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gnrh impairs diabetic wound healing through enhanced NETosis

Cellular & Molecular Immunology 2020年第8期17卷 856-864页

作者： Yun Sang Lee Sung Un Kang Myung-Hoon Lee Haeng-Jun Kim Chang-Hak Han Ho-Ryun Won Young Uk Park Chul-Ho Kim Department of Otolaryngology School of MedicineAjou UniversitySuwonRepublic of Korea Department of Molecular Science and Technology Ajou UniversitySuwonRepublic of Korea Department of Otolaryngology School of MedicineChungnam National UniversityDaejeonRepublic of Korea Department of Orthopedic Surgery Ajou UniversitySuwonRepublic of Korea

It has been reported that neutrophil extracellular traps(NETs)impair wound healing in diabetes and that inhibiting NET generation(NETosis)improves wound healing in diabetic ***-releasing hormone(gnrh)agonists are associated with a greater risk of ***,the role of gnrh in diabetic wound healing is *** determined whether gnrh-promoted NETosis and induced more severe and delayed diabetic wound healing.A mouse model of diabetes was established using five injections with *** with blood glucose levels>250 mg/dL were then used in the *** agonist treatment induced delayed wound healing and increased NETosis at the skin wounds of diabetic *** contrast,gnrh antagonist treatment inhibited gnrh agonist-induced delayed wound *** expression of NETosis markers PAD4 and citrullinated histone H3 were increased in the gnrh-treated diabetic skin wounds in diabetic mice and *** vitro experiments also showed that neutrophils expressed a gnrh receptor and that gnrh agonist treatment increased NETosis markers and promoted phorbol myristate acetate(PMA)-induced NETosis in mouse and human ***,gnrh antagonist treatment suppressed the expression of NETosis markers and PMA-induced NETosis,which were increased by gnrh *** results indicated that gnrh-promoted NETosis and that increased NETosis induced delayed wound healing in diabetic skin ***,inhibition of gnrh might be a novel treatment of diabetic foot ulcers.

关键词： Neutrophils gnrh Diabetic wound healing NETosis

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Immunization of Male Mice with a New Recombinant gnrh Fusion Protein Reduces the Testicular Function

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Agricultural Sciences in China 2009年第3期8卷 380-385页

作者： FANG Fu-gui YANG Ya-ping LIU Ya ZHANG Yun-hai TAO Yong WANG Suo-lu PU Yong ZHANG Xiao-rong Department of Animal Medicine College of Animal Science and Technology Anhui Agricultural University Hefei 230036 P.R.China Department of Animal Science College of Animal Science and Technology Anhui Agricultural University Hefei 230036 P.R.China

The objective of the present study was to evaluate the effect of an immunogenic maltose-binding protein-gonadotropin releasing hormone （gnrh-6-MBP） using genetic engineering. The synthetic mammalian tandem repeated gnrh hexamer gene was inserted into the expression plasmid pMAL-c2x. Recombinant gnrh-6-MBP protein was over- expressed in *** strain BL21. Amylose resin with affinity chromatograph was used to purify target protein. The reactiongenicity of fusion protein was identified by indirect enzyme linked immunosorbent assay （ELISA）, and the antigenicity and biological effects of gnrh-6-MBP were tested in mice. In the experiment, 20 male Kunming white mice of 20 d old were randomly divided into treatment and control group. Ten mice were immunized with 100 μg gnrh-6-MBP administered subcutaneously （s.c.） thrice at 2-week intervals with gnrh-6-MBP. Mice were sacrificed after 3 weeks following the booster injection, the testis was removed, weighed and measured, and the histological structure was observed. The reactiongenicity of fusion protein to gnrh antibody was much higher than the control. Active immunization against gnrh-6-MBP reduced remarkably （P 〈 0.01） the length and weight of the testis, and shortened the girth and width of the testis （P 〈 0.05）, and suppressed testicular spermatogenesis compared to the control mice. These results indicate that the recombinant gnrh-6-MBP acted as a strong immunogen and caused atrophy of the testis.

关键词： gnrh recombinant vaccine maltose-binding protein (MBP) immunocastration

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Effect of CD14/TLR4 antagonist on gnrh/LH secretion in ewe during central inflammation induced by intracerebroventricular administration of LPS

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Journal of Animal Science and Biotechnology 2018年第4期9卷 858-867页

作者： Karolina Haziak Andrzej Przemyslaw Herman Karolina Wojtulewicz Bartosz Pawlina Kamila Paczesna Joanna Bochenek Dorota Tomaszewska-Zaremba The Kielanowski Institute of Animal Physiology and Nutrition Polish Academy of Sciences05-110 JablonnaPoland

Background: Immune stress induced by lipopolysaccharide(LPS) influences the gonadotropin-releasing hormone(gnrh)/luteinizing hormone(LH) secretion. Presence of LPS interacting Toll-like receptor(TLR) 4 in the hypothalamus may enable the direct action of LPS on the gnrh/LH secretion. So, the aim of the study was to investigate the influence of intracerebroventricular(icv) injection of TLR4 antagonist on gnrh/LH secretion in anestrous ewes during LPS-induced central inflammation. Animals were divided into three groups icv-treated with: Ringer-Locke solution, LPS and TLR4 antagonist followed by ***: It was demonstrated that TLR4 antagonist reduced LPS-dependent suppression of gnrh gene expression in the preoptic area and in the medial basal hypothalamus, and suppression of receptor for gnrh gene expression in the anterior pituitary gland. It was also shown that TLR4 antagonist reduced suppression of LH release caused by icv injection of LPS. Central administration of LPS stimulated TLR4 gene expression in the medial basal ***: It was indicated that blockade of TLR4 prevents the inhibitory effect of centrally acting LPS on the gnrh/LH secretion. This suggests that some negative effects of bacterial infection on the hypothalamic-pituitary-gonadal axis activity at the hypothalamic level may be caused by central action of LPS acting through TLR4.

关键词： Ewe gnrh LH LPS TLR4 antagonist

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Characterization and differential expression of three gnrh forms during reproductive development in cultured turbot Schophthalmus maximus

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Journal of Oceanology and Limnology 2018年第4期36卷 1360-1373页

作者： ZHAO Chunyan XU Shihong FENG Chengcheng LIU Yifan YANG Yang WANG Yanfeng XIAO Yongshuang SONG Zongcheng LIU Qinghua LI Jun Key Laboratory of Experimental Marine Biology Institute of Oceanology Chinese Academy of Sciences Qingdao 266071 China University of Chinese Academy of Sciences Beijing 100049 China Laboratory for Marine Biology and Biotechnology Qingdao National Laboratory for Marine Science and Technology Qingdao 266237 China Weihai Shenghang Aquatic Product Science and Technology Co. Ltd. Weihai 264200 China

Turbots( Schophthalmus maximus), one of the most important economic marine flatfish species, fail to undergo final spawning and spermiation naturally under artificial farming conditions. In vertebrates, reproduction is regulated by the brain-pituitary-gonadal axis(BPG-axis), and gonadotropin releasing hormone(gnrh) is one of its key components. Therefore, to better understand the physiology of reproduction in the turbot, three of the genes encoding gnrh subtypes— sbgnrh, c gnrh-II and sgnrh —were cloned and sequenced by isolating the cDNA sequences. The localizations and patterns of expression of their mRNAs were also evaluated during seasonal gonadal development. All three mRNAs were expressed abundantly in the brain; sbgnrh and sgnrh mRNAs were also detected in the gonads and pituitary gland, and sbgnrh expression was much higher than that of sgnrh, indicating the critical role of sbgnrh in regulating the BPG-axis. Moreover, the brain expression patterns of sbgnrh and sgnrh mRNAs showed an increased trend during gonadal development, peaking in mature stages. This indicated the direct regulation of gonadal development by the gnrh system. In addition, c gnrh-II mRNA expression showed no significant variations, suggesting that c gnrh-II is not critically involved in the control of reproduction. Further, the mRNA abundances of the three gnrh forms in the breeding season were significantly higher than those in immature and post-breeding stages in all analyzed brain areas. Therefore, we propose that sbgnrh is the most important hormone for the regulation of reproduction in turbot via the BPG-axis. These results will help in better understanding the reproductive endocrine mechanisms of turbots and lay the groundwork for additional studies aimed at comparing the reproductive physiology of wild individuals with those raised under artificial conditions.

关键词： turbot gnrh brain gonad reproductive development

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Optimized functional and structural design of dual-target LMRAP,a bifunctional fusion protein with a 25-amino-acid antitumor peptide and gnrh Fc fragment

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Acta Pharmaceutica Sinica B 2020年第2期10卷 262-275页

作者： Meng Li Hanmei Xu Junzhi Wang Shenyang Pharmaceutical University Shenyang 110016China Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech Products National Institutes for Food and Drug ControlBeijing 102629China State Key Laboratory of Natural Medicines Ministry of Educationthe Engineering Research Center of Synthetic Polypeptide Drug Discovery and Evaluation of Jiangsu ProvinceDepartment of Marine PharmacyChina Pharmaceutical UniversityNanjing 211198China

To develop fusion protein of a gnrh Fc fragment and the integrin targeting AP25 antitumor peptide for gnrh receptor-expressing cancer *** LMRAP fusion protein was constructed.A transwell invasion assay was *** gene mRNA and protein levels of gnrhR-I,α5β1,andαvβ3 in different cancer cell lines were *** proliferation was measured using a cell counting *** antagonist assay was performed on gnrh ***-tumor activity was evaluated with a mouse xenograft tumor ***(IHC)was applied to detect CD31 and CD34 *** characteristics were determined with an indirect competition *** developed bifunctional fusion protein LMRAP not only inhibited HUVEC invasion,but also inhibited proliferation of gnrhR-I,α5β1,and high expression cancer *** IC50 for LMRAP in the gnrh receptor was 6.235 x 10-4 mol/*** significantly inhibited human prostate cancer cell line22 RV1 proliferation in vivo and in *** significantly inhibited CD31 and CD34 *** elimination half-life of the fusion protein LMRAP was 33 h in *** fusion protein made of a gnrh Fc fragment and the integrin targeting AP25 peptide retained the bifunctional biological activity of gnrhR blocking,angiogenesis inhibition,prolonged half-life and good tolerance.

关键词： Fusion protein gnrh Integrin Angiogenesis Prostate cancer

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Active immunization with gnrh-tandem-dimer peptide in young male rats reduces serum reproductive hormone concentrations, testicular development and spermatogenesis

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Asian Journal of Andrology 2016年第3期18卷 485-491,I0012页

作者： Xing-Fa Han Jun-Li Li Yu-Qin Zhou Xiao-Hua Ren Gong-Cheng Liu Xiao-Han Cao Xiao-Gang Du Xian-Yin Zeng Isotope Research Biological Engineering and Application Biology Department Laboratory Sichuan Agricultural University Ya'an 625014 People's Republic of China.

gnrh sterilization vaccines have been developed for various practical and clinical reasons. However, conjugation of gnrh peptide to carrier protein has many drawbacks, hampering the further commercialization of gnrh vaccines. In this study, a new nonconjugated gnrh vaccine, D-Lys6-gnrh-tandem-dimer peptide （TDK）, emulsified in Specol adjuvant was investigated for its immunocastration efficacy in young male rats. Prepubertal male rats were randomly allocated into three groups （n = 12）： control （no treatment）, surgically castrated or immunized against 100 μg TDK in Specol adjuvant at 6 weeks of age （with a booster 8 weeks later）. Blood samples （for antibody titers and hormone concentrations） were collected at 2-week intervals until rats were killed （18 weeks of age）. Compared to intact controls, active immunization against TDK reduced （P〈 0.05） serum concentrations of testosterone, inhibin B, LH and FSH, prevented the onset of spermatogenesis at puberty. Furthermore, mRNA expressions of gnrh receptor, LH-β and FSH-β in the pituitary, LH receptor, FSH receptor, inhibin α, βA and βB subunit in the testes were decreased in immunocastrated rats compared to intact controls （P 〈 0.05）. These results demonstrate for the first time that gnrh-tandem-dimer peptide emulsified in Specol is a promising veterinary sterilization medicine.

关键词： gnrh nonconjugation rat reproductive axis sterilization

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A COMPARATIVE STUDY ON INVOLVEMENT OF tPA ACTIVITY IN OVULATION INDUCED BY hCG AND gnrh AGONIST IN HYPOPHYSECTOMIZED RATS

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Science China Chemistry 1991年第10期34卷 1215-1224页

作者：刘以训冯强 CAJANDER STEFAN Laboratory of Reproductive Biology and Department of Endocrinology Institute of Zoology Academia Sinica Beijing 100080 PRC Department of Pathology Ume UniversityS-90187UmeSweden

A gnrh agonist (5-50 μg) is capable of inducing ovulation in PMSG-primed hypophy-sectomized immature rats, as is the case in hCG-induced ovulation, but 2-4 h earlier than hCG. gnrh-induced ovulation is effectively blocked by the concomitant administration of the gnrh-antagonist which failed to interfere with hCG-induced ovulation, indicating that gnrh and its agonists do not share a receptor with LH/hCG. Like hCG, gnrh is also capable of inducing tissue type (tPA), but not urokinase type (uPA) PA. The plasminogen activator activity in ovarian homogenates and the granulosa and theca-interstitial cells increase in a time-dependent manner, reaching maximum levels just prior to ovulation. Similar to hCG, gnrh also increases tPA activity in cumulus-oocyte complexes in a time-dependent fashion.

关键词： hCG gnrh agonist hypophysectomized rat ovulation tPA.

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A Second Dose of gnrha in Combination with Luteal gnrh Antagonist May Eliminate Ovarian Hyperstimulation Syndrome in Women with≥30 Follicles Measuring≥11 mm in Diameter on Trigger Day and/or Pre-trigger Peak Estradiol Exceeding 10 000 pg/mL

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Current Medical Science 2019年第2期39卷 278-284页

作者： Ling DENG Xue-Ian LI De-sheng YE Christophe Blockeel Xing-yu ZHOU Shi-ling CHEN Xin CHEN Center for Reproductive Medicine Department of Gynecology and ObstetricsNanfang HospitalSouthern Medical UniversityGuangzhou 510515China Department of Reproductive Medicine Jiangmen Central HospitalJiangmen 529030China Centre for Reproductive Medicine Universitair Ziekenhuis Brussel(UZ Brussel)Laarbeeklaan 101-1090 BrusselsBelgium

This observational study included 21 patients at remarkably high risk of ovarian hyperstimulation syndrome(OHSS),characterized by more than 30 follicles measuring≥11 mm in diameter on trigger day and/or pre-trigger peak estradiol exceeding 10 000 pg/*** was also the feature of women with established severe early OHSS followed by gonadotrophin-releasing hormone agonist(gnrha)trigger and freeze-all policy that previously have been *** patients received a second dose of gnrha 12 h after the first gnrha trigger combined with administration of gnrh antagonist at 0.25 mg/day for a period of 3 days from the day of oocyte retrieval *** in vitro fertilization(IVF)outcomes may be preferable compared with a bolus of gnrha trigger and none of the included patients developed moderate-to-severe ***,patients'symptoms,reproductive honnone levels and ultrasound findings were improved *** new strategy seems to be efficacious and could be a further supplement of gnrha trigger with or without applying freeze-all strategy to completely prevent early-onset moderate to severe OHSS,especially for the patients characterized by≥30 follicles measuring≥11 mm in diameter on trigger day and/or pre-trigger peak estradiol exceeding 10 000 pg/*** studies should be performed to compare this regimen with conventional methods of OHSS prevention.

关键词： in-vitro fertilization ovarian hyperstimulation syndrome gnrh agonist gnrh antagonist freeze-all

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PARADOXICAL EFFECT OF A gnrh AGONIST ON STEROIDOGENESIS IN CULTURED MONKEY GRANULOSA CELLS

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Science China Chemistry 1991年第12期34卷 1452-1460页

作者：刘以训胡召元冯强邹如金 Department of Endocrinology and Laboratory of Reproductive Biology Institute of ZoologyAcademia Sinica Beijing 100080 PRC Department of Endocrinology and Laboratory of Reproductive Biology Institute of ZoologyAcademia Sinica Beijing 100080 PRC Kunming Institute of ZoologyKunmingChina.

In this study we demonstrate: (i) The gnrh agonist exerts a direct dose-dependet stimulative effect on the aromatase activity and progesterone production in cultured monkey granulosa cells; (ii)the stimulative effect on steroidogenesis can be completely blocked by concomitant treatment with a gnrh antagonist, suggesting that the actions of gnrh are mediated through stringent stereospecific recongnition sites; (iii) in addition to the stimulative effect, the gnrh agonist in the presence of gonadotropins also exerts an inhibitory effect, even though the peptide by itself is more effective in the stimulation of steroidogenesis, and the stimulation of gonadotropin on steroidogenesis could be gradually restored by decreasing the concentration of the gnrh agonist in the culture; and (iv) paradoxical effect can also be observed in the presence of cAMP-inducing agents, suggesting that the inhibitory action of the peptide on gonadotropin-induced steroidogenesis is localized at a step distal to the stringent recognition sites.

关键词： gnrh agonist rhesus monkey granulosa cells paradoxical effect steroidogenesis.

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Flexible gnrh Antagonist Protocol versus Progestin-primed Ovarian Stimulation (PPOS) Protocol in Patients with Polycystic Ovary Syndrome: Comparison of Clinical Outcomes and Ovarian Response

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Current Medical Science 2019年第3期39卷 431-436页

作者： Zhuo-ni XIAO Jia-li PENG Jing YANG Wang-ming XU Center for Reproductive Medicine Renmin Hospital of Wuhan University Wuhan 430060China

Polycystic ovary syndrome (PCOS) is one of the most common causes of infertility in women. Progestin-primed ovarian stimulation (PPOS) protocol, which used oral progestin to prevent premature luteinizing hormone (LH) surges in ovarian stimulation, has been proved to be effective and safe in patients with PCOS. The aim of the present study was to compare the efficacy of PPOS protocol with that of the traditional gonadotropin-releasing hormone (gnrh) antagonist protocol in patients with PCOS. A total of 157 patients undergoing in-vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) were recruited into this study. The patients were divided into two groups by the stimulation protocols: the gnrh antagonist protocol group and the PPOS protocol group. There was no significant difference in the clinical characteristics between the two groups. Dose and duration of gonadotropin were higher in the PPOS protocol group. Estradiol levels on the day of human chorionic gonadotropin (hCG) administration were significantly lower in the PPOS protocol group. Fertilization rates and the number of good quality embryos were similar between the two groups. Remarkably, we found 6 patients with moderate ovarian hyperstimulation syndrome (OHSS) in the gnrh antagonist protocol group but 0 in the PPOS protocol group. A total of 127 women completed their frozen embryo transfer (FET) cycles. There were no significant differences between the two groups in terms of clinical pregnancy rate per transfer, implantation rate, first-trimester miscarriage rate and on-going pregnancy rate per transfer. To conclude, PPOS protocol decreased the incidence of OHSS without adversely affecting clinical outcomes in patients with PCOS.

关键词： polycystic ovary syndrome ovarian hyperstimulation syndrome progestin-primed ovarian stimulation gnrh antagonist protocol controlled ovarian hyperstimulation

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