| Objective: To study the effect of calmodulin on reactive astrocytosis and formation of glial scar after astrocytes were injured in vitro。Methods: Astrocytes were prepared from newborn rat cerebral cortex and evaluated the purity by immunostaining of glial fibrillary acidic protein (GFAP). The astrocyes injury model was made by means of surgical blade scraping. We observed the concentration of intracellular free calcium after injury in dynamic state .Then injured astrocytes were divided into two groups: the intervention group was added into suppressive agent of calmodulin—trifluoperazin (TFP) after injury; the control group was added into nothing after injury. At different times,we performed immunocytochemical staining for GFAP on each group, and analyzed immunocytochemical staining photographs by image analysis system, obtaining average optical densities.Results: (1)The purity of cultured astrocytes is above 95%;(2)Asreocytes injured by scratch injury model showed the same morphologic changes as other injury models in domestic and foreign studies.(3) After astrocyte injured the cellular free calcium concentration of arised quickly,and then drop down in 2 minutes。The concentration of the cellular free calcium fall-off maintenance in 15minutes ,then the concentration is nearly to the astrocyte in rest. (4)The immunocytochemical staining results indicated that there was statistical significance between the intervention group and the control group。Conclution: (1)The purity of cultured astrocytes is above 95%;(2)The astrocytes injury model had been achieved successfully by scraping;(3) After astrocytes injured the intracellular free calcium concentration obviously arised , activated the intracellular Ca2+ signal pathway and participate to the proliferation reaction,(4)After astrocyte injured the TFP can obviously inhibit this GFAP expression ,and that indicate the calmodulin regulates the reactive astrocytosis and formation of glial scar after astrocytes were injured in vitro. |
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