by the biological ion channels, various biomimetic nanofluidic devices have been constructed, featuring the characteristics of ion selectivity, ion gating, and ion rectification. [3] Polymer-based nanopore or nanochannel devices, such as polyethylene terephthalate (PET) and polyimide membranes, have been widely used in energy generator, [4] sensors, [5] ion gating, [6] and ion pump [7] due to their mature membrane building technology. In addition, 2D materials, such as graphene oxide, [8] MXene, [9] Kaolinite, [10] Vermiculite, [11] have also been used to fabricate layered nanofluidic systems. Nevertheless, most of these membranes suffer from insufficient ion transport flux and high membrane inner resistance due to their low pore density and disordered channels, which results in an unfavorable performance. Furthermore, the ion selectivity and energy harvesting are largely affected by the working area of the membrane. [12] Therefore, facile methods are urgently required to fabricate membranes with abundant ordered nanochannels and working area robustness for smart ion transport and multifunctional applications. Mesoporous materials are ideal in constructing nanofluidic devices owing to their ordered mesochannels, high pore density, and controllable thickness. [13] Based on evaporation-induced self-assembly (EISA) strategy, [14] a series of mesoporous materials, including mesoporous silica, [15] mesoporous carbon, [16] and mesoporous carbon-silica, [17] have been used to develop mesoporous nanofluidic systems. Mesoporous heterogeneous Asymmetric nanofluidic devices hold great potential in energy conversion applications. However, most of the existing asymmetric nanofluidic devices remain a single-level asymmetric structure and a single-ion selective layer, which results in weak ion selectivity and limited energy conversion efficiency. Herein, a multi-level asymmetric mesoporous carbon/anodized aluminum/ mesoporous silica (MC/AAO/MS) nanofluidic device with abundant and ordered mesochannels is constructed from super-assembly strategy. The resultant MC/AAO/MS exhibits diode-like ion transport and outstanding ion storage-release performance. Importantly, MC/AAO/MS couples the MC and MS dual-ion selective layers, which ensures a high ionic conductance and evidently enhances the cation selectivity. Thereby, the MC/AAO/MS demonstrates ascendant salinity gradient energy conversion performance. The power density and conversion efficiency can reach up to 5.37 W m −2 and 32.79%, respectively. Noteworthy, a good energy conversion performance of 63 mW m −2 can still be achieved upon high working area, outperforming 300% of the performance of MC/AAO and MS/AAO single-level asymmetric nanochannels. Theoretical calculation further verifies that the multi-level asymmetric structure and dual-ion selective transport are the reason for the enhanced cation selectivity and energy conversion efficiency. This work opens a new avenue for constructing multi-level asymmetric structured nanofluidic devices for various applications.