• Asian-Australasian Journal of Animal Sciences
• /
• v.24 no.9
• /
• pp.1204-1210
• /
• 2011

Objective of this study was to examine the effect of sodium nitroprusside (SNP), a nitric oxide (NO) donor on steroid synthesis, growth and apoptosis of buffalo granulosa cells (GCs) in vitro. Follicular fluid of antral follicles (3-5 mm diameter) was aspirated and GCs were cultured in 0 (control), $10^{-3}$, $10^{-5}$, $10^{-7}$, $10^{-9}\;M$ of SNP for 48 h. To evaluate whether this effect was reversible, GCs were cultured in presence of $10^{-5}\;M$ SNP+1.0 mM $N^{\omega}$-nitro-L-arginine methyl ester (L-NAME) a NO synthase (NOS) inhibitor or hemoglobin (Hb, $1.0{\mu}g$) as NO scavenger. Nitrate/nitrite concentration was evaluated by Griess method, progesterone and estradiol concentrations by RIA and apoptosis by TUNEL assay. SNP ($10^{-3}$, $10^{-5}$, $10^{-7}\;M$) significantly (p<0.05) inhibited estradiol and progesterone synthesis, growth, disorganized GCs aggregates and induced apoptosis in a dose dependent manner. However, $10^{-9}\;M$ SNP induced the progesterone synthesis and stimulated GCs to develop into a uniform monolayer. Combination of SNP $10^{-5}$ M+L-NAME strengthened the inhibitory effect while, SNP+Hb together reversed these inhibitory effects. In conclusion, SNP at greater concentrations ($10^{-3}$, $10^{-5}$ and $10^{-7}\;M$) has a cytotoxic effect and it may lead to cell death whereas, at a lower concentration ($10^{-9}\;M$) induced progesterone synthesis and growth of GCs. These findings have important implications that NOS derived NO are involved at physiological level during growth and development of buffalo GCs which regulates the steroidogenesis, growth and apoptosis.

• Journal of Animal Reproduction and Biotechnology
• /
• v.34 no.2
• /
• pp.75-85
• /
• 2019

Omega-3 α-linolenic acid and omega-6 linoleic acid are essential fatty acids for health maintenance of human and animals because they are not synthesized in vivo. The purpose of this study was to evaluate the effect of α-linolenic acid and linoleic acid supplementation on in vitro maturation and developmental potential of porcine oocytes. Various concentrations of α-linolenic acid and linoleic acid were added into in vitro maturation medium, and we evaluated the degree of cumulus expansion, oocyte nuclear-maturation rate, blastocyst rate, blastocyst quality, and levels of prostaglandin E₂, 17β-estradiol, and progesterone in the spent medium. High doses (100 μM) of α-linolenic acid and linoleic acid supplementation significantly inhibited cumulus expansion and oocyte nuclear maturation, and prostaglandin E₂ synthesis also significantly decreased compared with other groups (p < 0.05). Supplementation of 50 μM α-linolenic acid and 10 μM linoleic acid showed higher quality blastocysts in terms of high cell numbers and low apoptosis when compared with other groups (p < 0.05), and synthesis ratio of 17β-estradiol / progesterone also significantly increased compared with control group (3.59 ± 0.22 vs. 2.97 ± 0.22, 3.4 ± 0.28 vs. 2.81 ± 0.19, respectively; p < 0.05). Our results indicated that supplementation with appropriate levels of α-linolenic acid and linoleic acid beneficially affects the change of hormone synthesis (in particular, an appropriate increase in the 17β-estradiol / progesterone synthesis ratio) for controlling oocyte maturation, leading to improved embryo quality. However, high doses of α-linolenic acid and linoleic acid treatment results in detrimental effects.

• Journal of Animal Reproduction and Biotechnology
• /
• v.35 no.2
• /
• pp.183-189
• /
• 2020

The pregnancy rate in indigenous ewes inseminated with frozen-thawed Suffolk semen following natural and synchronized estrus was determined. The serum Progesterone and Estrogen concentration and vaginal electrical resistance (VER) of ewes at the time of Artificial Insemination (AI) were observed as successful pregnancy determinants. 21 healthy ewes were selected for this experiment during January-April, 2017. 10 ewes were inseminated in natural estrus. Whereas, 11 ewes were inseminated after estrus synchronization using intravaginal sponges containing 60 mg medroxyprogesterone acetate. Trans-cervical Al (TCAI) was performed in all ewes within 12-16 hours of observed heat. Prostaglandin E₁ analogue impregnated vaginal sponge was used for cervical relaxation 6-8 hours before insemination. Pregnancy was diagnosed through trans-abdominal ultrasonography after 40 days of AI. The pregnancy rate of ewes in synchronized estrus was higher (54.5%) than in natural estrus (30%). Higher serum Progesterone level (0.90 ± 0.02 ng/mL) and significantly (p < 0.001) lower VER (257.78 ± 10.11 ohm) were observed at the time of AI in ewes becoming pregnant. Results suggest that higher Progesterone concentration and lower VER could be considered as pregnancy indicators. Oestrous synchronization could be implemented to increase the pregnancy rate in ewes.

• The Journal of Korean Obstetrics and Gynecology
• /
• v.20 no.2
• /
• pp.43-59
• /
• 2007

Purpose : Astragalus membranaceus is a herb which has effectiveness of energy increase, protection against pathogenic and organ immune response control. Endometriosis has been thought to be related with immune response in uterus. Therefore in this study, the effects of Astragalus membranaceus on endometriosis were investigated. Methods : The endometrial tissue was autografted to rat's small intestine. Rats with surgically induced endometriosis was administerd with Astragalus membranaceus extracts for 40 days orally or rectally. We observed the size of ectopic uterine implants at the mesenteric with the naked eye and examined the concentrations of estradiol, progesterone, $TNF-{\alpha}$ and IL-2, IL-4, IL-6 and IL-10 in serum. Also, we inquired out the efficiency between oral and rectal administration groups. Results : The size of ectopic uterine implants of oral and rectal treated groups were much smaller than those of control groups. The concentration of estradiol, progesterone. $TNF-{\alpha}$ and IL-2 were significantly decreased in oral administration experimental group compared with the control group. The concentration of progesterone were significantly decreased in rectal administration experimental group compared with the control group. The efficiency of oral administration group was better than that of rectal administration group in the serum concentration of estradiol, $TNF-{\alpha}$ and IL-2. Conclusion : Our data suggest that Astragalus membranaceus reduce the size of ectopic uterine implants at the mesenteric wall and inhibit the growth of ectopic uterine implants. Judging from the above results, it can be suggested that Astragalus membranaceus should be a useful agent for inhibiting the proliferation of uterine endometrial tissue. May be, oral administration of Astragalus membranaceus is more effective than rectal administration for inhibiting the proliferation of endometriosis.

• Clinical and Experimental Reproductive Medicine
• /
• v.21 no.2
• /
• pp.191-200
• /
• 1994

The influence of hypoxanthine and ovarian steroids on the meiotic maturation process of mouse oocytes was investigated for the qualified application of culture medium in in vitro fertilization(IVF). Mouse oocytes were cultured in hypoxanthine and various ovarian steroids(progesterone, estradiol-17${\beta}$ and testosterone) and their effects on the oocyte maturation had been observed. When mouse oocytes were cultured in the various concentration(1-4mM) of hypoxanthine, meiotic maturation of cumulus cell-enclosed oocytes was inhibited by presence itself, which was a dose-dependent effect in meiotic arrest of mouse oocytes. The presence of progesterone, estradiol-17${\beta}$ and testosterone have made the mouse oocyte mature properly. Meanwhile maturation of cumulus cell-enclosed oocyte was severely inhibited by 3 hoursculture in the media of progesterone supplemented with hypoxanthine. However the continuous presence lasting 24 hours of progesterone even supplemented with hypoxanthine had got rid of the inhibition of oocytes maturation. Not only estradiol-17${\beta}$ supplemented with hypoxanthine but also testosterone supplemented with hypoxanthine exert the severe inhibition of the maturation of cumulus cell-enclosed oocytes for 3-hours culture. However the continuous presence lasting 24 hours of estradiol-17${\beta}$ and testosterone even supplemented with· hypoxanthine had relieved the inhibition of oocytes maturation. These results make us suggest that hypoxanthine inhibits the mouse oocyte maturation, particularly markedly in conjunction with ovarian steroids for short period, which indicated some sort of the synergistic inhibitory retationship between the ovarian steroids and hypoxanthine.

• Korean Journal of Veterinary Research
• /
• v.36 no.1
• /
• pp.101-108
• /
• 1996

This study was desinged to investigate the effect of estrogen(Est) on the proliferating of progesterone(Prog) target cells. The spayed 13 rats(Wistar, approximately 300gm) were randomly alloted into 3 groups. One group was the control group and another Prog-treated group was injected with 1mg of Prog/rat/day for 2 consecutive days, and Estand Prog-treated group was injected intramuscularly with $17{\beta}$-estradiol $20{\mu}g/rat/day$ for 3 consecutive days and then with Prog for 2 days as above from 4th day. Rats were administrated intraperitoneally with bromodeoxyuridinc(Brdur,0.2mg/BW once) befero 2 hours of exanguination. In gross finding, the groups with more level of dimension and weight on the uterus were ordered as Est- and Prog-treated group, Prog-treated group and control group. The investigation by immunohistochemical methods using paraffin sections of the uteri was performed by using anti-Brdur antibody for labeling proliferating cells of Prog target cells. The groups with higher labeling index(LI) were ordered as Prog-treated grop, Est- and Prog- treated group and control group. The number of proliferating cells from Prog target cells in the rats were rather deceased by Prog injection following Est injection than prog injection only. The cell types with higher LI in the wall layers of all 3 groups were ordered as endometrial stromal cells, glandular epithelial cells, luminal epithelial cells, myometrial muscle cells and serosa methodelial cells, and the region with highest LI was functional zone of the endometrium and the region with lower LI was muscular layer and then those with lowest LI was serosa and also the considerable different LI from individual rat were observed.

• The Korean Journal of Mycology
• /
• v.13 no.2
• /
• pp.83-87
• /
• 1985

The temperature dependency and the stability of enzyme systems for $11{\alpha}-hydroxyla­tion$ of progesterone were investigated using Aspergillus phoenicis. Though A. phoenicis conserves high enzyme activities for lactose hydrolysis even at high temperatures, the bioconversion reaction of progesterone by this strain was found to be very temperature sensitive. The compositions of reaction mediums of inside and outside of cells were analyzed using sonication technique. At early stage of reaction, the concentration of $11{\alpha}-hydroxyprogesterone$ of cell inside was higher than that of outside. But as the reaction proceeded further, the $11{\alpha}-hydroxyprogesterone$ existing inside of cells being converted into another products, its concentration was lower within the cells that in the bulk medium. Even in the reaction mediums containing organic solvents, A. phoenicis was founded to be able to metabolite, so that $11{\alpha}-hydroxyprogesterone$ can be produced continuously from fixed bed reactions packed with immobilized A phoenicis in vivo.

• Journal of Veterinary Clinics
• /
• v.28 no.3
• /
• pp.303-306
• /
• 2011

A 12-year-old female mixed breed dog receiving a progesterone drug was referred for evaluation of an abdominal mass. Abdominal radiography and ultrasonography revealed a swollen uterus and an associated mass. Serum chemistry revealed hyperglobulinemia consistent with acute inflammation based on the results of serum protein electrophoresis. Fine needle aspiration of the mass guided by ultrasonography was performed for cytological evaluation. The cytological impression was consistent with adenocarcinoma. Exploratory laparotomy identified a uterine body mass, which was surgically removed for histopathology. Histology of the mass identified a uterine adenocarcinoma. Immunochemistry using anti-estrogen and progesterone receptor antibodies was performed and neoplastic cells were negative to both antibodies while some normal elements were reactive to both of them. Computer tomography demonstrated evidence of metastatic disease in the lung one week after the surgery and the dog died about 40 days after surgery.

• The Korean Journal of Physiology and Pharmacology
• /
• v.20 no.4
• /
• pp.341-346
• /
• 2016

Membrane-bound HLA-G (mHLA-G) discovery on adipose derived stem cells (ADSCs) as a tolerogenic and immunosuppressive molecule was very important. Many documents have shown that HLA-G expression can be controlled via some hormones such as progesterone (P4) and estradiol (E2). Therefore, this study was designed to evaluate progesterone and estradiol effects on mHLA-G in ADSCs at restricted and combination concentrations. Three independent cell lines were cultured in complete free phenol red DMEM and subcultured to achieve sufficient cells. These cells were treated with P4, E2 and P4 plus E2 at physiologic and pregnancy concentrations for 3 days in cell culture conditions. The HLA-G positive ADSCs was measured via monoclonal anti HLA-G-FITC/ MEMG-09 by means of flow cytometry in nine groups. Data were analyzed by one way ANOVA and Tukey's post hoc tests. There were no significant values of the mean percentage of HLA-G positive cells in E2-treated and the combination of P4 plus $E_2-treated$ ADSCs compared to control cells (p value>0.05) but P4 had a significant increase on mHLA-G in ADSCs (p value<0.05). High P4 concentration increased mHLA-G but E2 and the combination of P4 plus E2 could not change mHLA-G on ADSCs.

• Asian-Australasian Journal of Animal Sciences
• /
• v.18 no.4
• /
• pp.470-474
• /
• 2005

The efficiency of a short-term treatment with an intravaginal device containing progesterone (CIDR) combined with pregnant mares' serum gonadotrophin (PMSG) and prostaglandin analogue ($PGF_{2\alpha}$) was evaluated for the induction of estrus, initiation of cyclic activity, and fertility in postpartum suckled yak cows. Seventy-five postpartum suckled yak cows were assigned to three treatments: (1) insertion of a CIDR intravaginal progesterone (1.9 g) (day 0), an administration of $PGF_{2\alpha}$ (0.2 mg i.m.) on day 6 and PMSG (1,000 IU i.m.) at the time of CIDR withdrawal on day 7 (CPP group, n=28); (2) an administration of $PGF_{2\alpha}$ (0.2 mg i.m.) on day 6 and PMSG (1,000 IU i.m.) on day 7 (PP group, n=21); (3) untreated animals served as the control (CG group, n=26). Seven yak bulls were placed in pastures with the cows for natural mating. Estrus rate in the CPP group (28/28) was higher (p<0.01) than in the PP group (6/21) and in the CG group (0/26) within 96 h after the end of treatment. The first service conception rate in the CPP group (21/28) was higher (p<0.01) compared with in the PP group (2/9) as judged by serum $P_4$ concentration $\geq$2.35 ng/ml on day 21 after breeding. It is concluded that a short-term progesterone treatment combined with PMSG and prostaglandin increased the proportion of yak cows that exhibited behavioral estrus with more synchronized estrus response and satisfactory conception rate in postpartum suckled yak cows.