Cryptotanshinone alleviates polycystic ovary syndrome in rats by regulating the HMGB1/TLR4/NF‑κB signaling pathway

Figures

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  Figure 1.. Effects of CRY and HMGB1 on (A) ovarian weight, (B) body weight, (C) ovaries quotiety, (D) Lee's index and (E) BMI in rats with human chorionic gonadotropin and insulin-induced PCOS. Data are presented as the mean ± SEM. *P<0.05, **P<0.01, ***P<0.005 vs. control; #P<0.05, ##P<0.01 vs. PCOS; &P<0.05, &&P<0.01, &&&P<0.005 vs. PCOS + HMGB1. CRY, cryptotanshinone; PCOS, polycystic ovary syndrome; HMGB1, high-mobility group box 1; BMI, body mass index.

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  Figure 2.. Effect of CRY on serum levels of (A) LH, (B) FSH, (C) LH/FSH ratio, (D) testosterone and (E) TNF-α. Data are presented as the mean ± SEM. *P<0.05, **P<0.01, ***P<0.005 vs. control; #P<0.05, ##P<0.01, ###P<0.005 vs. PCOS; &P<0.05, &&&P<0.005 vs. PCOS + HMGB1. CRY, cryptotanshinone; PCOS, polycystic ovary syndrome; LH, luteinizing hormone; FSH, follicle-stimulating hormone; TNF-α, tumor necrosis factor-α; HMGB1, high-mobility group box 1.

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  Figure 3.. Effects of CRY on pathological morphology of ovarian tissues analyzed under light microscopy. Differences in the numbers of granulosa cell layers (black arrow), cystic follicles (red arrow) and corpora lutea (white arrow) were observed. a: Primordial follicle (a single layer of flattened granulosa cells surrounding the oocyte); b: Primary follicle (a single layer of cuboidal granulosa cells surrounding the oocyte); c: Secondary follicle (two complete layers of cuboidal granulosa cells surrounding the oocyte). Magnification, ×100. CRY, cryptotanshinone; PCOS, polycystic ovary syndrome; HMGB1, high-mobility group box 1.

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  Figure 4.. Effects of CRY on HMGB1 expression in ovarian tissues. HMGB1 was primarily expressed in ovarian granulosa cells (black arrow). Magnification, ×100. Data are presented as the mean ± SEM. *P<0.05, **P<0.01 vs. control; ##P<0.01 vs. PCOS; &&P<0.001 vs. PCOS + HMGB1. CRY, cryptotanshinone; PCOS, polycystic ovary syndrome; HMGB1, high-mobility group box 1.

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  Figure 5.. Effects of CRY on HMGB1, TLR4 and NF-κB/p65 expression in ovarian tissues. (A) mRNA expression levels. (B) Protein expression levels. Data are presented as the mean ± SEM. *P<0.05, **P<0.01 vs. control; #P<0.05, ##P<0.01 vs. PCOS; &&P<0.001 vs. PCOS + HMGB1. CRY, cryptotanshinone; PCOS, polycystic ovary syndrome; HMGB1, high-mobility group box 1; TLR4, toll-like receptor 4; NF-κB, nuclear factor-κB.

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  Figure 6.. Effect of CRY on GC proliferation, and regulation of TNF-α and HMGB1 expression in vitro. (A) mRNA and (B) protein expression of HMGB1 in GCs transfected with siHMBG1. (C) Cell viability of insulin-resistant GCs following CRY treatment or siHMGB1 transfection. (D) Concentration of TNF-α and HMGB1 in culture medium. Data are presented as the mean ± SEM. *P<0.05, **P<0.01, ***P<0.005 vs. control; $P<0.05, $$P<0.01, $$$P<0.05 vs. si-NC, ##P<0.01 vs. IR. CRY, cryptotanshinone; IR, insulin resistance; GC, granulosa cell; HMGB1, high-mobility group box 1; TNF-α, tumor necrosis factor-α; si, small interfering; OD, optical density; si-NC, si-negative control.

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  Figure 7.. Effects of CRY on HMGB1, TLR4 and NF-κB/p65 expression in ovarian granulosa cells. (A) mRNA expression levels. (B) Protein expression levels. Data are presented as the mean ± SEM. *P<0.05, **P<0.01 vs. control; #P<0.05, ##P<0.01 vs. IR. CRY, cryptotanshinone; IR, insulin resistance; HMGB1, high-mobility group box 1; TLR4, toll-like receptor 4; NF-κB, nuclear factor-κB; si, small interfering; si-NC, si-negative control.