We identified the
inv gene that encodes left and right asymmetry and regulates kidney development based on the information of the
inv mutant mouse. However, functional properties and the modulator of gene expression of
inv have been unclear. We used the tissue injury model for assessing the functional roles of
inv in ischemia reperfusion injury (IRI). The kidney tissue taken from rats with IRI showed reciprocal changes in mRNA expression of
inv: a 0.25-fold decrease at 6 hours and then a gradual increase to a maximum 1.8-fold rise at 10 days of reperfusion. Next, oxidative stress was induced by exposing mouse inner medullary collecting duct (mIMCD-3) cells to hydrogen peroxide (H
2O
2) in the medium. Real-time PCR showed that mRNA expression of
inv decreased 0.52-fold at 3 hours with 0.2mM H
2O
2 in the medium, and then increased 3.1-fold at 24 hours with 0.1mM H
2O
2 in the medium.
RNA interference (RNAi) is a powerful tool to inhibit gene expression in experimental model systems. We knocked down
inv gene expression in mIMCD-3 cells using RNAi to investigate the function of the
inv gene. We designed a small interfering RNA (siRNA) to target the coding region of
inv (
inv-siRNA) and random-sequence scrambled siRNA (control siRNA). mIMCD-3 cells transfected with either the
inv-siRNA or control siRNA were observed by microscopy. The cells transfected with
inv-siRNA progressively lost cell-to-cell contact and the cell population significantly diminished approximately 48 hours post-transfection. The changes in gene expression profile were observed at time points (36 hours) using real-time PCR-based gene screening with categorized primer sets. Several genes related to structural protein of the matrix were downregulated. In contrast, repairing related genes were upregulated.
In conclusion, gene expression of
inv was modulated under oxidative stress and the
inv gene may play a role in repairing and regenerating renal epithelial cells.
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