Therachon Kamollerd

Morus alba L. fruit extract (MFE), which is rich in cyanidin 3-glucoside (C3G), demonstrates antioxidant properties and pharmacological effects, but its reproductive safety remains poorly understood. Polyphenols modulate steroidogenesis, spermatogenesis, and sperm acrosome integrity; therefore, toxicity assessment is needed for use. This study aimed to evaluate the antioxidant profiles and subchronic reproductive effects of MFE. MFE was standardized using high-performance liquid chromatography (HPLC), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and ferric reducing antioxidant power (FRAP) assays. Male rats were administered MFE (250 or 500 mg/kg BW) for 56 days. Assessments included computer-assisted sperm analysis (CASA), testosterone, seminal fructosamine, and testicular CYP11A1 and androgen receptor (AR) expression. Acrosomal status was determined using PNA lectin staining. The results showed that MFE contained C3G (119.42 mg/g), antioxidant capacity (DPPH IC₅₀: 0.101 mg/mL; FRAP: 465.01 µmol Fe (II)/g), and total contents (phenolics: 41.15 mg GAE/g; flavonoids: 3.15 mg CE/g; anthocyanidins: 11.04 mg C3G/g). MFE did not alter testicular histology and seminiferous stages VII-IX. High doses significantly increased sperm concentration, while both doses reduced sperm beat cross frequency. Testosterone, fructosamine, and CYP11A1/AR expressions showed increasing trends. Significantly, high doses induced a precocious acrosome reaction. In conclusion, MFE has no reproductive toxicity and pro-fertility effects on sperm quantity or androgenic markers, supporting safe subchronic use.

Chronic stress (CS) contributes to male infertility, reduced testosterone levels, and impaired semen quality. CS models induced by glucocorticoids, such as dexamethasone (DEX), negatively affect sperm parameters and testicular health, notably by promoting testicular apoptosis. While individual plant extracts have been studied for their ability to mitigate stress-induced reproductive dysfunction, the preventive effect of the Tri Garn Pis (TGP) polyherbal extract in DEX-induced CS (DexCS) has not previously been investigated. This study evaluated the effects of TGP extract on testicular function, sexual behavior, and sperm quality in DexCS male mice. Seventy-two ICR mice were randomly divided into six groups: control, DexCS, TGP (50, 100, and 200) + DexCS, and TGP200. Mice received TGP (50, 100, 200 mg/kgBW) for 14 days before DEX co-treatment for 28 days. Behavioral and reproductive assessments included depression-like behavior tests, sexual behavior, sperm quality, testicular histopathology, steroidogenesis proteins (AR, CYP11A1, StAR), and apoptosis markers (Hsp70, caspase-3, caspase-9). TGP extract—which is rich in phenolics and flavonoids with antioxidant activity—improved depressive behavior, sexual performance, testicular histology, and low sperm quality. TGP also upregulated testicular StAR expression while reducing caspase-3 and caspase-9 levels. TGP prevents testicular apoptosis, sexual dysfunction, and poor sperm motility induced by DexCS.

Dexamethasone (DEX), a synthetic glucocorticoid widely prescribed for chronic inflammatory conditions, autoimmune disorders, and acute pain management, has been implicated in male reproductive dysfunction. Although many studies have documented DEX-induced oxidative stress, apoptosis, and testosterone reduction in rodent models, the molecular mechanisms underlying steroidogenic disruption and tyrosine phosphorylation remain incompletely understood. Aim of this study was to investigate the effects of chronic DEX administration on key steroidogenic proteins and tyrosine phosphorylated (TyrPho) protein profiles in rat testis. Adult rats received either DEX (1.5 mg/kg body weight, intraperitoneally) or vehicle control daily for 21 consecutive days. Results showed that DEX treatment significantly reduced testicular weight, seminiferous tubule diameter, and epithelial height compared to those of control. Tubular atrophy and interstitial tissue degeneration were observed in DEX-treated testes. Western blot analysis demonstrated significant downregulation of steroidogenic acute regulatory protein (StAR) and cytochrome P450 family 11 subfamily A member 1 (CYP11A1), while heat shock protein 70 (HSP70) expression was not different. Immunofluorescence confirmed the reduced StAR localization in Leydig cells of DEX group. TyrPho protein profiling revealed differential responses, with DEX significantly increasing a 70 kDa protein band while decreasing a 25 kDa protein band; other Tyrpho proteins (50, 42, and 38 kDa, respectively) not affected. It was concluded that chronic DEX exposure compromised the testicular architecture and suppressed critical steroidogenic machinery, providing mechanistic insights into glucocorticoid-induced hypogonadism and reproductive toxicity in male mammals.

Chronic stress (CS) is known to induce testicular oxidative stress and apoptosis. Fruit of Momordica charantia (FMC) has antioxidant capacity to protect tissue damage, but its effect on sexual activity and testicular damage caused by CS has never been documented. This study aimed to investigate the impact of FMC extract against testicular damage and sexual dysfunction in chronic unpredictable stress (CUS) rats. Rats were divided into four groups and pretreated with vehicle or FMC (40 and 80 mg/kg) before CUS induction for 56 days. Sexual behaviors, serum hormones, and sperm quality were analyzed. Testes were collected to determine daily sperm production (DSP), malondialdehyde (MDA) level, and expressions of cleaved apoptotic proteins. Testicular DNA fragmentation, as revealed by TUNEL and morphometric analysis, was observed. FMC improved sexual activity, increased testosterone levels, and sperm count with improvements in DSP, testicular morphometrics, and MDA levels. Moreover, TUNEL-positive cells and expression of caspase 3 in the testis were decreased in FMC rats. FMC has antioxidant potential and could protect against male sexual dysfunction and testicular damage caused by stress-related apoptosis. It is a potential extract to be developed as a supplement in preventing CS-male subfertility. However, future studies about the optimal dose and clinical trials are required.

Chronic stress (CS) from physical stressors and glucocorticoid administrations has been shown to induce germ cell apoptosis and low sperm quality. Among CS animal models, the effects of dexamethasone induced CS (DexCS) on testicular apoptosis and epididymal sperm parameters have not been fully demonstrated. This study aimed to investigate the changes of male reproductive system after CS induction by Dex. Adult male mice were divided into control and DexCS groups. Control mice were injected with sodium phosphate while DexCS mice were injected with DEX (4 mg/kg BW) for 21 consecutive days. The stress tests (sucrose preference, tail suspension, and forced swimming) were used to confirm CS behaviors. Sperm concentration, motility, morphology, viability, and acrosome status were assessed using computer-assisted semen analysis (CASA) and special staining. Histopathology of testis, epididymis, and penis was observed. Apoptotic protein expressions (Hsp70, caspases 3 and 9) in the testicular lysate were also determined. The results revealed that DexCS mice had significant increase of the immobility periods with decrease of total sucrose intake. DexCS significantly decreased sperm quality parameters particularly progressive motility. Testicular damages and decreased sperm mass in epididymis were obviously found in DexCS group. The expressions of testicular caspases 3 and 9, but not Hsp 70 were significantly increased in DexCS group compared to the control. It was concluded that DEX is a potential drug to induce chronic stress in mouse model, affecting male reproductive system via testicular histopathology and apoptotic pathway. Such effect may cause sperm physiology impairments like low progressive motile patterns.

Mammalian prostate gland plays a role in alkaline substance synthesis including proteins. These functions are depending on glandular maturation and testosterone-androgen receptor (AR) dependent actions. Since tyrosine phosphorylated (TyrPho) proteins, also important for secreting pathways, have been localized in the androgen dependent organs, association between AR and TyrPho protein expressions in prostate is still unknown. This study aimed to investigate the changes of such proteins in prostate gland of male castrated rats. Nine prepubertal and adult twenty-two adult male rats were divided into the prepubertal (Pre, n=9), Sham (n=6), castrate for 3 (Cas-3, n=8) and for 7 (Cas-7, n=8) days groups, respectively. Serum testosterone level was determined. Histology and AR localization in each prostatic lobe were observed. TyrPho and AR protein expressions were also examined. The results showed undetectable testosterone level and low AR expression in Pre and Cas prostates with the decreased size. Few histopathologies were found in Cas groups. In ventral lobe, a Tyrpho protein was increased at the 48 kDa but the 52, 33, and 26 kDas were decreased in the Pre and Cas groups. For dorsolateral lobe, they were decreased at 33 and 30 kDas in Pre group and only 30 kDa was decreased in Cas-3 group. In the anterior lobe, the TyrPho proteins 57, 49, 39, 30, and 26 kDas were decreased in Pre group while 57, 30, and 26 kDas were decreased in Cas-3 group. In conclusion, the alterations of testosterone level and AR expressions associate with TyrPho protein expressions in prostate gland during development.