[Effects of soy isoflavones and major active component genistein on the expression of ovarian estrogen receptor-α in rats].

OBJECTIVE: To investigate the effects of soy isoflavones (SI) on the expression of estrogen receptor-α (ER-α) in senile rat ovaries and ovarian granulosa cell cultured in vitro treated with genistein, a major active component of SI. METHODS: The animal model of perimenopause rats was established by unforced aging. The animals were treated by intragastric administration (ig) with low (50 mg/kg), middle (158 mg/kg) and high (500 mg/kg) dose of SI for 8 weeks. The expressions of ER-α mRNA and protein were detected by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry respectively. The granulosa cells of rat ovaries were isolated and administered with genistein (0, 0.1, 1, 5, 10, 100 µmol/L) for 48 h and the expression levels of ER-α mRNA detected by RT-PCR. RESULTS: The ER-α mRNA expression levels of the low, middle and high dose groups of SI (0.207 ± 0.014, 0.316 ± 0.073 and 0.402 ± 0.170 respectively) were higher than those of the model group (0.671 ± 0.170) (all P < 0.01). The expression levels of ER-α protein for the low, middle and high dose groups of SI (7.35 ± 4.90, 13.90 ± 5.12 and 23.79 ± 10.31 respectively) were higher than those of the model group (2.74 ± 0.09) (all P < 0.01). The expression levels of ER-α mRNA in granulosa cells treated with 1, 5, 10 µmol/L genistein for 48 h were 0.927 ± 0.232, 1.067 ± 0.154, 1.118 ± 0.126 respectively (all P < 0.01). They were higher than those of the control group (0.671 ± 0.170). But the expression levels of 100 µmol/L genistein group were lower than those of the control group (P < 0.05). CONCLUSION: Soy isoflavones can up-regulate the expressions of ER-α mRNA and protein in senile rat ovaries. As a major active component of soy isoflavones, genistein can regulate the expressions ER-α mRNA in granulosa cells of rat ovaries. Such an effect is concentration-dependent. And 1-10 µmol/L genistein may up-regulate the expression of ER-α mRNA.

Expression of nucleophosmin in glandular epithelium of non-pregnant human endometrium during the menstrual cycle.

BACKGROUND: Nucleophosmin plays a critical role in embryonic development. This study aimed to examine the expression pattern of nucleophosmin in glandular epithelium of human endometrium during the menstrual cycle. METHODS: Endometrial tissues used for this study were obtained from 46 non-pregnant patients who underwent hysterectomy which had been performed to treat benign diseases. Nucleophosmin expression was assessed by in situ hybridization and immunohistochemistry. RESULTS: At the early-, mid- and late-proliferative phase, nucleophosmin mRNA was highly expressed in glandular epithelium of human endometrium. At the secretory phase, the expression of nucleophosmin mRNA was reduced in glandular epithelium in early-secretory phase, and the expression in mid- and late-secretory phases was not detected. Similarly, nucleophosmin protein was strongly expressed in endometrial glands throughout the proliferative phase, but was gradually reduced during secretory phase. CONCLUSION: Nucleophosmin mRNA and protein are expressed in glandular epithelium of human endometrium throughout the menstrual cycle.

Action mechanism of Zuo Gui Yin Decoction's promotion on estradiol production in rats during the peri-menopausal period.

AIM OF THE STUDY: Although Zuo Gui Yin Decoction has long been used in Traditional Chinese Medicine to treat menopausal symptoms, the underlying mechanism(s) by which these effects are induced remains to be defined. The aim of this study was to investigate the action mechanism of Zuo Gui Yin Decoction on estradiol production in the rat ovary during peri-menopause. MATERIALS AND METHODS: The peri-menopausal animal model was established by natural aging. Peri-menopausal rats were treated by intragastric administration (ig) with low (13.78gkg(-1)), middle (20.67gkg(-1)) or high (31gkg(-1)) dose of Zuo Gui Yin Decoction per day for 8 weeks. At the 8th weekend, the rats were sacrificed for sampling. Estradiol (E(2)) levels in rats' serum were evaluated by radioimmunoassay (RIA). RT-PCR, in situ hybridization and immunohistochemistry were used to determine mRNA and protein expression of relevant genes. RESULTS: Medium- and high-dose of Zuo Gui Yin Decoction could significantly increase serum estradiol concentration, ovarian CYP19 mRNA levels, and P450(arom) protein expression in rats during peri-menopause. Zuo Gui Yin Decoction at three different dosages all could promote FSHR expression and the effect of low-dose was the greatest. Zuo Gui Yin Decoction could elevate LRH-1 and ER(α) expression in a dose dependent manner. CONCLUSIONS: Taken collectively, these findings suggest that Zuo Gui Yin Decoction could promote estradiol production in rat serum during peri-menopausal period through ovarian ER(α)→LRH-1→CYP19 pathway as well as the ovarian classical FSHR→CYP19 mechanism.

[The expression of a novel estrogen receptor, GPR30, in epithelial ovarian carcinoma and its correlation with MMP-9].

The aim of the present study is to investigate the expression of a novel estrogen receptor, G protein-coupled receptor 30 (GPR30) and its correlation with matrix metalloproteinases-9 (MMP-9) in epithelial ovarian cancer (EOC). Ovary tissues were obtained from 39 female patients, including 30 cases of EOC and 9 cases of benign ovarian tumor. Four normal ovary tissues were used as control. Immunohistochemical staining was used to detect the expressions of GPR30 and MMP-9. Chi square test, Fisher's exact test and Spearman's rank correlation analysis were used for statistical analysis. The results showed that GPR30 overexpression rate in EOC cases was significantly higher than those in benign ovarian tumor and normal ovary cases. Whereas MMP-9 overexpression rate in EOC cases was significantly higher than that in normal ovary cases, without any difference to that in benign ovarian tumor cases. To demonstrate the relationship between GPR30 and clinicopathological variables of EOC, we further analyzed the pathology type, FIGO stage and age of patients sampled in our study. The analysis showed there were significant differences of GPR30 overexpression rate among various pathology types and different FIGO stages (P<0.05), and no significant difference of both GPR30 and MMP-9 among three age groups (P>0.05). Moreover, GPR30 expression was positively correlated with MMP-9 (r(s)=1.000, P=0.002). These results suggest that GPR30 may be involved in the invasion and metastasis of EOC, being a potential index of EOC early diagnosis and malignancy grade prediction.

[Effects of Yikunning on the apoptotic rate and expression of caspase-3 in rat ovaries during perimenopausal period].

OBJECTIVE: To investigate the effects of Yikunning (compound of Chinese traditional Medicine, YKN) on the apoptotic rate and expression of caspase-3 in rat ovaries during perimenopausal period. METHODS: Thirty female Wistar rats during perimenopausal period were selected by unforced aging. Then the rats were divided into 3 groups randomly: YKN group, livial control group and aged control group. Ten young female rats were selected as young control group. Intragastric administrations were conducted for 4 weeks once daily continuously. The apoptotic rate in rat ovaries were detected by TUNEL. The expression of caspase-3 mRNA and protein in rat ovaries were detected by RT-PCR and Western blot, respectively. RESULTS: The apoptotic rate in rat ovaries in YKN group was lower than that in aged control group, which showed difference between them (P < 0.01). The levels of caspase-3 mRNA and protein in rat ovaries in YKN group were lower than those in aged control group, which showed differences among them (P < 0.01). CONCLUSION: YKN can decrease the apoptotic rate and down-regulate the expression of caspase-3 mRNA and protein in rat ovaries of during perimenopausal period. It may be one of the molecular mechanisms of YKN postponed the ovarian failure and cured perimenopausal syndrome.

[Effects of yikunning on the expressions of bcl-2 and bax in rat ovaries during perimenopausal period].

OBJECTIVE: To investigate the effects of Yikunning (YKN, Chinese Traditional Medicine) on the expressions of bcl-2 and bax in rat ovaries during perimenopausal period. METHODS: Thirty female Wistar rats during perimenopausal period were selected by unforced aging. Then the rats were divided into 3 groups randomly: YKN group, Livial control group and Aged control group. Ten young female Wistar rats were selected as young control group. Intragastric administrations were conducted for 4 weeks once daily continuously. The expressions of Bcl-2 Bax mRNAs and proteins in rat ovaries were detected by RT-PCR and Western blot. RESULTS: The levels of Bcl-2, Bax mRNAs and proteins in rat ovaries in YKN group were higher than those in Aged control group, which showed differences among them (P < 0.01). The Bcl-2/Bax mRNA rate and protein rate in rat ovaries in YKN group were higher than those in Aged control group, which showed differences among them (P < 0.05 or P < 0.01). CONCLUSION: YKN could increase the expressions of Bcl-2, Bax mRNAs and proteins and up-regulate the Bcl-2/Bax mRNA rate, protein rate in rat ovaries during perimenopausal period, which might be one of the molecular mechanisms of YKN postponed the ovarian failure and cured perimenopausal syndrome.

[Effect of zuoguiyin on the expression of ovarian follicle-stimulating hormone receptor in rats during peri-menopausal period].

OBJECTIVE: To investigate the effect and mechanism of Zuoguiyin (ZGY, a Chinese medical remedy for regulating qi and nourishing yin) on expression of ovarian follicle-stimulating hormone receptor (FSHR) in rats during peri-menopausal period (PMP). METHODS: PMP rats were administered with low (13.78 g/kg), middle (20.67 g/kg) and high (31.00 g/kg) dose ZGY respectively by gastrogavage for eight weeks. FSHR mRNA and protein expressions were detected by RT-PCR and immunohistochemistry respectively. Besides, granulosa cells, collected from pregnant mare serum treated nonage rats, were incubated, and the level of FSHR mRNA expression in the cultured cells were detected after various disposals. RESULTS: (1) Ovarian levels of FSHR mRNA and protein expressions in PMP rats were significantly lower than those in young rats (P<0.01), but were up-regulated significantly by ZGY treatment (P<0.01). (2) As compared with the control, FSHR mRNA expression increased in cultured granulosa cells after treated by ZGY extract; the increasing effect of ZGY was enhanced by combined treatment with Forskolin and attenuated by Genistein (a tyrosine protein kinase inhibitor). CONCLUSION: ZGY could improve the ovarian functions during PMP by up-regulating the expression of FSHR and raise the ovarian responsibility to FSH, which may be possibly realized by activating cAMP-protein kinase and tyrosine protein kinase signal pathway.

[Changes of granulocyte distribution induced by thyroxin and epinephrine: experiment with rabbits].

OBJECTIVE: To investigate the distribution of granulocyte during thyrotoxicosis and the relation of granulopenia and the gathering of circulating granulocytes into the marginal pool. METHODS: Forty-five rabbits were randomly divided into 3 equal groups: epinephrine group fed with levo-thyroxin (LT4) for 14 days, injected subcutaneously with epinephrine 0.07 mg/kg, and undergoing collection of peripheral blood samples for white blood cell and granulocyte counts, free blood triiodothyroxine (FT3) and free blood thyroxine (FT4), electrocardiography, and body weight measurement; labeled group fed with LT4 for 14 days, injected subcutaneously with 131I-labeled anti-human granulocyte monoclonal antibody, and killed 6 hours later to calculate the ratio of radioactivity of heart, liver, spleen, and muscle tissues to blood (CPM/g), and control group, not fed with L-T4 but injected with 131I-labeled anti-human granulocyte monoclonal antibody on the day 14, and killed 6 hours later to calculate the ratio of radioactivity of tissues to blood. RESULTS: After 14-day feeding of L-T4, the heart rates of the epinephrine and labeled groups were significantly higher than those before injection, and the FT3 and FT4 levels were significantly increased;the venous WBC and granulocyte counts were significantly reduced. But the granulocyte count of 13 of the 15 rabbits in the epinephrine group increased 20 minutes after the injection of epinephrine, even becoming 2-4.8 times as high as those before the injection in 7 rabbits. The heart, liver, spleen, and muscle tissues to blood CPM/g ratios of the labeled group were all significantly higher than those of the control group (all P<0.01). CONCLUSION: During thyrotoxicosis the circulating granulocytes are reduced and the distribution of circulating granulocytes is abnormal with a gathering phenomenon of the granulocytes into the marginal pool.

[Role of platelet-activating factor in progesterone synthesis and vascular endothelial growth factor expression in rat luteal cells].

The present study aimed to investigate the role of platelet-activating factor (PAF) in progesterone synthesis and vascular endothelial growth factor (VEGF) expression in rat luteal cells. Immature (25-28 days old) female Sprague-Dawley rats were injected subcutaneously with 50 IU pregnant mare serum gonadotrophin (PMSG), and 25 IU human chorionic gonadotrophin (hCG) 48 h later, to induce follicular development and luteum formation. On day 6 after hCG administration (the day of hCG administration was the first day), the rats were killed by guillotine and the ovarian luteal cells were collected. After incubation for 24 h, luteal cells were incubated without or with different doses (0.1 µg/mL, 1 µg/mL, 10 µg/mL) of PAF at 37 °C (5% CO(2)) for 24 h, and then progesterone concentration was evaluated by radioimmunoassay (RIA); apoptotic rate and VEGF mRNA expression in luteal cells were assessed by flow cytometry and RT-PCR, respectively. The results showed that PAF promoted progesterone production, with a maximal effect at 1 µg/mL (P<0.05); PAF increased apoptotic rate but not in a dose-dependent manner, and 10 µg/mL PAF enhanced apoptotic rate significantly (P<0.05); furthermore, PAF stimulated VEGF mRNA expression in luteal cells, especially at 1 µg/mL (P<0.01). It is suggested that PAF regulates progesterone synthesis and VEGF mRNA expression in luteal cells to mediate corpus luteum formation in rat ovary.

[Effects of soy isoflavones on the expression of Bax mRNA and Ca(2+)-ATPase activity in ovaries of perimenopause rats].

AIM: To investigate the effects of soy isoflavones (SI) on the expression of Bax mRNA and Ca(2+) -ATPase activity in ovaries of perimenopause rats. METHODS: The animal model of perimenopause rats was established by unforced aging. 12 month-old presenilins female Wistar rats were administered by intragastric (ig) with low (500 mg/kg), middle (158 mg/kg) and high (500 mg/kg) does of SI for 8 weeks. The expression of Bax mRNA in ovaries were detected by RT-PCR. Ca(2+) -ATPase activity in ovaries and MDA content and SOD activity in serum were detected by chemi-chromatometry. RESULTS: Intervention of SI could significantly decrease the expression of Bax mRNA in ovaries and MDA content in serum, increase Ca(2+) -ATPase activity in ovaries and SOD activity in serum of presenilins rats (P < 0.05 or P < 0.01). CONCLUSION: Soy isoflavones could down-regulate the expression of Bax mRNA and increase Ca(2+) -ATPase activity in aged ovaries. It is probably one of the mechanisms to improve the function of aged ovaries in perimenopause rats.

[Genetic mapping of the intergenomic translocated "taigu" genic sterility gene (Ms2)].

The "Taigu" genic sterility gene Ms2 located on the short arm of the 4D chromosome of common wheat (AABBDD) originally incorporated into hexoploid triticale (AABBRR) and durum wheat (AABB) through intergenomic translocation in distant hybridization was introduced back into the genomes of common wheat. The dominant male sterility was expressed normally in the new "Taigu" genic sterile wheat carrying the intergenomically translocated Ms2, and the female fertility mechanism in its male sterile plants was normal as well. Observation of the chromosome configuration at meiosis of pollen mother cells (PMC) of the young ears of the sterile plants showed that they were euploid plants (2n = 42). No configurations different from those of the "Taigu" genic sterility gene located at the original locus were noticed of the Ms2 intergenomically translocated back into the common wheat. In systematic test crosses with marker genes the intergenomically translocated gene Ms2 was found to be linked with the dominant dwarf marker in common wheat Rht3 and, consequently, remapped and located on the short arm of the 4B chromosome of common wheat with a distance of 9.7 cM from Rht3. The new locus was designated as Ms2 (4BS). Discussions are given of the fate of Ms2 during translocation in the hexoploid triticale, the exchange of the names for 4A and 4B chromosomes in common wheat and the possible exploitation of the new locus Ms2 (4BS), and the following speculations are made: (a) In genic genes of allopolyploid organisms the donor chromosomes tend to be intergenomically translocated to their physiologically and evolutionarily close chromosomes with the same order number and the same arm; (b) it is confirmed that the 7th International Conference of Wheat Genetics was right to exchange the names between chromosomes 4A and 4B of common wheat in 1988; and (c) as a new genetic marker and a breeding tool for all the chromosome B-carrying species in the tribe of Triticeae, Ms2(4BS) may have wide application in building and expanding the gene pool of germplasm resources of various species of wheat.