A　self-assembled　peptides-calcium-Vitamin　D3　ternary　delivery　system　(CSPH-Ca-VD3)　was　prepared　to　investigate　the　promotion　of　cellular　calcium　transport.　The　constructed　CSPH-Ca-VD3　nanocomplex　exhibited　a　spherical　structure　with　a　size　of　135.2　±　10.2　nm.　Based　on　the　thermodynamic　calculation　of　fluorescent　spectra,　hydrophobic　interaction　was　shown　as　the　major　driving　force　for　this　nanocomplex　structure.　CSPH-Ca-VD3　nanocomplex　possessed　excellent　stability　during　simulated　gastrointestinal　digestion,　contributing　to　the　prevention　of　acid　degradation　of　VD3　and　the　enhancement　of　calcium　solubility.　Furthermore,　the　calcium　transport　efficiency　in　the　form　of　CSPH-Ca-VD3　(4　mg/mL)　across　a　Caco-2　cells　monolayer　was　significantly　increased　2.3-fold　compared　to　that　of　free　Ca2+,　mainly　attributed　to　the　upregulation　in　the　presence　of　CSPH-Ca-VD3　of　TRPV6,　calbindin　D9k　and　PMCA1b　expression　in　Caco-2　cells.　The　present　study　provided　a　basis　for　developing　a　novel　delivery　system　of　peptides-calcium　chelate　with　the　dual　effects　of　VD3　protection　and　calcium　uptake　promotion.　©　2023　Elsevier　Ltd