe senescent SD rats and corticosterone-treated rats were adopted for models of SDS and SYDS respectively, their syndrome was differentiated according to the therapeutic efficacy of treatment with epimedium flavonoids (EF). The gene expression profiles of hypothalamas, pituitary, adrenal gland and lymphocytes (HPAT axis) were detected before and after EF treatment using gene chip provided by Affymetrix company.

RESULTSAs compared with the young rats, the ageing rats and corticosterone-treated rats showed a significant down-regulation in highly consistent pattern, of various neurotransmitters of HPAT axis firstly, followed with that of growth and sex hormone related genes. EF could reverse the above genes expression in both models, and for SYDS model rats, it could also significantly up-regulate the gene expressions of heat shock protein, cytochrome P450 and thyroid stimulating hormone (TSH).

CONCLUSIONBoth SDS and SYDS model rats show connotation of Shen deficiency, and the substantial base of Shen-yang deficiency syndrome resides in the process of oxidative phosphorylation of energy metabolism accelerated by thyroid hormone.

Adrenal Glands ; drug effects ; metabolism ; Animals ; Diagnosis, Differential ; Epimedium ; chemistry ; Flavonoids ; therapeutic use ; Gene Expression Profiling ; Hypothalamus ; drug effects ; metabolism ; Lymphocytes ; drug effects ; metabolism ; Male ; Medicine, Chinese Traditional ; Oligonucleotide Array Sequence Analysis ; methods ; Pituitary Gland ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley ; Syndrome ; Time Factors ; Yang Deficiency ; diagnosis ; drug therapy ; genetics

Holistic concept is a characteristic contribution of Chinese medicine to medical science. Based on holistic concept, we investigated the correlation of Shenyang deficiency syndrome with the hypothalamus-pituitary-target gland axis; and revealed by gene chip technique the molecular network regulatory rule of intervention with Chinese drugs on hypothalamus-pituitary-adrenal-thymus axis in aged rats as a model for Shen deficiency syndrome; also the two major gene regulation pathways, namely, the neuro-endocrine-immune pathway and the neuro-endocrine-bone metabolic pathway, in hypothalamus, pituitary, adrenal, lymphocytes, kidney, liver, bone of aged rats, so as to advance the holistic research on Chinese medicine syndromes. Recently, based on the dynamic, intervention and multi-layer integration requirement of systematic biology, the common rule of aging was found at three different levels, the whole genomic transcription level, the metabolic level and the single ageing signaling pathway level. Meanwhile, it was demonstrated that the temporal and spatial association of genes will be weakened gradually along with aging. And Epimedium Flavonoids (EF), an extracts from Epimedium have been proved to be capable of effectively reversing both of these systemic changes. Thus, the transition of study from holistic concept to systematic biologic viewpoint could surely promote the progression of Chinese medicine to modern life science.
Aging ; genetics ; physiology ; Animals ; Holistic Health ; Kidney Diseases ; physiopathology ; Medicine, Chinese Traditional ; methods ; Rats ; Systems Biology ; methods ; Yang Deficiency ; physiopathology

Stem cells are the cells with capacities of self-renovation, multiplication and differentiation. By activating endogenous stem cells to promote regeneration response has provided a new thinking for the treatment of degenerative diseases. The authors found that epimedium flavonoids (EF) can promote the proliferation and migration of adrenocortical stem cells in corticosterone-treated rats (as a model of Shen-yang deficiency); and through gene-chip test it was showed that EF could significantly up-regulate the growth hormone (GH), growth hormone releasing hormone (GHRH) and other growth factors such as insulin-like growth factor binding protein (IGFBP) and nerve growth factor (NGF) in the model rats. In natural aging rats (as model for Shen deficiency), EF could make the gene expression of multiple tissues youthening, and up-regulate the lowered expressions of GH, GHRH, IGFBP and NGF, etc. Further study on the in vitro isolated and cultivated neuro-stem cells proved that EF and its components have direct promoting actions on stem cell proliferation. All the above-mentioned outcomes indicated that the actions of EF and its extracts on stem cells are possibly the cytological basis for their effects on counteracting the suppression of glucocorticoids on hypothalamus-pituitary-adrenal (HPA) axis and retarding aging; also illustrated that TCM could treat diseases by a way of activating endogenous stem cells through mobilizing and elevating hormones and cytokines levels, and bringing the reserved potential of organism into full play.
Adrenal Cortex ; cytology ; Animals ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Epimedium ; chemistry ; Flavonoids ; isolation & purification ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Stem Cells ; cytology

OBJECTIVETo investigate the effects and underlying molecular mechanisms of icariin (ICA) on self-renewal and differentiation of neural stem cells (NSCs).

METHODSNSCs were derived from forebrains of mice embryos by mechanical dissociation into single cell suspension. The self-renewal of NSCs was measured by neurosphere formation assay. The proliferation of NSCs was detected by water-soluble tetrazolium (WST) and 5-ethynyl-2'-deoxyuridine (EdU) incorporation assay. Protein expression of neuron-specific marker tubulin-βIII(TuJ1) and astrocyte-specific marker glial fibrillary acidic protein (GFAP) were measured by immunofluorescence and Western blotting. Using microarray, the differentially expressed genes (DEGs) were screened between NSCs with or without ICA treatment. The signaling pathways enriched by these DEGs and their role in mediating effects of ICA were analyzed.

RESULTSICA significantly promoted neurosphere formation of NSCs cultured in growth protocol in a dose-dependent manner and achieved the maximum effects at 100 nmol/L. ICA also increased optical absorbance value and EdU incorporation into nuclei of NSCs. ICA had no significant effects on the percentage of TuJ1 or GFAP-positive cells, and TuJ1 or GFAP protein expression in NSCs cultured in differentiation protocol. A total of 478 genes were found to be differentially regulated. Among signaling pathways significantly enriched by DEGs, mitogen activated protein kinase (MAPK) pathway was of interest. Blockade of extracellular signal-regulated kinase (ERK)/MAPK, other than p38/MAPK subfamily pathway partially abolished effects of ICA on neurosphere formation and EdU incorporation of NSCs.

CONCLUSIONICA can promote the selfrenewal of NSCs at least partially through ERK/MAPK signaling pathway.

Animals ; Cell Aggregation ; drug effects ; genetics ; Cell Differentiation ; drug effects ; genetics ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; genetics ; Deoxyuridine ; analogs & derivatives ; metabolism ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Female ; Flavonoids ; pharmacology ; Gene Expression Regulation ; drug effects ; MAP Kinase Signaling System ; drug effects ; genetics ; Mice ; Neural Stem Cells ; cytology ; drug effects ; enzymology