Simultaneous quantification of ginsenoside Rg1 and its metabolites by HPLC-MS/MS: Rg1 excretion in rat bile, urine and feces

Ginsenoside Rg1 (Rg1), the major effective component of ginseng, has been shown to have multiple bioactivities, but low oral bioavailability. The aim of this study was to develop a simple, sensitive and rapid high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method, which could be used to validate and quantify the concentrations of Rg1 and its metabolites in Sprague-Dawley rat bile, urine, and feces after oral administration (25 mg/kg). Calibration curves offered satisfactory linearity (>0.995) within the determined ranges. Both intra-day and inter-day variances were less than 15%, and the accuracy was within 80-120%. The excretion recoveries of Rg1, ginsenoside Rh1 (Rh1), and protopanaxatriol (Ppt) in bile, urine, and feces combined were all greater than 70%. The fecal excretion recoveries of Rg1, Rh1, and Ppt were 40.11%, 22.19%, and 22.88%, respectively, whereas 6.88% of Rg1 and 0.09% of Rh1 were excreted in bile. Urinary excretion accounted for only 0.04% of Rg1. In conclusion, the observed excretion profiles for Rg1 and its metabolites after oral administration are helpful for understanding the poor oral bioavailability of Rg1 and will aid further investigations of Rg1 as a pharmacologically active component.

Epigenetic mechanism in cognitive function / 中国药理学通报

The definition of epigenetics and its cellular basis are introduced firstly in the paper. Then, the research progress on the relationship between cognition and epigenetic changes is re-viewed in detail. In conclusion, epigenetic modifications occur-ring in hippocampus, cortex and other brain areas such as methy-lation , phosphorylation , ubiquitination , poly ( ADP-ribos ) poly-merases and DNA methylation may certainly change animal be-haviors including learning, memory, synaptic plasticity, depres-sion, drug abuse and so on. Long-term memory and long-term potentiation( LTP) , activation of AMPK-ERK signal transduction path-way and activation of key gene regulated by CREB-ABP transcriptional complex as well as transcription and expression of memory and synaptic plasticity related genes ( Zif/268, Creb, Bdnf, reelin ) are required. In contrast, epigenetic abnormal changes such as histone and DNA hypomethylation and increase of HDAC activity are observed in brains of aging and neurodegen-erative diseases. Therefore, the main epigenetic treatments for cognitive impairments are increasing histone and DNA methyla-tion, using HDAC inhibitors and RNA interference ( RNAi) to promote formation of long term memory and long term potentia-tion, block learning and memory decline.

Salvianolic acid B alleviate the disruption of blood-brain barrier in rats after cerebral ischemia-reperfusion by inhibiting MAPK pathway / 药学学报

The aim of the study is to investigate the effect of salvianolic acid B (SalB) on blood-brain barrier (BBB) in rats after cerebral ischemia-reperfusion, and to illustrate its possible mechanisms. Cerebral ischemia-reperfusion was induced by middle cerebral artery occlusion in rats. The break-down of BBB was indicated by extravasations of immunoglobulin (IgG) monitored with immunohistochemistry. The expression of MMP-9 and NOS2 in the brain was determined by immunohistochemistry, and the expression of p-p38 and p-ERK1/2 was detected by Western blotting. It was shown that on day 2 after ischemia-reperfusion the IgG accumulated around the vascular boundary zone, suggesting the break-down of BBB, and the expression of MMP-9 and NOS2 up-regulated at the same time. The result of Western blotting suggested that the expression of p-p38 and p-ERK1/2 increased. On day 7 after ischemia-reperfusion the. expression of MMP-9 and NOS2 was about the same level as day 2, the expression of p-p38 was higher than that on day 2 and the expression of p-ERK1/2 was slightly lower than that on day 2. SalB (1 and 10 mg x kg(-1)) significantly alleviated the extravasations of immunoglobulin induced by cerebral ischemia-reperfusion (P < 0.05). On day 2 and day 7 SalB attenuated the expression of MMP-9 and NOS2 (P < 0.05). SalB (10 mg x kg(-1)) reduced the expression of p-p38 and p-ERK1/2 apparently on day 2 and 7 after ischemia-reperfusion (P < 0.05). SalB (1 mg x kg(-1)) inhibited the expression of p-p38 on day 7 after ischemia-reperfusion (P < 0.05). The results indicate that SalB protects blood-brain barrier in rats after cerebral ischemia-reperfusion by inhibiting the MAPK pathway.

Nucleotide signals in the central nervous system and microglial movements / 中国药理学通报

Microglia cells are immune cells in the central nervous system.When the microenvironment of brain has changed,microglia will respond rapidly.ATP,UTP,or other nucleotide signals released by neurons from damaged site and their metabolites such as ADP,adenosine,UDP and so on will bind with the purinergic receptors on microglia to regulate the morphology and function of microglia,then the microglial cells activated by nucleotide signals are to regulate neural cells by phagocytosis or releasing cytokines.In this article,the function and corresponding mechanisms of nucleotide signals on chemotaxis,phagocytosis,and process retraction are reviewed.

The relationship between NO and gossypol in decreasing sperm quality in male rats / 中国药理学通报

Aim To elucidate the biochemical mechanism of gossypol in inducing the decline of sperm quality.Methods Gossypol was administered orally at the dose of 50 mg?kg-1/2 d for two weeks.Then,the sperm was collected from the left caudal epididymis and was analysed by CASA.The morphological changeand the concentration of nitric oxide(NO)in testes as well as the level of hormone〔follicle-stimulating hormone(FSH)〕,luteotrophic hormone(LH),testosterone(T)in serum were assayed.Results Gossypol could induce the decrease of sperm number and sperm quality.The concentration of NO in testes increased significantly.Among the three kind of hormone,only the concentration of T showed decrease after the oral administration of gossypol.NA1108 could antagonize the decline of sperm quality damaged by gossypol and decrease the content of NO in testes.Conclusions The concentration of NO in testes beyond normal value was one of the toxic mechanism of gossypol that contributed to the inhibition of spermatogenesis.Some drugs with the ability to reduce NO content in testes could also increase sperm quality.

Effects of salvianolic acids on endothelial cells against damage induced by cholestane-3beta-5alpha-6beta-triol / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the effects of salvianolic acids on human umbilical vein endothelial cells (HUVEC) against damage induced by cholestane-3beta-5alpha-6beta-triol (chol-triol).</p><p><b>METHODS</b>The viability of HUVEC was measured by MTT method. The apoptosis of HUVEC induced by chol-triol was detected by flow cytometry and TUNEL assay. The production of malondialdehyd (MDA) in HUVEC was tested by thiobarbaturic acid (TBA) assay.</p><p><b>RESULTS</b>The viability of HUVEC treated with chol-triol 100 micro mol/L decreased by 39.8% while salvianolic acids 100 micro g/ml increased by 27.9%. The apoptotic rate of HUVEC measured by PI staining increased from 6% - 8% to 17% - 20% after chol-triol treatment for 12 h. Salvianolic acids 100 micro g/ml reduced the apoptotic rate to 10% - 14% after treatment HUVEC for 1 h prior to chol-triol treatment. In another experiment, chol-triol increased the number of TUNEL-positive cells 5 times, but salvianolic acids 10 micro g/ml and 100 micro g/ml reduced the number of TUNEL-positive cells by 36.9% and 61.2%, respectively. The production of MDA in HUVEC increased by 120.7% after chol-triol treatment for 12 h. Salvianolic acids 10 micro g/ml and 100 micro g/ml also decreased the concentration of MDA by 28.7% and 39.8%, respectively.</p><p><b>CONCLUSION</b>Salvianolic acids has protective effect on endothelial cells against damage induced by chol-triol.</p>

Polysaccharide sulfate 916 inhibits neutrophil-endothelial adhesion / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To study the effect of polysaccharide sulfate 916 (PS916) on neutrophil-endothelial cell adhesion.</p><p><b>METHODS</b>Cell adhesion was evaluated by testing neutrophil myeloperoxidase activity. Expression of adhesion molecule in human umbilical vein endothelial cell (HUVEC) was measured by ELISA. The neutrophil activation rate induced by N-formyl-methionyl-leucyl-phenylalanine (fMLP) was tested by nitroblue tetrazolium (NBT) reduction.</p><p><b>RESULTS</b>Tumor necrosis factor alpha (TNFalpha, 50 - 800 U/ml) increased the adherence of neutrophil to TNFalpha-stimulated HUVEC in a concentration and time dependent manner. PS916 (0.01 - 1.0 mg/ml) dose-dependently inhibited the adherence of neutrophils to TNFalpha-stimulated HUVEC. fMLP increased the activation rate of neutrophils independent of concentration. PS916 also inhibited the adherence of fMLP-activated neutrophils to HUVEC. Moreover, PS916 inhibited adhesion molecule expression in TNFalpha-stimulated HUVEC.</p><p><b>CONCLUSIONS</b>PS916 inhibited neutrophil-endothelial adhesion. The mechanism of its action was partially related to suppressing the expressions of intercellular adhesion molecule-1 (ICAM-1) and vascular adhesion molecule-1 (VCAM-1).</p>

Stress and reproductive endocrine dystunction / 中国药理学通报

Physical and psychological stresses inhibit or harm reproductive endocrine especially female one at many levels of hypothalamo-pituitary-gonadal (HPG) axis, and result in subhealth conditions or clinic diseases in reproductive endocrine. The hypothalamic corticotropin-releasing factor (CRF) and adrenal glucocorticoids (GCS) overreleases and other neuroendocrine alterations induced by stress are implicated in the mechanisms responsible for the disturbance of reproductive endocrine. The declines in activities of synzyms for reproductive hormones cause sexual hormones reduces and disfunction in reproductive endocrine system.

EFFECT OF GINSENOSIDE Rg1 ON LEARNING AND MEMORY IMPAIRMENT INDUCED BY β-AMYLOID PEPTIDE(25-35) AND ITS MECHANISM OF ACTION / 药学学报

AIM　To study the effect of ginsenoside Rg1 on the learning and memory impairment in mice induced by aggregated β-AP(25-35). METHODS　Mice were administered Rg1 (5, 10 mg*kg-1, ip) for 10 d and control mice received daily ip injections of saline after the intracerebroventricular injection of aggregated β-AP(25-35). After the final treatment, passive avoidance and performance in the Morris water maze (MWM) were assessed. and the activity of cortical and hippocampal ChAT and AchE were detected after the final behavior test. RESULTS　Ginsenoside Rg1 (5, 10 mg*kg-1, ip) significantly ameliorated the learning and memory impairment induced by β-AP(25-35). Rg1 (5, 10 mg*kg-1) decreased the latencies and swim distances of mice to reach a hidden platform and improved the corresponding changes in search strategies occurred in the Morris water maze, and Rg1 (10 mg*kg-1, ip), increased step-through latencies also. Biochemical analysis showed that Rg1 (5, 10 mg*kg-1, ip) prevented the cortical and hippocampal ChAT activity decline induced by β- AP(25-35), and showed inhibition of the activity of AchE, although β-AP(25-35) showed no effect on the cortical and hippocampal AchE activity. CONCLUSION　These data showed that ginsenoside Rg1 significantly improved the learning and memory impairment induced by β-AP(25-35), and this effect could be attibuted to its inhibition of AchE and increase of ChAT activity.

INHIBITORY EFFECTS OF D-POLYMANNURONIC SULFATE ON PROLIFERATION OF RAT VASCULAR SMOOTH MUSCLE CELLS AND ITS RELATED MECHANISMS OF ACTION IN VITRO / 药学学报

AIM　To investigate the inhibitory effects of D-polymannuronic sulfate (DPS) on the proliferation of rat vascular smooth muscle cells (VSMC) induced by basic fibroblast growth factor (bFGF) or interleukin-1 (IL-1) and its related mechanisms. METHODS　Rat aortic smooth muscle cells pretreated with DPS in concentrations ranging from 0.001 μg*mL-1 up to 100 μg*mL-1 were incubated at 37℃ for 24 h, followed by addition of bFGF (50 ng*mL-1) or IL-1 (50 U*mL-1) for another 24 h. The effects of DPS on the proliferation of VSMC were evaluated by MTT assays. VSMC were pretreated with DPS in concentrations ranging from 0.001 μg*mL-1 up to 1 μg*mL-1, followed by addition of L-NAME (0.1 μg*mL-1) or bFGF (50 ng*mL-1) for 24 h. Supernatant nitric oxide (NO) was determined with NO assay kit, while supernatant angiotensin II (Ang II) and endothelin-1 (ET-1) were measured by radioimmunoassay. RESULTS　DPS exerted antiproliferative effects at concentrations ranging from 0.01 μg*mL-1 to 10 μg*mL-1, and its maximal effect was observed at the concentration of 1 μg*mL-1. Also, the suppressing actions of DPS on the proliferation of VSMC were diminished by increasing the concentrations of bFGF or IL-1. Furthermore, DPS increased NO synthesis and decreased Ang II and ET-1 contents released from VSMC in a concentration-dependent manner. CONCLUSION　DPS afforded the antiproliferative effects on bFGF- or IL-1-treated VSMC and its underlying mechanisms were associated with enhancement of NO synthesis and decrement of Ang II and ET-1 production/release in vitro.

The relationship between sulfated polysaccharides and atheroscle rosis / 中国药理学通报

The atherogensis was involved in a complex pathological process. Injury to endothelial cells of blood vessels was confirmed to be the in itial stage of this process. Migration to subendothelial layer and accumulation and proliferation of smooth muscle cells were attributed to various cytokines an d adhesive molecules secreted by activated endothelial cells, subsequently resul ting in aggregation of lymphocytes,platelets, monocytes and macrophages in the i ntima of artery. These cellular components ultimatedly led to the formation of a mature atherosclerotic plaque. Its quite acknowledged that a better understan ding of the atherogenic events might promise us the development of new chemical entities of anti-atherosclerotic therapies. A large body of evidence has demons trated that sulfated polysaccharides played a critical role in the development o f atherosclerosis. The underlying mechanisms of the anti-atherosclerotic activi ty of sulfated polysaccharides were reported to contribute to protecting against endothelial cells injury,inhibiting migration and proliferation of vascular smo oth muscle cells, and reducing the adhesion of inflammatory cells, platelets and lymphocytes. And also, the prevention of complement activation by sulfated poly saccharides could not be excluded. On the other hand,the promoting effects of su lfated polysaccharides atherosclerosis was also reported. Its therefore conclu ded that the relationships between atheriosclerosis and sulfated polysaccharides remained to be further elucidated.

Research of the activity and the inhibitors of cGMP specific PDE5 / 中国生化药物杂志

Purpose　The aim is to study the activity and the sildenafil selective inhibition of PDE5. Methods　The PDE isoenzymes were purified from bovine penis corpus cavernosum tissue by FPLC system. PDE activity was assayed by using 3　H-cGMP as substrate, the PDE isoenzymes hydrolyzed it to 3　H-GMP, and 3　H-GMP was further hydrolyzed to 3　H-guanosine by 5′-nuclease of snake venom. Add scintillation cocktail to observe the PDE isoenzymes activity. The selective inhibitor sildenafil of different concentrations were used to observe the inhibition of PDE5. Data replotted according to procedure of Dixon plots.Results　Three PDE isoenzyme peaks were purificated from bovine corpus cavernosum. The PDE of the third peak had the strongest activity of cGMP hydrolyzation which could be inhibited by sildenafil apparently.Conclusion　Since PDE5 was mainly found in corpus cavernosum tissue of mammalian, and sildenafil was a selective inhibitor of PDE5. It was suggested that the third peak was PDE5. The result was in agreement with the article reported.

Effect of Repeated Immobilization Stress on Ovarian Function in Female Rats / 中药新药与临床药理

Objective To observe the effects of repeated immobilization on rats ovarian function and to observe the changes of ovarian function in menopause animals.Methods Daily vaginal smears were permfored to follow the estrous cycle in adult female rats.The animals with regular cycle were exposed to the scheduled daily repeated immobilization for 15-day,which included 5 stress episodes with each episode consisting of three immobilizations (one time per day). The duration of immobilization was progressively increased with the subsequent episode. Serum levels of estradiol (E2) and progesterone (P) were measured throughout the estrous cycle before and after the stress.Results The repeated stress resulted in the disruption of the estrous cycle characterized by longer phase of diestrus or by fewer epithelium from vagina in proestrus and estrus.In the stressed rats, a significant decrease of serum E2 and an increase of P were shown throughout the cycle compared to that before stress;the normal secretion cycle was interrupted or even disappeared; changes of serum P level in some rats were similar to the menopause animals.Conclusion Repeated immobilization can cause the disruption or degeneration of ovarian function,indicating that rats with stress-induced ovarian dysfunction is an appropriate animal model for testing protective effects of agents against aging and for the diseases related to ovarian dysfunction.

Water soluble active ingredients of Danshen--review on the Salvianolic acids / 基础医学与临床

Danshen-Radix Salviae Miltiorrhizae，is one of the common traditional Chinese medicines，which has been used clinically in the treatment of cardiovascular diseases more than 20 centuries.The active ingredients of Danshen has been researched 5 decades by the modern methods.Many researchers investigated the pharmacological effects of water soluble ingredients of Danshen-salvianolic acids in vivo and in vitro.The results demonstrated that salvianolic acids have different pharmacological effects such as potent antioxidative effects，scavenging free radicals，protect neural cells against injuries caused by anoxia， etc.In present paper，the pharmacological effects of salvianolic acids and the mechanisms of their actions are reviewed based on the research results obtained in our laboratory and other authors.

Comparison of pharmacological effects of ginsenoside Rg1 and Rb1 / 基础医学与临床

Ginseng has long been used as a tonic and agent for prolonging life span in chinese traditional medicine． Using morden technology，ginsenoside Rgl and Rbl were proved to be main active principles of ginseng．Both conpounds showed the same effect in improving learning and memory， increasing Bmax of M-cholinergic receptors and accelerating cerebral protein and acetylcholine biosynthesis．However，Rgl but not Rbl had immunoregulatory action in aged rats and anti-osteoporosis effect in ovariectomized rats as well as enhanced basic synaptic transmission and magnitude of LTP induced by HFS． On the other hand，Rbl had anti-stress effects in antagonizing acute,chronic and repeated stress induced reduction of sexual behaviour and decrease of plasma andogen or estrogen．Rgl showed no such effect even aggravate stress induced damage．Rhl possessed anti-oxidant activity and prolong survival time of mice in cold(－10℃) condition．There was no any anti-cold effect with Rgl .These diference of biological activities between Rgl and Rbl may be arributed to their structures containing different number of glucoses．

The advance on study of melatonin atntioxidative effect / 中国药理学通报

Melatonin (MT) is secreted by the pineal gland and has obvious biological rhythmicity including circadian, season rhythm and life rhythm (aging clock). The reduction of MT secretion is related to body aging, particularly in close relation to brain aging. The hypothesis of aging is involved in pineal calcification, biological clock, neuro-en-docrinoimmunology, and free radical damage. MT is an endogenous free radical scavenger, may anto-gonize the attack of hydroxyl free radical ( ?OH)on organism and glutamate (Glu) excitotoxicity, and has a potent protective effect of central nervous system. In vivo studies showed that the food restriction and exogenous MT could obviously prolong life, postpone aging, and reduce the chances of age-related diseases. Investigating of MT anti-aging effect shows a vast prospect.

Clinical applications of misoprostol / 中国药理学通报

Misoprostol, a prostaglandin E1 analogue, have variety clinical applications. In early 80's, misoprostol is currently licensed primarily to prevent non-steroidal anti-inflammatory drug-associated gastric and duodenal ulcers. Further work shows the drugs have protection to hepatic and renal and prevention of inflammatory and allergic disorders. When combined with mifepristone,complete abortion rate is 95%. On the other hand, misoprostol is also an effective agent for cervical ripening and labor induction. It have no serious side effect, as a result, misoprostol is a drug of high effect and low toxicity, deserving popularization, but man and pregnant woman must prudent.

Protective effect of total salvianolic acids against cerebral ischemia reperfusion injuries in mice and rats / 中草药

Object It was reported that salvianolic acid A from Salvia miltiorrhiza Bge could improve memory dysfunction of mice with cerebral ischemia reperfusion injury The effects of total salvianolic acids (Sal), including salvianolic acid A and B, rosmaric acid and protocatechic aldehyde was now studied to observe its effect on cerebral ischemia reperfusion injury in hope of obtaining a preliminary view on its mechanism of action Methods The effects of Sal on lactic dehydrogenase (LDH), superoxide dismutase (SOD) and malondialdehyde (MDA) of cerebral ischemia reperfusion injured mouse and rat models were observed Results Sal 10, 20 mg/kg iv and 5, 10 mg/kg iv could markedly increase the activities of LDH and SOD and lower the production of MDA in the injured cerebral tissue On focal cerebral ischemia reperfusion injured rat, 24 h reperfusion after 2 h ischemic injury, 5, 10 mg/kg iv Sal could decrease infarct size, ameliorate neurological deficit with obvious inhibition of the decrease of LDH and SOD activities and the over production of MDA Conclusion Sal showed obvious protective effects on cerebral ischemia reperfusion injuries of mice and rats via attenuating oxygen free radical

Inhibitory Effect of (-)Clausenamide on Apoptosis of PC12 Cells Induced by Serum Deprivation and Its Related Mechanism / 中药新药与临床药理

Objective To study the protective effect of(-)clausenamide on the damage of PC12 cells induced by serum deprivation and to explore its related mechanism. Methods The cell viability was detected by MTT assay and morphological observation. The effect of(-)clausenamide on the PC12 cell apoptosis was analyzed by flow cytometry. Then western blotting and confocal microscope was used for the further study of effect of(-)clausenamide on the protein expression of GSK-3?,Bax and Bcl-2. Results(-)Clausenamide remarkably increased PC12 cell survival rate through inhibiting the PC12 cell apoptosis induced by serum deprivation at the concentration of 1 or 10 ? mol/L(P

Establishment of Female Adult Rat Models with Acute Stress Reproductive- endocrine Disorder / 中药新药与临床药理

Objective To observe the effects of acute stress on reproductive- endocrine function in female adult rats. Methods Acute immobilization stress was used to challenge the female adult rats and then the serum levels of progesterone (P), estradiol (E2) and corticosterone were measured 2 h after the stress.Results Acute stress significantly reduced E2 levels in each period of sexual cycle and increased the levels of P and corticosterone in the stressed rats.In addition, the ratio of estradiol and progesterone was disordered in stressed rats.Conclusion Acute stress can cause the disorder of estradiol and progesterone.Since stress is a major factor contributing to reproductive- endocrine disorder, this model will be useful for the research and development of drugs for reproductive- endocrine disorder. 