Journal: Frontiers in Pharmacology

Article Title: Salidroside derivative SHPL-49 enhances synaptic remodeling in BCCAO rats via the CDK5/p35/p25 signaling pathway

doi: 10.3389/fphar.2026.1727177

Figure Lengend Snippet: SHPL-49 promotes the expression of CDK5 at both mRNA and protein levels. (A,B) Correlation analysis among samples. (C,D) Differential expression analysis between experimental groups. (E) Venn diagram showing the overlap of differentially expressed genes. (F) GO enrichment analysis of differentially expressed genes. (G–J) qRT-PCR validation of selected differentially expressed genes in primary neurons. (K–N) qRT-PCR validation of differentially expressed genes in rat brain tissue. (O) Representative Western blot images of CDK5 expression in primary neurons. (P) Quantitative analysis of CDK5 protein levels in primary neurons. (Q) Representative Western blot images of CDK5 expression in rat brain tissue. (R) Quantitative analysis of CDK5 protein levels in rat brain tissue. Data are presented as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, OGD/R group vs. Control group, Model group vs. Sham group compared with the Control group, # P < 0.05, ## P < 0.01, ### P < 0.001, SHPL-49 group vs. OGD/R group. n = 3 for mRNA analysis in primary neurons and rat brain tissue. n = 6 for protein analysis in primary neurons and rat brain tissue.

Article Snippet: Membranes were blocked with 5% non-fat milk (Beyotime, China) in Tris-buffered saline containing 0.1% Tween-20 (TBST) and incubated overnight at 4 °C with the following primary antibodies: SYP (1:1,000, CST, United States), SYN1 (1:1,000, Boster, China), PSD95 (1:1,000, ABCAM, United States), CDK5 (1:1,000, Boster, China), p35/25 (1:1,000, CST, United States), p-PSD95 (1:1,000, CST, United States).

Techniques: Expressing, Quantitative Proteomics, Quantitative RT-PCR, Biomarker Discovery, Western Blot, Control

Journal: Frontiers in Pharmacology

Article Title: Salidroside derivative SHPL-49 enhances synaptic remodeling in BCCAO rats via the CDK5/p35/p25 signaling pathway

doi: 10.3389/fphar.2026.1727177

Figure Lengend Snippet: SHPL-49 inhibits the proteolytic conversion of p35 to p25. (A) Representative Western blot images showing p35 and p25 expression in primary neurons. (B) Quantitative analysis of p35 protein lev-els in primary neurons. (C) Quantitative analysis of p25 protein levels in primary neurons. (D) Representative Western blot images of p35 and p25 in rat brain tissue. (E) Quantitative analysis of p35 protein levels in rat brain tissue. (F) Quantitative analysis of p25 protein levels in rat brain tis-sue. (G) Representative Western blot images showing the expression levels of CDK5, p35, p25, and p-PSD95/PSD95 ratio in primary neurons following OGD/R treatment and co-treatment with SHPL-49 and CDK5 inhibitor Roscovitine. (H) Quantitative analysis of CDK5 protein levels in primary neurons under the same experimental conditions. (I) Quantitative analysis of p35 protein levels in primary neurons under the same experimental conditions. (J) Quantitative analysis of p25 protein levels in primary neurons under the same experimental conditions. (K) Quantitative analysis of p-PSD95/PSD95 ratio in primary neurons under the same experimental conditions. Data are presented as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, OGD/R group vs. Control group, Model group vs. Sham group, # P < 0.05, ## P < 0.01, ### P < 0.001, SHPL-49 group or PD151746 group vs. OGD/R group, SHPL-49 group or SAL group vs. Model group n = 6 per group.

Article Snippet: Membranes were blocked with 5% non-fat milk (Beyotime, China) in Tris-buffered saline containing 0.1% Tween-20 (TBST) and incubated overnight at 4 °C with the following primary antibodies: SYP (1:1,000, CST, United States), SYN1 (1:1,000, Boster, China), PSD95 (1:1,000, ABCAM, United States), CDK5 (1:1,000, Boster, China), p35/25 (1:1,000, CST, United States), p-PSD95 (1:1,000, CST, United States).

Techniques: Western Blot, Expressing, Control