Fig. 6. Rpph1 (ribonuclease P RNA component H1) promotes the expression of pro-inflammatory cytokines and cell proliferation via the Gal-3/Mek/Erk pathway.

a Forty-eight hours after transfection with Rpph1 over-expression plasmid or Rpph1 over-expression plasmid and Gal-3 small interfering RNA (siRNA) in L-MC, enzyme- linked immunosorbent assay (ELISA) was performed to analyze the effect of Rpph1 on the expression of tumor necrosis factor-α (Tnf-α) and monocyte chemoattractant protein-1 (Mcp-1) with Gal-3 knockdown. b Forty-eight hours after transfection with Rpph1 siRNA or Rpph1 siRNA and Gal-3 over-expression plasmid in H-MC, ELISA was performed to analyze the effect of Rpph1 on the expression of Tnf-α and Mcp-1 with Gal-3 over-expression. c Twenty-four hours after treatment with U0126 (10 μM) in L-MC, ELISA was performed to analyze the effect of over-expression of Rpph1 or Gal-3 on the expression of Mcp-1 and Tnf-α with U0126; U0126 inhibited the expression of inflammatory cytokines relative to the untreated group. In addition, the effect of over-expression of Rpph1 or Gal-3 on promotion of the expression of inflammatory cytokines was blocked by U0126 in L-MC. d Twenty-four hours after treatment with the U0126 (10 μM) in L-MC, EdU (5-ethynyl-2′-deoxyuridine) assay was performed to analyze the blocking effect of over-expression of Rpph1 or Gal-3 on cell proliferation with U0126; U0126 inhibited cell proliferation relative to the untreated group and blocked the effect of over-expression of Rpph1 or Gal-3 on cell proliferation. e The abridged general view of the mechanism whereby lncRNA Rpph1 promotes the expression of Mcp-1 and Tnf-α and the proliferation of mesangial cells (MCs) in DN via an interaction with Gal-3 and activation of Gal-3/Mek/Erk signaling. In all panels, the data are representative of three independent experiments. Data are presented as mean ± SD. *P < 0.05, **P < 0.01, NS not significant