Objectives: This study aimed to examine the safety, effects on proliferation of hair papilla cells, and anti-inflammatory and antioxidant mechanisms of Artemisia sieversiana Ehrh. ex Willd. (AS) extract. Methods: Safety tests through purity testing, acute toxicity tests, and repeated toxicity tests were performed using AS extract (ASE) which had been dried for over two years. Cell culture and proliferation tests were conducted; VEGF (vascular endothelial growth factor), bFGF (basic fibroblast growth factor), and EGF (epidermal growth factor) and protein expression analyses were performed for mechanistic evaluation; and inhibitory effects of ASE on the RNA expression of testosterone, 5𝛼-reductase, and aromatase was assessed. The anti-inflammatory and antioxidant efficacy of ASE was confirmed by measuring the levels of nitric oxide, inflammatory mediators (TNF-𝛼 and PGE2), inflammatory cytokines (IL-1𝛽, IL-6, and IL-8), and chemokine MCP-1. Results: The safety of ASE was confirmed. The mechanism of cell proliferation in human hair follicle dermal papilla cells involved the promotion of VEGF, bFGF, and EGF expression. ASE decreased mRNA expression of testosterone, 5𝛼-reductase, and aromatase-1 in a concentration-dependent manner. PGE2 and TNF-𝛼 production by inflammatory mediators was also significantly decreased in a concentration-dependent manner, and inflammatory cytokine and chemokine expression was inhibited. Conclusions: ASE is suggested to promote papillary cell growth at the cellular level, to suppress expression of various enzymes involved in hair cycle and cell death, and to inhibit hair loss through anti-androgen, anti-inflammatory, and antioxidant effects.
Soo Hyun Kim;Kyu-Sang Sim;Jung Yoon Cheon;Jae-Woong Jang;Su Jin Jeong;Ye Hei Seo;Hye Myoung Ahn;Bong-Geun Song;Gi-Seok Kwon;Jung-Bok Lee
Journal of Life Science
/
v.33
no.3
/
pp.234-241
/
2023
At present, many countries around the world are legalizing cannabis and its products, and research on various treatments using cannabis is being actively conducted. However, the cannabis plant contains other compounds whose biological effects have not yet been established. We investigated the effect of cannabidiol (CBD) on hair growth in human dermal papilla cells (HDPCs). 2,2'-Azino-bis (3-ethylbenzothiazolin-6-sulfonic acid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assays were performed to determine the antioxidant activity of CBD. The HDPCs viability of CBD was examined via water-soluble tetrazolium salt (WST-1) assay. The expression of hair-loss-related markers in HDPCs by CBD treatment was analyzed by real-time PCR and western blotting. The DPPH, ABTS radical scavenging activity assay showed that CBD had superior antioxidant activities. In HDPCs, CBD increased cellular proliferation at concentrations without cytotoxicity. It also increased the expressions of fibroblast growth factor 1 (FGF1), fibroblast growth factor 7 (FGF7), vascular endothelial growth factor (VEGF), and insulin-like growth factor (IGF). These results correlated with a decrease in the expression of inhibition-related factors, such as androgen receptor (AR) and transforming growth factor beta 1 (TGF-B1). Moreover, CBD resulted in a significant increase in the phosphorylation of AKT and extracellular signal-regulated kinase (ERK). Therefore, it is suggested that CBD may be a potential remedy for the treatment of alopecia.
Although the main geology of Korea consists of granite and gneiss, it Is not uncommon to encounter anisotropy Phenomena in crosshole radar tomography even when the basement is crystalline rock. To solve the anisotropy Problem, we have developed and continuously upgraded an anisotropic inversion algorithm assuming a heterogeneous elliptic anisotropy to reconstruct three kinds of tomograms: tomograms of maximum and minimum velocities, and of the direction of the symmetry axis. In this paper, we discuss the developed algorithm and introduce some case histories on the application of anisotropic radar tomography in Korea. The first two case histories were conducted for the construction of infrastructure, and their main objective was to locate cavities in limestone. The last two were performed In a granite and gneiss area. The anisotropy in the granite area was caused by fine fissures aligned in the same direction, while that in the gneiss and limestone area by the alignment of the constituent minerals. Through these case histories we showed that the anisotropic characteristic itself gives us additional important information for understanding the internal status of basement rock. In particular, the anisotropy ratio defined by the normalized difference between maximum and minimum velocities as well as the direction of maximum velocity are helpful to interpret the borehole radar tomogram.
Changes in the fine structure of testicular Leydig cell from hatching to adulthood were studied in Korean native chickens of 1, 2, 4, 6, 8, 10, 12, 14, 16, 18, 21, 24, 28, 32, 44, 52 and 64 weeks (n=13 chickens per group) of age. The objective of this study were to elucidate Leydig cell ultrastructure during testicular development. Testes of chickens were fixed by whole body perfusion using a fixative containing 2.5% glutaraldehyde in cacodylate buffer, processed and embedded in Epon-araldite. Using $1{\mu}m$ sections stained with methylene blue-azure II, qualitative and quantitative(stereological) morphological studies were performed. The ultrastructural changes of the Leydig cell were investigated by ultrathin section with the transmission electron microscope. The stages of the Leydig cell development described focus on mitochondria, endoplasmic reticulum, and lipid droplets which are involved in androgens as fullows. 1) Approaching puberty. The closely packed Leydig cells and sparse intercellular space. The nucleus occupied a large portion of the Leydig cell volume. The population of Leydig cells contained two types of cells that differed in the appearance of their nuclei which were either highly electron-opaque or relatively electron-lucid. The cytoplasm was characterized by large amounts of lipid droplets, relatively few spherical mitochondria, and sparse smooth endoplasmic reticulum. 2) Puberty to adult. The Leydig cells which display features compatible with significant androgen synthesis: large volume of cytoplasm containing extended smooth endoplasmic reticulum, abundant mitochondria, and reduction of lipid droplets.
Vinclozolin (VCZ) is a systemic fungicide commonly used in fruits, vegetables and the wine industry. VCZ and its metabolites, butenoic acid (M1) and enanilide (M2) derivatives, act as anti-androgens through actions on the androgen receptor. Although there is growing body of evidence that VCZ's action as an endocrine disrupting chemical (EDC) in male reproductive physiology and pathphysiology, no evidence on the VCZ's EDC action in female is available yet. Previously we found that the prepubertal VCZ exposures could effectively delay the onset of puberty in female rats, suggesting the postponed or weakened activities of hypothalamus-pituitary-ovary (H-P-O) reproductive hormonal axis. The present study was performed to examine whether the VCZ administration affects the transcriptional activities of reproductive hormone-related genes in the same animal model. VCZ (10 mg/kg/day) was administered daily from postnatal day 21 (PND 21) through the day when the first vaginal opening (V.O.) was observed. To determine the transcriptional changes of reproductive hormone-related genes in hypothalamus and pituitary, total RNAs were extracted and applied to the semiquantitative reverse transcription polymerase chain reaction (RT-PCR). As a result, treatment with VCZ significantly lowered the transcriptional activity of nitric oxide synthase-2 (NOS-2) which is known to adjust gonadotropin-releasing hormone (GnRH) secretion in the hypothalamus (p<0.01). Similarly, the mRNA levels of KiSS-1, G protein-coupled receptor 54 (GPR54) and GnRH were significantly decreased in hypothalamus (p<0.01) from VCZ-treated group. As expected, the transcriptional activities of luteinizing hormone-${\beta}$ (LH-${\beta}$) and follicle stimulating hormone-${\beta}$ (FSH-${\beta}$) in the anterior pituitary from VCZ-treated group were also significantly lower than those from the control group. The present study indicates that(i) the inhibitory effect of VCZ exposure on the onset of puberty in immature female rats could be derived from the reduced transcriptional activities of gonadotropin subunits and their upstream modulators such as GnRH and KiSS-1 in hypothalamus-pituitary neuroendocrine axis, and (ii) these inhibitory effects could be mediated by NO signaling pathway.
Background: The CREST syndrome is an indolent form of progressive systemic sclerosis. Although its clinical progress is indolent, pulmonary hypertension(PH) associated with CREST syndrome have grave prognosis with over 40 percent mortality rate at 2 year follow-up. But the pathogenesis of pulmonary hypertension in this disease is not known, and classified as either primary or secondary PH. Clonality of endothelial cell proliferation in plexiform lesion is a molecular marker which allows distinction between primary and secondary PH. We performed this study to know whether the PH associated with CREST syndrome is a variant of primary PH or is a secondary PH. Methods: We assessed the X-chromosome inactivation based on the methylation pattern of the human androgen-receptor gene by PCR(HUMARA). Endothelial cells in plexiform lesions from female patients(n=3) with PH associated with CREST syndrome were microdissected from paraffin blocks. Vascular smooth muscle cells and lung parenchyma were also microdissected for clonality studies. Results: The proliferating endothelial cells in 14 plexiform lesions were all polyclonal. Similarly proliferated smooth muscle cells from 5 vessels with medial hypertrophy were also polyclonal. Conclusion: These results suggest that the pulmonary hypertension associated with CREST syndrome has different pathogenesis from primary PH and to be classified as secondary PH.
Mifepristone (MIF) and Tamoxifen (TAM) have been used in the treatment of prostate cancer and breast cancer for more than a decade. MIF can induce apoptosis in both AR-positive and negative prostate cancer cells. Because of its pleiotropic ligand-receptor properties, TAM exerts cytotoxic activity in estrogen (ER)-positive and various ER.negative cancer cells. However, the molecular mechanisms of these two substances are not yet clear. In the present work, we report that the cytotoxic effects of MIF and TAM are due to the modulation of intracellular $Ca^{2+}$ level in DU-145, androgen-insensitive cells. When the cells were treated with micromolar concentrations of either MIF or TAM, the growth and viability were significantly decreased in a dose- and time-dependent manner. The apoptosis induced by MIF or TAM was further proved and analyzed by confocal laser scanning microscopy (CLSM) and fluorescence-activated cell sorting (FACS). In the cells cultivated in a normal 1.5 mM $Ca^{2+}$ medium, both MIF and TAM also induced an increase of the intracellular $Ca^{2+}$ level in a dose-dependent fashion. Since a change in calcium level could not be found in cells of the $Ca^{2+}$-free medium, the increase of intracellular $Ca^{2+}$ level might be due to an increase in extracellular calcium uptake. Our results show that the apoptotic effect was more prominent in TAM treatment compared to MIF treatment in DU-145 cells. The above findings might be due to the difference in the uppermost pathways of apoptosis induced by either MIF or TAM. When we checked the level of procaspase-8 activation, TAM showed minor level of activation, as opposed to MIF, which exerted strong activation. In both treatments, the levels of anti-apoptotic protein Bcl-2 decreased, and pro-apoptotic protein Bax level increased more than 2-fold. The activation of caspase-3, a key protease enzyme in the downstream pathway of apoptosis, was much higher in the cells treated with TAM, compared to the MIF treatment. The overall apoptotic activity shown in the present work was closely related to intracellular $Ca^{2+}$ concentration levels. Therefore, the cytotoxic activity induced by MIF and TAM might have been due to intracellular calcium modulation.
Sex steroids are known to be involved in skeletal muscle development (anabolic effect) and are frequently used in medicines. It has been known that pork contains a variety of steroids that are mainly synthesized in the gonads (testis and ovary). Thus, the present study was conducted to evaluate the effects of anabolic steroids of pork on the proliferation and differentiation of myogenic satellite cells (MSC). Three different methods (M1, M2, and M3) were developed for the isolation and purification of steroids from porcine tissues. Among three extraction methods that we developed, M3 was the best method with respect to the quantities of steroids and the induction of MSC proliferation. Hormonal analysis showed that the steroid hormone levels were the highest in muscle and fat of intact male than those of castrated males and females. In addition, the highest serum levels of nandrolone and testosterone were detected in intact males, whereas estrone and $17{\beta}$-estradiol levels were similar in the entire experimental serum samples. Expression of androgen receptor (AR), myoD, desmin, and myogenin in bovine muscle cells were significantly up-regulated by the treatment of steroid extracts. The highest increas of myogenin and AR mRNA abundance were observed in the MSCs treated with M3 extract (p<0.001). Altogether, the present research showed the positive effect of steroids on MSC proliferation and differentiation in vitro. These results would certainly imply a beneficial effect of pork consumption on human muscle development.
Vinclozolin(VCZ), a systemic dicarboximide fungicide, has been used in the control of diseases caused by microorganism of some species in fruits, vegatables and ornamental plants. Although VCZ itself is a very weak antagonist for androgen receptor binding, both melabolites M1 and M2 are effective antagonists. The present study was undertaken to examine whether prepubertal exposure to VCZ affects on the onset of puberty and the associated reproductive parameters such as hormone receptor expressions in female rats. VCZ(10 mg/kg/day) was administered daily from postnatal day 21(PND 21) through the day when the first vaginal opening(V.O.) was observed. Gross anatomy and weight of reproductive tissues were compared to test the VCZ's effects on the cell proliferation. Furthermore, histological studies were performed to assess the structural alterations in the tissues. To determine the transcriptional changes in progesterone receptor(PR), total RNAs were extracted and applied to the semi-quantitative reverse transcription polymerase chain reaction(RT-PCR). As a result, delayed V.O. was shown in the VCZ group(PND $34.00{\pm}1.22$) compared to the control group(PND $38.20{\pm}1.92$; p<0.01). VCZ treatment significantly decreased the wet weight of ovaries and uteri compared to the control group(p<0.01). Graafian follicles and corpora lutea were observed only in the ovaries from the control animals, while numerous primary, secondary follicles and small atretic follicles were observed in the ovaries from VCZ group. Similarly, hypotrophy of luminal and glandular uterine epithelium was found in the VCZ group. In the semi-quantitative RT-PCR studies, the transcriptional activity of PR in ovary(p<0.01) from VCZ group were significantly lower than those from the control group while in uterus were similar compared with the control group. The present studies demonstrated that the acute exposure to VCZ during the critical period of prepubertal stage could inactivate the reproductive system resulting delayed puberty in female rats.
Colored potatoes are an excellent source of dietary polyphenols including anthocyanins. Generally, anthocyanins from fruits and vegetables exhibit anti-carcinogenesis and anti-cancer properties in vitro test. This experiment was conducted to know the effects of colored potato extracts contained anthocyanins on antimutagenic activity and anticancer activity to six human cancer cell lines containing LNCaP (androgen-dependent) prostate cancer cells. Extracts of three colored potatoes ('Hongyoung', 'Jayoung' and 'Jasim') and the white potato ('Superior') cultivars were used in this study. The extracts of three colored potatoes inhibited the mutagenicities induced by direct mutagen such as 4-nitro-quinoline-1-oxide (4-NQO) and another indirect mutagens of bezo(a)pyrene (BaP). Also, the extracts of 'Hoyoung' and 'Jayoung' showed higher antimutagenic activity than 'Jasim' and 'Superior' against to direct or indirect mutagen on both strains of TA98 and TA100. The activity of growth-inhibitory of extract of four potato cultivars were screened by SRB (sulphorhodamine B) method on diverse human cancer cells representing different types of cancers. Among the extract of four potato cultivars, the extract of 'Jasim' showed moderate inhibition on proliferation of LNCaP, ACHN and MOLT-4F cells and did not inhibit the proliferation of other cancer cells. On the other hand, extract of 'Superior' did not inhibit the proliferation of any tested cancer cell lines. However, the extracts of 'Hongyoung and Jayoung' inhibited the proliferation of cancer cells with $GI_{50}$ values ranging from 2.5 to $30\;{\mu}g/mL$. On the basis of the $GI_{50}$ values, it is clear that LNCaP cells were more sensitive to extracts of colored potato cultivars than other cancer cells. The extract of 'Jayoung' at $30\;{\mu}g/mL$ were more active and inhibited cell proliferation, and induced apoptosis in LNCaP cells. This result revealed that the extracts of colored potatoes are expected to be good candidate for development into source of antimutagenic and anticancer agent.
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