• Clinical and Experimental Reproductive Medicine
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• v.33 no.1
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• pp.35-35
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• 2006

Objective: The purposes of the present study were to investigate the effect of ${\gamma}$-radiation on the expression of inhibin-${\alpha}$ proteins and genes for inhibin ${\alpha}$, ${\beta}A$, and ${\beta}B$ in the ovary. Methods: Immature mice were whole-body ${\gamma}$-irradiated with 25% of a lethal dose. At time 0, 3, 6, 12, and 24 hours after the irradiation, the ovaries were collected and used for immunohistochemistry for inhibin-${\alpha}$, and RT_PCR for inhibin-${\alpha}$, ${\beta}A$, and ${\beta}B$. Results: The expression of the immunoreactive inhibins-${\alpha}$ was maintained at 12 hours post-irradiation and reduced thereafter. The expression of inhibin-${\alpha}$ mRNA was significantly increased with the time after the irradiation. However there were no significant changes in the expression of ${\beta}A$ and ${\beta}B$ mRNAs. Conclusion: It might be thought that inhibin acts as one of the regulatory factors in the ${\gamma}$-radiation-induced follicular atresia in mice

• Journal of Acupuncture Research
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• v.36 no.2
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• pp.92-99
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• 2019

Background: This study was conducted to evaluate the effects of pharmacopuncture and dermal application of Sebalgukhwa-san extracts on hair growth in an alopecia mouse model. Methods: Twenty-one C57BL/6 mice were divided into 3 groups; control group-normal saline injection or vehicle solution application, positive control group-minoxidil (MNXD), experimental group-pharmacopuncture and applied Sebalgukhwa-san (SGS) extract. The effects of the treatment on hair growth, were determined through photographs, and phototrichogram analysis by folliscope. Hair follicle morphometry by hematoxylin-eosin staining was performed, and hair growth-related protein expression of vascular endothelial growth factor, insulin like growth factor-1, and transforming growth factor-beta 1 were monitored by Western blotting. Serum levels of aspartate aminotransferase and alanine aminotransferase were measured for liver function test. Results: Body weight increased consistently in all groups. Hair growth was improved in the MNXD and SGS groups compared with the control. Hair density and thickness improved statistically significantly in the MNXD and SGS groups compared with the control p < 0.05. The number of hair follicles improved in the MNXD and SGS groups compared with the control but the size did not. The expression of vascular endothelial growth factor and insulin like growth factor-1 increased, and there was a decrease in the expression of transforming growth factor-beta 1 in the MNXD and SGS groups compared with the control, however, there was no significant difference. Sebalgukhwa-san treatment had no toxicity in liver function tests. Conclusion: Pharmacopuncture and dermal application of Sebalgukhwa-san extract may be therapeutically beneficial for the treatment of alopecia.

• Natural Product Sciences
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• v.29 no.1
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• pp.31-37
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• 2023

In this study, we investigated the efficacy of Celtis choseniana Nakai (C. choseniana) as complementary herbal medicine to ameliorate androgenic alopecia (AGA). The effects of C. choseniana on AGA were evaluated using testosterone propionate-induced AGA mouse model and dihydrotestosterone-treated human hair follicle dermal papilla cells. In vivo, C. choseniana treatment deactivated androgen signaling by reducing the concentration of serum dihydrotestosterone level and expressions of 5α-reductase 2 and androgen receptor. Next, C. choseniana treatment increased the hair regrowth rate. Histological studies demonstrated that C. choseniana induced the anagen phase in testosterone propionate-induced AGA mouse model. Cellular proliferation was promoted by C. choseniana treatment via increasing the expression of proliferation factors, such as proliferating cell nuclear antigen and cyclin D1. Furthermore, C. choseniana treatment increased the expression of proteins related to the Wnt/β-catenin signaling pathway. In addition, dickkopf-1, a Wnt inhibitor, was downregulated with C. choseniana treatment. Likewise, C. choseniana treatment promoted cellular proliferation in vitro. This study demonstrated the inhibitory effect of C. choseniana on androgen-induced AGA. Moreover, C. choseniana induced activation of Wnt/β-catenin signaling, resulting in prolonged anagen and cellular proliferation. Therefore, we suggest that C. choseniana can be used as a therapeutic agent to alleviate AGA.

• Development and Reproduction
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• v.7 no.1
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• pp.15-22
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• 2003

The Fas antigen (Fas) as a cell-surface receptor protein which mediates apoptosis-inducing signals plays an important role in the immune system. Expression of Fas mRNA is detected not only in lymphoid organs but also in the nonlymphoid organs. In the ovary, most of the follicles is known to undergo atreisa through apoptosis. However, the exact mechanism of atresia was not elucidated yet. Therefore, the purposes of the present study were to investigate the expression of Fas and Fas ligand in mouse ovary and to clarify the relationship between expression of Fas and Fas ligand and atresia of follicle. The result of RT-PCR demonstrated that Fas and Fas ligand mRNA was expressed in ovary, especially granulosa cells and oocytes. The immunohistochemistry showed that the granulosa cells and oocytes in growing follicles were stained for Fas and Fas ligand, but primordial follicles were not. Furthermore, Fas and Fas ligand were intensively stained in the atretic follicles As results of TUNEL staining to detect apoptotic cells in the ovaries, the number of TUNEL-positive (apoptotic) granulosa cells and oocytes increased in the atretic follicles compared to the healthy normal follicles. These results demonstrate that there is the positive relationship between expression of Fas and Fas ligand in granulosa cells and oocyies and apoptosis of them leading to atresia of follicles. It suggests that expression of Fas and Fas ligand could be associated with atresia of follicles in mouse ovary.

• Development and Reproduction
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• v.8 no.1
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• pp.27-33
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• 2004

To investigate the mechanism of germ cell death in postnatal stage of mouse, the involvement of apoptotic executioners, caspase-3 and caspase-activated DNase(CAD), and apoptotic initiators, Bax Fas and Fas ligand, in the germ cell death has been studied. Immune-labels of active caspase-3 and CAD were located in TUNEL-positive, apoptotic, oocytes as well as normal oocytes of primary or secondary follicles. CAD immune-labels were also detected in the nucleus of TUNEL-positive oocytes. Most of oocytes showing positive immune-labeling of active caspase-3 or CAD had vacuoles in their cytoplasm, which is the morphological characteristic of oocyte during folliclar atresia. Bax immune-stains were detected in the atretic oocytes which showed the vacuole in their cytoplasm. Positive immune-labels for Fas ligand was localized in TUNEL-positive or atretic oocytes. Presence of immunoreactivity of active caspase-3 and CAD in TUNEL-positive germ cells implicate that active raspase-3 and CAD might play a role in germ cell apoptosis during early development of mouse ovarian follicle. Immunohistochemical localization of Bax and Fas ligand in TUNEL-positive oocytes suggests that these might be the most plausible modulator of oocyte apoptosis.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.15 no.1
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• pp.96-126
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• 2002

In the course of screening natural extracts for hair growth, we found that the extract of dried root of Sophora flavescens has the prominent hair growth promoting effect. After topical application of Sophora flavescens extract to the back of C57BL/6 mice, the earlier conversion of telogen-to-anagen phase was induced. In addition, the Sophora flavescens extract revealed to possess potent inhibitory effect on $5{\alpha}$-reductase Ⅰ and Ⅱ activity. The growth of dermal papilla cells and mouse vibrissae hair follicle cultured in vitro, however, was not affected by Sophora flavescens extract treatment. RT-PCR analysis showed that Sophora flavescens extract induced mRNA levels of growth factors such as insulin-like growth factor-Ⅰ and keratinocyte growth factor in dermal papilla cells, suggesting hair growth promoting effect of Sophora flavescens extract is mediated through inhibition of $5{\alpha}$-reductase type Ⅱ activity and the regulation of growth factors in dermal papilla cells. Furthermore, Sophora flavescens extract also showed anti-bacterial effect on Propionibacterium acnes. These results suggest that Sophora flavescens can be used as a potent treatment agent for helping hair growth stimulation and acne inhibition.

• Biomolecules & Therapeutics
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• v.29 no.6
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• pp.643-649
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• 2021

Literature has revealed that the delta opioid receptor (DOR) exhibited diverse pharmacological effects on neuron and skin. In the present study, we have investigated whether the activation of DOR has hair-growth promotion effects. Compared with other opioid receptor, DOR was highly expressed in epidermal component of hair follicle in human and rodents. The expression of DOR was high in the anagen phase, but it was low in the catagen and telogen phases during mouse hair cycle. Topical application of UFP-512, a specific DOR agonist, significantly accelerated the induction of the anagen in C₃H mice. Topical application of UFP-512 also increased the hair length in hair organ cultures and promoted the proliferation and the migration of outer root sheath (ORS) cells. Similarly, pharmacological inhibition of DOR by naltrindole significantly inhibited the anagen transition process and decreased hair length in hair organ cultures. Thus, we further examined whether Wnt/β-catenin pathway was related to the effects of DOR on hair growth. We found that Wnt/β-catenin pathway was activated by UFP-512 and siRNA for β-catenin attenuated the UFP-512 induced proliferation and migration of ORS cells. Collectively, result established that DOR was involved in hair cycle regulation, and that DOR agonists such as UFP-512 should be developed for novel hair-loss treatment.

• Applied Microscopy
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• v.39 no.4
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• pp.301-309
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• 2009

Estrogens induce pronounced structural and functional changes in male and female reproductive system, but the exact mechanisms of estrogen are not fully understood. In relation to estrogen's function, the present study was designed to identify effects of estrogen receptor agonist, 4,4',4"- (4-propyl-[1H]-pyrazole-1,3,5-triyl)tris phenol (PPT) in the reproductive organ of the female mouse. The PPT was subcutaneously given to adult female mice at a weekly dosage of 3 mg in a volume 0.06 mL of vehicle for 3, 5 or 8 weeks whereas controls received weekly injections of the castor oil vehicle. Effects of PPT on reproductive organs were analyzed using a light microscope. PPT induced decreases of body, ovary and adipose tissue weights with experimental time. Ovary diameter of PPT treatment group was reduced as compared with control group. The number of Graffian follicle and corpus luteum was reduced in PPT treatment group. The luminal diameter of uterus was increased in relation with decrease of myometrium and endometrium height by PPT administration. The number of uterine glands was decreased by PPT treatment. These data indicate that PPT treatment induced morphological change of female reproductive organs resulting in alteration of fertility.

• Applied Microscopy
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• v.33 no.2
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• pp.117-130
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• 2003

This research investigates the fine structural as well as the morphological changes of the mouse ovarian tissue after irradiation of various dose rates of 6 MeV LINAC radiation. The normal structure of the ovarian tissue is consisted of various stages of follicles including primordial and growing follicles, and ovarian stromal connectives. When we observed the ovarian tissues irradiated with a dose rate of 200 cGy/min using light and electron microscopes, granular cells in growing follicles are in irregular shape unlike normal follicles. Small segments of cells scattered in follicular antrum among granular cells. We could observe neutrophils and macrophages around the segments, which means the cells already got in the process of decease owing to the effects radiation. With coincident to the increase of the dose rate of x-ray irradiation as 400 or 600 cGy/min, the mature follicles appeared as an irregular form and the granular cells surrounding oocyte also deformed comparing to their normal counterparts. The granulosa cells within mature follicle are already occurred necrotic change and apoptosis. The nuclei in some cells got so fragmented that the segments formed the shape of a horseshoe or scattered in small and condensed pieces. All the cells at a granular layer irradiated with a dose rate of 600 cGy/min show typical characteristics of apoptosis. The neutrophils involved in inflammatory reaction appear evidently in follicular antrum of growing follicles, and macrophage scattered with residual and apoptotic bodies.

• Development and Reproduction
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• v.3 no.1
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• pp.95-100
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• 1999

Growth differentiation factor-9 (GDF-9) is a member of the transforming growth factor $\beta$ (TGF-$\beta$) superfamily. It has been known that GDF-9 is a growth factor having a crucial role in normal folliculogenesis and its expression is oocyte-specific. The present study was aimed to elucidate the expression of GDF-9 mRNA in the mouse primordial follicles as well as in the other developmental stages. The semiquantitative analysis of GDF-9 mRNA expression was conducted. Total RNA was extracted from the ICR mice ovaries at gestational day 19, postnatal day 1, day 10, day 21, and day 28, and RT-PCR was performed to measure GDF-9 and $\beta$-actin mRNA levels. Level of GDF-9 mRNA were normalized against the level of $\beta$-actin mRNA, and compared among different stages. GDF-9 mRNA was detected in all samples including the fetal ovaries that mainly consists of primordial follicles. The highest level of mRNA was observed in ovaries obtained at day 10 that mainly consists of growing follicles. The present result suggests that GDF-9 may play an important role in the early stage of folliculogenesis.