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G protein-gated inward rectifier potassium(GIRK)channels are widely distributed in the central nervous system and play important roles in maintaining the resting membrane potential of neurons,adjusting neuronal excitability,and regulating the release of neurotransmitter.Studies have shown that addictive behavior is closely related to the expression and activity of the GIRK channels in the brain reward system and the GIRK channels may be a potential target for addiction treatment.This article summarizes the recent research advances in GIRK channels in terms of structure,intracranial tissue distribution,and especially substance addiction.
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Objective To explore the effects and regulation mechanism of miR-489 on the stem cell phenotype and epithelial-mesenchymal transition of colon cancer cells in vitro.Methods The miR-489 expression of colon cancer cell lines ( HT29, SW480 and SW620, HCT116) and normal intestinal epithelial cells HIEC were detec-ted by RT-qPCR, the miR-489 expression in colon cancer cells was raised by gene transfection technology, the colon cancer stem cell markers CD133, CD44, EpCAM, ALDH1 and epithelial marker E-cadherin, mesenchy-mal cells markers vimentin and N-cadherin were detected by Western blot.The expression of TWIST1 mRNA and protein were detected by RT-qPCR and Western blot.Results The miR-489 relative expression in four kinds of colon cancer cells ( HT29, SW480 and SW620, HCT116) were significantly lower than that of normal HIEC in-testinal epithelial cells, the difference was statistically significant ( P<0.05) .Up-regulation of miR-489 expres-sion in HT29 and HCT116 cells leaded to lower expression of colon cancer stem cell marker CD133, CD44, EpCAM ALDH1 and mesenchymal cells markers Vimentin , N-cadherin, higher expression of epithelial markers ;E-cadherin (P<0.05).Also, up-regulation of miR-489 expression in HT29 and HCT116 cells leaded to lower ex-pression of TWIST 1 mRNA and protein ( P<0.05 ) .Conclusions miR-489 is down-regulated in colon cancer cells and up-regulation of miR-489 expression inhibits stem cell phenotype and epithelial-mesenchymal transition through targeting TWIST1 in colon cancer .
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<p><b>OBJECTIVE</b>To observe the effect of natural type ginsenoside Rg2 (Rg2) and its stereoisomers [20 (R)-Rg2 and 20 (S)-Rg2] at different concentrations on oxygen-glucose deprivation/ reperfusion (OGD/R) induced cortical neuronal injury model in vitro, and to explore the mechanism, and compare their differences of action.</p><p><b>METHODS</b>Cortical neurons after 7-day culture were randomly divided into 5 groups, i.e., the control group, the model group, the Rg2 group, 20 (R) -Rg2 group, and 20 (S) - Rg2 group. Cortical neurons in the Rg2 group, 20 (R)-Rg2 group, and 20(S)-Rg2 group were pretreated with 20, 40, and 80 μmol/L Rg2, 20 (R) -Rg2, and 20 (S) -Rg2 for 24 h to prepare OGD/R model. The cell survival rate, the activity of Caspase-3, the intracellular Ca2+ concentration, contents of superoxide dismutase (SOD) and malondialdehyde (MDA) were detected 24 h later.</p><p><b>RESULTS</b>Compared with the control group, cell survival rates and activities of SOD obviously decreased, the activity of Caspase-3, Ca2+ fluorescent optical gray value, and contents of MDA significantly increased with statistical difference (P < 0.05). Compared with the model group, cell survival rates and activities of SOD obviously increased, the activity of Caspase-3, Ca2+ fluorescent optical gray value, and contents of MDA significantly decreased in 20 μmol/L Rg2 group, 40 μmol/L 20 (R) -Rg2 group, and 80 μmol/L 20 (S) -Rg2 group (P < 0.05). Compared with 20(S)-Rg2 group, cell survival rates increased and contents of MDA significantly decreased in 20, 40, and 80 μmol/L Rg2 and 20 (R)-Rg2 groups (P < 0.05). The activity of Caspase-3 decreased and contents of SOD increased in 80 μmol/L 20 (R)-Rg2 group, and 40, 80 μmol/L Rg2 groups (P < 0.05). Ca2+ fluorescent optical gray value decreased in 40, 80 μmol/L Rg2 and 20 (R)-Rg2 groups (P < 0.05). Compared with 20 (R)-Rg2 group, Ca2+ fluorescent optical gray value decreased in 80 μmol/L Rg2 group (P < 0.05); contents of SOD increased in 40 and 80 μmol/L Rg2 groups (P < 0.05); contents of MDA decreased in 20, 40, and 80 μmol/L Rg2 groups (P < 0.05).</p><p><b>CONCLUSIONS</b>Rg2 and its stereoisomers could improve cell vitality of cortical neurons against OGD/R induced injury. This might be related to improving anti-apoptotic capacities and antioxidant abilities, and reducing Ca2+ inflow. Besides, the neuroprotective effect of 20 (R) -Rg2 was better than that of 20 (S) -Rg2, but inferior to that of Rg2.</p>
Subject(s)
Humans , Antioxidants , Metabolism , Apoptosis , Calcium , Metabolism , Caspase 3 , Metabolism , Cell Survival , Cells, Cultured , Ginsenosides , Pharmacology , Glucose , Malondialdehyde , Metabolism , Neurons , Neuroprotective Agents , Pharmacology , Oxygen , Random Allocation , Reperfusion Injury , Stereoisomerism , Superoxide Dismutase , MetabolismABSTRACT
We developed a method that allows us to label nociceptive neurons innervating tooth-pulp in rat trigeminal ganglion neurons using a retrograde fluorescence-tracing method, to record ATP-activated current in freshly isolated fluorescence-labeled neurons and to conduct single cell immunohistochemical staining for P2X1 and P2X3 subunits in the same neuron. Three types of ATP-activated current in these neurons (F, I and S) were recorded. The cells exhibiting the type F current mainly showed positive staining for P2X3, but negative staining for P2X1. The results provide direct and convincing evidence at the level of single native nociceptive neurons for correlation of the characteristics of ATP-activated currents with their composition of P2X1 and P2X3 subunits and cell size. The results also suggest that the P2X3, but not P2X1, is the main subunit that mediates the fast ATP-activated current in nociceptive neurons.
Subject(s)
Animals , Rats , Action Potentials , Physiology , Adenosine Triphosphate , Metabolism , Dental Pulp , Physiology , Nociceptors , Cell Biology , Physiology , Rats, Sprague-Dawley , Receptors, Purinergic P2X1 , Metabolism , Receptors, Purinergic P2X3 , Metabolism , Tissue Distribution , Trigeminal Nerve , Cell Biology , MetabolismABSTRACT
We developed a method that allows us to label nociceptive neurons innervating tooth-pulp in rat trigeminal ganglion neurons using a retrograde fluorescence-tracing method, to record ATP-activated current in freshly isolated fluorescence-labeled neurons and to conduct single cell immunohistochemical staining for P2X1 and P2X3 subunits in the same neuron. Three types of ATP-activated current in these neurons (F, I and S) were recorded. The cells exhibiting the type F current mainly showed positive staining for P2X3, but negative staining for P2X1. The results provide direct and convincing evidence at the level of single native nociceptive neurons for correlation of the characteristics of ATP-activated currents with their composition of P2X1 and P2X3 subunits and cell size. The results also suggest that the P2X3, but not P2X1, is the main subunit that mediates the fast ATP-activated current in nociceptive neurons.
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<p><b>OBJECTIVE</b>To explore the effect of composite salvia injection (CSI) on platelet parameters in children with anaphylactoid purpura (AP) and its clinical significance.</p><p><b>METHODS</b>One hundred and fifty children with AP were assigned to two groups, 80 in Group A and 70 in Group B. They were treated, respectively, with conventional therapy only or conventional therapy combined with CSI. Their platelet parameters, including blood platelet count (BPC), mean platelet volume (MPV), platelet distribution width (PDW) and plateletcrit (PCT) were determined at the acute stage and convalescent stage, respectively.</p><p><b>RESULTS</b>At the acute stage, the BPC in AP children of both groups was in the normal range, but significant abnormality was shown in the levels of MPV, PDW and PCT. As for comparisons of these parameters at the convalescent stage, significant difference between the two groups was also shown in terms of MPV, PDW and PCT (P<0.05 or P<0.01).</p><p><b>CONCLUSION</b>Although platelet shows no quantitative change in the pathogenic process of AP, important functional changes are surely existent. CSI could promote the normalization of platelet function.</p>