Simultaneous determination of five constituents of areca nut extract in rat plasma using UPLC-MS/MS and its application in a pharmacokinetic study.

Areca nuts have been used as a traditional Chinese medicine (TCM) for thousands of years. Recent studies have shown that it exhibits good pharmacological activity and toxicity. In this study, the pharmacokinetics of five major components of areca nut extract in rats were investigated using a highly sensitive ultra-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UPLC-MS/MS) method. Arecoline, arecaidine, guvacoline, guvacine, and catechin were separated and quantified accurately using gradient elution with mobile phases of (A) water containing 0.1 % formic acid-10 mM ammonium formate, and (B) methanol. The constituents were detected under a timing switch between the positive and negative ion modes using multiple reaction monitoring (MRM). Each calibration curve had a high R2 value of >0.99. The method accuracies ranged -7.09-11.05 % and precision values were less than 14.36 %. The recovery, matrix effect, selectivity, stability, and carry-over of the method were in accordance with the relevant requirements. It was successfully applied for the investigation of the pharmacokinetics of these five constituents after oral administration of areca nut extract. Pharmacokinetic results indirectly indicated a metabolic relationship between the four areca nut alkaloids in rats. For further clarification of its pharmacodynamic basis, this study provided a theoretical reference.

BDE-209 disrupted the blood-testis barrier integrity by inhibiting estrogen receptor α signaling pathway in Sprague-Dawley rats.

Deca brominated diphenyl ether (BDE-209) is a widely used flame retardant with endocrine-disrupting activity which reportedly caused sperm quality decline and damaged blood-testis barrier (BTB). However, whether BDE-209 exposure led to BTB integrity dysfunction through affecting microtubule cytoskeletal organization and junctions was not well-elucidated. This study aimed to investigate the role of estrogen receptor α (ERα) in BDE-209-mediated perturbation of BTB integrity. Male rats and primary culture Sertoli cells were co-treated with BDE-209 and propylpyrazoletriol (PPT). The data demonstrated that BDE-209 impaired BTB integrity by reducing crucial tight junction-related proteins with ZO-1 and Occludin. Furthermore, the data suggested that BDE-209 diminished the apical ectoplasmic specialization markers with Eps8 and Formin1. In addition, BDE-209 damaged BTB ultrastructure including tight junctions and ectoplasmic specialization structures with broken tight junctions and the absence of actin microfilaments. Further experiments revealed that ERα was triggered in BDE-209-treated Sertoli cells. Unexpectedly, we found that PPT rescued BDE-209-mediated disruption of BTB integrity including tight junction and apical ectoplasmic specialization by activating ERα in Sertoli cells. Taken together, these findings indicated that intratesticular BDE-209 exposure perturbed BTB integrity and destroyed BTB structure by blocking ERα pathway. Our findings provide a new therapeutic target for male reproductive dysfunction.

N-acetylcysteine rescues meiotic arrest during spermatogenesis in mice exposed to BDE-209.

Deca-bromodiphenyl ethers (BDE-209) has been widely used in electronic devices and textiles as additives to flame retardants. Growing evidence showed that BDE-209 exposure leads to poorer sperm quality and male reproductive dysfunction. However, the underlying mechanisms of BDE-209 exposure caused a decline in sperm quality remains unclear. This study aimed to evaluate the protective effects of N-acetylcysteine (NAC) on meiotic arrest in spermatocytes and decreased sperm quality in BDE-209-exposed mice. In the study, mice were treated with NAC (150 mg/kg BW) 2 h before administrated with BDE-209 (80 mg/kg BW) for 2 weeks. For the in vitro studies, spermatocyte cell line GC-2spd cells were pretreated with NAC (5 mM) 2 h before treated with BDE-209 (50 µM) for 24 h. We found that pretreatment with NAC attenuated the oxidative stress status induced by BDE-209 in vivo and in vitro. Moreover, pretreatment with NAC rescued the testicular histology impairment and decreased the testicular organ coefficient in BDE-209-exposed mice. In addition, NAC supplement partially promoted meiotic prophase and improved sperm quality in BDE-209-exposed mice. Furthermore, NAC pretreatment effectively improved DNA damage repair by recovering DMC1, RAD51, and MLH1. In conclusion, BDE-209 caused spermatogenesis dysfunction related to the meiotic arrest medicated by oxidative stress, decreasing sperm quality.

BDE-209 disturbed proliferation and differentiation of spermatogonia during mitotic process through estrogen receptor α.

Deca-bromodiphenyl ether (BDE-209) exposure caused spermatogenesis disorder resulting in poor sperm quality has become a public concern in recent years. Spermatogenesis refers to the process by which the division of spermatogonia stem cells (SSCs) produces haploid spermatozoa, including mitosis, meiosis, and spermiogenesis. However, the mechanism of mitosis including proliferation and differentiation of spermatogonia dysfunction induced by BDE-209 remains largely unclear. Here, our data showed that BDE-209 exposure caused a decline in sperm quality with seminiferous tubule structure disorder in rats. In addition, BDE-209 exposure damage spermatogonia proliferation and differentiation with decreasing level of PLZF and cKit in testis. Moreover, rats exposed to BDE-209 decreased the expression of ERα, whereas an elevated expression of Wnt3a and Wnt5a. Mechanistically, supplementation with propipyrazole triol (PPT, a selective ERα pathway agonist) rescued sperm quality and attenuated impairment of proliferation and differentiation of spermatogonia in BDE-209-induced rats. Therefore, ERα plays a crucial role in the proliferation and differentiation of spermatogonia during mitotic process. In conclusion, our study clarified the role of ERα in BDE-209-induced spermatogonia proliferation and differentiation in rats and provides a potential therapeutic application on poor sperm quality caused by BDE-209 exposure.

Vitamin C supplementation rescued meiotic arrest of spermatocytes in Balb/c mice exposed to BDE-209.

Deca-brominated diphenyl ether (BDE-209) is a ubiquitous industrial chemical as brominated flame retardant (BFRs). Exposure to BDE-209 has been clearly associated with male reproductive disorders. However, the meiotic arrest mechanism of spermatocytes exposed to BDE-209 is still unclear. The present work aimed to explore the protective effect of vitamin C on BDE-209-induced meiotic arrest of spermatocytes and its possible mechanism. Vitamin C (100 mg/kg BW) was administered to BDE-209-exposed (80 mg/kg BW) male Balb/c mice once daily by intraperitoneal injection for 2 weeks. Our results showed that vitamin C played male reproductive protection effects as showed by attenuated BDE-209-induced testicular damage, and reduced sperm abnormality rate. Vitamin C also attenuated BDE-209-induced increase in SOD and MDA in testes and GC-2 spd cells. Moreover, vitamin C promoted meiotic prophase in BDE-209-induced mice, with suppressed γ-H2AX, restored DMC1, RAD51, and crossover marker MLH1 levels, and prevented BDE-209-induced DNA impairment. In addition, vitamin C supplementation also interfered with BDE-209-induced upregulation of testicular H3K4me3 through inhibition of KDM5s capacity and decreasing ferrous ion concentration. Furthermore, ferrous sulfate pretreatment could partially restore the expression of H3K4me3 via maintaining the concentration of ferrous ions. Taken together, vitamin C exerts a potential therapeutic agent for preventing BDE-209-induced reproductive toxicity with meiotic arrest, which is attributed to its antioxidant and electron donor properties, as well as, modulation of ferrous ion levels and demethylation of H3K4me3.

Effects of perinatal exposure to endocrine-disrupting chemicals on the reproductive system of F3 generation male rodents: a meta-analysis.

To explore the relationship between perinatal exposure to endocrine-disrupting chemicals and the male reproductive system of F3 generation, and to evaluate the toxicological effects of endocrine-disrupting chemicals on the reproductive system of F3 generation male rodents. PubMed and Web of Science databases were searched to obtain the studies; overall risk ratios (RRs) with 95% confidence intervals (95% CIs) were used to evaluate the relationship between exposure to endocrine-disrupting chemicals and reproductive system damage in F3 generation male rodents. Nine studies were included for analysis. Endocrine-disrupting chemicals are significantly associated with the reproductive system of male rodents of F3 generation, especially the testis (RR = 3.13, 95% CI: 2.05, 4.76), prostate (RR = 2.26, 95% CI: 1.27, 4.00), and kidney (RR = 2.83, 95% CI: 1.77, 4.52), but the current analysis does not prove that EDCs are the adverse factors for puberty abnormalities. The results indicated that the overall associations between atrazine (RR = 3.06, 95% CI: 1.10, 8.51, P = 0.032), DDT (RR = 6.26, 95% CI: 1.56, 25.08, P = 0.010), pesticide and insect repellent mixture (permethrin and DEET) (RR = 2.23, 95% CI: 1.34, 3.69, P = 0.002), and vinclozolin (RR = 4.71, 95% CI: 2.74, 8.10, P = 0.000) and reproductive system damage in F3 generation male rodents were statistically significant. Our study indicated that EDCs have an atavistic effect on the male reproductive system, and we should pay attention to the long-term effects of environmental exposure to endocrine disruptors in future generations.

BDE-209 induce spermatocytes arrest at early-pachytene stage during meiotic prophase I in mice.

Deca-brominated diphenyl ether (BDE-209) is a common flame retardant utilized in electronic products, textiles, furniture, and upholstery materials. Environmental BDE-209 exposure results in spermatogenesis disorder, because of the characteristics of bioaccumulation, persistence, and probably toxicity. Meiotic prophase I is a crucial phase during spermatogenesis which is a key influential factor of normal sperm production. However, the effects of BDE-209 on meiotic prophase I during spermatogenesis are poorly understood. The present study aimed to evaluate whether BDE-209 exposure impairs meiotic prophase I during spermatogenesis of spermatocytes. We validated the effects of BDE-209 on alternations of meiotic prophase I in Balb/c male mice. Firstly, we analyzed sperm quality in cauda epididymis with decreasing sperm count, increasing abnormal sperm, and male reproductive dysfunction after exposure to BDE-209. Then, reactive oxygen species (ROS) and malondialdehyde (MDA) levels in testis and GC-2spd cells were significant increased after treated with BDE-209. Furthermore, we found that meiotic prophase I arrest at early-pachytene stage during spermatogenesis with increasing of DSBs damage and trimethylated histone H3 at lysine-4 (H3K4me3) in spermatocytes exposed to BDE-209. Finally, we conducted homologous recombination (HR) analyses to identify the progression of meiosis. The recombination markers, including DMC1 and RAD51, and crossover marker MLH1 were decreased during spermatogenesis after exposure to BDE-209. Collectively, our data indicated that BDE-209 has detrimental impacts on meiotic prophase I of spermatocytes in mice.

Study on the association between ambient air pollution and daily cardiovascular death in Hefei, China.

Cardiovascular disease has always been the most serious public health problem in China. Although many studies have found that the risk of death caused by cardiovascular disease is related to air pollutants, the existing results are still inconsistent. The aim of this study was to investigate the effects of air pollutants on the risk of daily cardiovascular deaths in Hefei, China. Daily data on cardiovascular deaths, daily air pollutants, and meteorological factors from 2007 to 2016 were collected in this study. A time-series study design using a distributed lag nonlinear model was employed to evaluate the association between air pollutants and cardiovascular deaths. First, a single air pollutant model was established based on the minimum value of Akaike information criterion (AIC), and the single day lag effects and multi-day lag effects were discussed separately. Then, two-pollutant models were fitted. Subgroup analyses were conducted by gender (male and female), age (< 65 age and ≥ 65 age), and disease type (ischemic heart disease and cerebral vascular disease). There were 34,500 cases of cardiovascular deaths during the period 2007-2016, and the average concentrations of air pollutants (PM10, SO2, NO2, PM2.5, CO, O3) were 106.11, 20.34, 30.49, 72.59, 958.7, and 67.88 µg/m3, respectively. An increase of interquartile range (IQR) in PM10, SO2, NO2, PM2.5, CO, and O3 were associated with an increase of 4.34% (95%CI 1.54~7.23%) at lag 0-6, 5.79% (95%CI 2.43~9.27%) at lag 0-5, 4.47% (95%CI 1.64~7.37%) at lag 0-5, 3.14% (95%CI 0.03~6.36%) at lag 0-4, 3.11% (95%CI 0.21~6.10%) at lag 0-3, and 8.17% (95%CI 1.89~14.84%) at lag 0-5 in cardiovascular deaths, respectively. Females, older group (≥ 65 years) and deaths from cerebral vascular disease were more vulnerable to air pollution than males, younger individuals (< 65 years) and deaths from ischemic heart disease. Our results suggest that air pollution increased the risk of cardiovascular deaths in Hefei. These findings can provide evidence for effective air quality interventions in Hefei.