Flow cytometric analysis of the in situ hybridization of cyclooxygenase isoforms in mesangial cells treated with cyclosporine A

Abstract

Background Cyclosporine A increases oxidative stress in kidney and we hypothesized that cyclooxygenase (COX) may be involved in this effect. Material and Methods Mesangial cells of Cyclosporine A‐treated (4, 7 or 10 days) rats were obtained to evaluate mRNA expression of COX‐isoforms (COX‐1, constitutive and COX‐2, inducible) by “in situ” hybridization. Probes were labelled using “Gene Image Random Prime Labelling Protocol” and COX expression was measured by flow cytometry. Results and Discussion “In situ” hybridization by flow cytometry is an useful method to detect mRNA. We observed an increased COX‐2 expression in a time‐dependent manner in parallel with Reactive Oxygen Species synthesis. COX‐1 expression increased only at 10 days. © 2006 International Society for Analytical Cytology

Novel aspects of systems biology and clinical cytomics

“…Th1/Th2 cytokine analysis with the multiplex array technique in mitogen‐stimulated whole blood provides the possibility to predict immunosuppression in heart‐transplant recipients (33) or other immunology‐related complications (34). Novel flow and image cytometric assays were developed to assess the effects of immunosupressants or chemotherapy on leukocyte or tumor‐cell apoptosis (35–37) or cyclooxygenase mRNA expression by quantitative “in situ” hybridization (38). The Th1/Th2 reaction monitoring shows a transient immunosupression with an increased proportion of Th2 cells induced by cardiac surgery with cardiopulmonary bypass.…”