cytochrome-c-t and daidzein

Exercise training has a neuroprotective effect against ischaemic injury, but the underlying mechanism is not completely clear. This study explored the potential mechanisms underlying the protective effects of treadmill training and caveolin-1 regulation against mitochondrial dysfunction in cerebral ischaemic injury.. After middle cerebral artery occlusion (MCAO) surgery, rats were subjected to treadmill training and received daidzein injections and combined therapy. A series of analyses, including neurological function scoring; body weight measurement; Nissl, haematoxylin and eosin staining; cerebral infarction volume assessment; mitochondrial morphology examination; caveolin-1, cytoplasmic and mitochondrial cytochrome C (CytC), and translocase of outer membrane 20 (TOM20) expression analysis; apoptosis index analysis; and transmission electron microscopy were conducted.. Treadmill training increased caveolin-1 expression, reduced neurobehavioral scores and cerebral infarction volumes, improved tissue morphology, reduced neuronal loss, inhibited mitochondrial outer membrane permeabilization (MOMP) through the caveolin-1 pathway, prevented excessive Cyt-C release from mitochondria, and reduced the degrees of apoptosis and mitochondrial damage. In addition, treadmill training increased the expression of TOM20 through the caveolin-1 pathway and maintained import signal function, thereby protecting mitochondrial integrity.. Treadmill exercise protected mitochondrial integrity and inhibited the endogenous mitochondrial apoptosis pathway. The damage of cerebral ischaemia was alleviated in rats through enhancement of caveolin-1 by treadmill exercise.

Topics: Animals; Apoptosis; Body Weight; Brain Ischemia; Caveolin 1; Cytochromes c; Exercise Test; Isoflavones; Male; Microscopy, Confocal; Microscopy, Fluorescence; Mitochondria; Neuroprotective Agents; Physical Conditioning, Animal; Rats; Rats, Sprague-Dawley; Signal Transduction

Daidzein, which belongs to the group of isoflavones from soybeans, has been extensively researched prostate, cervix, brain, breast, and colon cancer cell lines. However, daidzein has not been thoroughly investigated in human hepatic cancer cells; therefore, we investigated whether it inhibits hepatic cancer cell growth. Decreased cell proliferation was measured in daidzein-treated hepatic cancer cells (SK-HEP-1) upon real-time cell electronic sensing analysis however, it was not affected on normal human hepatocytes (Chang). Daidzein-induced apoptosis was demonstrated by comet and TUNEL assay. Moreover, we conducted two-dimensional electrophoresis to study the mechanism of daidzein-induced apoptosis in daidzein-treated SK-HEP-1 cells. Expression of peroxiredoxin-3 (Prdx-3), which modulates redox homeostasis of cells, was increased in protein analysis. Additionally, we measured the levels of reactive oxygen species and it was decreased in daidzein-treated SKHEP-1 cells. Daidzein-induced apoptosis in SK-HEP-1 cells was also associated with the up-regulation of Bak and down-regulation of Bcl-2 and Bcl-xL proteins. Moreover, daidzein treatment increased in the release of mitochondrial cytochrome c and activation of APAF-1, caspase 9 and caspase 3. Overall, these result indicate that daidzein is a potent inducer of apoptosis in hepatic cancer cells via mitochondrial pathway.

Topics: Apoptosis; Apoptosis Regulatory Proteins; Apoptotic Protease-Activating Factor 1; bcl-2 Homologous Antagonist-Killer Protein; bcl-X Protein; Caspase 3; Caspase 9; Cell Line, Tumor; Cell Proliferation; Comet Assay; Cytochromes c; Down-Regulation; Humans; In Situ Nick-End Labeling; Isoflavones; Liver Neoplasms; Mitochondria; Peroxiredoxin III; Reactive Oxygen Species; Signal Transduction; Up-Regulation

Cytochrome c (CytC) released from mitochondria induces apoptosis in both normal and tumor cells. Expression of Fas ligand (FasL) helps maintain tumor cell survival by inducing apoptosis of Fas-bearing anti-tumor immune cells. A risk of endometrial cancer has been reported to associate with phytoestrogen consumption. Therefore the effects of phytoestrogens, genistein and daidzein, on FasL and CytC protein expression were examined in primary cultured porcine endometrial cells (PE) and human cancerous endometrial cells (RL95-2) by Western blot analysis. Both cells were cultured in standard medium (SM) and switched to estrogen-deprived medium (SF) with or without 17beta-estradiol (E, 1 nM), genistein (10 microM) or daidzein (10 microM) for 48 h. FasL (25 kDa) which was found only in RL95-2 cells was upregulated in SF compared to SM. Treatment of RL95-2 cells with E, daidzein or genistein significantly increased the FasL expression by 7-10 folds. In the present study, low level of CytC was detected in both cells cultured in SM but markedly increased in SF by 1.5-2 folds. The SF-induced increase in CytC level was reversed by genistein or daidzein while E suppressed CytC in PE cells, but not in RL95-2 cells. The findings suggest that genistein and daidzein appear to act as a survival factor by inhibiting intracellular apoptogenic initiator in both normal and cancer endometrial cells. In addition, estrogen and phytoestrogens inducing the death signal FasL expressed by cancerous endometrial cells may cause the tumor progression. Thus, consuming phytoestrogen as a supplement should be awareness in patient with endometrial cancer.

Topics: Analysis of Variance; Animals; Apoptosis; Blotting, Western; Cells, Cultured; Cytochromes c; Endometrial Neoplasms; Estradiol; Fas Ligand Protein; Female; Genistein; Humans; Isoflavones; Phytoestrogens; Swine

We have investigated the ability of certain dietary flavonoids, known to exert beneficial effects on the central nervous system, to affect neuronal apoptosis. We used cerebellar granule cells undergoing apoptosis due to potassium deprivation in a serum-free medium in either the absence or presence of the flavonoids genistein and daidzein, which are present in soy, and of catechin and epicatechin, which are present in cocoa. These compounds were used in a blood dietary concentration range. We found that genistein and daidzein, but not catechin and epicatechin, prevented apoptosis, with cell survival measured 24h after the induction of apoptosis being higher than that of the same cells incubated in flavonoid free medium (80% and 40%, respectively); there was no effect in control cells. A detailed investigation of the effect of these compounds on certain mitochondrial events that occur in cells en route to apoptosis showed that genistein and daidzein prevented the impairment of glucose oxidation and mitochondrial coupling, reduced cytochrome c release, and prevented both impairment of the adenine nucleotide translocator and opening of the mitochondrial permeability transition pore. Interestingly, genistein and daidzein were found to reduce the levels of reactive oxygen species, which are elevated in cerebellar granule cell apoptosis. These findings strongly suggest that the prevention of apoptosis depends mainly on the antioxidant properties of genistein and daidzein. This could lead to the development of a flavonoid-based therapy in neuropathies.

Topics: Animals; Apoptosis; Cell Survival; Cells, Cultured; Cerebellum; Cytochromes c; DNA Fragmentation; Genistein; Glucose; Isoflavones; Mitochondria; Mitochondrial Membrane Transport Proteins; Mitochondrial Permeability Transition Pore; Neurons; Oxidation-Reduction; Potassium; Protein Kinase Inhibitors; Rats; Rats, Wistar; Superoxides

Early stage cervical cancer is treated with surgery or radiation with equivalent results. Radiation is used for curative therapy of locally advanced disease and is combined with additional anti-tumor agents to improve control. We determined the potential role of genistein as a radiosensitizer for cervical cancer cells.. Human cervical cell lines (CaSki and ME180) were used. Sensitivity of cells to genistein, radiation and the combination was determined by colony assays. Western blotting was used to study the expression of cell-response-related gene products.. Genistein results in the dose-dependent inhibition of all cell lines (2.5-40.0 microM). Effect of genistein on the radiosensitivity of the two cervical tumor cells was variable. Me180 cells were more sensitive at 20 and 40 microM of genistein. At 40 microM, less than 5% of Me180 cells survived the radiation (200-800 cGy). Potentiation of the radiation effect in CaSki cells was seen (500-800 cGy). The most significant enhancement of radiosensitivity was seen at 20 and 40 microM genistein at 500 and 800 cGy. G(2)M arrest was demonstrated only in ME180 cells with genistein. There was significant inhibition of Mcl-1 by genistein that correlated with increase in radiosensitivity in Me180 cells. Activated pAKT (Thr 308) was inhibited with genistein and radiation in CaSki cells. CONCLUSIONS.: Genistein inhibits growth of cervical cancer cells. Genistein results in variable and significant enhancement of the radiation effect that may be partially mediated by G(2)M arrest, Mcl-1 and activation of the AKT gene.

Topics: Antineoplastic Agents; Antineoplastic Combined Chemotherapy Protocols; Cell Cycle; Cell Growth Processes; Cell Line, Tumor; Combined Modality Therapy; Cytochromes c; Dose-Response Relationship, Drug; Female; Genistein; Humans; Isoflavones; Myeloid Cell Leukemia Sequence 1 Protein; Neoplasm Proteins; Protein Serine-Threonine Kinases; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-akt; Proto-Oncogene Proteins c-bcl-2; Radiation-Sensitizing Agents; Uterine Cervical Neoplasms