Neuropeptide Y promotes myocardial fibrosis post myocardial infarction reperfusion through Neuropeptide Y receptor type 1

Figures

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  Fig. 1. Expression of NPY in mouse myocardial cell lines and heart tissue after myocardial infarction reperfusion. A. Expression of NPY in heart tissue after MI/ R; B. Expression of NPYR1 in heart tissue after MI/ R; C-D. Statistical analysis of the number of positive cells expressing NPY and NPYR1; E. The relative mRNA expression of NPY in heart tissue after MI/ R; F. Western blot detecting the expression of proteins containing NPY and NPYR1 in heart tissue after MI/ R; G. Relative quantitative statistical analysis of NPY and NPYR1protein in heart tissue after MI/ R. H. The relative mRNA expression of NPY in mouse myocardial cell lines; cell lines. I. Western blot detecting the expression of proteins containing NPY and NPYR1in mouse myocardial. J. Relative quantitative statistical analysis of NPY and NPYR1protein in HL-1 cell. (N = 3, ** represents P<0.01). Original blots/gels are presented in Supplementary material

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  Fig. 2. Knockdown of NPYR1 modulates myocardial fibrosis myocardial fibrosis after I/R injury. A. Expression of NPY and NPYR1 in heart tissue after I/ R while injection of NPYR1 RNA interference lentivirus and overexpression lentivirus; B. Western blot were used to detect the expression of NPYR1, HIF1-α and fibrotic factor α-SMA, Col1, Col3 in the sham group, MI/ R group, I/ R+NPYR1 KD group, I/ R+NPYR1 OE respectively; Original blots/gels are presented in Supplementary material. C. Relative quantitative statistical analysis of proteins in B. D-H. RT-PCR detecting relative mRNA expression of NPY, HIF-1α and fibrotic factor α-SMA, Col1, Col3; I-J. Masson staining of cardiac paraffin sections after I/R in four groups. K. Statistical analysis of the fibrosis areas. (N = 3, ** represents P<0.01, * represents P<0.05)

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  Fig. 3. The fibrosis of HL-1 cell was ameliorated under hypoxic condition while knocking down NPYR1. A. NPYR1 gene shRNA lentivirus and overexpression lentivirus were successfully transfected into HL-1; B. Western blot was used to detect the expression of NPYR1, HIF-α and fibrotic factor α-SMA, Col1, Col3 in control group, H/R group, H/R+ NPYR1 KD group, H/R+ NPYR1 OE group; Original blots/gels are presented in Supplementary material. C. Relative quantitative statistical analysis of proteins. D-F. RT-PCR detecting relative mRNA expression of NPY, HIF-α and fibrotic factor α-SMA, Col1, Col3 in control group, H/R group, H/R+ NPYR1 KD group, H/R+ NPYR1 OE group; G. Scratch test of (Control) H/R group, H/R+ NPYR1 KD group, H/R+ NPYR1 OE group. (N = 3, *** represents P<0.001, ** represents P<0.01, * represents P<0.05)

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  Fig. 4. The cardiac function post MI/R 4weeks while knocking down NPYR1. A. Cardiac color Doppler ultrasound images of mice in sham group, MI/ R group, MI/ R+NPYR1 KD group, MI/ R+NPYR1 OE group; B Left ventricular ejection fraction (LVEF) of sham group, MI/ R group, MI/ R+NPYR1 KD group, MI/ R+NPYR1 OE group; C Left ventricular short axis shortening rate (LVFS) ofsham group, MI/ R group, MI/ R+NPYR1 KD group, MI/ R+NPYR1 OE group; D. Left ventricular end systolic diameter (LVIDs) of sham group, MI/ R group, MI/ R+NPYR1 KD group, MI/ R+NPYR1 OE group; E. Left ventricular end diastolic diameter (LVIDd) sham group, MI/ R group, MI/ R+NPYR1 KD group, MI/ R+NPYR1 OE group. (N = 3, **** represents P<0.0001, *** represents P<0.001, ** represents P<0.01, * represents P<0.05)