The mechanism of painful diabetic neuropathy (PDN) is still unclear. This study investigates whether the mechanism involves astrocytes activation and phosphorylation of extracellular signal-regulated kinase (ERK) and modulation of Downstream regulatory element antagonist modulator (DREAM), and Brain derived neurotrophin factor (BDNF) proteins expression that affects the neuropathic pain responses in PDN rats model. Eighty-four Sprague-Dawley male rats were assigned into 6 groups (n=14 per group) consisting of normal control (C), PDN-diabetic (PDN), PDN+ astrocytes inhibitor (LAA) treated: L100 (low dose) and L200 (high dose) and PDN+ ERK inhibitor (U0126) treated: U5 (low dose) and U10 (high dose). After streptozotocin (STZ) was injected, 2 weeks period was allowed for all diabetes mellitus (DM) rats to develop PDN conditions. Then, an intrathecal injection of LAA and U0126 were given for seven days. The neuropathic pain responses were performed by Von Frey, hot plate and formalin test. The rats were sacrificed, and spinal cord were collected for histology and quantitative PCR (qPCR) analyses. The PDN group showed significant increment in GFAP, ERK, DREAM and BDNF mRNA expression level in the spinal cord with increased neuropathic pain responses compared to control group. LAA and U0126 significantly modulated the GFAP, ERK, DREAM and BDNF mRNA expression and alleviated the neuropathic pain responses in the PDN treated group. It suggested that the mechanism of PDN involves astrocytes activation and phosphorylation of ERK to modulate the expression of DREAM and BDNF mRNA levels in the spinal cord that affects the neuropathic pain responses in PDN rats model.