Acute pancreatitis (AP) is an inflammatory disease primarily detected through the estimation of amylase and lipase as conventional serum markers that have a short half-life. MicroRNAs have emerged as biomarkers of different diseases including AP, cardiovascular disorders, and cancer. This study was designed to evaluate the possible ameliorating impact and anti-inflammatory role of kynurenic acid (KYNA) in AP-induced in rats and identify the possible use of miR-216a and miR-217 as diagnostic biomarkers for AP. Forty male albino rats were divided into 4 equal groups. Group 1 (control group) rats injected (i.p) with physiological saline. Group 2 (AP group) rats were injected with a single dose of L-ornithine-HCl (ORN) (3 g/kg b.wt/i.p). Group 3 (KYNA+ORN group) rats were injected with a single dose of KYNA (300 mg/kg b.wt/i.p) 1 hour before ORN injection. Group 4 rats were injected with a single dose of KYNA (300 mg/kg b.wt/i.p). The ORN-induced AP group showed significant elevation in serum amylase and lipase activities, miR-216a and miR-217 expressions, glucose, IL-1β and IL-10 levels with a significant decrease in insulin level. Moreover, oxidative stress markers (MPO and MDA) were significantly increased, while the antioxidants SOD and GSH were markedly reduced. Interestingly, the severity of these alterations was reduced and ameliorated in KYNA+ORN group compared to AP group. Rats treated with KYNA alone showed non-significant changes. These data suggest that KYNA can alleviate the severity of ORN-induced AP in rats and serum miR-216a and miR-217 could be potential biomarkers for the diagnosis of AP and could be associated with inflammatory conditions in rats with AP.