促卵泡激素受体结合抑制剂抑制小鼠卵巢发育
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摘要
目的探讨不同浓度的促卵泡激素受体结合抑制剂(FRBI)对小鼠卵巢发育、生殖激素分泌及其蛋白表达的影响。方法给FRBI组小鼠分别肌肉注射20、30和40 mg/kg FRBI;促卵泡激素(FSH)组注射10 IU/kg FSH;对照组(CG)注射0.2 mL 0.9%氯化钠溶液,每组均连续注射5 d。精确称量每只小鼠体质量和双侧卵巢质量;测定卵巢组织切片中皮质厚度(OCT)和次级卵泡形态大小;ELISA测定血清FSH浓度;Western blot检测卵巢FSH受体(FSHR)表达。结果实验第15天起,FRBI组小鼠卵巢质量低于CG和FSH组(P<0.05),以FRBI-30 mg组小鼠卵巢质量最低;FRBI组OCT、次级卵泡最大纵径(MLD)和最大横径(MTD)低于CG组和FSH组(P<0.05),FRBI-40 mg组小鼠卵巢的MLD和MTD值最低;第15天时,FRBI-30 mg组小鼠血清FSH的浓度最低;随着FRBI注射剂量的增加,卵巢中FSHR表达水平逐渐降低,FRBI组的FSHR蛋白水平明显低于FSH组(P<0.05)。结论 FRBI能抑制卵巢和卵泡发育,降低血清FSH浓度,下调卵巢FSHR蛋白表达水平。
Objective To evaluate the effects of different concentrations of follicle stimulating hormone receptor binding inhibitor(FRBI) on ovarian development, reproductive hormone secretion and protein expression in mice. Methods The mice of FRBI groups were intramuscularly injected with 20, 30 and 40 mg/kg FRBI, respectively. Mice of FSH group were intramuscularly injected with 10 IU/mL FSH. Mice of the control group(CG) were injected with 0.20 mL normal saline. Each group was injected daily for 5 days. The body weight and ovarian weights of five groups mice were accurately weighted. Ovarian cortex thickness(OCT)and secondary follicles were observed and measured under the light microscope. ELISA was used to determine the serum concentrations of FSH. Expression levels of FSH receptor(FSHR) in the ovarian tissues were detected using Western blot. Results There was no significant differences in body weights among all groups. Ovarian weights of FRBI groups were significantly lower than that of control group(P<0.05) after day 15 on, with the least value of FRBI-30 mg group. The OCT,maximum longitudinal diameter(MLD) and maximum transverse diameter(MTD) of three experiment groups were less than those of CG and FSH group( P < 0. 05). The smallest values of MLD and MTD were found in the FRBI-40 mg group. Serum concentration of FSH FRBI-20 mg group was the lowest. Expression level of FSHR proteins in the ovaries was gradually reduced along with the increase of FRBI doses. They were significantly less than that of FSH group( P<0. 01). Conclusions FRBI may dose-dependently inhibit the growth and development of mice ovaries,and reduce serum FSH concentration,additionally attenuate expression levels of FSHR protein in ovaries.
Objective To evaluate the effects of different concentrations of follicle stimulating hormone receptor binding inhibitor(FRBI) on ovarian development, reproductive hormone secretion and protein expression in mice. Methods The mice of FRBI groups were intramuscularly injected with 20, 30 and 40 mg/kg FRBI, respectively. Mice of FSH group were intramuscularly injected with 10 IU/mL FSH. Mice of the control group(CG) were injected with 0.20 mL normal saline. Each group was injected daily for 5 days. The body weight and ovarian weights of five groups mice were accurately weighted. Ovarian cortex thickness(OCT)and secondary follicles were observed and measured under the light microscope. ELISA was used to determine the serum concentrations of FSH. Expression levels of FSH receptor(FSHR) in the ovarian tissues were detected using Western blot. Results There was no significant differences in body weights among all groups. Ovarian weights of FRBI groups were significantly lower than that of control group(P<0.05) after day 15 on, with the least value of FRBI-30 mg group. The OCT,maximum longitudinal diameter(MLD) and maximum transverse diameter(MTD) of three experiment groups were less than those of CG and FSH group( P < 0. 05). The smallest values of MLD and MTD were found in the FRBI-40 mg group. Serum concentration of FSH FRBI-20 mg group was the lowest. Expression level of FSHR proteins in the ovaries was gradually reduced along with the increase of FRBI doses. They were significantly less than that of FSH group( P<0. 01). Conclusions FRBI may dose-dependently inhibit the growth and development of mice ovaries,and reduce serum FSH concentration,additionally attenuate expression levels of FSHR protein in ovaries.
引文
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[8] Chitnis SS, Selvaakumar C, Jagtap DD, et al. Interaction of Follicle-stimulating hormone (FSH) receptor binding inhibitor-8: a novel fsh-binding inhibitor, with fsh and its receptor [J]. Chem Biol Drug Design, 2009, 73:637-643.
[9] Zhang H, Cheng GH, Li YJ, et al. Superovulation and expression of follicle-stimulating hormone receptor in young rabbit females [J]. World Rabbit Sci, 2017, 25:167-172.
[10] Ghezelayagh Z, Totonchi M, Zarei-Moradi S, et al. The impact of genetic variation and gene expression level of the follicle-stimulating hormone receptor on ovarian reserve [J]. Cell J, 2018, 19: 620-626.
[11] Kong D, Guan Q, Li G, et al. Expression of FSHR in chondrocytes and the effect of FSH on chondrocytes [J]. Biochem Biophy Res Commun, 2018: 495: 587-593.
[12] Poljak Z, Hulin I, Maruscakova L, et al. Are GnRH and FSH potentially damaging factors in the cardiovascular system? [J].Pharmazie. 2018, 73: 187-190.
[13] Gong Z, Liang H, Deng Y, et al. FSH receptor binding inhibitor influences estrogen production, receptor expres-sion and signal pathway during in vitro maturation of sheep COCs [J]. Theriogenology. 2017, 101:144-150.
[2] Alfredo PP, Nikolaos S, Melanie RR, et al. Follicle stimulating hormone receptor is expressed by most ovarian cancer subtypes and is a safe and effective immunotherapeutic target [J]. Clinic Cancer Res, 2017, 23: 441-453.
[3] Donga C, Chen J, Zhang X, et al. Follicle stimulating hormone receptor mediated gene delivery to target ovarian carcinoma in vitro [J]. Asian Biomedicine, 2017, 6:185-194.
[4] Patel H. Bhartiya D. Testicular stem cells express follicle-stimulating hormone receptors and are directly modulated by FSH [J]. Reprod Sci, 2016, 23: 1493-1508.
[5] Navalakhe RM, Jagtap DD, Nayak SU, et al. Effect of FSH receptor-binding inhibitor-8 on FSH-mediated granulosa cell signaling and proliferation [J]. Chem Biol Drug Design, 2013, 82:178-188.
[6] Bramble MS, Goldstein EH, Lipson A, et al. A novel follicle-stimulating hormone receptor mutation causing primary ovarian failure: a fertility application of whole exome sequencing [J]. Hum Reprod, 2016, 31:905-914.
[7] Perales-Puchalt A, Svoronos N, Rutkowski MR, et al. Follicle-stimulating hormone receptor is expressed by most ovarian cancer subtypes and is a safe and effective immunotherapeutic target [J]. Clinic Cancer Res, 2017, 23:441-453.
[8] Chitnis SS, Selvaakumar C, Jagtap DD, et al. Interaction of Follicle-stimulating hormone (FSH) receptor binding inhibitor-8: a novel fsh-binding inhibitor, with fsh and its receptor [J]. Chem Biol Drug Design, 2009, 73:637-643.
[9] Zhang H, Cheng GH, Li YJ, et al. Superovulation and expression of follicle-stimulating hormone receptor in young rabbit females [J]. World Rabbit Sci, 2017, 25:167-172.
[10] Ghezelayagh Z, Totonchi M, Zarei-Moradi S, et al. The impact of genetic variation and gene expression level of the follicle-stimulating hormone receptor on ovarian reserve [J]. Cell J, 2018, 19: 620-626.
[11] Kong D, Guan Q, Li G, et al. Expression of FSHR in chondrocytes and the effect of FSH on chondrocytes [J]. Biochem Biophy Res Commun, 2018: 495: 587-593.
[12] Poljak Z, Hulin I, Maruscakova L, et al. Are GnRH and FSH potentially damaging factors in the cardiovascular system? [J].Pharmazie. 2018, 73: 187-190.
[13] Gong Z, Liang H, Deng Y, et al. FSH receptor binding inhibitor influences estrogen production, receptor expres-sion and signal pathway during in vitro maturation of sheep COCs [J]. Theriogenology. 2017, 101:144-150.