Background: C-peptide assumes a basic part in the pathogenesis of vascular intricacy indiabetic patients. Objective: This study aimed to monitor the production of pro-inflammatory )1L-1-ß, IL-6, IFN-?, TNF-?, IL-8 and Rantes) and anti-inflammatory (IL-10) mediators in response to C-peptide in vitro. Materials and Methods: This study involved 131 Bahraini T2DM patients and 81 healthy subjects. C peptide levels were measured by Enzyme- Linked Immunoassay (ELISA). Cytokines expression on the RNA level in PBMCs culture were estimated. Polymerase Chain Reaction (PCR) products were resolved by gel electrophoresis. The quantitative cytokines were estimated by ELISA. Results: Measurements of C peptide levels were significantly low in diabetic patients with peripheral neuropathy. In PBMCs obtained after treatment with C-peptide in all studied groups, there were significantly lower levels of mRNA  for (TNF-?, IL-6, IL-8 and IFN-?) compared to non-stimulated cells (NC) (p<0.05). While, the exposure to C-peptide treatment resulted in a significant increase in mRNA expression of IL-1-ß, Rantes and IL-10 in all groups compared to (p<0.5). However, the variations of cytokine levels were significantly low in diabetic neuropathy patients' cells (TNF-?, IL-6, IL-8 and IFN-?) and also significantly high (IL-1-ß, IL-10 and Rantes) compared to both healthy subjects and diabetic patients without neuropathy (p<0.05). Interestingly, there was no significant differences when the effects of C-peptide in the production of the cytokines were compared to the positive control (PHA) in all studied groups. Three selected cytokines (IL-1-ß, IL-10 and Rantes) were studied at a protein level using ELISA to measure their levels in supernatant of cultured cells. The data showed similar cytokine protein patterns as for mRNA for the detected cytokines. Conclusion: This work demonstrated the effect of C peptide in regulating the immune response by stimulating certain cytokines and inhibition of others depending on their roles as pro or anti-inflammatory mediators. Keywords:  T2DM, C-Peptide, cytokines, mRNA, PCR, ELISA

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