The opioid receptor-libel receptor (ORL), an orphan receptor whose human and murine complementary DNAs,has been characterized recently. ORL transcripts are particularly abundant in the central nervous system. We demon...The opioid receptor-libel receptor (ORL), an orphan receptor whose human and murine complementary DNAs,has been characterized recently. ORL transcripts are particularly abundant in the central nervous system. We demonstrated that ORL expressed in human neuroblastoma SK-N-SH and SH-SY5Y cell lines by radioligand binding assay, reverse transcription polymerase chain reaction (RT-PCR) and Northern analysis in the present study. Stimulation with ORL1 specific agonist, nociceptin/orphanin Fo, increased [34S]GTPrγS binding to SK-N-SH cell membranes (EC50 = 14 ±0.45 nM), and attenuated forskolin-stimulated accumulation of cellular cAMP (EC50= 0.80 ±0.45 nM, indicative that activation of ORL1 activates G proteins and inhibits adenylyl cyclase. Activation of ORL1 receptor was also accessed using CHO:hORL1 cell line by microphysiometer. Treatment of nociceptin/orphanin FQ increased extracellular acidification rate significantly.展开更多
We evaluated the effects of pre-germinated brown rice extract (PGBR ex) with enhanced levels of GABA on proliferation and apoptosis of neuronal SK-N-SH cells line. Firstly, we used HPLC methods to study the level of ...We evaluated the effects of pre-germinated brown rice extract (PGBR ex) with enhanced levels of GABA on proliferation and apoptosis of neuronal SK-N-SH cells line. Firstly, we used HPLC methods to study the level of γ-aminobutyric acid (GABA) in all rice extracts. We found that the concentration of GABA in the PGBR ex were 3 and 8 times higher than the GABA concentration in non-germinated brown rice (BR ex) and white rice (WR ex) compared with the stan- dard GABA respectively. Next we study the protective effects of brown rice extract by investigating various methods, we found that the effects of dose-dependent study by treated with PGBR ex, BR ex and WR ex at (0 - 4000 μg/ml). The data from MTT assay showed that the higher concentration of all rice extracts were not induced toxicity to SK-N-SH cells. To test the protective effect by study the viability of SK-N-SH cells. These results showed that PGBR ex and BR ex can protect cells by significantly increase cells survival up to 29.3% ± 0.01% and13.4% ± 0.07 % (p 56.9% ± 0.02 % (p < 0.05), compared with un- treated cells (control). Next study we test the effect of cells apoptotic by ROS assay and DNA fragmentation. The results showed that PGBR ex were definitely decrease the amount of ROS formation and had a little of DNA ladders comparable with condition that induced by 150 ?M H2O2. Our data indicating that PGBR ex with enhanced levels of GABA effectively inhibit SK-N-SH cells proliferation and apoptosis. These present results suggest that intake of PGBR and BR instead of WR is effective to protect cell proliferation and apoptosis which may be useful nutritional to prevent neuronal cells from neurodegenerative disease.展开更多
文摘The opioid receptor-libel receptor (ORL), an orphan receptor whose human and murine complementary DNAs,has been characterized recently. ORL transcripts are particularly abundant in the central nervous system. We demonstrated that ORL expressed in human neuroblastoma SK-N-SH and SH-SY5Y cell lines by radioligand binding assay, reverse transcription polymerase chain reaction (RT-PCR) and Northern analysis in the present study. Stimulation with ORL1 specific agonist, nociceptin/orphanin Fo, increased [34S]GTPrγS binding to SK-N-SH cell membranes (EC50 = 14 ±0.45 nM), and attenuated forskolin-stimulated accumulation of cellular cAMP (EC50= 0.80 ±0.45 nM, indicative that activation of ORL1 activates G proteins and inhibits adenylyl cyclase. Activation of ORL1 receptor was also accessed using CHO:hORL1 cell line by microphysiometer. Treatment of nociceptin/orphanin FQ increased extracellular acidification rate significantly.
文摘We evaluated the effects of pre-germinated brown rice extract (PGBR ex) with enhanced levels of GABA on proliferation and apoptosis of neuronal SK-N-SH cells line. Firstly, we used HPLC methods to study the level of γ-aminobutyric acid (GABA) in all rice extracts. We found that the concentration of GABA in the PGBR ex were 3 and 8 times higher than the GABA concentration in non-germinated brown rice (BR ex) and white rice (WR ex) compared with the stan- dard GABA respectively. Next we study the protective effects of brown rice extract by investigating various methods, we found that the effects of dose-dependent study by treated with PGBR ex, BR ex and WR ex at (0 - 4000 μg/ml). The data from MTT assay showed that the higher concentration of all rice extracts were not induced toxicity to SK-N-SH cells. To test the protective effect by study the viability of SK-N-SH cells. These results showed that PGBR ex and BR ex can protect cells by significantly increase cells survival up to 29.3% ± 0.01% and13.4% ± 0.07 % (p 56.9% ± 0.02 % (p < 0.05), compared with un- treated cells (control). Next study we test the effect of cells apoptotic by ROS assay and DNA fragmentation. The results showed that PGBR ex were definitely decrease the amount of ROS formation and had a little of DNA ladders comparable with condition that induced by 150 ?M H2O2. Our data indicating that PGBR ex with enhanced levels of GABA effectively inhibit SK-N-SH cells proliferation and apoptosis. These present results suggest that intake of PGBR and BR instead of WR is effective to protect cell proliferation and apoptosis which may be useful nutritional to prevent neuronal cells from neurodegenerative disease.
