Reverse of β-Amyloid Induced Apoptosis in PC12 Cells by Nattokinase: Role of SIRT1-ROCK1 Pathway

Background and Objective: It is well-established fact that Alzheimer's disease (AD) is a neurodegenerative disease of CNS with progressive nature. The available treatment for AD unable to treat the pathology of disease. Due to this limitation urgent need the newer treatment of AD. The current investigation was aimed to scrutinize the neuro-protective effect of nattokinase against beta-amyloid peptide(25-35) (A beta(25-35)) induced neurotoxicity and explored the underlying mechanism. Materials and Methods: The PC12 cells were damaged by A beta(25)(-)(35) and different concentration of nattokinase was added into the culture medium. Lactate dehydrogenase (LDH) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay were performed for the estimation of cell viability and also estimation the hydrogen peroxide (H2O2) generation for estimation the intracellular signalling pathways. Furthermore, determination of mitochondrial membrane potential changes and caspase-3 effect of this compound occurred in this study. Additionally, ROS intracellular and expression of silent information regulator 1 (SIRT1) and Rho-associated kinase 1 (ROCK1) were also estimated. Results: The concentration-dependent treatment of nattokinase down-regulated the cell viability counts. The result exhibited that cell death induced via A beta(25)(-35) was mediated through an up-regulation of ROS production. It is estimated that there was enhanced caspase-3 and caspase-8 activity attributing to depolarization of mitochondrial membrane and reduced by the nattokinase. Nattokinase reduced the reactive oxygen species and down-regulated the overproduction of IL-1 beta, IL-6 and TNF-alpha, respectively. Nattokinase reduced the cell apoptosis and expression of ROCK1 and increased the expression of SIRT1. Conclusion: On the basis of the result, concluded that SIRT1-ROCK1 pathway plays a significant role in the AD pathology and nattokinase reduced the AD complication via SIRT1-ROCK1 pathway.