Study On The Mechanism Of Mitomycin C In The Prevention Of Peridural Adhesion After Laminectomy

Objective: To elucidate the mechanism of mitomycin C prevention of peridural adhesion after laminectomy on inducing apoptosis of fibroblastMethods The cultured fibroblasts were exposed for 12h with 0mg/ml,0.001 mg/ml,0.01mg/ml,0.05 mg/ml,0.1 mg/ml,0.2 mg/ml,0.3 mg/ml mitomycin C, respectively. The cell proliferation was detected by CCK-8 assay and Apoptotic cell death was analyzed by Hoechst 33342 stain. The expression of caspase-3 and the activation of ERK1/2, Akt ,Bad were analyzed by Western blotResults Treatment of fibroblast with MMC induced a significant decreased of proliferation. fibroblasts exposed for 0.001 mg/ml to 0.2 mg/ml MMC, Staining with Hoechst 33342 MMC displayed nuclei with abnormal morphologies:"bean"-shaped nuclei with highly condensed chromatin or nuclei with irregular clumps of dense chromatin. 0.2mg/ml MMC,PD98059(ERK kinase inhibitor),LY294002(AKT kinase inhibitor) induced a significant increased of caspase-3 than control, accompanied by an increase in cell apoptosis.0.2mg/ml MMC PD98059,LY294002 caused decrease of p-ERK1/2 and p-Akt,at the same time, 0.2mg/ml MMC increased Bad than PD98059,LY294002 and control.Conclusion 0.2 mg/ml MMC can induce fibroblast apoptosis by activation of caspase-3 in which requires inactivation of p-ERK1/2, p-Akt and increase the Bad. MMC can inhibit fibroblast proliferation and induce its apoptosis, which may be one of mechanism of that MMC prevent peridural adhesion after laminectomy...