The effects of secondary metabolites from marine halophilic bacteria against okadaic acid induced alzheimers disease in the zebrafish

Abstract

Abstract newlineMarine-derived bioactive molecules are rich sources of structurally and chemically diverse molecules for industrial and biomedical applications. Bacillus enclensis-CARE-V7 and Halomona spacifica CARE-V15, two halophilic bacteria, are isolated from South East of Coast of India, and their gene sequences are analyzed. The anti-SMASH tool is used to find gene clusters linked to the synthesis of secondary metabolites. GC-MS analysis is used to identify compounds in the ethyl acetate extract of selected halophilic stains, including heptasiloxane, hexadecamethyl,3-benzyl-6-isobutyl-2,5-dioxo-piperazine, 3,6-diisobutyl-2,5-piperazinedione(DIP), 2-Benzenedicarboxylic acid, dibutyl ester, and 3-Isobutylhexahydropyrrolo [1,2-A] Pyrazine-1,4-Dione. The bioactive metabolites are fractionated by gel column chromatography, and the collected active fractions from the ethyl acetate extract s antibacterial activity is studied using human pathogenic bacterial strains Moreover, HPLC is used to further purify the active fractions. FTIR and NMR spectroscopy are used to characterize the structure of purified DIP. In silico molecular docking predicts oral efficacy for the purified metabolite DIP, which has a higher affinity for protein receptors like apoE (-6.86), Tau (-6.41), GSK3and#42933; (-6.27), AAP (-5.83), GPx (and#8722; 5.37), GST (and#8722; 5.27) and GSR (and#8722; 4.70). Hence, the purified DIP from the H. pacifica CARE-V15 compound is an alternative source of the neuroprotective agent. In vitro studies indicated that the DIP offers neuroprotection due to its antioxidant, mitochondrial protective, and anti-apoptotic properties. The DIP (50 and#956;M) exhibited commendable neuroprotective activity against the SH-SY5Y cells, verified using the MTT cell viability assay, ROS, mitochondrial membrane potential, and anti-apoptosis activity. The in vivo studies showed that 50 and#956;M of DIP bacterial metabolite concentrations had no cytotoxic or toxic effects on the zebrafish larvae model. The compound pre-treated experimental groups demonstrated reduced lipid peroxidation and cell death brought on by OA. The in vivo results supported the protection of DIP ability to inhibit behavioral impairments. However, pre-treated adult zebrafish exhibited elevated antioxidant enzyme activity, and upregulated GST, GPx, GSR, GSK3and#42933; and downregulated tau gene expression. The DIP from H. pacifica CARE-V15 could be an excellent way to treat diseases caused by oxidative stress and AD. newline newline