Research Paper Volume 13, Issue 10 pp 13859—13875

Curcumin inhibits the proliferation and migration of vascular smooth muscle cells by targeting the chemerin / CMKLR1 / LCN2 axis

Downregulation of CMKLR1 inhibit VSMCs proliferation and migration. (A, B) Percentage of 5-Ethynyl-2'-deoxyuridine (EdU)-incorporated cells following the application of two independent shRNAs targeting CMKLR1 (scramble control, SC; shCMKLR1#2, shCM#2; shCMKLR1#3, shCM#3) to knock down CMKLR1 expression in mice vascular smooth muscle cells (VSMCs). *, ppC, D) Percentage of cell migration as determined by wound healing assay. *, ppE) Western blots showing CMKLR1, MMP-9 and PCNA expressions in cells treated with PDGF (48 h) for different durations. (F) Western blots showing CMKLR1, matrix metalloproteinase-9 (MMP-9) and proliferating cell nuclear antigen (PCNA) expression in cells treated with chemerin-9 (48 h) for different durations. (G) Western blots showing CMKLR1, MMP-9 and PCNA expressions with or without CMKLR1 knockdown. (H) Western blots showing CMKLR1, MMP-9 and PCNA expressions following the application of plasmids encoding CMKLR1 to overexpress CMKLR1 expression in VSMCs with or without CMKLR1 knockdown. (I) Analysis of the percentage EdU-incorporated cells with or without LCN2 depletion. ****, ppJ) Percentage of cell migration as determined by wound healing assay. ****, pp

Figure 2. Downregulation of CMKLR1 inhibit VSMCs proliferation and migration. (A, B) Percentage of 5-Ethynyl-2'-deoxyuridine (EdU)-incorporated cells following the application of two independent shRNAs targeting CMKLR1 (scramble control, SC; shCMKLR1#2, shCM#2; shCMKLR1#3, shCM#3) to knock down CMKLR1 expression in mice vascular smooth muscle cells (VSMCs). *, p< 0.05; **, p< 0.01. (C, D) Percentage of cell migration as determined by wound healing assay. *, p< 0.05; **, p< 0.01. (E) Western blots showing CMKLR1, MMP-9 and PCNA expressions in cells treated with PDGF (48 h) for different durations. (F) Western blots showing CMKLR1, matrix metalloproteinase-9 (MMP-9) and proliferating cell nuclear antigen (PCNA) expression in cells treated with chemerin-9 (48 h) for different durations. (G) Western blots showing CMKLR1, MMP-9 and PCNA expressions with or without CMKLR1 knockdown. (H) Western blots showing CMKLR1, MMP-9 and PCNA expressions following the application of plasmids encoding CMKLR1 to overexpress CMKLR1 expression in VSMCs with or without CMKLR1 knockdown. (I) Analysis of the percentage EdU-incorporated cells with or without LCN2 depletion. ****, p< 0.0001; **, p< 0.01; ns, no significant difference. (J) Percentage of cell migration as determined by wound healing assay. ****, p< 0.0001; **, p< 0.01.