LncRNA H19 Drives Proliferation of Cardiac Fibroblasts and Collagen Production via Suppression of the miR-29a-3p/miR-29b-3p-VEGFA/TGF-β Axis

Fig. 1. Clinical significance of lncRNA H19, miR-29a-3p, and miR-29b-3p in atrial fibrillation (AF). (A) Serum levels of lncRNA H19, miR-29a-3p, and miR-29b-3p were compared between AF patients and healthy volunteers. *P < 0.05 when compared with healthy control. (B) Serum level of lncRNA H19 was negatively correlated with that of miR-29a-3p and miR-29b-3p among AF patients.

Fig. 2. Correlations between expression of RNAs and clinical traits of AF patients. Serum levels of lncRNA H19 (A), miR-29a-3p (B), and miR-29b-3p (C) were correlated with clinical traits of AF patients.

Fig. 3. Cell activities were impacted by lncRNA H19, miR-29a-3p, and miR-29b-3p in CFs. (A) LncRNA H19 expression was determined after transfection of pcDNA3.1-H19, siRNA-H19-1, siRNA-H19-2, and siRNA-H19-3. *P < 0.05 when compared with negative control (NC). (B) MiR-29a-3p expression in CFs was detected after transfection of miR-29a-3p mimic and miR-29a-3p inhibitor. *P < 0.05 when compared with NC. (C) MiR-29a-3p expression was detected after transfection of miR-29b-3p mimic and miR-29b-3p inhibitor into CFs. *P < 0.05 when compared with NC. (D-G) Viability and proliferation of CFs was evaluated among NC, pcDNA3.1-H19, si-H19, miR-29a-3p mimic and miR-29a-3p inhibitor group or NC, pcDNA3.1-H19, si-H19, miR-29b-3p mimic, and miR-29b-3p inhibitor group. *P < 0.05 when compared with NC.

Fig. 4. Cell apoptosis were regulated by lncRNA H19, miR-29a-3p, and miR-29b-3p in CFs. (A and B) apoptosis were evaluated separately among negative control (NC), pcDNA3.1-H19, si-H19, miR-29a-3p mimic, and miR-29a-3p inhibitor group or NC, pcDNA3.1-H19, si-H19, miR-29b-3p mimic, and miR-29b-3p inhibitor group. *P < 0.05 when compared with NC. PI, propidium iodide; FITC, fluorescein isothiocyanate. (C and D) apoptins were determined between NC, pcDNA3.1-H19, si-H19, miR-29a-3p mimic, and miR-29a-3p inhibitor group and NC, pcDNA3.1-H19, si-H19, miR-29b-3p mimic, and miR-29b-3p inhibitor group. *P < 0.05 when compared with NC.

Fig. 5. ECM-related proteins of CFs were affected by lncRNA H19, miR-29a-3p, and miR-29b-3p. (A) ECM-related proteins were evaluated separately among negative control (NC), pcDNA3.1-H19, si-H19, miR-29a-3p mimic, and miR-29a-3p inhibitor group. *P < 0.05 when compared with NC. (B) ECM-related proteins were detected among negative NC, pcDNA3.1-H19, si-H19, miR-29b-3p mimic, and miR-29b-3p inhibitor group. *P < 0.05 when compared with NC.

Fig. 6. Sponging relationships between H19 and miR-29a-3p/miR-29b-3p in CFs. (A) LncRNA H19 targeted miR-29a-3p, and luciferase activity of CFs was compared between pmiRGLO-H19-Wt+miR-29a-3p mimic group and pmiRGLO-H19-Wt+miR-NC group. NC, negative control. *P < 0.05 when compared with pmiRGLO-H19-Wt+miR-NC group. (B) MiR-29a-3p expression was determined after transfection of NC, pcDNA3.1-H19 and si-H19. *P < 0.05 when compared with NC. (C) MiR-29b-3p was targeted by lncRNA H19, and luciferase activity of CFs was compared between pmiRGLO-H19-Wt+miR-29b-3p mimic group and pmiRGLO-H19-Wt+miR-NC group. *P < 0.05 when compared with pmiRGLO-H19-Wt+miR-NC group. (D) MiR-29b-3p expression was measured after transfection of NC, pcDNA3.1-H19, and si-H19. *P < 0.05 when compared with NC.

Fig. 7. Cell activities were impacted by miR-29a-3p and si-VEGFA in CFs. Viability (A), proliferation (B), apoptosis (C), apoptins (D), and ECM-related proteins (E) of CFs were evaluated among miR-NC, miR-29a-3p inhibitor, and miR-29a-3p inhibitor+si-VEGFA group. NC, negative control; PI, propidium iodide; FITC, fluorescein isothiocyanate. *P < 0.05 when compared with miR-NC, #P < 0.05 when compared with miR-29a-3p inhibitor.

Fig. 8. Cell activities were regulated by miR-29b-3p and SB431542 in CFs. Viability (A), proliferation (B), apoptosis (C), apoptins (D), and ECM-related proteins (E) of CFs were evaluated among miR-NC, miR-29b-3p inhibitor and miR-29a-3p inhibitor+SB431542 group. NC, negative control; PI, propidium iodide; FITC, fluorescein isothiocyanate. *P < 0.05 when compared with miR-NC, #P < 0.05 when compared with miR-29b-3p inhibitor.