TWIK1 is a potassium leak channel of the K2P family that contributes to the resting membrane potential and excitability in a variety of neurons. It is expressed in the dorsal root ganglion (DRG), where it participates in pain processes, although its regulation remains poorly understood. It is believed that Angiotensin-II (Ang-II) can have analgesic effects in chronic pain models by the activation or inhibition of its main receptors, AT1R and AT2R, possibly involving regulation of K2P channel expression. Thus, we evaluated whether selective inhibition or activation of AT1R and AT2R modulates TWIK1 expression in primary DRG neuron cultures from P6 rats. Cells were maintained in vitro for 1 and 2 days under different conditions: Control, Ang-II, Ang-II + Azilsartan (AT1R antagonist), Ang-II + PD123319 (AT2R antagonist), and Ang-II + Azilsartan + PD123319. Both TWIK1 and AT1/AT2R expressions were assessed by qRT-PCR, immunofluorescence, and Western blot. Ang-II significantly increased TWIK1 expression at 1 DIV, an effect mainly mediated by AT2R. At 2 DIV, however, TWIK1 expression was markedly reduced across all groups. Moreover, Ang-II treatment altered the expression of its own receptors, suggesting a feedback regulation of the RAS in DRG neurons. These findings demonstrate that Ang-II regulates TWIK1 expression in sensory neurons, providing a previously unknown link between angiotensinergic signaling and neuronal excitability in models of neuropathic pain.