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Bis(2-butoxyethyl) Phthalate Delays Puberty Onset by Increasing Oxidative Stress and Apoptosis in Leydig Cells in Rats

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Biomedical and Environmental Sciences 2023年第1期36卷 60-75页

作者： LIU Miao Qing CHEN Hai Qiong DAI Hai Peng LI Jing Jing TIAN Fu Hong WANG Yi Yan CHEN Cong De LI Xiao Heng LI Jun Wei LI Zhong Rong GE Ren Shan Department of Pediatric Surgery the Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical UniversityWenzhou 325027ZhejiangChina Key Laboratory of Structural Malformations in Children of Zhejiang Province Wenzhou 325000ZhejiangChina

Objective This study investigated the effects of bis(2-butoxyethyl) phthalate(BBOP) on the onset of male puberty by affecting Leydig cell development in rats.Methods Thirty 35-day-old male Sprague-Dawley rats were randomly allocated to five groups mg/kg bw per day that were gavaged for 21 days with BBOP at 0, 10, 100, 250, or 500 mg/kg bw per day. The hormone profiles;Leydig cell morphological metrics;mRNA and protein levels;oxidative stress;and AKT,mTOR, ERK1/2, and GSK3β pathways were assessed.Results BBOP at 250 and/or 500 mg/kg bw per day decreased serum testosterone, luteinizing hormone, and follicle-stimulating hormone levels mg/kg bw per day(P < 0.05). BBOP at 500 mg/kg bw per day decreased Leydig cell number mg/kg bw per day and downregulated Cyp11a1, Insl3, Hsd11b1,and Dhh in the testes, and Lhb and Fshb mRNAs in the pituitary gland(P < 0.05). The malondialdehyde content in the testis significantly increased, while Sod1 and Sod2 mRNAs were markedly downregulated, by BBOP treatment at 250–500 mg/kg bw per day(P < 0.05). Furthermore, BBOP at 500mg/kg bw per day decreased AKT1/AKT2, mTOR, and ERK1/2 phosphorylation, and GSK3β and SIRT1levels mg/kg bw per day(P < 0.05). Finally, BBOP at 100 or 500 μmol/L induced ROS and apoptosis in Leydig cells after 24 h of treatment in vitro(P < 0.05).Conclusion BBOP delays puberty onset by increasing oxidative stress and apoptosis in Leydig cells in rats.The graphical abstract is available on the website ***.

关键词： Bis(2-butoxyethyl)phthalate Leydig cells Testosterone Oxidative stress gonadotropin

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Testosterone undecanoate supplementation together with human chorionic gonadotropin does not impair spermatogenesis in males with isolated hypogonadotropic hypogonadism:a retrospective study

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Asian Journal of Andrology 2019年第4期21卷 413-418页

作者： Yin-Wei Chen Yong-Hua Niu Hao Xu Dao-Qi Wang Hong-Yang Jiang Gaurab Pokhrel Tao Wang Shao-Gang Wang Ji-Hong Liu Department of Urology Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 China Institute of Urology Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 China

Gon adotropin therapy is comm only used to in duce virilizati on and spermatoge nesis in male isolated hypog on adotropic hypog on adism (IHH) patients. In clinical practice, 5.6%-15.0% of male IHH patients show poor responses to gonadotropin treatment;therefore, testosterone (T) suppleme ntation can serve as an alter native therapy to no rmalize serum T levels and promote virilization. However, treatment with exogenous T impairs spermatogenesis and suppresses intratesticular T levels. This retrospective study aimed to determine whether oral testosterone undeca noate (TU) suppleme ntation together with human chorionic gonadotropin (hCG) would negatively affect spermatogenesis in IHH patients compared with hCG alone. One hundred and seven IHH patients were included in our study. Fifty-four patients received intramuscular hCG and oral TU, and 53 patients received intramuscular hCG alone. The median follow-up time was 29 (range: 12-72) mon ths in both groups. Compared with the hCG group, the hCG/TU group required a shorter median time to normalize serum T levels (P < 0.001) and achieve Tanner stage (III and V) of pubic hair and genital development (P < 0.05). However, there were no significant differences in the rate of seminal spermatozoa appearance, sperm concentration, or median time to achieve different sperm concentration thresholds between the groups. In addition, there were no significant differences in side effects, such as acne and gynecomastia, observed in both groups. This study indicates that oral TU supplementation together with hCG does not impair spermatogenesis in treated IHH patients compared with hCG alone, and it shortens the time to normalize serum T levels and promote virilization.

关键词： human chorionic gonadotropin isolated hypogonadotropic hypogonadism spermatogenesis testosterone undecanoate virilization

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Association of Thr307Ala and Asn680Ser of Follicle-Stimulating Hormone Receptor Gene Polymorphisms with gonadotropin Administration during Controlled Ovarian Hyperstimulation

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Reproductive and Developmental Medicine 2018年第2期2卷 81-87页

作者： Qing-Xia Meng Dan Song Hong Li Wei Wang Jian Ou Yong-Le Xu Ai-Yan Zheng Center for Reproduction and Genetics Suzhou Municipal HospitalNanjing Medical University Affiliated Suzhou HospitalSuzhouJiangsu 215002China

Objective:This study is to investigate the effect of different single-nucleotide polymorphisms of follicle-stimulating hormone receptor(FSHR)gene on gonadotropin(Gn)administration dosage during controlled ovarian hyperstimulation(COH)protocol of in vitro fertilization and embryo transfer.Methods:This retrospective study included 184 Chinese infertile women in Center for Reproduction and Genetics of Suzhou Municipal Hospital from June 2012 to 2014.All of the enrolled patients were homogeneous in some basal characteristics,and they all met the eligibility criteria.Blood tests were conducted on day 3 of menstrual cycle or the day of human chorionic gonadotropin administration for hormonal profile analysis and DNA extraction.DNA sequencing was performed for polymorphism analysis.The participants were classified into threonine(Thr)/Thr,Thr/alanine(Ala),and Ala/Ala groups according to genotype at position 307,and asparagine/asparagine(Asn/Asn),Asn/serine(Ser),and Ser/Ser groups according to genotype at position 680.Logistic regression and correlation analyses were performed to identify the effect of these two polymorphisms on Gn consumption.Results:The frequency of Thr307Ala and Asn680Ser distribution was consistent with Hardy-Weinberg equilibrium(P>0.05).No significant difference was found in age,basal hormone levels for different genotype groups.Logistic regression analysis results revealed that patients with Ser680Ser genotype have a higher risk of requiring a high dose of Gn compared with patients with Asn680Asn genotype,while polymorphism of Thr307 Ala has no such effect.Conclusion:This study suggested that FSHR genotype Asn680Ser would be helpful in determining the dosage of Gn in COH;patients with Ser680Ser genotype may require higher dose of Gn.

关键词： Controlled Ovarian Hyperstimulation Follicle‑Stimulating Hormone Receptor gonadotropin Single‑Nucleotide Polymorphism

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Kisspeptin stimulates oocyte maturation,and food deprivation modulates the abundance of kisspeptin system in zebrafish gonads

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Aquaculture and Fisheries 2022年第5期7卷 484-493页

作者： Azadeh Hatef Jithine Jayakumar Rajeswari Suraj Unniappan Laboratory of Integrative Neuroendocrinology Department of Veterinary Biomedical SciencesWestern College of Veterinary MedicineUniversity of SaskatchewanSaskatoonSaskatchewanS7N 5B4Canada Toxicology Centre University of SaskatchewanSaskatoonSaskatchewanS7N 5B4Canada

Kisspeptin is a hormone involved in the neuroendocrine control of reproduction in fish.We hypothesized that kisspeptin stimulates oocyte maturation and modulates other reproductive hormones in zebrafish;and the gonadal kisspeptin system in zebrafish is affected during energy unavailability.The main goals of this research were to test in vitro effects of kisspeptin on oocyte maturation and mRNA abundance in zebrafish ovarian follicles and determine how short-term feed restriction affects kisspeptin and its receptors in zebrafish testis and ovary.This study demonstrates the presence of kisspeptin and its receptors in zebrafish(Danio rerio)gonads and its direct action on ovarian follicles in vitro.Kisspeptin(10 ng/mL)induced oocyte maturation,as indicated by germinal vesicle break down at 18-and 24-hours post-treatment.Kisspeptin significantly increased the abundance of mRNAs encoding reproductive hormones and its receptors in zebrafish oocytes.This suggests that kisspeptin-10 affects ovarian functions by modulating other hormones.Reproduction is a process that requires energy.Therefore,whether energy availability affects the kisspeptin system in zebrafish gonads was determined.Food deprivation modulated kisspeptin expression differently in zebrafish testis and ovary.Kiss2 and kiss1ra were upregulated while kiss1rb was downregulated in the testis post-food deprivation.Meanwhile,no changes in kiss in the ovary were found after food deprivation.However,kiss1rb was downregulated in unfed fish at 3-and 7-days post-food deprivation.Overall,our results suggest sex-and tissue-specific changes in the gonadal abundance of the kisspeptin system in zebrafish.The fine tuning of reproduction during energy fluctuations in fish is likely mediated via changes in hormones,including kisspeptin as shown in this research.

关键词： Oocyte Kisspeptin Food deprivation GVBD gonadotropin Testis Ovary Zebrafish Oocyte maturation

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Heshouwu decoction, a Chinese herb for tonifying kidney, ameliorates hypothalamic-pituitary-testicular axis secretion in aging rats

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Neural Regeneration Research 2012年第21期7卷 1611-1617页

作者： Siyun Niu Suru Kou Xiaochun Zhou Liang Ding School of Basic Medical Science Hebei University Baoding 071000 Hebei Province China Institute of Basic Medical Sciences Chengde Medical College Chengde 067000 Hebei Province China

An increasing amount of evidence demonstrates the anti-aging effect of Heshouwu in pill form. In this study, a subacute aging rat model was established by continuous intraperitoneal injection of D-galactose and treated with Heshouwu decoction （a Chinese herb for tonifying the kidney, comprising Heshouwu pill, Herba Epimedii, Radix Salviae Miltiorrhiae, and Poria）. Heshouwu pill treated rats were the positive control group. Radioimmunoassay, immunohistochemical staining, and western blot assay showed hypothalamic gonadotropin-releasing hormone, hypothalamic substance P, and serum gonadotropin levels to be significantly increased in the model rats; the concentrations of hypothalamic ~3-endorphin, and serum levels of insulin-like growth factor I and testosterone were significantly decreased. 1713- and 3[3-hydroxysteroid dehydrogenase expression in testicular tissue was also decreased. Intragastric administration of Heshouwu decoction at high （9.6 g/mL/100 g）, medium （4.8 g/mL/100 g）, and low （2.4 g/mlJ100 g） doses, Heshouwu decoction pretreatment at a medium dose （4.8 g/mL/100 g）, and Heshouwu pill （2.06 g/mL/100 g） significantly reversed these changes. Heshouwu decoction pretreatment and high-dose Heshouwu decoction had the greatest anti-aging effects. These experimental findings indicate that Heshouwu decoction can improve hypothalamic-pituitary-testicular axis secretion in a subacute aging rat model, and prevent and delay gonadal axis aging, with an effect superior to that of Heshouwu pill.

关键词： Heshouwu decoction pituitary gonadal axis aging gonadotropin-releasing hormone gonadotropin hydroxysteroid dehydrogenase hypothalamic-pituitary-testicular axis neural regeneration

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gonadotropin and Testosterone hormone’s serum levels and partial deletions in the AZFc region in Iranian oligozoospermia infertile males

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Health 2011年第9期3卷 566-570页

作者： Nasser Salsabili Reza Mirfakhraei Maryam Montazeri Mitra Ataei Paricheher Yaghmaei Gholamreza Pourmand 不详

To investigate the relation of luteinizing hormone (LH), follicle-stimulating hormone (FSH) and Testosterone serum levels with partial deletions in the AZFc region in Iranian oligozoospermia males. Material and methods: thirty infertile oligozoospermia and 52 Iranian fertile men included. The hormonal assays were measured by the Radioimmunoassay (RIA). Multiplex polymerase chain reaction (M-PCR) using eight sequence-tagged site (STS) markers were measured on the Yq11 chromosome. Results: The mean of FSH and LH levels in all oligozoospermia males were higher than fertile men (p < 0.001) and testosterone was lower significantly (p < 0.001). Five patients showed partial deletions in AZFc region (four had gr/gr and one had b2/b3 deletions). Six fertile men showed partial deletions (five gr/gr and one b2/b3) with higher level of FSH, LH in their group (p < 0.05). Conclusion: According to high incidence of partial deletions in the AZFc region among Iranian oligozoospermia males, hormonal assay and molecular screening should be advised before considering for ART treatments.

关键词： Oligozoospermia gonadotropin Hormone AZFc Partial Deletion

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The effectiveness of zinc supplementation in men with isolated hypogonadotropic hypogonadism

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Asian Journal of Andrology 2017年第3期19卷 280-285页

作者： Yan-Ling Liu Man-Na Zhang Guo-Yu Tong Shou-YueSun Yan-Hua Zhu Ying Cao Jie Zhang Hong Huang Ben Niu Hong Li Qing-Hua Guo Yan Gao Da-Long Zhu Xiao-Ying Li Shanghai Institute of Endocrinology and Metabolism Department of Endocrinology and Metabolism Ruijin Hospital Shanghai Jiao Tong University School of Medicine Shanghai 200025 China Department of Endocrinology Drum Tower Hospital Affiliated to Nanjing University Medical School Nanjing 210008 China Department of Endocrinology The Third Affiliated Hospital Sun Yat-sen University Guangzhou 510630 China Department of Endocrinology and Metabolism Nanfang Hospital Southern Medical University Guangzhou 510515 China Department of Endocrinology Guangxi Medical University and First Affiliated Hospital Nanning 530021 China Department of Endocrinology The First People's Hospital of Yunnan Province Kunming 650032 China Department of Endocrinology The Affiliated Hospital of Guiyang Medical College Guiyang 550004 China 00853 China.

A multicenter, open-label, randomized, controlled superiority trial with 18 months of follow-up was conducted to investigate whether oral zinc supplementation could further promote spermatogenesis in males with isolated hypogonadotropic hypogonadism （IHH） receiving sequential purified urinary follicular-stimulating hormone/human chorionic gonadotropin （uFSH/hCG） replacement. Sixty-seven Chinese male IHH patients were recruited from the Departments of Endocrinology in eight tertiary hospitals and randomly allocated into the sequential uFSH/hCG group （Group A, n = 34） or the sequential uFSH plus zinc supplementation group （Group B, n = 33）. In Group A, patients received sequential uFSH （75 U, three times a week every other 3 months） and hCG （2000 U, twice a week） treatments. In Group B, patients received oral zinc supplementation （40 mg day-1） in addition to the sequential uFSH/hCG treatment given to patients in Group A. The primary outcome was the proportion of patients with a sperm concentration 〉1.0 × 106 ml-1 during the 18 months. The comparison of efficacy between Groups A and B was analyzed. Nineteen of 34 （55.9%） patients receiving sequential uFSH/hCG and 20 of 33 （60.6%） patients receiving sequential uFSH/hCG plus zinc supplementation achieved sperm concentrations ≥1.0 × 106 ml-1 by intention to treat analyses. No differences between Group A and Group B were observed as far as the efficacy of inducing spermatogenesis （P = 0.69）. We concluded that the sequential uFSH/hCG plus zinc supplementation regimen had a similar efficacy to the sequential uFSH/hCG treatment alone. The additional improvement of 40 mg day-1 oral zinc supplementation on spermatogenesis and masculinization in male IHH patients is very subtle.

关键词： gonadotropin isolated hypogonadotropic hypogonadism masculinization spermatogenesis zinc supplementation

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Emergency irradiation with 3.4 Gy/2f in sellar/suprasellar germinoma patients with rapid visual acuity decline

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Chinese Medical Journal 2019年第17期132卷 2073-2078页

作者： Bo Li You-Qi Li Chun-De Li Yao Chen Yan-Wei Liu Shuai Liu Xiao-Guang Qin Shi-Qi Luo Department of Radiation Oncology Beijing Tiantan Hospital Capital Medical University Beijing 100070 China Department of Neurosurgery Beijing Tiantan Hospital Capital Medical University Beijing 100070 China Department of Ophthalmology Beijing Tiantan Hospital Capital Medical University Beijing 100070 China

Background: Rapid visual acuity (VA) decline was a common complaint in patients with sellar/suprasellar germinoma. In our hospital, 3.4 Gy/2f of emergency irradiation was applied to save patient VA and enable subsequent chemoradiotherapy. This study aimed to investigate the efficacy of emergency irradiation with 3.4 Gy/2f in patients with sellar/suprasellar germinoma who had rapid VA decline. Methods: From January 2014 to December 2017, 33 patients with sellar/suprasellar germinoma who complained of VA decline within 3 months received 3.4 Gy/2f of emergency irradiation in Beijing Tiantan Hospital. The best-corrected VA (BCVA) and mean deviation (MD) were measured. Correlations between visual function change and clinical factors, including age at diagnosis, duration of VA decline, extent of tumor regression, serum level of tumor markers, were analyzed. Results: Among 33 patients with sellar/suprasellar germinoma, the median diameter and volume of sellar/suprasellar lesions were 32 mm (range: 5–55 mm) and 12.9 cm3 (range 0.6–58.5 cm3), respectively. Data on pre- and post-emergency-irradiation BCVA were obtained in 32 patients. For the right eyes, BCVA was improved in 23 patients (71.9%), unchanged in 7 (21.9%), and worsened in 2 (6.2%);and for the left eyes, these numbers were 27 (84.4%), 4 (12.5%), and 1 (3.1%), respectively. In terms of the logarithm of the minimum angle of resolution (logarithm of the minimum angle of resolution = Log (1/BCVA) score, the improvement was significant in both eyes (P < 0.001). In terms of MD, six patients had paired data and the improvement was marginal in the right eyes (P = 0.068) and significant in the left eyes (P = 0.043). However, no clinical factor was found to have correlation with visual function improvement. Conclusion: In sellar/suprasellar germinoma patients with VA decline, 3.4 Gy/2f of emergency irradiation was effective in improving visual function.

关键词： Germinoma Sellar/suprasellar Radiotherapy Chemotherapy Visual acuity Beta-human chorionic gonadotropin

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Hormonal Regulation of Oocyte Development and Maturation of Amphioxus

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Science China Chemistry 1994年第7期37卷 842-850页

作者：方永强赵维信林秋明 Third Institute of Oceanography SOA Shanghai Fisheries University

Using the electron microscopic technique and radioimmunoassay,the hormonal regulationof the oocyte development and maturation of the amphioxus has been investigated.The results indicate thatthe exogenous GnRH-A can stimulate the oocyte development and maturation as well as the spawning of theamphioxus.It has further been revealed by radioimmunoassay that GnRH-A can increase the content of sexsteroid hormone,especially,estradiol-17β,by 1—3 times.The increase of estradiol-17β coordinates withthe synthesis of oocyte into yolk substance.The results further justify that the hormonal regulation in theoocyte development and maturation of amphioxus is similar to that of vertebrates.This is of great theoreticalsignificance for reproduetive endocrine physiology of amphioxus.

关键词： amphioxus oocyte gonadotropin releasing hormone analogue radioimmunoassay

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Stability of Human Follicle-Stimulating Hormone Receptor mRNA in Stably Transfected Cells

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2001年第1期21卷 8-12页

作者：朱长虹田红 Family Planning Research Institute Tongji Medical College Huazhong University of Science and Technology Wuhan

In order to assess the impact of mRNA degradation on steady state levels of follicle stimulating hormone receptor (FSHR) mRNA and on regulation of FSHR gene expression, the stability and half life of FSHR mRNA were determined in transfected cells expressing recombinant FSHR. Time dependent changes in FSHR mRNA content were determined by nuclease protection solution hybridization assay (NPA) or by qualitative reverse transcription competitive polymerase chain reaction (RT PCR) in cultured hFSHR YI cells, cell lines stably transfected with a human FSHR cDNA. FSHR mRNA content remained constant during 8 h control incubations of hFSHR Y1 cells (NPA, 2.9±0.3 μg/mg RNA; RT PCR, 2.7±0.3 μg/mg RNA). Actinomycin D (ActD, 5 μg/ml) inhibited mRNA synthesis, as assessed by incorporation of uridine into total RNA, by 90 % within 1 h in hFSHR Y1 cells. No effect of ActD on cellular morphology or viability was observed. ActD caused a time dependent decrease in FSHR mRNA content in hFSHR Y1 cell lines with a lag time of 1 h. There were no significant differences in the rate of FSHR mRNA degradation between the two methods of mRNA quantification. The half life of hFSHR mRNA was 3.6±0.2 h by NPA and 3.1±0.1 h by RT PCR. The results indicated that degradation of mRNA was an important process in maintenance of steady state expression of the FSHR gene in cells stably expressing recombinant receptor.

关键词： gonadotropin granulosa cell folliculogenesis follicle stimulating hormone mRNA

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