Aldosterone-induced galectin-3 in THP-1 and RAW 264.7 cells enhanced fibrosis-related factor in fibroblasts.

<p>(A) Effect of galectin-3 siRNA treatment on inhibiting aldosterone-induced galectin-3 expression. THP-1 cells were pretreated with galectin-3 siRNA or control siRNA for 24 h and then serum-starved for 24 h prior to 10⁻⁶ M aldosterone or vehicle treatment for 24 h. The conditional medium was collected, and the level of galectin-3 was detected by enzyme immunoassay. (B) Effect of conditional medium from aldosterone-treated THP-1 cells on fibroblast cell growth. The conditional medium was collected and cultured with the human fibroblast cell line 293T. Cell growth was determined by MTT assay. Data were recorded as relative cell growth in the indicated conditions with the vehicle-treated conditional medium in lane 1 defined as 100%. *, <i>P</i><0.05 (n = 5). (C) Effect of conditional medium from aldosterone-treated THP-1 cells on fibronectin and procollagen (I) mRNA expressions. Conditional medium as described in (A) was used to treat 293T cells. After 24 hours, the expressions of fibronectin and procollagen (I) mRNA were determined by real-time quantitative RT-PCR. Data were compared between the indicated groups. A paired Student's <i>t</i>-test was used to evaluate statistically significant differences between the indicated groups. ^*<i>P</i><0.05. (D) Effect of galectin-3 siRNA treatment on inhibiting aldosterone-induced galectin-3 expression. Raw 264.7 cells were pretreated with galectin-3 siRNA or control siRNA for 24 h, and then serum-starved for 24 h prior to 10⁻⁶ M aldosterone or vehicle treatment for 24 h. The conditional medium was collected, and the level of aldosterone was detected by enzyme immunoassay. (E) Effect of conditional medium from aldosterone-treated RAW 264.7 cells on fibroblast cell growth. The conditional medium was collected and cultured with the mouse fibroblast cell line NIH/3T3. The cell growth was determined by MTT assay. Data were recorded as relative cell growth in the indicated conditions with the vehicle-treated conditional medium in lane 1 defined as 100%. *, <i>P</i><0.05 (n = 5). (F) Effect of conditional medium from aldosterone-treated RAW 264.7 cells on fibronectin and procollagen (I) mRNA expressions. Conditional medium as described in (E) was used to treat NIH/3T3 cells. After 24 h, the expressions of fibronectin and procollagen (I) mRNA were determined by real-time quantitative RT-PCR. Data were compared between the indicated groups. A paired Student's <i>t</i>-test was used to evaluate statistically significant differences between the indicated groups. *<i>P</i><0.05. AD = aldosterone.