Among autoimmune disorders, rheumatoid arthritis is one of the most prevalent. The IL-1 family and the IL36 subfamily both include interleukin-38. IL-38 is anti-inflammatory cytokine. IL-38 is essential for inflammation control and host defense. In the Pakistani population, this research intends to evaluate the relationship flanked by IL-38 levels and its polymorphism (rs 6743376) C>A and rheumatoid arthritis. Eighty samples in total were included and divided into sick and control groups. IL-38 levels were measured through ELISA. For the SNP of IL-38, an allele-specific polymerase chain reaction assay was applied. All subjects’ serum levels of CRP, RF, and ACCP were assessed. 70% of patients had negative CRP tests, while 30% had positive CRP tests. All of the patients’ RF and ACCP tests came back positive. As the p-value was smaller than (0.001), a significant difference was observed in the mean ESR test values between patients and controls. IL-38 concentration was higher in the RA having a mean value of 26.8621. In comparison to homozygous allele (CC) and heterozygous allele (AC), patients with homozygous allele (AA) are more common. Contrarily, the control group revealed that the homozygous allele (CC) was more prevalent than the homozygous allele (AA) and heterozygous allele (AC). When compared to homozygous allele (CC) and heterozygous allele (AC), IL-38 rs6743376 (A/A) is related with a higher risk of developing RA illness. In contrast to heterozygous (A/A) allele, homozygous (C/C) allele and heterozygous (A/C) allele are protective in RA.  According to this study, IL-38 homozygous alleles (CC) and heterozygous alleles (AC) are associated with a lower probability of developing RA than heterozygous alleles (AA).