ABSTRACT
OBJECTIVE@#To investigate the effect of anesthesia on the cognitive status damage and MMP-2 expression in rats.@*METHODS@#A total of 120 healthy rats were selected and randomly divided into the control group, CF3-CH(OCH2F)-CF3 (Sevoflurane) group and CF3-CH2-O-CHF-CF3 group (Sevoflurane) (n=40). After training for 3 d by the Morris water maze, the control group were injected with fentanyl for analgesia, the CF3-CH(OCH2F)-CF3 group and the CF3-CH2-O-CHF-CF3 group were anesthesia with CF3-CH (OCH2F)-CF3 and CF3-CH2-O-CHF-CF3 on the basis of fentanyl, then rats in three groups underwent open surgery and suture conventional incision. Morris water maze was used to measure the rats' cognitive ability in three groups on the 1st d, 3rd d, 5th d and 7th d, and the brain tissue MMP-2 expression was detected.@*RESULTS@#After 1 d/7 d of the surgery, Morris water maze performance and MMP-2 expression were not significantly different among three groups (P>0.05); After 3 d/5 d of the surgery, compared with the control group, the Morris water maze test result was significantly worsened, MMP-2 expression levels were significantly increased (P<0.05); After 3 d/5 d of the surgery, compared with the CF3-CH2-O-CHF-CF3 group, Morris water maze test result of CF3-CH(OCH2F)-CF3 group was significantly worsened, MMP-2 expression levels were significantly increased (P<0.05).@*CONCLUSIONS@#Anesthesia can cause some injury on cognitive status, different anesthetic drugs may cause different injury, and the cognitive status injury is related to the MMP-2 expression.
Subject(s)
Animals , Male , Rats , Anesthetics, Inhalation , Pharmacology , Behavior, Animal , Brain , Metabolism , Brain Chemistry , Cognition , Matrix Metalloproteinase 2 , Metabolism , Maze Learning , Methyl Ethers , Pharmacology , Postoperative Complications , Random Allocation , SevofluraneABSTRACT
<p><b>OBJECTIVE</b>To analyze the change of the neuronal restricted silencing factor (NRSF) gene as well as the NRSF regulation genes in beta-mercaptoethanol induction of the marrow mesenchymal stem cells (MSCs) to neurons, and to discuss the function of NRSF in neural induction of the MSCs and the mechanism of the differentiation from MSCs to neurons.</p><p><b>METHOD</b>We used beta-mercaptoethanol, serum-free DMEM, and dimethyl sulfoxide to induce rat MSCs to differentiate to neurons, and then analyzed the changes of the expressions of NRSF gene and NRSF-regulated genes through real-time PCR.</p><p><b>RESULTS</b>The rat MSCs were successfully induced to differentiate into neuron-like cells. The induced neuron marker, neuron-specific enolase, was positive. Real-time PCR showed that the expression of NRSF gene remarkably declined. The expressions of neurotrophic tyrosine kinase receptor, type 3, synaptosomal-associated protein 25, L1 cell adhesion molecular,neuronal pentraxin receptor in the NRSF-regulated genes also increased at varied extents.</p><p><b>CONCLUSIONS</b>The differentiation from MSCs to neurons is relevant with the decline of NRSF expression and the increase of the expressions of NRSF-regulated genes. The NRSF may be the key gene during the differentiation from MSCs to neurons.</p>
Subject(s)
Animals , Rats , Bone Marrow Cells , Cell Biology , Metabolism , Cell Differentiation , Physiology , Cells, Cultured , Mesenchymal Stem Cells , Cell Biology , Metabolism , Neurons , Cell Biology , Metabolism , Phosphopyruvate Hydratase , Genetics , Metabolism , Rats, Wistar , Repressor Proteins , Metabolism , Physiology , Synaptosomal-Associated Protein 25 , Genetics , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To construct a lentiviral vector of repressor element-1/neuron-restrictive silencer element (RE-1/NRSE) double-stranded RNA (dsRNA).</p><p><b>METHODS</b>The RE-1/NRSE cDNA containing both sense and antisense oligo DNA fragments of the targeting sequence was synthesized and cloned into the pGC-LV vector. The obtained lentiviral vector containing RE-1/NRSE dsRNA was confirmed by PCR and sequencing. A total of 293T cells were cotransfected with lentiviral vector of L-smNRSE/RE-1, pHelper 1.0, and pHelper 2.0. The titer of virus was measured based on the expression level of green fluorescent protein. The transfection efficiency of green fluorescent protein into rat mesenchymal stem cells was calculated.</p><p><b>RESULTS</b>PCR and DNA sequencing demonstrated that the constructed lentivirus vector of L-smNRSE/RE-1 produced RE-1/NRSE dsRNA.The titer of the concentrated virus was 4x108 TU/m1. The virus was stably transfected into rat mesenchymal stem cells, and the infection efficiency reached 100% when the multiplicity of infection was 80.</p><p><b>CONCLUSION</b>The lentivirus vector of RE-1/NRSE dsRNA is successfully constructed.</p>