Involvement of Autophagy and Oxidative Stress-Mediated DNA Hypomethylation in Transgenerational Nephrotoxicity Induced in Rats by the Mycotoxin Fumonisin B1.

Fumonisin B1 (FB1), a mycotoxin produced by Fusarium verticillioides, is one of the most common pollutants in natural foods and agricultural crops. It can cause chronic and severe health issues in humans and animals. The aim of this study was to evaluate the transgenerational effects of FB1 exposure on the structure and function of the kidneys in offspring. Virgin female Wistar rats were randomly divided into three groups: group one (control) received sterile water, and groups two and three were intragastrically administered low (20 mg/kg) and high (50 mg/kg) doses of FB1, respectively, from day 6 of pregnancy until delivery. Our results showed that exposure to either dose of FB1 caused histopathological changes, such as atrophy, hypercellularity, hemorrhage, calcification, and a decrease in the glomerular diameter, in both the first and second generations. The levels of the antioxidant markers glutathione, glutathione S-transferase, and catalase significantly decreased, while malondialdehyde levels increased. Moreover, autophagy was induced, as immunofluorescence analysis revealed that LC-3 protein expression was significantly increased in both generations after exposure to either dose of FB1. However, a significant decrease in methyltransferase (DNMT3) protein expression was observed in the first generation in both treatment groups (20 mg/kg and 50 mg/kg), indicating a decrease in DNA methylation as a result of early-life exposure to FB1. Interestingly, global hypomethylation was also observed in the second generation in both treatment groups despite the fact that the mothers of these rats were not exposed to FB1. Thus, early-life exposure to FB1 induced nephrotoxicity in offspring of the first and second generations. The mechanisms of action underlying this transgenerational effect may include oxidative stress, autophagy, and DNA hypomethylation.

Effect of Acute Melatonin Injection on Metabolomic and Testicular Artery Hemodynamic Changes and Circulating Hormones in Shiba Goats under Sub-Tropical Environmental Conditions.

The beneficial effects of melatonin were investigated to mitigate various detrimental effects and toxicity on reproductive performance. The present study aimed, for the first time, to explore the effect of intravenous melatonin injection on testicular artery hemodynamics (TH) and metabolomic changes, reproductive hormones in heat-stressed bucks. Ten bucks were randomly split into two groups (five each): (1) the melatonin group, treated with a single intravenous dose of melatonin solution containing 10 mg melatonin each, and (2) the control group, which was treated with 10 mL of the vehicle without melatonin. Changes in the TH at the level of the supra testicular artery (STA) were assessed by triplex ultrasonography just before (0 h) and at 0.5, 2, 7, 24, and 168 h after melatonin or vehicle administration. Doppler velocity parameters of peak systolic velocity (PSV; cm/s), end-diastolic velocity (EDV; cm/s), and time average maximum velocity (TAMAX; cm/s) were measured. Doppler indices (resistive index; RI and pulsatility index; PI), systole/diastole (S/D) ratio and total arterial blood flow volume (TABFV; ml/minute) were measured. Peripheral concentrations of FSH, LH, inhibin, melatonin, testosterone (T), estradiol (E2), and cortisol were measured just before injection (0 h) and at 0.5, 2, 7, and 24 h and daily up to day 7 post administration in both groups. Results revealed reductions in the RI values and increases in the TABFV in the melatonin group compared to the control one, especially 2 h after administration. Significant increases in concentrations of FSH, T, E2, and melatonin and decreases in cortisol and inhibin in the melatonin group compared to the control one. Plasma metabolomic analysis at 2 h indicated the up-regulation of L-glutamine, L-arginine, sorbitol, D-glucose, ascorbic acid, and ornithine and the down-regulation of D-xylose, D-arabitol, ribitol, and oleic acid in the melatonin versus the control group. In conclusion, acute administration of melatonin (10 mg IV) enhanced testicular artery blood flow and plasma reproductive hormones in the Shiba goat under heat-stress circumstances.

Metabolic Syndrome and Cardiometabolic Risk Factors in the Mixed Hypercholesterolemic Populations with Respect to Gender, Age, and Obesity in Asir, Saudi Arabia.

This record study aimed to investigate the prevalence of metabolic syndrome (MetS) profiles regarding sex, age, and obesity for the riskier factor of cardiovascular diseases in a general population in Saudi Arabia. Laboratory and anthropometric measurements were performed on non-specific participants with variant ages and BMI in either sex. Serobiochemical changes were measured for metabolic profiles, i.e., A1C/FSG, TC, TGC, HDLC/LDLC, Vit.D, TSH/T4, Hb, and Cr. The study was applied in a Polyclinic, Abha, Saudi Arabia in 2020 G. The general population showed variable incidences of MetS profiles, such as 69.4% diabetes, 85.5% hypothyroidism, and 92.2% obesity. Hypothyroidism showed a higher incidence in women rather than in men, but men were more dyslipidemic, with higher TGC and LDLC but low HDLC, compared to women. Men <40 Y. showed diabetes and hypothyroidism, but elders were dyslipidemic. Women <40 Y. showed anemia and hypovitaminosis-D but were suffering from hypothyroidism at all ages. Diabetes, hypothyroidism, hypovitaminosis-D, and dyslipidemia were the main MetS components in both overweight and obese participants, and an incidence of more than 50% in each profile was recorded. Diabetes with hypertension was characteristic of obese participants rather than those overweight. About 66.1% of the mixed-hypercholesterolemic cases were diabetic, but 18.9% of the mixed-diabetic participants were hypercholesterolemic. Castelli's risk factors, CRI-I and CRI-II, and atherogenic indices, AIP and AC, were measured for evaluating the cardiac risk in different populations based on the AUC-ROC and cut-off values. Insulin-resistance marker (TyG) was also measured, showing considerable cut-off values for diabetic susceptibility in the lipidemic participants with higher TGC and TC rather than HDLC or LDLC. In conclusion, MetS showed higher susceptibility to sex and age with increased incidence in women rather than men. However, the cardiac risk was more susceptible to men of higher TGC and low HDLC than women. Type 2 Diabetes mellitus (T2DM) was more prominent in both elders (≥40 Y.) than younger ages of either sex. Anemia and deficiency of Vit. D was characteristic of young women (<40 Y.). Hypothyroidism affects young men <40 Y. but was recorded in women of all ages. Both dyslipidemia and diabetes could trigger CVD, showing higher cardiac risk in mixed-hypercholesterolemic men rather than women. Our study strongly suggests that the consumption of unhealthy junk food, tobacco smoking, lack of exercise, and physical inactivity could be conclusive evidence of MetS in the Saudi population.

Fetal programming: in utero exposure to acrylamide leads to intergenerational disrupted ovarian function and accelerated ovarian aging.

In this study we investigated the effects of multigenerational exposures to acrylamide (ACR) on ovarian function. Fifty-day-old Wistar albino female rats were divided into the control and ACR-treated groups (2.5, 10, and 20 mg/kg/day) from day 6 of pregnancy until delivery. The obtained females of the first (AF1) and second generation (AF2) were euthanized at 4 weeks of age, and plasma and ovary samples were collected. We found that in utero multigenerational exposure to ACR reduced fertility and ovarian function in AF1 through inducing histopathological changes as evidenced by the appearance of cysts and degenerating follicles, oocyte vacuolization, and pyknosis in granulosa cells. TMR red positive cells confirmed by TUNEL assay were mostly detected in the stroma of the treated groups. Estradiol and IGF-1 concentrations significantly decreased as a result of decreased CYP19 gene and its protein expression. However, ACR exposure in AF2 led to early ovarian aging as evidenced by high estradiol and progesterone levels among all treated groups compared to control group, corresponding to the upregulation of the CYP19 gene and protein expression. The apoptotic cells of the stroma were greatly detected compared to that in the control group, whereas no significant difference was reported in ESR1 and ESR2 gene expression. This study confirms the developmental adverse effects of ACR on ovarian function and fertility in at least two consecutive generations. It emphasizes the need for more effective strategies during pregnancy, such as eating healthy foods and avoiding consumption of ACR-rich products, including fried foods and coffee.

Upregulation of Nrf2/HO-1 Signaling and Attenuation of Oxidative Stress, Inflammation, and Cell Death Mediate the Protective Effect of Apigenin against Cyclophosphamide Hepatotoxicity.

Liver injury is among the adverse effects of the chemotherapeutic agent cyclophosphamide (CP). This study investigated the protective role of the flavone apigenin (API) against CP-induced liver damage, pointing to the involvement of Nrf2/HO-1 signaling. Rats were treated with API (20 and 40 mg/kg) for 15 days and received CP (150 mg/kg) on day 16. CP caused liver damage manifested by an elevation of transaminases, alkaline phosphatase (ALP), and lactate dehydrogenase (LDH), and histological alterations, including granular vacuolation, mononuclear cell infiltration, and hydropic changes. Hepatic reactive oxygen species (ROS), malondialdehyde (MDA), and nitric oxide (NO) were increased and glutathione (GSH) and antioxidant enzymes were decreased in CP-administered rats. CP upregulated the inflammatory markers NF-κB p65, TNF-α, IL-6, and iNOS, along with the pro-apoptotic Bax and caspase-3. Pre-treatment with API ameliorated circulating transaminases, ALP, and LDH, and prevented histopathological changes in CP-intoxicated rats. API suppressed ROS, MDA, NO, NF-κB p65, iNOS, inflammatory cytokines, oxidative DNA damage, Bax, and caspase-3 in CP-intoxicated rats. In addition, API enhanced hepatic antioxidants and Bcl-2 and boosted the Nrf2 and HO-1 mRNA abundance and protein. In conclusion, API is effective in preventing CP hepatotoxicity by attenuating oxidative stress, the inflammatory response, and apoptosis. The hepatoprotective efficacy of API was associated with the upregulation of Nrf2/HO-1 signaling.

Ethylbenzene exposure disrupts ovarian function in Wistar rats via altering folliculogenesis and steroidogenesis-related markers and activating autophagy and apoptosis.

Ethylbenzene is a hydrocarbon that is extensively used in both industry and in the home and has been reported as toxic to various tissues. Nevertheless, its effect on ovarian function remains unclear. For this purpose, we assessed ovarian tissue morphology, evaluated protein and gene expression related to folliculogenesis and steroidogenesis, and investigated the involvement of both apoptosis and autophagy processes in this effect. Female Wistar albinos rats were treated with 2000, 4000 and 8000 ppm doses of ethylbenzene by inhalation for 30 min daily for one month. Ovaries were then removed and proceeded for histopathological and molecular analyses. We found that ethylbenzene affected folliculogenesis by decreasing the number of growing follicles and increasing the number of abnormal follicles, leading to faster female reproductive aging. Interestingly, it disrupted female reproductive hormone balance, including progesterone, estradiol, testosterone and IGF-1 plasma levels. The latter protein, along with GDF-9, significantly decreased in all ethylbenzene-treated groups, leading to the disruption of follicular cell proliferation and development. TUNEL assay study showed that ethylbenzene exposure significantly increased the number of apoptotic cells. The mRNA levels of genes involved in granulosa cell proliferation and differentiation, such as INSL3, CCND2 and ACTB, were significantly decreased. In addition, LC3 protein expression increased, and its encoding gene was upregulated, suggesting that ethylbenzene treatment induced autophagy. In summary, ethylbenzene exposure caused structural and functional disorders of the ovary by disrupting the normal growth of follicles, altering reproductive hormone balance, inhibiting the expression of key reproductive proteins and triggering autophagy as well as apoptosis.

Toluene Can Disrupt Rat Ovarian Follicullogenesis and Steroidogenesis and Induce Both Autophagy and Apoptosis.

Toluene has been shown to be highly toxic to humans and animals and can cause damage to various tissues. However, studies reporting its effects on ovarian function are still limited. In this study, we investigated the in vivo effect of toluene using female Wistar rats. We found that toluene exposure decreased ovarian weight and affected ovarian structure by increasing the number of abnormally growing follicles. Moreover, it significantly increased progesterone and testosterone levels. We also showed that toluene exposure decreased GDF-9 protein and its encoding gene. In addition, it inhibited the expression of most of the genes involved in granulosa cell proliferation and differentiation, such as Insl3, ccnd2 and actb. The TUNEL assay showed that apoptosis occurred at the middle and high doses only (4000 and 8000 ppm, respectively), whereas no effect was observed at the low dose (2000 ppm). Interestingly, we showed that toluene exposure induced autophagy as LC3 protein and its encoding gene significantly increased for all doses of treatment. These results may suggest that the activation of autophagy at a low dose of exposure was to protect ovarian cells against death by inhibiting apoptosis, whereas its activation at high doses of exposure triggered apoptosis leading to cell death.

High-fat diet-induced aggravation of cardiovascular impairment in permethrin-treated Wistar rats.

This study characterized the impact of post-weaning high-fat diet (HFD) and/or permethrin (PER) treatment on heart dysfunction and fibrosis, as well as atherogenic risk, in rats by investigating interactions between HFD and PER. Our results revealed that HFD and/or PER induced remarkable cardiotoxicity by promoting cardiac injury, biomarker leakage into the plasma and altering heart rate and electrocardiogram pattern, as well as plasma ion levels. HFD and/or PER increased plasma total cholesterol, triacylglycerols, and low-density lipoprotein (LDL) cholesterol levels but significantly reduced high-density lipoprotein (HDL) cholesterol. Cardiac content of peroxidation malonaldehyde, protein carbonyls, and reactive oxygen species were remarkably elevated, while glutathione levels and superoxide dismutase, catalase and glutathione peroxidase activities were inhibited in animals receiving a HFD and/or PER. Furthermore, cardiac DNA fragmentation and upregulation of Bax and caspase-3 gene expression supported the ability of HFD and/or PER to induce apoptosis and inflammation in rat hearts. High cardiac TGF-ß1 expression explained the profibrotic effects of PER either with the standard diet or HFD. Masson's Trichrome staining clearly demonstrated that HFD and PER could cause cardiac fibrosis. Additionally, increased oxidized LDL and the presence of several lipid droplets in arterial tissues highlighted the atherogenic effects of HFD and/or PER in rats. Such PER-induced cardiac and vascular dysfunctions were aggravated by and associated with a HFD, implying that obese individuals may be more vulnerable to PER exposure. Collectively, post-weaning exposure to HFD and/or PER may promote heart failure and fibrosis, demonstrating the pleiotropic effects of exposure to environmental factors early in life.

The flavonoid, kaempferol-3-O-apiofuranosyl-7-O-rhamnopyranosyl, as a potential therapeutic agent for breast cancer with a promoting effect on ovarian function.

It is widely known that breast cancer cells eventually develop resistance to hormonal drugs and chemotherapies, which often compromise fertility. This study aimed to investigate the effect of the flavonoid, kaempferol-3-O-apiofuranosyl-7-O-rhamnopyranosyl (KARP), on 1) the viability of MCF-7 breast cancer cells and 2) ovarian function in rats. A dose-dependent decrease in MCF-7 cell survival was observed, and the IC50 value was found to be 48 µg/ml. Cells in the control group or those exposed to increasing concentrations of KARP experienced a similar generation of reactive oxygen species and induction of apoptosis. For the rats, estradiol levels correlated negatively to KARP dosages, although a recovery was obtained at administration of 30 mg/kg per day. Noteworthily, when compared against the control, this dosage led to significant increases in mRNA levels for CYP19, CYP17a, CCND2, GDF9, and INSL3 among the treatment groups, and ER1 and ER2 mRNA levels decreased in a dose-dependent manner. KARP shows great promise as an ideal therapy for breast cancer patients since it induced apoptosis and autophagy in cancerous cells without harming fertility in our animal model. Future investigations on humans are necessary to substantiate these findings and determine its efficacy as a general line of treatment.

HPLC-ESI-QTOF-MS/MS profiling and therapeutic effects of Schinus terebinthifolius and Schinus molle fruits: investigation of their antioxidant, antidiabetic, anti-inflammatory and antinociceptive properties.

The aim of the current work was to study the phytochemical variability among Schinus terebinthifolius (STE) and Schinus molle (SME) fruit extracts. The in vitro antioxidant, antihemolytic, antidiabetic, and macromolecule damage protective activities, as well as, the in vivo anti-inflammatory and antinociceptive capacities were assessed. Using the HPLC-ESI-QTOF/MS analysis, the chemical profile of fruit extract varied between S. terebinthifolius (30 compounds) and S. molle (16 compounds). The major compound was masazino-flavanone (5774.98 and 1177.65 µg/g sample for STE and SME, respectively). The investigations highlighted significant antioxidant proprieties when using ABTS radical (IC50; 0.12 and 0.14 mg/ml for STE and SME, respectively), superoxide (IC50; 0.17 and 0.22 mg/ml for STE and SME, respectively) and hydrogen peroxide (IC50; 014 and 0.17 mg/ml for STE and SME, respectively). In addition, STE and SME proved preventive effects against H2O2-induced hemolysis (IC50; 0.22 and 0.14 mg/ml for STE and SME, respectively). The in vitro antidiabetic effect revealed that STE and SME exhibited important inhibitory effects against α-amylase (IC50; 0.13 and 0.19 mg/ml for STE and SME, respectively) and α-glycosidase (IC50; 0.21 and 0.18 mg/ml for STE and SME, respectively) when compared with acarbose. Furthermore, the extracts showed potent inhibitory activity against AAPH-induced plasmid DNA damage, and protein oxidation. In vivo study revealed that STE and SME presented interesting antinociceptive and anti-inflammatory capacities. All observed effects highlighted the potential application of Schinus fruit extract in food and pharmaceutical industries against ROS-induced damage.

Acrylamide impairs ovarian function by promoting apoptosis and affecting reproductive hormone release, steroidogenesis and autophagy-related genes: An in vivo study.

Acrylamide (ACR) toxicity is quite common due to its widespread use in industry and due to the Maillard browning reaction that occurs in foods containing high concentrations of hydrocarbons subjected to high temperatures. This study aimed to elucidate the female reproductive toxicity of ACR in vivo. Fifty-day-old Wistar-Albino female rats were treated with different dosages of ACR (2.5, 10, and 50 mg/kg/day). After treatment, the animals were sacrificed, and serum and ovary samples were collected for histological examination, hormone analysis, TUNEL analysis, and RT-PCR studies. We found that ACR acts by significantly reducing ovarian weight and serum progesterone and estradiol concentrations. In addition, ACR treatment led to pyknotic, heterochromatic characteristics and nuclear fragmentation, as evidenced by hematoxylin staining. The TUNEL assay revealed that granulosa cells were affected after the oral administration of ACR, leading to the apoptosis of follicles at different stages of growth. Compared with the control condition, high doses of ACR (50 mg/kg/day) significantly induced the overexpression of INSL3, CYP17a, IGF1, ESR1, ESR2, ATG5, ATG12 and LC3 in the ovary. Moreover, LC3 mRNA levels significantly increased with increasing doses of ACR (2.5, 10 and 50 mg/kg/day), suggesting that ACR treatment induced autophagy. In conclusion, ACR induced ovarian dysfunction by affecting steroid hormone release, increasing apoptosis and mRNA levels of autophagy-related genes. The eventual correlation between apoptotic granulosa cell death and autophagy needs to be further explored.