• The Journal of Korean Medicine
• /
• v.20 no.1 s.37
• /
• pp.91-105
• /
• 1999

The effects of Yinjin and Yinjinsaryongsangagambang on a DNA damaging agent, etoposide-induced apoptosis, cell viability, cell cycle progression, and mRNA expression of apoptosis-related genes of human hepatocyte cell line HepG2 were investigated using tryphan blue exclusion assay, MTT assay, flow cytometry, immunocytometric analysis of PCNA, and quantitative RT-PCR analysis. MTT assay showed that Yinjin and Yinjinsaryongsangagambang increases cellular viability of HepG2 cells in a dosage-dependent manner. Stimulation of cell cycle progression by Yinjin or Yinjinsaryongsangagambang was detected by flow cytometric analysis of the DNA content and immunocytometric analysis of PCNA expression. A significant reduction of a DNA-damaging agent, etoposide-induced apoptosis were found in both Yinjin and Yinjinsaryongsangagambang-treated cells in dosage-dependent manner. In overall, 3-fold reduction of apoptosis was recognized in $10.0\;{\mu}g/ml$ of Yinjin or Yinjinsaryongsangagambang-treated cells compared to untreated cells. Although the difference is not significant, Yinjinsaryongsangagambang showed slightly higher effect on the inhibition of apoptosis than Yinjin. From flow cytometric analysis of apoptosis, while 39.9% of untreated cells showed etoposide-induced apoptotic cell death, only 19.6% or 17.4% of Yinjin or Yinjinsaryongsangagambang-treated cells were fond at apoptotic sub G1 phase, respectively. Interestingly, strong induction of Gadd45-mRNA was observed from Yinjin or Yinjinsaryongsangagambang-treated cells. However, no changes in expression levels of p53 and Waf1 were detected, demonstrating that induction of Gadd45 mRNA expression by Yinjin or Yinjinsaryongsangagambang occurs by p53-independent mechanism. Marked mRNA inductions of two apoptosis-inhibiting genes, Bcl-2 and Bcl- XL, were found in both Yinjin or Yinjinsaryongsangagambang-treated HepG2 cells while no changes was detected in expression levels of an apoptosis-promoting gene, Bax.

• Reproductive and Developmental Biology
• /
• v.28 no.1
• /
• pp.1-6
• /
• 2004

This study was conducted to investigate the effects of donor cell type, individual, passage number and trypsinization time on the in vitro development of bovine somatic cell nuclear transfer embryos. Three cell types (skin, muscle and cumulus cells) and cells from 3 individuals were used for nuclear transfer. Cell were passaged by 5, 15 or 30 times, and cell were trypsinized for 1 or 3 min before injection. Nuclear transfer were performed by conventional fusion method. Development rates to the blastocyst stage were not significantly different among three cell types (16.5∼23.9%) and individuals (16.4∼19.5%). Blastocyst formation rate of cloned embryos reconstituted with cells at passage 30 (5.8%) was significantly lower than those of embryos reconstituted with 5- and 15-passaged cells (25.3 and 23.5%, respectively, P＜0.05). The rate of embryos developed to the blastocyst stage was higher in embryos reconstituted with cells trypsinized for 1 min (30.7%) compared to embryos reconstituted with cells trypsinized for 3 min (P＜0.05). The result of the present study indicates that different donor cell types and individuals used in this study did not affect the development of cloned bovine embryos. However, passage number and trypsinization time of donor cells affect the in vitro development of cloned bovine embryos.

• The Journal of Korean Medicine
• /
• v.29 no.4
• /
• pp.68-82
• /
• 2008

Objective: The purpose of our study was to show the effects of Hwalhyulmaksung-bang (Huoxiemoxing-fang, HHMSB) treatment on cBSA-induced in MN Mouse Model. Methods: We divided the 20 mice into 4 groups. One group, named NR, was not treated. The second group, named CT, was treated with cBSA (7mg/kg i.p) only. The third group, named HH-250, was treated with cBSA (7mg/kg i.p) and HHMSB extract (250mg/kg, p.o). The fourth group, named HH-500, was treated with cBSA (7mg/kg i.p) and HHMSB extract (500mg/kg, p.o). 4 weeks after cBSA, proteinuria, serum albumin, total cholesterol, serum creatinine, BUN, total cell number of spleen and kidney of all groups were measured. CD3e+/CD19+ and CD4+/CD8 cells ratio of peripheral blood, kidney and spleen of all groups were analyzed. $IL-1{\beta}$ and TNF-$\alpha$, IL-6, IgG, IgM, and IFN-$\gamma$ levels of all groups were gauged. Histological analysis of kidney tissue and immunohistochemical staining (CD4, CD8) of kidney were observed. Results: The level of proteinuria significantly decreased and serum albumin increased in the group treated with cBSA and HHMSB extract compared with the control. Total cholesterol decreased but not significantly. CD3e+/CD19cells ratio of peripheral blood is decreased, but CD4+/CD8cells ratio has no significancy. CD3e+/CD19+ and CD4+/CD8 cells percentage of kidney and spleen has no significancy. Level of $IL-1{\beta}$, IL-6 is significantly decreased, and IFN-$\gamma$ is significantly increased on HHMSB compared with control. Total IgG level significantly decreased on HHMSB compared with the control. Thickness of GBM decreased on histological analysis of kidney. Deposition of CD4 and CD8 decreased on immunohistochemical staining of kidney. Conclusions: We conclude that Hwalhyulmaksung-bang treatment may could be a useful remedy agents for treating Membranous Neuropathy(MN) induced by cationized bovine serum albumin.

• Journal of Animal Science and Technology
• /
• v.63 no.6
• /
• pp.1397-1410
• /
• 2021

The present study was designed to determine the influence of all-trans retinoic acid (ATRA) on adipogenesis-related gene regulation in bovine intramuscular (IM) and subcutaneous (SC) adipose cells during differentiation. Bovine IM and SC adipocytes were isolated from three 19-mo-old, crossbred steers. Adipogenic differentiation was induced upon cultured IM and SC preadipocytes with various doses (0, 0.001, 0.01, 0.1, 1 µM) of ATRA. After 96 h of incubation, cells were harvested and used to measure the gene expression of CCAAT/Enhancer binding protein β (C/EBPβ), peroxisome proliferator-activated receptor (PPAR) γ, glucose transporter 4 (GLUT4), stearoyl CoA desaturase (SCD), and Smad transcription factor 3 (Smad3) relative to the quantity of ribosomal protein subunit 9 (RPS 9). Retinoic acid receptor (RAR) antagonist also tested to identify the effect of ATRA on PPARγ -RAR related gene expression in IM cells. The addition of ATRA to bovine IM decreased (p < 0.05) expression of PPARγ. The expression of PPARγ was also tended to be downregulated (p < 0.1) in high levels (10 µM) of ATRA treatment in SC cells. The treatment of RAR antagonist increased the expression of PPARγ in IM cells. Expression of C/EBPβ decreased (p < 0.05) in SC, but no change was observed in IM (p > 0.05). Increasing levels of ATRA may block adipogenic differentiation via transcriptional regulation of PPARγ. The efficacy of ATRA treatment in adipose cells may vary depending on the location.

• Journal of the Korean Chemical Society
• /
• v.67 no.2
• /
• pp.120-128
• /
• 2023

Osteoporosis is a disease that causes the weakening of bone by increasing porosity, which often results in fractures. Osteoporosis treatment measures include the use of Bisphosphonates and estrogen. However, these treatments cannot be used in the long term as these treatments have adverse side effects. Therefore, there is a need to identify better and safer treatment options. For this, 63 plant extracts were screened and among them, six extracts showed high anti-osteoclastic activity with low cytotoxicity. Of these six extracts, three extracts, Cudrania tricuspidata (P371), Ulmus davidiana var. japonica (P401), and Torilis japonica (P411), showed more than 50 percent osteoclast inhibition. While the remaining, Stewartia pseudocamellia extracts I and II (P370, P397) and Cuscuta chinensis (P418), showed moderate or between 40-50 percent osteoclast inhibition. Among all the extracts, Torilis japonica (P411) showed the highest inhibitory action against osteoclast development. Torilis japonica (P411) primary components include Kaempferol, Quercetin, and Luteolin, all proven to inhibit osteoclastogenesis. Stewartia pseudocamellia extracts I and II (P370 and P397) showed moderate or 44% osteoclast inhibition. Stewartia pseudocamellia extract II (P397) enhanced the growth of RAW 264.7 cells by 19%. Torilis japonica (P411) and Stewartia pseudocamellia extract II (P397) suppressed the expression of osteoclast-specific genes in RANKL-induced osteoclastogenesis in RAW 246.7 cells. Torilis japonica (P411) extracts even increased osteoblast-specific RUNX2 gene expression. This results provide that six extracts could be used as a potential treatment option for osteoporosis disease with the extracts of Torilis japonica (P411) and Stewartia pseudocamellia (P397) as an ideal candidates. However, the combination of the extract with higher osteoclastic inhibition and less toxic effects with further analysis should be recommended.

• Natural Product Sciences
• /
• v.19 no.2
• /
• pp.192-200
• /
• 2013

As part of the research for the natural products about prostate-related disease, this study screened 159 plant species from 46 families, which included a total of 213 different kinds of local native plants and these plants were tested for the ability to inhibit LNCaP proliferation, an androgen-sensitive prostate cancer cell line, and DU145 proliferation, which is a more aggressive androgen-insensitive prostate cancer cell line. The results indicated that nineteen of 213 types of plants exhibited antiproliferative activity (cell viability < 30%, $500{\mu}g/mL$) on the growth of androgen-sensitive LNCaP cell lines, and five of them exhibited DU145 cell antiproliferative activity (cell viability < 30%, $500{\mu}g/mL$). The methanol extracts of Eurya emarginata (stems), Gleditsia japonica var. koraiensis (leaves), Photinia glabra (leaves) and Elaeagnus macrophylla (leaves) showed antiproliferative activity on both the androgen-sensitive LNCaP cells (cell viability < 30%) and androgen-insensitive DU145 cells (cell viability > 100%). The study also found that the methanol extracts of Styrax japonica (fruits), Aralia continentalis (leaves), Fagus crenata var. multinervis (stems), Thuja orientalis (stems) and Poncirus trifoliate (branches) presented the strongest activity and demonstrated potent antiproliferative activity on both cell lines (LNCaP and DU145 cell viability < 30%).

• Clinical and Experimental Reproductive Medicine
• /
• v.28 no.2
• /
• pp.131-140
• /
• 2001

Objective: To know the effects of xenoestrogen on spermatogenesis, we investigated the expression of cytochrome P450s enzymes (CYPscc, $CYP_{17{\alpha}}$, CYP19) and $3{\beta}$-HSD genes involved in steroidogenesis. Methods: Mouse testicular cells were prepared from 15-day-old ICR mice which had only pre-meiotic germ cells by enzyme digestion using collagenase and trypsin. Testicular cells were cultured in DMEM supplemented with FSH (0.1 IU/ml) and 10% FBS or medium with estrogen ($E_2$), bisphenol-A (BPA), octylphenol (OP; $10^{-9},\;10^{-7},\;10^{-6},\;10^{-5},\;10^{-4}M$, respectively) and aroclor 1254 (A1254) known as PCBs for 48 hours. The gene expression of cytochrome P450 enzymes were examined by semi-quantitive RT-PCR. The production of estrogen and testosterone was examined by RIA. Results: As results, expression of CYPscc mRNA was not significantly decreased, but $3{\beta}$-HSD and $CYP_{17{\alpha}}$. mRNA were significantly dose-dependent decreased. And production of testosterone and estrogen were not different except BPA and OP group ($10^{-5}M$). Conclusion: BPA, OP and A1254 might inhibit steroidogenesis by decreasing CYPscc, $3{\beta}$-HSD and $CYP_{17{\alpha}}$. mRNA expression in the mouse testis. These results suggest that BPA, OP and PCBs like as an endocrine disruptors inhibit the productions of steroidogenic enzymes and decrease the production of T and E by negative feedback mechanism. Therefore, these might disrupt steroidogenesis in Leydig cells of testis and would disturb testicular function and subsequently impair spermatogenesis.

• Molecules and Cells
• /
• v.46 no.7
• /
• pp.420-429
• /
• 2023

Age-related macular degeneration (AMD) is one of the leading causes of blindness in elderly individuals. However, the currently used intravitreal injections of anti-vascular endothelial growth factor are invasive, and repetitive injections are also accompanied by a risk of intraocular infection. The pathogenic mechanism of AMD is still not completely understood, but a multifactorial mechanism that combines genetic predisposition and environmental factors, including cellular senescence, has been suggested. Cellular senescence refers to the accumulation of cells that stop dividing due to the presence of free radicals and DNA damage. Characteristics of senescent cells include nuclear hypertrophy, increased levels of cell cycle inhibitors such as p16 and p21, and resistance to apoptosis. Senolytic drugs remove senescent cells by targeting the main characteristics of these cells. One of the senolytic drugs, ABT-263, which inhibits the antiapoptotic functions of Bcl-2 and Bcl-xL, may be a new treatment for AMD patients because it targets senescent retinal pigment epithelium (RPE) cells. We proved that it selectively kills doxorubicin (Dox)-induced senescent ARPE-19 cells by activating apoptosis. By removing senescent cells, the expression of inflammatory cytokines was reduced, and the proliferation of the remaining cells was increased. When ABT-263 was orally administered to the mouse model of senescent RPE cells induced by Dox, we confirmed that senescent RPE cells were selectively removed and retinal degeneration was alleviated. Therefore, we suggest that ABT-263, which removes senescent RPE cells through its senolytic effect, has the potential to be the first orally administered senolytic drug for the treatment of AMD.

• Journal of Ginseng Research
• /
• v.47 no.1
• /
• pp.23-32
• /
• 2023

Coronavirus disease 2019 (COVID-19) is a highly infectious respiratory disease caused by a severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2). SARS-CoV-2 infection may cause clinical manifestations of multiple organ damage, including various neurological syndromes. There are currently two oral antiviral drugs-Paxlovid and molnupiravir-that are recognized to treat COVID-19, but there are still no drugs that can specifically fight the challenges of SARS-CoV-2 variants. Nucleotide-binding oligomerization domain-like receptor pyrin domain-containing-3 (NLRP3) inflammasome is a multimolecular complex that can sense heterogeneous pathogen-associated molecular patterns associated with neurological disorders. The NLRP3 activation stimulates the production of caspase-1-mediated interleukin (IL)-1β, IL-18, and other cytokines in immune cells. Panax (P.) ginseng is a medicinal plant that has traditionally been widely used to boost immunity and treat various pathological conditions in the nervous system due to its safety and anti-inflammatory/oxidant/viral activities. Several recent reports have indicated that P. ginseng and its active ingredients may regulate NLRP3 inflammasome activation in the nervous system. Therefore, this review article discusses the current knowledge regarding the pathogenesis of neurological disorders related to COVID-19 and NLRP3 inflammasome activation and the possibility of using P. ginseng in a strategy targeting this pathway to treat neurological disorders.

• Journal of Life Science
• /
• v.19 no.7
• /
• pp.892-898
• /
• 2009

MIF is a progesterone analogue and is known as a potent progesterone antagonist. Although MIF has been known to inhibit prostate cancer cell growth, its molecular mechanisms are not yet clear. In the present study, when the cells were treated for 2-4 days with 5-40 $\mu$M of MIF, the growth and viability of LNCaP cells were significantly decreased in a dose- and time-dependent manner. When the cells, cultivated in a normal 2 mM calcium concentration medium, were treated with 15 $\mu$M MIF for 1 day, the intracellular calcium level increased by 26% compared to the control. Similar results were also found in cells located in the calcium-free reaction buffer, indicating that MIF induced the increase of intracellular Ca$^{2+}$ levels, regardless of the presence of calcium in the surrounding medium. In the cells treated with various concentrations of MIF, the intracellular calcium levels increased in a dose dependent manner. Cells treated with MIF revealed typical early apoptotic signs, i.e., chromosome condensation and nuclei fragmentation. In cells treated with 40 11M MIF, Bcl-2 decreased to 19% of the control. The expression of Bax increased to almost 2 fold of the control. These results demonstrated very clearly that MIF treatment blocks the expression of Bcl-2 but stimulates the expression of Bax. According to the results of the present investigation, the apoptotic mechanism of MIF is triggered by intracellular modulation.