Background： In the current society, infertility related to age has become a social problem. The in vitro fertilization （IVF） success rate in women with poor ovarian response （POR） is very low. Dandelion extract T...Background： In the current society, infertility related to age has become a social problem. The in vitro fertilization （IVF） success rate in women with poor ovarian response （POR） is very low. Dandelion extract T-1 （DE-T1） is an effective component of the extract from the leaves and stems of Traxacum officinale, which is one of the medicines used in some patients with POR, but its molecular mechanism remains unclear. Methods： Following IVF, ovarian granulosa cells （GCs） of sixty patients were extracted and divided into normal ovarian response （NOR） and POR groups. GCs were cultured in a dose-dependent and time-dependent manner with DE-TI, proliferation of GCs was determined by Cell Counting Kit-8 assay, and mRNA levels of insulin-like growth factor 1 receptor （IGF-1R）, luteotropic hormone receptor （LHR）, follicle-stimulating hormone receptor （FSHR）, LHR, and CYP19A1 （aromatase） were determined by quantitative polymerase chain reaction. Progesterone and estradiol （E2） concentrations were determined by enzyme-linked immunosorbent assay. Results： The cell viability gradually increased with the progressive increase in the DE-T1 concentration. Compared with the control group （without DE-T1）, the mRNA expressions of FSHR, LHR, IGF-IR, and CYPIgAI were upregulated after the addition of DE-T l, especially in the 2.5% DE-T 1 group （P 〈 0.01 ）. The expression of IGF- 1R was upregulated approximately 25 times （24.97 ± 4.02 times） in the POR group with 2.5% DE-T1. E2 and progesterone levels increased with the increasing DE-T1 concentration. There were highly significant differences in the E2 and progesterone secretion between the NOR and POR groups （P 〈 0.01）. Conclusion： DE-TI may promote steroid hormone synthesis by promoting GC proliferation and upregulating GC receptor expression, thereby improving ovarian endocrine function.展开更多
Follicle-stimulating hormone receptor （FSHR）, which is expressed only on Sertoli cells and plays a key role in spermatogenesis, has been paid attention for its potential in male contraception vaccine research and de...Follicle-stimulating hormone receptor （FSHR）, which is expressed only on Sertoli cells and plays a key role in spermatogenesis, has been paid attention for its potential in male contraception vaccine research and development. This study introduces a method for the preparation and purification of human FSHR 57-amino acid protein （FSHR-57aa） as well as determination of its immunogenicity and antifertility effect. A recombinant pET-28a（＋）-FSHR-57aa plasmid was constructed and expressed in Escherichia coil strain BL21 StarTM （DE3） and the FSHR-57aa protein was separated and collected by cutting the gel and recovering activity by efficient refolding dialysis. The protein was identified by Western blot and high-performance liquid chromatography analysis with a band of nearly 7 kDa and a purity of 97.4%. Male monkeys were immunized with rhFSHR-57aa protein and a gradual rising of specific serum IgG antibody was found which reached a plateau on day 112 （16 weeks） after the first immunization. After mating of one male with three female monkeys, the pregnancy rate of those mated with males immunized against FSHR-57aa was significantly decreased while the serum hormone levels of testosterone and estradiol were not disturbed in the control or the FSHR-57aa groups. By evaluating pathological changes in testicular histology, we found that the blood-testis barrier remained intact, in spite of some small damage to Sertoli cells. In conclusion, our study demonstrates that the rhFSHR-57aa protein might be a feasible male contraceptive which could affect sperm production without disturbing hormone levels.展开更多
文摘Background： In the current society, infertility related to age has become a social problem. The in vitro fertilization （IVF） success rate in women with poor ovarian response （POR） is very low. Dandelion extract T-1 （DE-T1） is an effective component of the extract from the leaves and stems of Traxacum officinale, which is one of the medicines used in some patients with POR, but its molecular mechanism remains unclear. Methods： Following IVF, ovarian granulosa cells （GCs） of sixty patients were extracted and divided into normal ovarian response （NOR） and POR groups. GCs were cultured in a dose-dependent and time-dependent manner with DE-TI, proliferation of GCs was determined by Cell Counting Kit-8 assay, and mRNA levels of insulin-like growth factor 1 receptor （IGF-1R）, luteotropic hormone receptor （LHR）, follicle-stimulating hormone receptor （FSHR）, LHR, and CYP19A1 （aromatase） were determined by quantitative polymerase chain reaction. Progesterone and estradiol （E2） concentrations were determined by enzyme-linked immunosorbent assay. Results： The cell viability gradually increased with the progressive increase in the DE-T1 concentration. Compared with the control group （without DE-T1）, the mRNA expressions of FSHR, LHR, IGF-IR, and CYPIgAI were upregulated after the addition of DE-T l, especially in the 2.5% DE-T 1 group （P 〈 0.01 ）. The expression of IGF- 1R was upregulated approximately 25 times （24.97 ± 4.02 times） in the POR group with 2.5% DE-T1. E2 and progesterone levels increased with the increasing DE-T1 concentration. There were highly significant differences in the E2 and progesterone secretion between the NOR and POR groups （P 〈 0.01）. Conclusion： DE-TI may promote steroid hormone synthesis by promoting GC proliferation and upregulating GC receptor expression, thereby improving ovarian endocrine function.
基金This study was supported by the National Key Technologies R and D Program （No. 2012BAI31B07 and No. 2006BAI03B12）, the National Science Foundationof China （No. 81172694）. This project was also funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions.
文摘Follicle-stimulating hormone receptor （FSHR）, which is expressed only on Sertoli cells and plays a key role in spermatogenesis, has been paid attention for its potential in male contraception vaccine research and development. This study introduces a method for the preparation and purification of human FSHR 57-amino acid protein （FSHR-57aa） as well as determination of its immunogenicity and antifertility effect. A recombinant pET-28a（＋）-FSHR-57aa plasmid was constructed and expressed in Escherichia coil strain BL21 StarTM （DE3） and the FSHR-57aa protein was separated and collected by cutting the gel and recovering activity by efficient refolding dialysis. The protein was identified by Western blot and high-performance liquid chromatography analysis with a band of nearly 7 kDa and a purity of 97.4%. Male monkeys were immunized with rhFSHR-57aa protein and a gradual rising of specific serum IgG antibody was found which reached a plateau on day 112 （16 weeks） after the first immunization. After mating of one male with three female monkeys, the pregnancy rate of those mated with males immunized against FSHR-57aa was significantly decreased while the serum hormone levels of testosterone and estradiol were not disturbed in the control or the FSHR-57aa groups. By evaluating pathological changes in testicular histology, we found that the blood-testis barrier remained intact, in spite of some small damage to Sertoli cells. In conclusion, our study demonstrates that the rhFSHR-57aa protein might be a feasible male contraceptive which could affect sperm production without disturbing hormone levels.
