Journal: MedComm
Article Title: Spatially Characterized Aortic Proteome Reveals Novel Regional Signatures and Glucocorticoid Receptor/Dipeptidase 1 Axis in Diabetic Vasculopathy
doi: 10.1002/mco2.70714
Figure Lengend Snippet: Vascular inflammation and endothelial dysfunction driven by diabetes‐related Dpep1 upregulation. (A) Western blotting on Dpep1 expression in TA and AA from C57BL/6 mice following ex vivo treatment with HG and LPS, and corresponding quantification. (B) Western blotting on DPEP1 expression in HG‐ and LPS‐treated HAECs, and corresponding quantification. (C) Schematic overview of chronic cilastatin treatment in diabetic mice. (D) Western blotting on Dpep1 expression in TA and AA from nondiabetic and diabetic mice following chronic cilastatin treatment ( n = 4 per group). (E) Dpep1 activity in TA, and (F) MPO activity in TA and AA from db/m + and db/db mice following chronic cilastatin treatment. (G) Heatmap on the RT‐PCR results of inflammatory markers in mouse aortas after cilastatin treatment. (H) Functional assay on EDRs of aortas from cilastatin‐treated mice by wire myograph ( n = 5 per group). (I) Schematic overview of intravenous AAV1‐ICAM2‐shDpep1 injection in diabetic mice. (J) Western blotting on Dpep1 expression in isolated endothelial cells from TA and AA of nondiabetic and diabetic mice subjected to AAV‐shDpep1 injection, and corresponding quantification ( n = 4 per group). (K) MPO activity in TA and AA from AAV‐shDpep1‐injected db/m + and db/db mice. (L) Heatmap on the RT‐PCR results of inflammatory markers in mouse aortas from AAV‐shDpep1‐injected mice. (M) Functional assay on EDRs of aortas from AAV‐shDpep1‐injected mice by wire myograph ( n = 5 per group). (N) Schematic overview of diabetes‐related endothelial Dpep1 upregulation and neutrophil‐released MPO in promoting vascular inflammation and endothelial dysfunction. Data are presented as mean ± SD. * p < 0.05 (Brown–Forsythe and Welch ANOVA, and unpaired t with Welch's correction). AA, abdominal aorta; AAV, adeno‐associated virus; ACh, acetylcholine; DPEP1, dipeptidase 1; EC, endothelial cell; HAEC, human aortic endothelial cell; HG, high glucose; IV, intravenous; LPS, lipopolysaccharide; MPO, myeloperoxidase; Phe, phenylephrine; TA, descending thoracic aorta.
Article Snippet: For AAV‐mediated Dpep1 knockdown in vivo, some db/m + and db/db mice were intravenously injected with AAV1‐scramble or AAV1‐ICAM2‐shDpep1 (2 × 10 11 vg per mouse), generated by Vigene Biosciences (Maryland, USA).
Techniques: Western Blot, Expressing, Ex Vivo, Activity Assay, Reverse Transcription Polymerase Chain Reaction, Functional Assay, Injection, Isolation, Virus
Addgene inc is a verified supplier 
