图 1 

Fig.1 CRKL promotes the secretion of milk in postpartum hypogalactia rats ($ {\overline x} \pm {s}$) Hourly milk production (A). Weight gain of offspring (B). PRL levels in MT was detected by ELISA (C). CCK8 cell proliferation activity (D). The mRNA expression of PRLR in RMECs was detected by RT-qPCR (E) (A-B, n=8; C-E, n=3). cP<0.01, compared with normal; eP<0.05, fP<0.01, compared with model. iP<0.01, compared with 1.0%.

图 2 

Fig.2 CRKL promotes gene expression of milk lactose, milk fat synthesis, and milk protein in RMECs ($ {\overline x} \pm {s}$) The mRNA expression of FASN, CSN2, and GLUT1 in RMECs were detected by RT-qPCR (A, B, C). The protein expression of FASN, CSN2, and GLUT1 in RMECs were detected by immunofluorescence (D, E, F) (scale bar=50 µm) (n=3). cP<0.01, compared with normal; eP<0.05, fP<0.01, compared with model.

图 3 

Fig.3 CRKL target predicted by network pharmacology Screening of CRKL components (A). Obtained 464 OGEs from the Venn diagram tool (B). Core target geneswere obtained by PPI network analysis (C, D). GO function analysis (E). KEGG enrichment analysis (F). Compound-Target-Pathway network (G).

图 4 

Fig.4 CRKL promotes the expression of Wnt/β-catenin signaling pathway in RMECs ($ {\overline x} \pm {s}$) The mRNA expression of CCND1 and c-Myc in RMECs were detected by RT-qPCR (A, B). The protein expression of CCND1, c-Myc, and β-catenin in RMECs were detected by WB (C, D, E) (n=3). cP<0.01, compared with normal; eP<0.05, fP<0.01, compared with model.

图 5 

Fig.5 Molecular docking and molecular dynamics Molecular docking of WTAP with Picraquassioside C, 7- (beta Xylosyl) cephalomannine, Wallichilide, and Wallichilide (A, B, C, D). Stability analysis of WTAP molecular dynamics complex with four components (Picraquassiosis C, 7- (beta Xylosyl) Cephalomanine, Wallichilide, and Wallichilide) (E, F, G, H, I, J). Gibbs free energy analysis of WTAP molecular dynamics complexes with four components (Picraquassiosis C, 7- (beta Xylosyl) cephalomanine, Wallichilide, and Wallichilide) (K, L, M, N).

图 6 

Fig.6 CRKL inhibits WTAP expression ($ {\overline x} \pm {s}$) The mRNA expression of WTAP in MT and RMECs were detected by RT-qPCR (A, B, C, D) (n=3). cP<0.01, compared with normal; fP<0.01, compared with model.

图 7 

Fig.7 WTAP inhibits the expression of Wnt5a/β-catenin signaling pathway ($ {\overline x} \pm {s}$) The mRNA expression of Wnt5a in MT and RMECs were detected by RT-qPCR (A, B). RIP experiment (C). The mRNA expression of CCND1 and c-Myc in RMECs were detected by RT-qPCR (D, E). The protein expression of β-catenin in RMECs was detected by WB (F). The mRNA expression of FASN, CSN2, and GLUT1 in RMECs were detected by RT-qPCR (G, H, I) (n=3). cP<0.01, compared with normal; fP<0.01, compared with WTAP vector; iP<0.01, compared with anti-IgG group; lP<0.01, compared with anti-WTAP group.

图 8 

Fig.8 Cuiru Keli improves bromocriptine-induced postpartum hypogalactia in rats via WTAP-Wnt5a-β-catenin pathway ($ {\overline x} \pm {s}$) The mRNA expression of CCND1 and c-Myc in RMECs were detected by RT-qPCR (A, B). The protein expression of β-catenin in RMECs was detected by WB (C). The mRNA expression of FASN, CSN2, and GLUT1 in RMECs were detected by RT-qPCR (D, E, F) (n=3). cP<0.01, compared with normal; fP<0.01, compared with CRKL.