Suppressing the activity of down-regulates the expression of renal fibrosis related genes in primary glomerular cells.

BACKGROUND

C-X-C chemokine receptor type 4 () has a certain effect on renal fibrosis, and there are few specific studies in cells. We want to investigate the impact of suppressing activity on the expression of renal fibrosis-related genes in primary glomerular endothelial cells, mesangial cells, and podocytes.

METHODS

Immunofluorescence assays were used to determine the purity of isolated glomerular endothelial cells, mesangial cells, and podocytes. knockdown cell lines were established by transfecting the short hairpin (sh)RNA against CXCR4. T140 and AMD3100 were used to inhibit the activity of . LY294002 was used to inhibit the activity of phosphoinositide 3-kinase (). The mRNA expression of was determined by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). The protein expression of , , matrix metallopeptidase , and vascular cell adhesion protein 1 () was evaluated by Western blot analysis.

RESULTS

High purity was observed on isolated primary glomerular endothelial cells and podocytes. However, the purity of isolated mesangial cells was relatively low. The mRNA expression of was significantly suppressed by the transfection of shRNA. Compared to control cells, the expression of , collagen , and were dramatically downregulated in cell lines transfected with shRNA against . Furthermore, cell lines treated with T140, AMD3100, or LY294002 also showed downregulated expression of these proteins compared to untreated cells. No significant differences were observed in the protein expression of these proteins between control cells and cells transfected with the shRNA negative control (NC).

CONCLUSIONS

Suppressing the activity of downregulated the expression of renal fibrosis-related genes in primary glomerular cells, even under a non-inflammatory state.