Anti-BAX monoclonal antibody, clone 7B8 [Biotin]

The marine environment has been represented as the most vital source of bioactive constituents. In the present work, we intended to purification, characterization, and investigation of the apoptotic effect of the bioactive molecule from edible mollusc Donax variabilis on NSCLC cell lines. The bioactive molecules were purified using Anion Exchange Chromatographic method with different millimolar of NaCl concentrations. The fractions were further performed to check the proliferative effect on A549 and NCI-H23 cells by MTT assay. The molecular mass of the active purified protein fraction (PPF-V) was 40 kDa, determined by SDS-PAGE. Furthermore, MALDI-TOF/MS was disclosed that it shared a 91% protein sequence similarity with FMRFamide peptide. Meanwhile, the apoptotic effect of PPF-V on A549 and NCI-H23 cells were investigated. Immunoblot analysis found that the PPF-V treatment groups showed the upregulation of Bax, Cytochrome-c, Cleaved Caspase-9 and 3 expression and downregulation of Bcl-2 protein expression on both NSCLC cell lines. In addition, purified protein fraction induced the loss of mitochondrial membrane potential and generation of reactive oxygen species (ROS) on lung cancer cells. Altogether, our results proved that the PPF-V from Donax variabilis inducing apoptosis against NSCLC cell lines, which could be a potential natural candidate for lung cancer therapy.

Our previous studies showed that Astragaloside IV derivative (LS-102) exhibited potent protective function against ischemia reperfusion (I/R) injury, but little is known about the mechanisms. Mitochondrial fission regulated by dynamin-related protein1 (Drp1) is a newly recognized determinant of mitochondrial function. This study aimed to investigate the protection of LS-102 on mitochondrial structure and function by regulating the activity of Drp1 using models of H9c2 cardiomyocyte injury induced by hypoxia-reperfusion (H/R), and rat heart injury induced by I/R. The results showed that LS-102 significantly decreased apoptosis, levels of ROS, CK, LDH, and calcium, upregulating MMP, and the Bax/Bcl-2 ratio in cardiomyocytes during I/R injury. Furthermore, LS-102 prevented I/R-induced mitochondrial fission by decreasing Drp1's mitochondrial localization through decreasing the phosphorylation of Drp1 at Ser616 (Drp1(Ser616)) and increasing the phosphorylation of Drp1 at Ser637 (Drp1(Ser637)) in H9c2 cells. Importantly, we also robustly confirmed Drp1(Ser616) as a novel GSK-3 beta phosphorylation site. GSK-3 beta-mediated phosphorylation at Drp1(Ser616) may be associated with mitochondrial fission during I/R of cardiomyocytes. In conclusion, LS-102 exerts cardio protection against I/R-induced injury by inhibiting mitochondrial fission via blocking GSK-3 beta-mediated phosphorylation at Ser616 of Drp1.