ABSTRACT
<p><b>OBJECTIVE</b>To observe the neuroprotective and neurotrophic effects ofliquiritin (LQ) on rats primary cultured hippocampal neuronal damage induced by Abeta25-35.</p><p><b>METHOD</b>The rats hippocampal neuronal damage model by Abeta25-35 was established. The protective effects of LQ to the cells were observed through the MTU assay. The apoptosis of neurons was detected by flow cytometer and the concentration of intracellular calcium by fluorescence probe dying. LQ' s neurotrophic effects were observed through measuring the neurite outgrowth induced by LQ in primary cultured hippocampal neurons and the differentiation of LQ on hippocampal stem cells to cholinergic neurons was assayed by flow cytometry.</p><p><b>RESULT</b>Treatment of the cells with 10 micromol x L(-1) Abeta25-35 could induce a significant decrease of cell viability, enhance the level of intracellular [Ca2+] and increase the percentage of apoptosis to 28%. However, pretreatment with LQ (0.1, 1, and 10 micromol x L(-1)) for 6 hours exhibited cytoprotective effects, inhibited the cells' s death induced by Abeta25-35, prevented the accumulation of [Ca2+], and decreased the apoptosis neurons significantly to 10%, 15% and 9%, which meaned that LQ could antagonize Abeta25-35 induced apoptosis. LQ together with NGF had a dramatic prolonged effect on the neurite of the primary cultured hippocampal neurons, which was blocked by a specific MAPK kinase inhibitor to some extent. In addition, LQ could induce the differentiation of hippocampal stem cells to cholinergic neurons in vitro.</p><p><b>CONCLUSION</b>These results demonstrate that LQ has the neuroprotective capacity to cell damage iduced by Abeta25-35 in primary cultured hippocampal neurons, and also has the neurotrophic effects.</p>
Subject(s)
Animals , Female , Male , Rats , Amyloid beta-Peptides , Apoptosis , Calcium , Metabolism , Cell Differentiation , Cell Movement , Cell Survival , Cells, Cultured , Flavanones , Pharmacology , Glucosides , Pharmacology , Hippocampus , Cell Biology , Pathology , Neurites , Pathology , Neurons , Cell Biology , Metabolism , Pathology , Neuroprotective Agents , Pharmacology , Peptide Fragments , Stem Cells , PathologyABSTRACT
This study is to investigate if madecassoside can protect against myocardial reperfusion injury in rabbit heart in vivo. The ischemia reperfusion model was established. Left ventricular function and ECG were monitored at the ischemia and reperfusion period. The infarct areas were expressed as percentage. The levels of LDH, CK, MDA and SOD were measured and C-reactive protein (CRP) in serum was measured by ELISA kit. Cardiomyocyte apoptosis were measured by TUNEL staining. A monoclonal rabbit anti-goat Bcl-2 proteins as primary antibody was used for Bcl-2 immunohistochemical staining. Treatment with madecassoside (3.2, 1.6 and 0.8 mg x kg(-1)) i.v. during ischemia reperfusion injury attenuated myocardial damage, that is, characteristic of decreasing infarct size, decreasing LDH and CK release. Activities of SOD were diminished and MDA level increased obviously in control group whereas pretreatment with madecassoside significantly blunted the decrease of SOD activity, markedly reduced the levels of MDA, CRP and cardiomyocyte apoptosis, and upregulated the expression of Bcl-2. Madecassoside has the protective effect against myocardial ischemia reperfusion injury, and effects of anti-lipid peroxidation, enhancement of SOD activity, anti-inflammation and anti-apoptosis.
Subject(s)
Animals , Male , Rabbits , Apoptosis , C-Reactive Protein , Metabolism , Cardiotonic Agents , Pharmacology , Centella , Chemistry , Creatine Kinase , Blood , Electrocardiography , L-Lactate Dehydrogenase , Blood , Lipid Peroxidation , Malondialdehyde , Blood , Myocardial Reperfusion Injury , Metabolism , Pathology , Myocardium , Metabolism , Pathology , Myocytes, Cardiac , Pathology , Plants, Medicinal , Chemistry , Proto-Oncogene Proteins c-bcl-2 , Metabolism , Random Allocation , Superoxide Dismutase , Blood , Triterpenes , PharmacologyABSTRACT
The effect of recombinant microplasmin (micro-plasmin) on acute cerebral infarction was evaluated in rats, and compared with recombinant tissue plasminogen activator (rt-PA). After the model of middle cerebral artery occlusion (MCAO) was established by autologous blood clots, different doses of micro-plasmin (2.5, 5, and 10 mg x kg(-1)) were administered into the thrombus intra-arterial. Twelve hours after administration of micro-plasmin, the neurological deficit score of rats was recorded and the infarct volumes were determined. Bleeding time (BT), fibrin degradation product (FDP) concentration in serum and thrombin time (TT), prothrombin time (PT) and fibrinogen (FIB) concentration in plasma were tested after administration. Intra-arterial administration of micro-plasmin could reduce significantly neurological deficit score and infarct volumes in MCAO rats. FDP concentration increased significantly as compared with model group. There were no significant differences in TT, PT and BT. FIB concentration reduced markedly as compared with model group, but had no significant difference as compared with sham group. The results suggest that micro-plasmin is effective in treatment of rat acute cerebral infarction, and has no significant influence on fibrinolytic system and blood clotting system, indicating that micro-plasmin may be useful for treatment of acute cerebral infarction, and not lead to hemorrhage. Micro-plasmin seems to be distinguished from clinical used rt-PA by its no hemorrhage effect.
Subject(s)
Animals , Male , Rats , Bleeding Time , Brain , Pathology , Cerebral Hemorrhage , Metabolism , Pathology , Cerebral Infarction , Drug Therapy , Pathology , Fibrin Fibrinogen Degradation Products , Metabolism , Fibrinogen , Metabolism , Fibrinolysin , Pharmacology , Infarction, Middle Cerebral Artery , Peptide Fragments , Pharmacology , Prothrombin Time , Random Allocation , Rats, Sprague-Dawley , Recombinant Proteins , Pharmacology , Thrombin Time , Tissue Plasminogen Activator , PharmacologyABSTRACT
<p><b>AIM</b>To establish a sensitive and efficient reporter gene-based screening model for finding agonists of estrogen receptor beta subtype.</p><p><b>METHODS</b>A recombinant vector pTAL-ERE-SEAP was constructed by inserting a synthetic sequence composed of five estrogen responsive elements in front of promoter of pTAL-SEAP vector. pTAL-ERE-SEAP was then transfected into human embryonic kidney (HEK293) cells. G418 (200 microg x mL(-1)) was added to select positive clones that can be induced by E2 to express reporter gene SEAP. The speciality was tested by several ligands of relative nuclear receptors of the same family. The stability of the model, the time-effect relationship, the dose-response relationship, and the immunocytochemistry staining of ERbeta expression after transfection were observed. 2 622 compounds were screened by using this model.</p><p><b>RESULTS</b>Stably transfected clones were obtained. The expression levels of reporter gene SEAP of positive clones was induced by E2 in a dose-response and time-effect relationship manners. The Z' factor value was 0.7. The expression levels of dexamethasone and other ligands were low. The result of immunocytochemistry staining showed the expression of ERbeta. E2 had no proliferating effects on stably transfected clones.</p><p><b>CONCLUSION</b>Stably transfected positive clones transfected with recombinant vector pTAL-ERE-SEAP were obtained. The positive clones may be used to screen for agonists of estrogen receptor beta subtype by measurement of luminescent value of expressed SEAP in wells of microlitre plate.</p>