Physis is a complex cartilaginous structure that is critical for longitudinal bone growth. As one of the endocrine-disrupting chemicals, bisphenol A (BPA) can interfere with the physis by deranging the complex networks of nutritional, cellular, paracrine, and endocrine factors, and this affects longitudinal bone growth, leading to different growth outcomes. However, the exact mechanisms underlying these phenomena remain unclear. Therefore, understanding the molecular pathways involved in the physis after neonatal exposure to low-dose BPA may permit the identification of research targets for therapeutics, which may aid in modulating the process of growth plate closure. In the present study, female Sprague-Dawley rats were exposed to 0.05 mg·kg-1·day-1 of BPA and corn oil vehicle from postnatal day 1 (PND1) to 15 via subcutaneous injection. Next-generation RNA sequencing was performed for the mRNA isolated from the physis. The levels of osteocalcin (OC), growth hormone (GH), and insulin-like growth factor 1 (IGF-1) were measured on PND30 (BPA0.05mg vs. Vehicle; n = 5 for each group). We observed statistically significant enrichment of gene sets in the BPA0.05mg tissues compared with the Vehicle tissues. Further analysis of the differentially expressed genes (DEGs) identified BPA0.05mg-specific networks of secreted proteins (LEP, NPY, AGT, ACE2, C4B, and C4BPA) as well as those of local matrix and protease proteins (FBN2, LAMC2, ADAMTS16, and ADAMTS19). Furthermore, the levels of OC and GH were affected by BPA exposure. Our results revealed the specific molecular characteristics of physis contaminated with BPA and may provide new clues for physis maturation and supervision of industrial products and occupational exposure.

OBJECTIVE: To observe the effect of electroacupuncture on sexual development and ovarian estrogen receptor β(ER-β) expression in female adolescent obese rats induced by high-fat diet, so as to explore its underlying mechanisms of improving adolescent obesity.
METHODS: Female SD rats (age of 21 days) were randomly divided into control, model and acupuncture groups, with 6 rats in each group. The obese model was established by feeding high-fat diet for 6 weeks. Rats of the acupuncture group received electroacupuncture(2 Hz, 0.5-1.2 mA)stimulation at bilateral \"Sanyinjiao\"(SP6), \"Fenglong\"(ST40) and \"Zusanli\"(ST36) for 30 min, once a day for 14 days. The body mass and abdominal circumference of rats were measured before and after treatment. The contents of serum follicle stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E2) were detected by ELISA. The number of corpus luteum and follicle were observed by HE staining. The expression levels of ER-β mRNA and protein in ovary were detected by fluorescence quantitative PCR and Western blot, respectively.
RESULTS: Compared with the control group, the body mass and abdominal circumference, the contents of serum FSH and E2, and the expression levels of ER-β mRNA and protein in ovary were significantly increased (P<0.05)in the model group, while the number of mature follicles and corpus luteum increased significantly. Compared with the model group, the body mass and abdominal circumference, the contents of serum FSH and E2, and the expression levels of ER-β mRNA and protein in ovary were significantly decreased (P<0.05) in the acupuncture group, while the number of mature follicles and corpus luteum decreased significantly.
CONCLUSIONS: Electroacupuncture can effectively improve the levels of sex hormone and the development of ovary, down-regulate the expression levels of ER-β mRNA and protein in ovary, so as to regulate the process of sexual development of female adolescent obese rats induced by high-fat diet.
目的:观察电针对高脂饮食诱导的雌性生长期肥胖大鼠性发育水平和卵巢雌激素受体β(ER-β)表达的影响, 探讨电针治疗青少年肥胖的部分作用机制。方法:21日龄雌性SD大鼠随机分为对照组、模型组和针刺组, 每组6只。采用高脂饲料喂养6周建立肥胖大鼠模型。针刺组给予双侧“三阴交”“丰隆”“足三里”电针治疗, 连续波, 频率2 Hz, 强度0.5~1.2 mA, 30 min/d, 连续干预14 d。治疗前、后测量大鼠体质量和腹围;ELISA法检测大鼠血清卵泡刺激素(FSH)、黄体生成素(LH)、雌二醇(E2)含量;HE染色法观察大鼠卵巢黄体及卵泡数量;荧光定量PCR法检测大鼠卵巢ER-β mRNA的表达水平;Western blot法检测大鼠卵巢组织ER-β蛋白的表达水平。结果:与对照组比较, 模型组大鼠体质量、腹围明显升高(P<0.05), 血清FSH、E2含量和卵巢ER-β mRNA、蛋白的表达明显升高(P<0.05), 卵巢组织空卵泡及黄体数量明显增加;与模型组比较, 针刺组大鼠体质量、腹围明显降低(P<0.05), 血清FSH、E2含量及卵巢ER-β mRNA、蛋白的表达明显降低(P<0.05), 卵巢组织空卵泡数目及黄体数量降低, 可见不同发育阶段的卵泡。结论:电针可以改善高脂饮食诱导的雌性生长期肥胖大鼠的性激素水平, 改善卵巢组织发育, 调节卵巢ER-β mRNA及蛋白的表达水平, 从而调节雌性生长期肥胖大鼠性发育水平。.

Objective: To investigate the effect of chronic stress of pregnant rats on the gut microbiota of female rats and offspring, and explore the role of intestinal microbiota in chronic stress during pregnancy. Methods: In November 2019, SPF-grade healthy adult SD rats were selected. 16 female rats were randomly divided into control group and model group, with 8 in each group; 12 male rats were randomly divided into model mating group (8) and control mating group (4) . A model of chronic unpredictable mild stress (CUMS) during pregnancy was established. Blood samples were collected from the iliac vein of the female rats 1 day before and 1, 7, and 14 days after the CUMS protocol, and measured for plasma corticosterone content by radioimmunoassay. After the stress was completed, fresh feces of the female rats were collected for testing. The offspring\'s fresh stool samples were collected on postnatal day 20 (PND20) , and they were divided into control offspring group and model offspring group samples. The sequence of 16S rRNAV3-V4 regions of microorganisms in the feces of offspring was determined by Illumina MiSeq technique; and the interaction between microbial community structure and diversity were analyzed. Results: The content of plasma corticosterone in the model group was higher than that in the control group on the 7th and 14th day of stress (P<0.05) . Compared with the control group, the Sobs index, Chao index, ACE index and Shannon index of the model group were decreased (P<0.05) . The number of unique species abundance (OTU) in the control group was 130, and 91 in the model group. The relative abundance of female Firmicutes in the control group (64.87%) was higher than that in the model group, and the relative abundance of Bacteroides (31.72%) was lower than that of the model group (46.35%) . The Sobs index, Chao index, ACE index, Simpson index and Shannon index of the control offspring group were higher than those of the model offspring group (P<0.05) . The number of unique OTUs in the model offspring group was 75, and 93 in the control offspring group. The relative abundance of Firmicutes (60.24%) in the control offspring group was higher than that of the model offspring group (52.95%) . Conclusion: Chronic stress during pregnancy can not only lead to the disorder of intestinal flora in female rats, but also lead to the change of intrauterine environment, thus affecting the diversity of intestinal flora in offspring.
目的： 观察孕期慢性应激雌鼠及子代肠道菌群的变化，探索肠道菌群在孕期慢性应激中发挥的作用。 方法： 于2019年11月，选择SPF级健康成年SD大鼠，16只雌鼠随机分为对照组和模型组，每组8只；12只雄鼠随机分为对照交配组（4只）和模型交配组（8只）。建立孕期慢性不可预知温和应激（CUMS）模型，雌鼠在应激前1天和应激第1、7、14、21天进行眼内眦静脉采血，利用放射免疫法测定血浆皮质酮含量；应激完成后，收集雌鼠新鲜粪便待测。两组子鼠在出生后20 d（PND20）收集新鲜粪便待测，分为对照子鼠组和模型子鼠组样品。采用Illumina Miseq测序技术，测定大鼠粪便中微生物16S rRNA V3-V4区序列，并对群落结构和多样性进行分析。 结果： 模型组雌鼠在应激第7、14天血浆皮质酮含量均高于同期对照组雌鼠（P<0.05）；与对照组比较，模型组Sobs指数、Chao指数、Ace指数、Shannon指数均降低（P<0.05）；对照组所特有的物种丰度（OTU）数目为130种，模型组91种；对照组雌鼠厚壁菌门的相对丰度（64.87%）高于模型组（50.83%），而拟杆菌门的相对丰度（31.72%）低于模型组（46.35%）；对照子鼠组Sobs指数、Chao指数、Ace指数、Shannon指数和Simpson指数均高于模型子鼠组（P<0.05）；模型子鼠组所特有的OTU数目为75种，对照子鼠组93种；对照子鼠组厚壁菌门相对丰度（60.24%）高于模型子鼠组（52.95%）。 结论： 孕期慢性应激不仅会引起雌鼠肠道菌群紊乱，而且可能会导致宫内环境改变，进而对子代肠道菌群的群落多样性产生影响。.

G-protein coupled estrogen receptor 30 (GPR30) was proved the specific estrogen receptor relating to mechanical hyperalgesia. Studies have shown that the GABAA receptor subunits α4, β1, and δ in the periaqueductal gray (PAG) neurons promote the descending facilitation system. This study inquired into whether and how GPR30 and GABAA-α4β1δ in the PAG promote preoperative anxiety-induced postoperative hyperalgesia in female rats.
All the female rats were subjected to the single prolonged stress (SPS) to stimulate preoperative anxiety. Subsequently, mechanical allodynia was evaluated before and after the incision, based on the paw withdrawal mechanical threshold (PWMT). The selective GPR30 agonist G1 and antagonist G15 were locally microinjected into the PAG. The expression of GPR30, protein kinase A (PKA), and GABAA receptor subunits α4, β1, and δ in the PAG neurons were detected using western blotting and immunofluorescence.
Behavioral testing revealed that Group S and Group I decreased the nociceptive threshold levels of PWMT in female rats. PWMT in Group S + I decreased more than that of Group S and Group I. Further, results of western blotting showed the expression of GPR30, PKA, and GABAA α4, β1, and δ subunits significantly up-regulated in Group S + I, and immunofluorescence indicated that the neurons of PAG in Group S + I appeared simultaneously immunopositive for GPR30 and GABAA α4, β1, and δ receptors. After microinjection of G1 into the PAG, female rats with plantar incision continued to exhibit significant hyperalgesia until postoperative 48 h. On the other hand, microinjection of G15 with SPS and plantar incision procedure relieved postoperative hyperalgesia in female rats. Western blotting demonstrated that intra-PAG injection of G15 markedly decreased the GPR30, PKA, and GABAA α4, β1, and δ levels in Group G15 + I.
Our results indicate that the GPR30-PKA-GABAAα4β1δ pathway in the PAG promotes preoperative anxiety-induced postoperative hyperalgesia in female rats. This mechanism might be a potential novel therapeutic target for hyperalgesia in females.

Neonatal exposure to bisphenol A (BPA) is hypothesized to advance pubertal development. However, the effects of neonatal BPA exposure on pubertal development has not been described. In this study, female Sprague-Dawley rats were exposed to 0.05, 0.5, 5, or 10 mg·kg-1 ·day-1 BPA, or corn oil vehicle alone from postnatal day 1 (PND1) to PND10 via subcutaneous injection. We evaluated day of vaginal opening (DVO), ovarian morphology, serum hormone concentrations, and hypothalamic expression of Gnrh1 and Kiss1 in female rats at PND35. DVO was significantly advanced in rats exposed to 5 and 10 mg·kg-1 ·day-1 BPA. Serum hormone concentrations increased as BPA dose increased. Additionally, hypothalamic Gnrh1 and Kiss1 expression were increased with BPA exposure; rats exposed to 10 mg·kg-1 ·day-1 BPA had significantly upregulated hypothalamic Gnrh1 and Kiss1 expressions in terms of both messenger RNA and protein levels. Our results suggest that exposure to a 10 mg·kg-1 ·day-1 dose of BPA might advance pubertal development significantly. In addition, within the range of 0 to 10 mg·kg-1 ·day-1 , neonatal exposure to BPA may affect pubertal development in a dose-dependent manner.

Acrylamide (AA), a food contaminant, caused islet remodeling and increased hepatic glycogen content in male rats, but the effect of AA on glucose homeostasis in female rats remains unclear. In this study, female SD rats were orally treated with 0, 15, or 30 mg/kg·bw AA for 3 weeks. The levels of fasting blood glucose (FBG), blood glucose after oral administration of glucose, plasma insulin and hepatic glycogen were measured. The histology of the pancreas was observed, and the transcription of key genes involved in glucose metabolism and insulin signaling in liver were determined. Compared with the control, exposure to 30 mg/kg·bw of AA significantly increased FBG level, reduced hepatic glycogen content and impaired glucose tolerance. Moreover, damaged islets were observed at 15 and 30 mg/kg·bw AA-exposed groups. In addition, AA exposure significantly promoted gluconeogenesis and glycogenolysis (up-regulation of pc, g6p and gp) and decreased glycolysis (down-regulation of gck and pfk). Alternations in these processes may be associated with decreased plasma insulin levels and inhibited insulin-regulated IRS/PI3K/Akt/Foxo1 signaling transduction under AA exposure. Overall, our findings demonstrated that AA disrupted glucose homeostasis and elevated FBG level in female rats possibly by interfering with glucose metabolism and hampering the physiological effect of insulin.

The purpose of the study was to study the activity of the phytoestrogen genistein (GEN) act- ing on FSHR and LHR in rat ovaries with polycystic ovary syndrome (PCOS). Sixty rats were di- vided into six groups. Rats in the dose group received genistein at a concentration of either 5 (low genistein dose group, L-gen), 10 (middle genistein dose group, M-Gen) or 20 (high genistein dose group, H-Gen) mg per kg of body weight per day. Estrogen group (EG, received 0.5 mg/kg Dieth- ylstilbestrol). Concentration of sex hormones in serum was quantified by enzyme-linked immuno- sorbent assay (ELISA). Expressions of follicle-stimulating hormone receptor (FSHR) and lutein- izing hormone receptor (LHR) protein were determined by immunohistochemistry. Treatment with genistein resulted in a strong stimulation of the concentration of sex hormone in serum. The concentration of progesterone and FSH was significantly higher in H-Gen when compared to the PCOS model control group (MG) (P ⟨ 0.01). In contrast, the concentration of testosterone, LH and the ratio of LH/FSH decreased in GEN treatment groups compared to MG, the effect was statistically significant, tested by the ANOVA test (p⟨0.01). For hormone receptor activity, treat- ment with genistein resulted in an improvement of ovarian function with LHR protein expression being enhanced and FSHR protein expression being suppressed. Our results demonstrate that Genistein played a significant role in regulating FSH and LH receptor expression to improve perimenopausal ovarian and uterine function.

Growing evidence has revealed the deleterious influence of environmental and food contaminants on puberty onset and development in both animals and children, provoking an increasing health concern. T-2 toxin, a naturally-produced Type A trichothecene mycotoxin which is frequently found in cereal grains and products intended for human and animal consumption, has been shown to impair the reproduction and development in animals. Nevertheless, whether this trichothecene mycotoxin can disturb the onset of puberty in females remains unclear. To clarify this point, infantile female rats were given a daily intragastric administration of vehicle or 187.5 μg/kg body weight of T-2 toxin for five consecutive days from postnatal day 15 to 19, and the effects on puberty onset were evaluated in the present study. The results revealed that the days of vaginal opening, first dioestrus, and first estrus in regular estrous cycle were delayed following prepubertal exposure to T-2 toxin. The relative weights of reproductive organs uterus, ovaries, and vagina, and the incidence of corpora lutea were all diminished in T-2 toxin-treated rats. Serum levels of gonadotropins luteinizing hormone, follicle-stimulating hormone, and estradiol were also reduced by T-2 toxin treatment. The mRNA expressions of hypothalamic gonadotropin-releasing hormone (GnRH) and pituitary GnRH receptor displayed significant reductions following exposure to T-2 toxin, which were consistent with the changes of serum gonadotropins, delayed reproductive organ development, and delayed vaginal opening. In conclusion, the present study reveals that prepubertal exposure to T-2 toxin delays the onset of puberty in immature female rats, probably by the mechanism of disturbance of hypothalamic-pituitary-gonadal (HPG) axis function. Considering the vulnerability of developmental children to food contaminants and the relative high level of dietary intake of T-2 toxin in children, we think the findings of the present study provide valuable information for the health risk assessment in children.