Due to the complicated environment and high tissue hydraulic pressure in tumors that easily pumps the nanomedicines back to the systemic circulation, the concentration of released photosensitizers (PSs) retained in a tumor by a traditional nano-delivery system is very low, causing an unsatisfactory photodynamic therapy (PDT) effect. Therefore, we prepared a pH/H2O2-responsive nano-system (ZnP-OC-M) through modified porphyrin PS units with a long-unsaturated oleoyl chloride chain, and by the further introduction of hydrophilic hydroxyl groups and MnO2 through a cis-addition reaction between the unsaturated double bonds of oleoyl chloride and dilute KMnO4 solution. Making use of the sensitivity of MnO2 to the H2O2 in the acid environment of tumor cells, ZnP-OC-M selectively realized responsive disintegration and O2 generation. More importantly, the rich amphiphilic PS units were shedded simultaneously and spontaneously completed the self-assembly into nanofibers in situ by helical stacking, which displayed a 1.85-fold higher retention effect of PSs in vivo compared with free PS groups and showed a great tumor inhibition effect in enhancing PDT. This nanosystem effectively solves the problem of the low retention abilities leading to a poor PS concentration in a tumor, prolonging the treatment time window efficiently after only a single administration and achieving the purpose of PDT enhancement.