2D nanofluidic membranes are capable of regulating ion transport toward various applications concerning energy and environment, which is primarily contributed by the excess charge on the interior surface of narrow nanoscale pores. However, there is still a lack of comprehensive summaries and discussions on the surface charge modification principles and strategies of 2D nanofluidic membranes, as well as the practical applications of charge‐modified 2D nanofluidic membranes for regulating ion transport. In this review, the surface charge modification principles and charge modification methods of 2D nanofluidic membranes are first introduced in detail, which is of great significance for improving the ion regulation capability of membranes and realizing the design of nanochannel materials. Next, recent advances in the two typical applications of concentration cells and water treatment based on charge‐modified 2D nanofluidic membranes are summarized. Finally, some challenges and prospects related to charge‐modified 2D nanofluidic membranes are discussed to indicate directions for future research in this field. It is anticipated that this review will provide valuable strategies for the development of high‐performance charge‐modified 2D nanofluidic membranes toward energy and environment applications.