Momordica charantia fruit extract with antioxidant capacity improves the expression of tyrosine-phosphorylated proteins in epididymal fluid of chronic stress rats.

OBJECTIVE: Although the protective effects of Momordica charantia L. (MC) extract on chemical-induced testicular damage have been studied, the preventive effects of MC extract on functional proteins in the epididymis under chronic stress have never been reported. This study investigated the protective effects of MC fruit extract on protein secretion, especially tyrosine-phosphorylated proteins, in the epididymis of rats exposed to chronic unpredictable stress (CUS). METHODS: Total phenolic compounds (TPC), total flavonoid compounds (TFC) and antioxidant capacities of MC extract were measured. Adult male rats were divided into 4 groups: control group, CUS group, and 2 groups of CUS that received different doses of MC extract (40 or 80 mg/kg). In treated groups, rats were given MC daily, followed by induction of CUS (1 stressor was randomly applied from a battery of 9 potential stressors) for 60 consecutive days. Plasma corticosterone and testosterone levels were analyzed after the end of experiment. Expressions of heat-shock protein 70 (HSP-70) and tyrosine-phosphorylated proteins present in the fluid of the head and tail of the epididymis were quantified using Western blot. RESULTS: MC extract contained TPC of (19.005 ± 0.270) mg gallic acid equivalents and TFC of (0.306 ± 0.012) mg catechin equivalents per gram, and had 2,2-diphenyl-1-picrylhydrazyl antioxidant capacity of (4.985 ± 0.086) mg trolox equivalents per gram, radical 50% inhibitory concentration of (2.011 ± 0.008) mg/mL and ferric reducing antioxidant power of (23.697 ± 0.819) µmol Fe(II) per gram. Testosterone level in the epididymis was significantly increased, while the corticosterone level was significantly improved in groups treated with MC extract, compared to the CUS animals. Particularly, an 80 mg/kg dose of MC extract prevented the impairments of HSP-70 and tyrosine-phosphorylated protein expressions in the luminal fluid of the epididymis of CUS rats. CONCLUSION: MC fruit extract had antioxidant activities and improved the functional proteins secreted from the head and tail of the epididymis. It is possible to develop the MC fruit extract as a male fertility supplement for enhancing functional sperm maturation in stressed men.

Seed extract of Thai Mucuna pruriens (L.) DC. var. pruriens enhances sexual performance and improves male reproductive damages in ethanol-induced rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Thai Mucuna pruriens (L.) DC. var. pruriens (T-MP) has been traditionally used in treating depressive disorders, dysuria and enhancing male sexual desire. Although T-MP seed is demonstrated to have antioxidant capacity, its aphrodisiac and protective tissue damage properties have never been documented. Recently, ethanol (Eth) is known to cause sexual behavior dysfunction and damage reproductive system. This study aimed to investigate the protective effects of T-MP seed extract on sexual behavior dysfunction and reproductive damages in male rats admisted with Eth. MATERIALS AND METHODS: T-MP possessing antioxidant activity was determined for L-DOPA content using NMR analysis. Thirty-six male rats were divided into four groups (9 animals/group). Control rats received DW and the ethanol (Eth) group was given with Eth (3 g/kgBW; 40%v/v). In preventive groups (T-MP150 + Eth and MP300 + Eth groups), animals were treated with T-MP extract at a dose of 150 and 300 mg/kgBW before Eth administration for consecutive 56 days. Sexual behaviors including mounting frequency (MF), intromission frequency (IF), mounting latency (ML), intromission latency (IL), ejaculation latency (EL), post-ejaculatory interval (PEI), and ejaculation frequency (EF) were evaluated. Epididymal sperm quality and daily sperm production (DSP) were examined. Testicular histology was observed using Masson's trichrome staining. The malondialdehyde (MDA) levels and expressions of androgen receptor (AR), heat shock protein 70 (HSP70), steroidogenic acute regulatory (StAR), and tyrosine-phosphorylated (TyrPho) proteins in testis were also determined. RESULTS: T-MP extract contained L-DOPA and improved sexual behaviors including increased MF and IF and decreased ML and IL in Eth treated rats. Significantly, sperm quality, DSP, and testicular histopathology observed in Eth group were improved after T-MP treatment. T-MP also decreased the testicular MDA levels. Additionally, T-MP could correct testicular functional proteins of AR and StAR except HSP70 expression in Eth group. Expressions of TyrPho proteins in testicular and sperm lysates were improved in co-administered groups. CONCLUSIONS: T-MP seed extract possessing L-DOPA could enhance the sexual behaviors and protect reproductive damages via improvement of testicular functional proteins.

Chronic stress increases the tyrosine phosphorylation in female reproductive organs: An experimental study.

BACKGROUND: Changes in tyrosine-phosphorylated (TyrPho) protein expressions have demonstrated stress in males. In females, chronic stress (CS) is a major cause of infertility, especially anovulation. However, the tyrosine phosphorylation in the female reproductive system under stress conditions has never been reported. OBJECTIVE: To investigate the alteration of TyrPho protein expression in ovary, oviduct, and uterus of CS rats. MATERIALS AND METHODS: In this experimental study, 16 female Sprague-Dawley rats (5 wk: 220-250 gr) were divided into control and CS groups (n = 8/group). Every day, the CS animals were immobilized within a restraint cage and individually forced to swim in cold water for 60 consecutive days. Following the stress induction, the ovary, oviduct, and uterus of all rats were observed for their morphologies. The total protein profiles of all tissues were revealed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) before detecting TyrPho proteins using western blot. Intensity analysis was used to compare the expression of proteins between groups. RESULTS: The results showed that the morphology and weights of ovary and oviduct in the CS group were not different from control. In contrast, the CS significantly increased the uterine weight as compared to control. Moreover, the expressions of TyrPho proteins in the ovary (72, 43, and 28 kDas), oviduct (170, 55, and 43 kDas), and uterus (55, 54, and 43 kDas) were increased in CS group as compared to those of control. CONCLUSION: The increased expressions of TyrPho proteins in ovary, oviduct, and uterus could be potential markers used to explain some machanisms of female infertility caused from chronic stress.

Chronic stress affects tyrosine phosphorylated protein expression and secretion of male rat epididymis.

Chronic stress (CS) is shown to decrease the semen quality with changed expression of tyrosine phosphorylated (TyrPho) proteins in testicular and seminal tissues. However, the alterations of such proteins and fluid contents in the epididymis, producing sperm maturation factors, have never been reported. Sixteen adult rats were randomly divided into 2 groups (n = 8). The control animals were not subjected to stressors whereas CS rats were immobilised within restraint cage (4 hr/day) before cold forced-water swimming (15 min/day) for 60 days. Corticosterone, testosterone, blood glucose level (BGL), malondialdehyde (MDA) and biochemical components in epididymal fluid were assayed. Expressions of heat shock protein 70 (HSP-70), androgen receptor (AR) and TyrPho protein were investigated in epididymal tissue and fluid. Significantly, CS increased the corticosterone and BGL but decreased testosterone and epididymal substance levels. MDA level in tail epididymal fluid and HSP-70 expression in both regions of epididymal tissues and fluids, except in head epididymal fluid of CS were increased. Epididymal tissues showed the decrease of AR expression. Presence and changes of many TyrPho proteins were observed in CS. In conclusion, CS could affect functional proteins particularly TyrPho in epididymis, resulted in low semen quality.

The effect of Dolichandrone serrulata (wall. ex DC.) Seem. flower extract containing antioxidant capacity and terpenoids on the male reproductive system.

Although the fruit extract of Dolichandrone genus was shown to inhibit spermatogenesis, the reproductive toxicity of Dolichandrone serrulata flowers (DSFs) is not documented. Recent study aimed to evaluate the sub-chronic toxicity of DSF on male reproductive system. Antioxidant capacity and total phenolic contents of DSF extract were determined using Folin-Ciocalteu's, 2,2-diphenyl-1-picrylhydrazyl and ferric reducing antioxidant power assays. The terpenoid components were determined using nuclear magnetic resonance spectrum. Adult male rats were treated orally with DSF (100, 300 or 600 mg/kg) for 48 days. Histopathology of testis and epididymis was observed. Sperm concentration, viability, acrosome status and morphology were also examined. Expressions of heat shock protein 70 (Hsp70), tyrosine-phosphorylated (TyrPho) proteins, androgen receptor (AR) and steroidogenic acute regulatory (StAR) protein in testis were investigated. Results showed that DSF contained phenolic compounds and terpenoids (phytoandrogens; rengyolone and cleroindicin B). No reproductive histopathology was observed in DSF-treated rats. Although DSF decreased the serum testosterone level, the sperm qualities were not affected. Particularly, sperm concentration of DSF-treated animals was significantly increased. DSF changed the testicular TyrPho proteins but the expression of AR, StAR or Hsp70 was not altered. In conclusion, DSF possesses antioxidant capacity with no toxicity on male reproductive system.

Localization (and profiles) of tyrosinephosphorylated proteins in female reproductive organs of adult rats.

OBJECTIVE: Tyrosine phosphorylation is an essential process in many biological systems, including the male reproductive system. The presence of tyrosine-phosphorylated (TyrPho) proteins has been well documented in male reproductive organs, but research in fertile females is still limited. METHODS: The ovary, oviduct, and uterus of adult female Sprague-Dawley rats in the estrus phase were used to localize TyrPho proteins using an immunohistochemical technique. These proteins were separated and their expression patterns were examined by sodium dodecyl sulfatepolyacrylamide gel electrophoresis and Western blot analysis, respectively. RESULTS: TyrPho proteins were localized in the cytoplasm of the oocyte except the antral fluid; in the granulosa cells, theca cells, and stromal cells of the ovary; at the apical surface of oviductal epithelial cells; and in the basal epithelium and submucosa of the uterine wall. Moreover, we found that 72-, 43-, and 28-kDa TyrPho proteins were localized in the ovary, while 170-, 55-, and 43-kDa proteins were localized in the oviduct. In the uterus, we detected four major bands, corresponding to 61-, 55-, 54-, and 43-kDa TyrPho proteins. CONCLUSION: Given that these TyrPho proteins were found in major reproductive organs in the estrus phase, these proteins may play important roles in female fertility.

Changes of tyrosine phosphorylation in liver and kidney of polycystic ovarian rats induced by letrozole / Cambios de fosforilación de tirosina en hígado y riñón de ratas ováricas poliquísticas inducidas por letrozol

Letrozole (Letro) is a drug commonly used for breast cancer treatment since it can decrease estrogen level. In experimental animal, the Letro has been used to induce the polycystic ovarian syndrome (PCOS) model. Tyrosine phosphorylation (TyrPho) is an essential process in various biological functions both normal and abnormal conditions especially reproduction. Although some side effects of Letro are reported, the alterations of TyrPho responsible for liver and kidney functions have never been demonstrated. In this study, the blood serum, liver, and kidney of control and PCOS rats induced with Letro (orally, 1 mg/ KgBW) for consecutive 21 days were used to determine the serum biochemical components and to investigate the TyrPho expression using western blot analysis. Histopathology of such tissues was observed by Masson's trichrome staining. The results showed that Letro did not affect histological structures but significantly increased the serum levels of urea nitrogen, cholesterol, triglyceride, HDL, LDL, ALT, AST, and alkaline phosphatase. Additionally, the TyrPho protein expressions of 32 and 27 kDas in liver and of 55 and 43 kDas in kidney were increased while of a kidney 26 kDa was decreased as compared to those of control. In conclusion, this recent study indicated that the changes of TyrPho proteins in liver and kidney induced with Letro associated with their functions by alteration of serum biochemical levels.

El letrozol (Letro) es un medicamento utilizado comúnmente para el tratamiento del cáncer de mama, debido a que puede disminuir el nivel de estrógeno. En animales de experimentación, el Letro se ha utilizado para inducir el modelo de síndrome de ovario poliquístico (PCOS). La fosforilación de tirosina (TyrPho) es un proceso esencial en diversas funciones biológicas, tanto en condiciones normales como anormales, especialmente en la reproducción. A pesar de informes que indican algunos efectos secundarios de Letro, no se han demostrado las alteraciones de TyrPho responsables de las funciones hepáticas y renales. En este estudio, el suero sanguíneo, el hígado y el riñón control y las ratas PCOS inducidas con Letro (por vía oral, 1 mg / KgBW) durante 21 días consecutivos se usaron para determinar los componentes bioquímicos del suero y para investigar la expresión de TyrPho usando análisis de transferencia Western. La histopatología de los tejidos se observó mediante la tinción tricrómica de Masson. Los resultados mostraron que Letro no afectó las estructuras histológicas, pero aumentó significativamente los niveles séricos de urea, colesterol, triglicéridos, HDL, LDL, ALT, AST y fosfatasa alcalina. Además, las expresiones de la proteína TyrPho de 32 y 27 kDas en el hígado y de 55 y 43 kDas en el riñón aumentaron mientras que en un riñón disminuyeron 26 kDa en comparación con el control. En conclusión, este estudio indicó que los cambios de las proteínas TyrPho en el hígado y los riñones inducidos con Letro se asociaron con sus funciones mediante la alteración de los niveles bioquímicos en suero.

Evaluation of antioxidant capacity and reproductive toxicity of aqueous extract of Thai Mucuna pruriens seeds.

OBJECTIVE: In traditional medicine, the seeds of Thai Mucuna pruriens (T-MP) are used to treat male dysuria and are believed to enhance fertility. However, information pertaining to the toxicity of T-MP and its interaction with other properties is limited. This study was thus conducted to evaluate the antioxidant capacity and subacute toxicity of T-MP in the reproductive system. METHODS: Total phenolic content and antioxidant capacity of T-MP seed extract were determined using total phenolic content, 2,2-diphenyl-1-picrylhydrazyl and ferric reducing antioxidant power assays. Male and female adult rats were treated orally with T-MP at a dosage of 150 or 300 mg/kg body weight for 14 consecutive days. Sex hormones and functional parameters in the liver and kidney were evaluated. Histopathology of all tissue was conducted using Masson's trichrome staining. Sperm parameters, including concentration, morphology, acrosome reaction status and DNA damage, were also examined. Expression of tyrosine phosphorylated protein (TyrPho), androgen receptor and A-kinase-anchoring protein 4 (AKAP4) were investigated using the Western blot technique. RESULTS: T-MP seed extract contained phenolic compounds and exhibited high antioxidant capacity with no toxicity at the tested doses. It did not affect liver or kidney function parameters in the male rats, but increased estradiol, aspartate aminotransferase and alanine aminotransferase levels in the females. Additionally, it decreased serum progesterone and alkaline phosphatase levels in female rats. Serum and intratesticular testosterone levels were significantly lower in male rats that received a high dosage of T-MP. Histopathological changes were not observed in any tissue treated with T-MP. T-MP also significantly increased sperm concentration (but did not affect sperm parameters), and enhanced testicular TyrPho protein and androgen receptor and expression of AKAP4 in sperms. CONCLUSION: T-MP seed extract exhibited antioxidant capacity and was not harmful to reproductive tissues. It also had a phytoestrogenic effect on females and increased the expression of testicular and sperm markers of male fertility.