Morphine’ Effect on Inflammation and Gls1 gene in IEC18 Cells.

Introduction: IEC18 Cells are epithelial cells that represent the innermost lining of the colon. We are using these cells to study Inflammatory Bowel Disease (IBD). IBD is an excessive inflammatory response in the colon. The Gls1 and MeCP2 genes are an important part of the inflammatory response in cells. Morphine is known to have immunosuppressive and anti-inflammatory effects in many cell-types, macrophages for example. While morphine has many unwanted side-effects, there may be some circumstances where treating IBD with morphine is practical. A meta-analysis in 2021 found that 21% of outpatients with IBD and 62% of hospitalized IBD patients use opioids. In this experiment, we looked at how effectively morphine counters inflammation in IEC18 cells.

DNA methylation is one of the epigenetic mechanisms that our cells use to regulate gene transcription. MeCP2 and other proteins bind to methylated promoter regions and inhibit gene expression. Previous experiments in the lab have shown that Gls1 is an exception where MeCP2 interacts with co-activators and increases gene expression.

Methods: Intraepithelial cells-18 were used in this study. Cells were treated with 2,4,6 trinitrobenzenesulfonic acid (TNBS). Azacytidine is used in this study as a demethylating agent. DNA and RNA were extracted from the treated cells and were analyzed for methylation status of the Gls 1 gene.

Results: Azacytidine treatment reduced DNA methylation and thus can be used as a possible treatment for IBD patients.