Lonicera Caerulea(Honey berry) has been used in medical treatment in Russia, Japan, China and Korea. It has high level of vitamin C and polyphenolics. Polyphenolics can improve anti-inflammatory effect and prevent cancer, diabetes mellitus type 2. Also, Vitamin C is a representative anti- oxidant. however, It is still unknown what effect it will have on the oxidation stress of the reproductive system. In previous studies, ROS can be produced when it is exposed to heat stress and has negative effect on sperm's maturation, capacitation, hyperactivation, acrosome reaction and fusion of egg and sperm. Therefore, The purpose of this study is to investigate the antioxidant effects of L.caerulea on the sperm and egg cells of mice. At first, it conducted using ICR mouse(n=20) during 4 weeks. There are four groups of mouse(n=5 per group). Also, L.caerulea was taken by oral gavage. Group Ⅰ(control) kept at 23℃~27℃ and administer D.W(0.5ml/day), Likewise, Group Ⅱ(HB) kept at room temperature but gave HB(0.5ml/day), Group Ⅲ (HB+HS) received heat stress (40℃) using hyperthermia induction chamber and gave HB at same dose. and Group Ⅳ(HS) exposed heat stress only. Mainly, we showed degree of gene expression using Western blot in SOD, HSP 70, 17β-HSD and Real time PCR. It can find correlation between intracellular activity like steroid hormone, apoptosis under ROS and antioxidant activity of L.caerulea.

Previous studies have shown that Lonicera caerulea has a chemical protective effect. Phenolic and vitamin C contained in Lonicera caerulea prevent cancer, diabetes and cardiovascular disease, lower blood pressure and delay the aging process. However, the antioxidant mechanism of male reproductive system to heat stress is still unknown. Male reproductive system is very sensitive to heat. When scrotum temperature increase, oxidative stress can occur. Oxidative stress affects sperm motility and spermatogenesis, resulting in infertility. Therefore, we investigated the antioxidant effect of L. Caerulea in male genitalia by inducing oxidative stress by artificially exposing the testicles to heat at 42 ° C. The experiment was performed by dividing the ICR mouse into four groups. Each group is n = 5. Control group (C) and heat stress group (HS) were oral gavage administered D.W. Honey berry group (HB) and Honeyberry / heat stress group (HB + HS) were oral gavage administered honey berry (250mg / kg / day). HS groups (n=5, per n=5) received heat stress by exposing their lower bodies in the water bath at 42℃ for 30 minutes. We confirmed that there was a significant difference in the motility, morphology and the number of sperms using CASA(computer-assisted semen analysis). Lipid peroxidation assay results showed heat causes oxidative stress in serum. This study is conducting to investigate the antioxidant effect of L. Caerulea. Histologically analyzed the testicular form of each group, the activity level of heat shock protein and the level of reactive oxygen species were measured by Western blot and the level of catalase and HSP-90 was examined by RT-PCR analysis. Thus, studies of testicular morphology, sperm kinetics, hormone levels, heat shock protein expression and antioxidant enzymes under heat stress have shown that L. Caerulea ingestion has Anti-oxidant and thermal protective activity on the testis by heat damage.

Embryo transfer is one of the important process of assisted reproductive technology (ART) and that is associated with uterus endometrial receptivity. Recently, mouse endometrial stimulation by artificial injury had shown the favorable effect on conception. In this experiment, we used uterus stimulation method that injury the endometrium to increase implantation rate for spontaneous Diabetes Mellitus (sDM) rat. Rats are divided into several groups involved a control group. We performed the surgical method to Experimental group bilaterally or unilaterally After that, we investigated morphological change and calculated implanted embryos respective sides of the uterus. The number of implanted embryo in the experimental group was significantly higher and there were lots of morphological changes including glands and endometrial cells that support implantation. Our results showed that rat uterus endometrial injury in ART help enhancing implantation rate.

The purpose of this study is to evaluate the effects of astaxanthin added to freezing buffer on semen parameters, total sperm oxidation stress after post-thawing of boar sperm and lipid peroxidation (LPO) which is caused by reactive oxygen species (ROS) in sperm membrane. Varying concentrations of astaxanthin (0, 10, 50, 100 and 500 μM) were used in the freezing buffer during cryopreservation to protect the DNA of thawed miniature pig sperm. Semen parameter was measured using computer assisted sperm analysis (CASA) for sperm motility and determine ROS rate, oxidative stress of boar sperm using fluorescence-activated cell sorting (FACS). Sperm motility was higher (p<0.05) in the astaxanthin group than in the control group. Sperm motility and the number of progressive motile sperm was higher (p<0.05) in the astaxanthin 500 μM group (66±1.7%) than in the control group (49.8±4%). In ROS evaluation, the astaxanthin group lowered intracellular O2 and H2O2 in viable sperm. The Yo-Pro-I/HE and PI/H2DCFDA staining as revealed using flow cytometry was lower in astaxanthin groups than in the other groups. As the result, we found that astaxanthin could protect the sperm plasma membrane from free radical and LPO during boar sperm post-thawing.

Superovulation is a technique to acquire rats to produce many eggs than normal rats. Superovulated eggs were used to make cloned animals through somatic cell nuclear transfer technology (SCNT). Healthy and valuable oocyte retrieval is essential for successful somatic cell nuclear transfer. Superovulation is also essential to maximize to the yield of IVF-derived rat eggs. Osmotic pumps (Alzet®) are miniature in order to provide research with a convenient, and reliable alternative to chronic injections. Acquiring superovulated oocytes through osmotic pump in minimizing irritation to the uterus and ovaries are competitive. We investigated the effects of pregnant mare serum gonadotropin (PMSG) and human Chorionic Gonadotropin (hCG) using osmotic pump in sDM rat. Adult female rats at 11 wks of age were used for superovulation. The response to PMSG and hCG were examined by osmotic pump of 150 IU/kg PMSG + 75 IU/kg hCG or 150 IU/kg PMSG + 150 IU/kg hCG or 300 IU/kg PMSG + 150 IU/kg hCG or 300 IU/kg PMSG + 300 IU/kg hCG. HCG was administered 48 hrs later after administration of PMSG. Oocytes were collected from the oviducts 16–18 hrs after hCG administration. Superovulation was significantly higher in rats administrated 150 IU/kg PMSG + 75 IU/kg hCG. This study demonstrated that healthy oocytes were produced in DM rat by PMSG and hCG that flowed through the osmotic pump ameliorate on uterus and ovaries.

Several species show low sensitivity to IVM, and the development of optimized medium possible oocyte quality and stable growth. Furthermore, adding additive to the medium can effectively reducing development cost and leads to easy handling of oocytes. Isoliquritigenin and formononetin are extracts found in licorice. Previous studies reported that isoliquritigenin and formononetin affected the activity of sperm, but the oocytes are unknown. This study adds isoliquritigenin or formononetin to αMEM to mature oocytes under simple IVC conditions. Recovered oocytes are cultured in αMEM, isoliquritigenincontaining medium and formononetin-containing medium. In study we proved that in addition to the medium, above the quality of oocytes cultured when specific additives were added, more stable growth is possible. collection and IVM of oocyte. SD rats at 6 to 8 wks of age are injected is intraperitoneal with 30 IU/mL of PMSG and 48 hrs later, HCG 50 IU/mL is intraperitoneal injected. Oocytes are collected ovary after 17 hrs. Collected oocytes are cultured for 16 hrs with 200 μL αMEM and 200 μL αMEM containing isoliquritigenin or formononetin at 0, 0.01, 0.02, 0.04, 0.1 mg/mL. Also, isoliquritigenin and formononetin were mixed with 200 μL αMEM at a ratio of 0.25: 0.75, 0.50: 0.50, and 0.75: 0.25 mg/mL respectively. Oocytes supplemented with isoliquritigenin and Formononetin had high quality than oocytes cultured with αMEM and showed an increase in the IVF fertility rate. Our experimental results indicate that using isoliquritigenin, formononetin when cell culture, rather than used only in medium, more effective oocyte quality and stable growth.

Our objective was to evaluate the function of treahlose and erythritol in reducing ROS concentrations, which is associated with a general improvement in the quality of frozen-thawing miniature pig sperm. Semen was mixed in modified Modena B extender, added to cooling media and freezing media, followed by the supplement of 100 mM trehalose and/or 100 mM erythritol with spermatozoa (1000x 109cells/straw). The trehalose plus erythritol (TE) added group had less intracellular H2O2 than did control and trehalose (36.6±1.6 vs. 49.0±5.8 and 48.8±7.9; P<0.05). The percentage of viable acrosome-intact sperm (FITC-PNA-/PI-) was higher in erythritol and TE than controls (57.0±5.5% and 62.5±4.3% vs. 45.4±5.4%; P<0.05 and P<0.001). The percentage of sperm with high fragmented DNA was observed in control group when compared with erythritol and TE also trehalose (65.5±1.3% vs 59.3±0.7% and 59.0±0.3% vs 62.2± 0.8%; P<0.001). The percentage of sperm LPO was higher in control and trehalose than erythritol (4.4±0.5% and 5.0±0.5% vs. 3.5±0.2; P<0.01 and P<0.001), and was lowest in the TE (control and trehalose vs. TE: P<0.001, erythritol vs. TE: P<0.05). Also, we performed that surgical insemination based on above data to evaluate the function of new cryoprotectant such as trehalose plus erythritol in vivo. Finally, 1 pregnant gilt showed natural estrus was allowed to go to term and 8 live piglets were born. In conclusion, miniature pig sperm was successfully cryopreserved with trehalose plus erythritol provided the increasing the sperm quality and reducing the ROS.