Evaluation of the effect of Berberine-loaded plant-derived nanovesicles on

Abstract

Berberine (BBR), a naturally occurring isoquinoline alkaloid, has shown significant anticancer activity in the treatment of various cancers. Yet, its low bioavailability, water solubility, and intestinal absorption have limited its clinical application. The present study is primarily concerned with the bioformulation of grape-derived nanoparticles (GDN) as a carrier for BBR and the investigation of whether GDNs induce anticancer activity against acute lymphoblastic leukemia (ALL). Method: Grape nanoparticles were isolated and purified by differential ultracentrifugation steps. The structure, size, and zeta potential of nanoparticles were studied using scanning electron microscopy (SEM) and dynamic light scattering (DLS). BBR was loaded into GDNs through passive method and Entrapment Efficiency, in vitro release, and cellular uptake were evaluated. MTT assay was used to determine the cytotoxicity of free BBR and BBR-loaded GDNs on Jurkat T-ALL cell line. Following treatment with 30 and 60 M of BBR-loaded GDNs or free BBR, the gene expressions of caspase-3, 8, 9, BAX, and BCL-2 were quantified by qRT-PCR. Furthermore, flow cytometry was performed to analyze apoptosis. Results: SEM images and DLS data revealed that GDNs were spheres with an average size of 278 nm and a polydispersity index (PDI) of 0.4. Also, encapsulation efficiency was and 66%. Over a 72-hour period, the drug release kinetics indicated a slower and more sustained release of BBR. The obtained data from MTT assay showed that BBR-loaded GDNs exerted higher cytotoxic effect compared to free BBR solution. Flow cytometry results showed that BBR-loaded GDNs significantly triggered apoptosis in leukemic cells in a dose- and time-dependent manner, as well as exhibiting a higher anti-apoptotic activity than free BBR. Furthermore, analysis of the gene expression involved in apoptosis and cell survival revealed that BBR-loaded GDNs greatly increased the mRNA expression of caspase 3, 8, 9, and Bax genes while decreasing the expression of the anti-apoptotic protein BCL-2.