This work aimed to develop a novel formulation based on human milk (HM) enriched with a probiotic formulation comprised of Lactobacillus acidophilus CL1285, Lactobacillus casei LBC80R and Lactobacillus rhamnosus CLR2 and to evaluate in vitro and in vivo anticancer properties and its impact on the prevention of risk factors of metabolic syndrome (MetS). In vitro studies were conducted on Hepa-1c1c7, HT-29, and CHO-K1 cell lines to assess the antiproliferative and anticancer properties of HM and probiotics. The in vivo study was conducted with 28 male Wistar rats fed a high-fat diet (HFD). Rats were daily force-fed with HM and HM + probiotics or water for 8 weeks. Inflammation and oxidative stress were monitored through blood cytokines and kidney investigation. Body weight and blood glucose were recorded throughout the study. The results showed that HM had antiproliferative properties and caused apoptosis of intestinal tumoral cells. Quinone reductase (QR) was induced up to 1.8x in Hepa-1c1c7 cell line demonstrating chemotherapeutic potential. HM enriched with probiotics reduced weight gain by 10% while renal oxidative damages were reduced by up to 97%. Blood insulin and Plasminogen Activator Inhibitor-1 (PAI-1) levels were also significantly reduced. The study showed that HM had antiproliferative and chemopreventive properties on cancer cell lines. The in vivo experiment suggests that the development of a formulation based on HM powder enriched with probiotics could be used to modulate the rate of inflammation and oxidative damage to tissues, thus reducing the risk of obesity-related diseases.
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