This study aimed to investigate the anatomical substrates involved in pain processing, focusing on the anterior cingulate cortex (ACC) and insular cortex (INS) and their interactions with other pain-related brain regions. Utilizing functional magnetic resonance imaging (fMRI), we analyzed brain activation patterns and connectivity in response to nociceptive and neuropathic pain, considering varying pain intensities and durations. The study included 455 participants with a range of pain conditions. Major findings revealed that the ACC and INS exhibited distinct activation patterns depending on the type of pain, with neuropathic pain leading to greater activation compared to nociceptive pain. Additionally, increased pain intensity correlated with heightened activation in these regions. Functional connectivity analyses showed strong integration between the ACC, INS, primary somatosensory cortex (S1), and thalamus during pain processing. These results underscore the role of the ACC and INS in encoding pain intensity and emotional responses. The study highlights the need for personalized pain management strategies and suggests further research into chronic pain’s long-term effects on brain function.