ABSTRACT
In this study, one immortalized human normal prostatic epithelial cell line (BPH) and four human prostate cancer cell lines (LNCaP, 22Rv1, PC-3, and DU-145) were treated with Ganoderma Lucidum triterpenoids (GLT) at different doses and for different time periods. Cell viability, apoptosis, and cell cycle were analyzed using flow cytometry and chemical assays. Gene expression and binding to DNA were assessed using real-time PCR and Western blotting. It was found that GLT dose-dependently inhibited prostate cancer cell growth through induction of apoptosis and cell cycle arrest at G1 phase. GLT-induced apoptosis was due to activation of Caspases-9 and -3 and turning on the downstream apoptotic events. GLT-induced cell cycle arrest (mainly G1 arrest) was due to up-regulation of p21 expression at the early time and down-regulation of cyclin-dependent kinase 4 (CDK4) and E2F1 expression at the late time. These findings demonstrate that GLT suppresses prostate cancer cell growth by inducing growth arrest and apoptosis, which might suggest that GLT or Ganoderma Lucidum could be used as a potential therapeutic drug for prostate cancer.
Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , G1 Phase Cell Cycle Checkpoints/drug effects , Gene Expression Regulation, Neoplastic , Prostate/drug effects , Reishi/chemistry , Triterpenes/pharmacology , Antineoplastic Agents, Phytogenic/isolation & purification , Apoptosis/drug effects , Caspase 3/genetics , Caspase 3/metabolism , Caspase 9/genetics , Caspase 9/metabolism , Cell Line, Tumor , Cell Survival/drug effects , Cyclin D1/genetics , Cyclin D1/metabolism , Cyclin-Dependent Kinase 4/genetics , Cyclin-Dependent Kinase 4/metabolism , Cyclin-Dependent Kinase Inhibitor p21/genetics , Cyclin-Dependent Kinase Inhibitor p21/metabolism , Dose-Response Relationship, Drug , E2F1 Transcription Factor/genetics , E2F1 Transcription Factor/metabolism , G1 Phase Cell Cycle Checkpoints/genetics , Humans , Male , Nucleosomes/drug effects , Nucleosomes/metabolism , Nucleosomes/pathology , Plant Extracts/chemistry , Prostate/metabolism , Prostate/pathology , Signal Transduction , Triterpenes/isolation & purificationABSTRACT
Oxidative stress is implicated in male infertility and significantly higher reactive oxygen species are detected in 25% of infertile males. Although different agents of various alternative medicines, including traditional Chinese medicine, have been tried with varying success, evidence remains limited on whether and how much herbs or supplements might help increase the anti-oxidant ability of the sperm. This study examined the anti-oxidative effects of icariin, a flavonoid isolated from Herba Epimedii, on the human sperm. We prepared the FeSO4/H2O2-damaged human sperms, which were co-cultured with icariin in vitro, and then observed the changes of the sperm by employing Raman micro-spectroscopy. The results showed that Raman mapping with a 514 nm excitation laser allowed clear differentiation of the nucleus, neck, and, in particular, the mitochondria-rich middle piece of a human sperm cell. The effect of icariin on different organelles of the sperm was quantified by localized spectral Raman signatures obtained within milli-seconds, and icariin could keep the "Raman fingerprint" of the human sperm the same as the control groups, suggesting that icariin could protect the human sperm from being damaged by FeSO4/H2O2. Icariin may serve as a tonifying and replenishing agent of herbal origin for enhancing reproductive functions.
Subject(s)
Ferrous Compounds/pharmacology , Flavonoids/pharmacology , Hydrogen Peroxide/pharmacology , Oxidative Stress/drug effects , Protective Agents/pharmacology , Spermatozoa/drug effects , Cells, Cultured , Drugs, Chinese Herbal/pharmacology , Flow Cytometry , Humans , L-Lactate Dehydrogenase/metabolism , Male , Malondialdehyde/metabolism , Oxidants/pharmacology , Spectrum Analysis, Raman , Spermatozoa/cytology , Spermatozoa/metabolism , Superoxide Dismutase/metabolism , Time FactorsABSTRACT
AIM: To further investigate the relaxation mechanism of neferine (Nef), a bis-benzylisoquinoline alkaloid extracted (isolated) from the green seed embryo of Nelumbo nucifera Gaertn in China, on rabbit corpus cavernosum tissue in vitro. METHODS: The effects of Nef on the concentrations of cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) in isolated and incubated rabbit corpus cavernosum tissue were recorded using 125I radioimmunoassay. RESULTS: The basal concentration of cAMP in corpus cavernosum tissue was 5.67 +/- 0.97 pmol/mg. Nef increased the cAMP concentration in a dose-dependent manner (P < 0.05), but this effect was not inhibited by an adenylate cyclase inhibitor (cis-N-[2-phenylcyclopentyl]azacyclotridec-1-en-2-amine, MDL-12, 330A) (P > 0.05). The accumulation of cAMP induced by prostaglandin E1 (PGE1, a stimulator of cAMP production) was also augmented by Nef in a dose-dependent manner (P < 0.05). The basal concentration of cGMP in corpus cavernosum tissue is 0.44 +/- 0.09 pmol/mg. Nef did not affect this concentration of cGMP, either in the presence or in the absence of a guanyl cyclase inhibitor (1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one, ODQ) (P > 0.05). Also, sodium nitroprusside (SNP, a stimulator of cGMP production)-induced cGMP production was not enhanced by Nef (P > 0.05). CONCLUSION: Nef, with its relaxation mechanism, can enhance the concentration of cAMP in rabbit corpus cavernosum tissue, probably by inhibiting phosphodiesterase activity.
Subject(s)
Benzylisoquinolines/pharmacology , Drugs, Chinese Herbal , Nelumbo , Penis/drug effects , Phytotherapy , Vasodilator Agents/pharmacology , Animals , Benzylisoquinolines/therapeutic use , Cyclic AMP/metabolism , Cyclic GMP/metabolism , Erectile Dysfunction/drug therapy , In Vitro Techniques , Male , Muscle Relaxation/drug effects , Muscle, Smooth/drug effects , Penile Erection/drug effects , Penis/metabolism , Plant Extracts , Rabbits , Radioimmunoassay , Seeds , Vasodilator Agents/therapeutic useABSTRACT
OBJECTIVE: To study the relaxants effects of six extractions from Chinese Herbs (neferine, tetrandrine, kakonein, scutellarin, ginsenoside Rgl and ginsenoside Rb1) on the corpus cavernosum tissue of rabbit in vitro. METHODS: Isolated stripes of rabbit corpus cavernosum tissue were precontracted with 10(-5) mol/L phenylephrine (PE). Relaxation in response to cumulative doses of six extracts at (10(-8) - 10(-3)) mol/L was determined. RESULTS: On rabbit cavernosal muscle stripes precontracted with PE, neferine, tetrandrine, kakonein and scutellarin showed dose dependent relaxation. IC50 values were 4.60 x 10(-6), 3.73 x 10(-5), 8.03 x 10(-4) and 3.33 x 10(-3) mol/L, respectively. However, in the meantime, it was found that the relaxant effects of ginsenoside Rgl and ginsenoside Rbl less significant to stripes precontracted with PE. When the final concentration was 10(-3) mol/L, the relaxations were only (16.32 +/- 5.45)% and (11.21 +/- 3.10)%. CONCLUSION: Among the six extracts which showed relaxant effects to rabbit cavernosal muscle stripes precontracted with PE, neferine had greater functions than the other five extracts.