Journal: Frontiers in Endocrinology

Article Title: Jinkui Shenqi Pill accelerates osteoporotic fracture healing by promoting bone formation through neurosensory PGE2/EP4/p-CREB axis

doi: 10.3389/fendo.2025.1570685

Figure Lengend Snippet: JKSQP promotes EP4 in DRG and p-CREB in hypothalamus during OPF healing. (A) Immunofluorescence (IF) staining of EP4 in the fractured callus of OPF mice at the 14 days post-fracture. (B) Histomorphology of mouse dorsal root ganglion (DRG) at 14 days post-fracture. (C, D) IF staining and quantitative analysis of EP4 and CGRP in DRG at 14 days post-fracture. (E) Histomorphology of mouse brain at 14 days post-fracture. Black boxes: hypothalamus region. (F) IF staining and quantitative analysis of p-CREB in hypothalamus. (G) The flowchart of primary dorsal root ganglion (DRG) neurons cultivation and intervention. Free-CM: JKSQP-free serum treated BMSCs-conditioned medium; High-CM: high JKSQP-containing serum treated BMSCs-conditioned medium; OPF-MS: the serum from OPF mice; JKSQP-MS: the serum from JKSQP-treated mice. (H) Phase-contrast microscopy of DRG neurons. (I) IF staining of β3-tubulin and glial fibrillary acidic protein (GFAP) in DRG neurons. (J) Heat map of EP1-EP4 mRNA levels in DRG neurons post-intervention. (K) IF staining of EP4 and CGRP in DRG neurons post-intervention. *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: After fixation with 4% paraformaldehyde and blocking with 3% bovine serum albumin for each 30 minutes, the coverslips were incubated with primary antibodies including β3-tubulin (diluted 1:300, wanleibio, wl03860, CN), GFAP (diluted 1:100, wanleibio, wl0836, CN), EP4 (diluted 1:200, abcam, ab92763, UK) and CGRP (diluted 1:200, Abcam, ab81887, UK) overnight at 4°C.

Techniques: Immunofluorescence, Staining, Microscopy