Journal: Frontiers in Endocrinology

Article Title: Amelioration of polycystic ovarian morphology by Tokishakuyakusan in a PCOS rat model: association with bone morphogenetic protein 4

doi: 10.3389/fendo.2025.1649124

Figure Lengend Snippet: Bmp4 , Bmp5 , and Inhba expression in the ovaries. (A–C) Relative mRNA expression quantified by real-time PCR. Expression levels are relative to 18S ribosomal RNA. Data are shown as mean ± standard deviation with individual data points. P -values were obtained by one-way analysis of variance (ANOVA) with Tukey’s post hoc test (vehicle: n = 6; DHT: n = 9; DHT+TSS: n = 10). (D–G) Correlation analysis between gene expression and ovarian follicle counts in the DHT and DHT+TSS groups ( n = 6 per group). The correlation coefficient ( r 2 ) and P -value were calculated. DHT, 5α-dihydrotestosterone; TSS, Tokishakuyakusan; Bmp, bone morphogenetic protein; Inhba, inhibin-βa; PCR, polymerase chain reaction.

Article Snippet: The following TaqMan primer–probe sets (Thermo Fisher Scientific) were used: Bmp4 , Rn00432087_m1; Bmp5 , Rn01447676_m1; inhibin-βa ( Inhba ), Rn01538592_m1; steroidogenic acute regulatory protein ( Star ), Rn00580695_m1; cytochrome P450 11A1 ( Cyp11a1 ), Rn00568733_m1; 3β hydroxysteroid dehydrogenase ( Hsd3b ), Rn01789220_m1; 18s , Hs99999901_s1; and Gapdh , Rn01775763_g1.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Standard Deviation, Gene Expression, Polymerase Chain Reaction

Journal: Frontiers in Endocrinology

Article Title: Amelioration of polycystic ovarian morphology by Tokishakuyakusan in a PCOS rat model: association with bone morphogenetic protein 4

doi: 10.3389/fendo.2025.1649124

Figure Lengend Snippet: BMP4 and inhibin βA protein expression in the ovaries. (A) Representative Western blot images. (B, C) Quantification of BMP4 and inhibin βA protein levels relative to β-actin. Data are shown as mean ± standard deviation with individual data points (vehicle: n = 6; DHT: n = 9; DHT+TSS: n = 10). * P < 0.05, ** P < 0.01, one-way analysis of variance (ANOVA) followed by Tukey’s post hoc test; P < 0.0167 by Bonferroni correction. BMP, bone morphogenetic protein; DHT, 5α-dihydrotestosterone; TSS, Tokishakuyakusan.

Article Snippet: The following TaqMan primer–probe sets (Thermo Fisher Scientific) were used: Bmp4 , Rn00432087_m1; Bmp5 , Rn01447676_m1; inhibin-βa ( Inhba ), Rn01538592_m1; steroidogenic acute regulatory protein ( Star ), Rn00580695_m1; cytochrome P450 11A1 ( Cyp11a1 ), Rn00568733_m1; 3β hydroxysteroid dehydrogenase ( Hsd3b ), Rn01789220_m1; 18s , Hs99999901_s1; and Gapdh , Rn01775763_g1.

Techniques: Expressing, Western Blot, Standard Deviation

Journal: Frontiers in Endocrinology

Article Title: Amelioration of polycystic ovarian morphology by Tokishakuyakusan in a PCOS rat model: association with bone morphogenetic protein 4

doi: 10.3389/fendo.2025.1649124

Figure Lengend Snippet: Effect of Tokishakuyakusan (TSS) on target gene expressions in granulosa cells (GCs) derived from prenatally 5α-dihydrotestosterone (DHT)-treated rats. (A) Timeline of ovarian GC collection. (B, C) Relative mRNA expression of Bmp4 and Inhba in primary cultured GCs after 24 h of TSS treatment (125–500 μg/mL). Expression levels are relative to Gapdh . Data are shown as mean ± standard deviation with individual data points ( n = 3 per group). ** P < 0.01, P -values for the untreated control were obtained using one-way analysis of variance (ANOVA) followed by Dunnett’s post hoc test. PMSG, pregnant mare serum gonadotropin; Bmp, bone morphogenetic protein; Inhba, inhibin-βa; Gapdh, glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: The following TaqMan primer–probe sets (Thermo Fisher Scientific) were used: Bmp4 , Rn00432087_m1; Bmp5 , Rn01447676_m1; inhibin-βa ( Inhba ), Rn01538592_m1; steroidogenic acute regulatory protein ( Star ), Rn00580695_m1; cytochrome P450 11A1 ( Cyp11a1 ), Rn00568733_m1; 3β hydroxysteroid dehydrogenase ( Hsd3b ), Rn01789220_m1; 18s , Hs99999901_s1; and Gapdh , Rn01775763_g1.

Techniques: Derivative Assay, Expressing, Cell Culture, Standard Deviation, Control

Journal: Frontiers in Endocrinology

Article Title: Amelioration of polycystic ovarian morphology by Tokishakuyakusan in a PCOS rat model: association with bone morphogenetic protein 4

doi: 10.3389/fendo.2025.1649124

Figure Lengend Snippet: Effect of Tokishakuyakusan (TSS) on progesterone synthesis in granulosa cells (GCs) derived from prenatally 5α-dihydrotestosterone (DHT)-treated rats. (A–D) Relative mRNA expression of Bmp4 (A) , Star (B) , Cyp11a1 (C) , and Hsd3b (D) in primary cultured GCs after 24 h of TSS (500 μg/mL) treatment with or without FSH (3 ng/mL). Expression levels are relative to Gapdh. (E) Progesterone concentration in GC culture medium after 24 h of treatment. Data are shown as mean ± standard deviation with individual data points ( n = 3 per group). * P < 0.05, ** P < 0.01, two-way analysis of variance (ANOVA) followed by Tukey’s post hoc test. FSH, follicle-stimulating hormone; Star, steroidogenic acute regulatory protein; Cyp11a1, cytochrome P450 11A1; Hsd3b, 3β hydroxysteroid dehydrogenase; Gapdh, glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: The following TaqMan primer–probe sets (Thermo Fisher Scientific) were used: Bmp4 , Rn00432087_m1; Bmp5 , Rn01447676_m1; inhibin-βa ( Inhba ), Rn01538592_m1; steroidogenic acute regulatory protein ( Star ), Rn00580695_m1; cytochrome P450 11A1 ( Cyp11a1 ), Rn00568733_m1; 3β hydroxysteroid dehydrogenase ( Hsd3b ), Rn01789220_m1; 18s , Hs99999901_s1; and Gapdh , Rn01775763_g1.

Techniques: Derivative Assay, Expressing, Cell Culture, Concentration Assay, Standard Deviation