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. Author manuscript; available in PMC: 2022 Feb 15.
Published in final edited form as: Neuropharmacology. 2021 Jan 16;186:108463. doi: 10.1016/j.neuropharm.2021.108463

Fig. 3. 3α,5α-THP administration decreased CRF peptide in hippocampus and central nucleus of amygdala of male rats.

Fig. 3.

Hippocampus: A. 3α,5α-THP (15 mg/kg) IP administration increased 3α,5α-THP levels in hippocampus of male and female rats (males VEH = 14.52 ± 1.8 ng/g tissue vs 3α,5α-THP = 938.3 ± 128.2 ng/g tissue, p < 0.0001; females VEH = 26.08 ± 4 ng/g tissue vs 3α,5α-THP = 1112 ± 138.3 ng/g tissue, p < 0.0001) (n = 6–8 rats per group). B. qPCR analysis did not show any sex or treatment differences in CRF mRNA expression (n = 6–14 rats per group). C. Western blotting analysis did not show any sex differences in baseline CRF peptide levels; however, 3α,5α-THP injection reduced CRF protein expression in male (VEH CRF ratio = 1.12 vs 3α,5α-THP CRF ratio = 0.7906, p = 0.0254), but not in female rats. Male rats treated with 3α,5α-THP exhibited lower CRF levels than the same group of female rats (male = 0.7906 ± 0.06 vs. female = 1.273 ± 0.05, p = 0.0017) (n = 5–6 per group). Central nucleus of amygdala: D. 3α,5α-THP (15 mg/kg) IP administration increased 3α,5α-THP levels in male and female rats, but the increase in female rats is higher than male rats (males VEH = 90.48 ± 37.24 ng/g tissue vs 3α,5α-THP = 778.1 ± 51.39 ng/g tissue, p < 0.0001; females VEH = 22.4 ± 1.96 ng/g tissue vs 3α,5α-THP = 1011 ± 95.73 ng/g tissue, p < 0.0001; males 3α,5α-THP = 778.1 ± 51.39 ng/g tissue vs females 3α,5α-THP = 1011 ± 95.73 ng/g tissue, p = 0.0386) (n = 6–8 rats per group). E. qPCR analysis did not show any sex or treatment differences in CRF mRNA expression (n = 6–14 rats per group). F. The results of western blotting analysis did not show any sex differences in CRF peptide levels under basal condition. However, 3α,5α-THP injection significantly reduced CRF protein expression in male (males VEH = 0.1658 ± 0.01 vs 3α,5α-THP = 0.0632 ± 0.02, p = 0.0033), but not in female rats. Male rats treated with 3α,5α-THP exhibited lower CRF levels than female rats (males = 0.0632 ± 0.02 vs females = 17.36 ± 0.02, p = 0.0017) (n = 5–6 per group). Significant effect was found using Two-Way ANOVA, followed by Tukey HSD test, *p < 0.05, **p < 0.01, ****p < 0.0001. Data are represented as mean ± SEM. VEH = rats treated with vehicle; 3α,5α-THP = rats treated with 3α,5α-THP.