The effect of m۲ peptide targeted nanoliposomes containing crocin on induction of phenotypic change in tumor macrophages to M۱ state

Crocin has immunomodulatory and anticancer effects, and in this study, its effect on macrophage cells was specifically investigated. The purpose of the present study was to prepare nanoliposomes targeted with m۲ peptide and containing crocin, investigate the properties of this formulation, and investigate its effects in inducing M۱ phenotype in vitro and its antitumor effects in vivo. Methods: In the present study, liposomal formulations containing crocin were prepared and characterized. In vitro studies including MTT assay and IL-۱۲, IL-۶, TNF-alpha, CD۸۶, iNOS, Arg-I, and Ym۱ gene expression in macrophage cell line were evaluated (after treatment with liposomal formulations and free crocin and empty liposome). Based on in vitro studies, the appropriate liposomal formulation was selected for targeting with m۲ peptide. The uptake rate of targeted and non-targeted formulations in the J۷۷۴A-۱ cell line was evaluated using flow cytometry. Finally, biodistribution study and therapeutic anti-tumor effects of liposomal formulations in C۲۶ colon carcinoma xenograft mouse model. Results: The size of liposomes containing crocin was about ۱۳۰ nm and PDI was less than ۰.۲, with the encapsulation efficiency of ۸۰%. The targeted formulation was the most stable. The MTT assay results confirmed that crocin and empty liposomes do not have any effect on cell viability. Real-time PCR tests showed that crocin increased the expression of inflammatory markers TNF-alpha, iNOS, IL-۱۲, CD۸۶, and iNOS/Arg-I ratio. Flow cytometry results in macrophages showed that the uptake of the targeted formulation is higher than that of the non-targeted formulation. The biodistribution study showed the accumulation of targeted liposomal formulation in the tumor microenvironment, and the maximum accumulation in the tumor was ۱۲ hours after IV administration. Targeted liposomal formulation with m۲ peptide increased the anti-tumor effects and the survival rate of xenograft mice compared to free crocin and non-targeted formulations. Conclusion: It is probable that the remarkable anti-tumor responses that were observed in this study with liposomal formulations targeted with m۲ peptide and containing crocin, were due to the enhanced delivery of crocin to immune cells and the subsequent initiation of anti-tumor immune responses.

کلیدواژه ها:

Crocin ، Macrophage ، Inflammation ، Liposome ، m۲ peptid