• The Korea Journal of Herbology
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• v.29 no.2
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• pp.61-67
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• 2014

Objectives : Aconiti Lateralis Preparata Radix (Aconitum Carmichaeli, AC) and Cinnamomi Cortex (Cinnamomi Cortex, CC) have been treated to elderly for kidney yang enhancement in Korean traditional medicine. In this study, the effects of water extract of AC and CC on RANKL (Receptor Activator for Nuclear Factor ${\kappa}B$ Ligand)-induced osteoclast differentiation were evaluated in culture system. Methods : MTT assay was used to evaluate the potential cytotoxicity of AC and CC extracts in bone macrophage marrows (BMMs) stimulated with M-CSF. TRAP (tartrate-resistant acid phosphatase) staining and TRAP activity were performed to know the inhibitory effect on osteoclast differentiation. The protein expression levels of nuclear factors such as activated T cell(NFAT)c1, c-Fos, MAPKs and ${\beta}$-actin in cell lysates treated with AC and CC extracts were analysed by western blotting. Results : AC, CC extracts and their co-administration inhibited significantly RANKL-induced osteoclast differentiation in BMMs in a dose dependent manner without toxicity. Each AC and CC extracts inhibited the phosphorylation of p38. Also, AC and CC extracts, respectively, inhibited the protein expression of c-Fos and NFATc1 more than Co-administration of AC and CC even if all treatments did. It was observed that RANKL-induced degradation of I-${\kappa}B$ is significantly suppressed by all treatments. Conclusions : Taken together, It was concluded that AC and CC have beneficial effect on osteoporosis by inhibition of osteoclast differentiation. Thus, Atractylodis AC and CC could be a treatment option for osteoporosis.

• Journal of Animal Science and Technology
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• v.65 no.4
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• pp.767-778
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• 2023

The aim of the research is to identify that porcine oocytes can function as recipients for interspecies cloning and have the ability to develop to blastocysts. Furthermore each mitochondrial DNA (mtDNA) in interspecises cloned embryos was analyzed. For the study, mouse-porcine and porcine-porcine cloned embryos were produced with mouse fetal fibroblasts (MFF) and porcine fetal fibroblasts (PFF), respectively, introduced as donor cells into enucleated porcine oocytes. The developmental rate and cell numbers of blastocysts between intraspecies porcine-porcine and interspecies mouse-porcine cloned embryos were compared and real-time polymerase chain reaction (PCR) was performed for the estimate of mouse and porcine mtDNA copy number in mouse-porcine cloned embryos at different stages.There was no significant difference in the developmental rate or total blastocyst number between mouse-porcine cloned embryos and porcine-porcine cloned embryos (11.1 ± 0.9%, 25 ± 3.5 vs. 10.1 ± 1.2%, 24 ± 6.3). In mouse-porcine reconstructed embryos, the copy numbers of mouse somatic cell-derived mtDNA decreased between the 1-cell and blastocyst stages, whereas the copy number of porcine oocyte-derived mtDNA significantly increased during this period, as assessed by real-time PCR analysis. In our real-time PCR analysis, we improved the standard curve construction-based method to analyze the level of mtDNA between mouse donor cells and porcine oocytes using the copy number of mouse beta-actin DNA as a standard. Our findings suggest that mouse-porcine cloned embryos have the ability to develop to blastocysts in vitro and exhibit mitochondrial heteroplasmy from the 1-cell to blastocyst stages and the mouse-derived mitochondria can be gradually replaced with those of the porcine oocyte in the early developmental stages of mouse-porcine cloned embryos.

• Biomolecules & Therapeutics
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• v.32 no.4
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• pp.432-441
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• 2024

Systemic sclerosis is an autoimmune disease characterized by inflammatory reactions and fibrosis. Myofibroblasts are considered therapeutic targets for preventing and reversing the pathogenesis of fibrosis in systemic sclerosis. Although the mechanisms that differentiate into myofibroblasts are diverse, transforming growth factor β (TGF-β) is known to be a key mediator of fibrosis in systemic sclerosis. This study investigated the effects of extracellular vesicles derived from human adipose stem cells (ASC-EVs) in an in vivo systemic sclerosis model and in vitro TGF-β1-induced dermal fibroblasts. The therapeutic effects of ASC-EVs on the in vivo systemic sclerosis model were evaluated based on dermal thickness and the number of α-smooth muscle actin (α-SMA)-expressing cells using hematoxylin and eosin staining and immunohistochemistry. Administration of ASC-EVs decreased both the dermal thickness and α-SMA expressing cell number as well as the mRNA levels of fibrotic genes, such as Acta2, Ccn2, Col1a1 and Comp. Additionally, we discovered that ASC-EVs can decrease the expression of α-SMA and CTGF and suppress the TGF-β pathway by inhibiting the activation of SMAD2 in dermal fibroblasts induced by TGF-β1. Finally, TGF-β1-induced dermal fibroblasts underwent selective death through ASC-EVs treatment. These results indicate that ASC-EVs could provide a therapeutic approach for preventing and reversing systemic sclerosis.

• Clinical and Experimental Reproductive Medicine
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• v.50 no.4
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• pp.292-298
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• 2023

Objective: Ovarian torsion is a gynecological disorder that causes ischemia-reperfusion injuries in the ovary. Our study investigated berberine's short- and long-term effects on ovarian ischemia-reperfusion injuries. Methods: This study included 28 Wistar albino female rats weighing 180 to 220 g, which were divided into four groups: sham (S), torsion/detorsion (T/D), torsion/ detorsion+single dose berberine (T/D+Bb), and torsion/detorsion+15 days berberine (T/D+15Bb). The torsion and detorsion model was applied in all non-sham groups. In the T/D+Bb group, a single dose of berberine was administered, while in the T/D+15Bb group, berberine was administered over a period of 15 days. After the rats were euthanized, their ovaries were excised. The left ovaries were used for histopathologic evaluation, which included ovarian injury scoring and follicle count, while the right ovaries were used for biochemical analyses (tissue transforming growth factor-β [TGF-β] and alpha-smooth muscle actin [α-SMA] levels). Results: The histopathologic evaluation scores for the ovaries were significantly lower in the T/D+B group (p<0.05) and the T/D+15B group (p<0.005) than in the T/D group. The follicle counts in the T/D group were lower than those in both the sham and treated groups (p<0.005). The TGF-β levels were significantly lower in the T/D+15B group (p<0.005), whereas the α-SMA levels did not show a significant difference. Conclusion: Both short- and long-term berberine use could potentially have therapeutic effects on ovarian torsion. Long-term berberine use exhibited anti-inflammatory effects by reducing TGF-β levels, thereby preventing ischemia-reperfusion injuries. Therefore, we suggest that long-term berberine use could be beneficial for ovarian torsion.

• The Korean Journal of Mycology
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• v.51 no.3
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• pp.179-190
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• 2023

Grapevine downy mildew, caused by Plasmopara viticola, significantly damages vineyards and is one of the most devastating diseases affecting cultivated grapes worldwide. In this study, we characterized the phenotypic and molecular traits of 11 P. viticola isolates from four grape-growing regions in South Korea. Additionally, we investigated the diversity of pathogenicity among these isolates and conducted an assay to evaluate the response of grape cultivars to P. viticola infection. Lemon-shaped sporangia were identified in the collected isolates, which released zoospores into the suspension at room temperature. Within a few hours of inoculation, the zoospores developed germ tubes. We tested 11 P. viticola isolates for pathogenicity in 845 grape cultivars to screen for grape host resistance to downy mildew infection. Among the tested isolates, JN-9 showed the highest virulence. Grape cultivars displayed varying phenotypic reactions to P. viticola infection: approximately 7% were highly susceptible, 41% were susceptible, 20% were moderately susceptible, 8% were resistant, and 24% exhibited extreme resistance. Phylogenetic analysis based on four genomic regions (internal transcribed spacer 1 [ITS1], actin, beta-tubulin, and cytochrome c oxidase II) revealed a close evolutionary relationship among all the Korean isolates, forming a single monophyletic lineage. Notably, these isolates showed greater similarity to European isolates than to American isolates. This comprehensive study contributes to a deeper understanding of the identity and behavior of P. viticola, which is crucial for developing effective resistance strategies against this pathogen in grape cultivars cultivated in South Korea.

• Herbal Formula Science
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• v.32 no.2
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• pp.141-153
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• 2024

Objective : Ephedrae Herba (Ephedra) has been frequently used in the East Asian traditional medicine including Korea, China and Japan in the clinical treatment of asthma, cold and influenza etc. This study was performed to explore an anti-fibrogenic potential of Ephedra Herba water extract (EHE) using immortalized human hepatic stellate cell line, LX-2 cells. Methods : We examined the anti-fibrogenic effects of EHE on canonical pathway of transforming growth factor-β1 (TGF-β1) signaling in LX-2 cells. Cell viability was measured using the MTT assay. mRNA levels were detected by real-time PCR. Proteins expression were detected by Western blot. Results : Treatment of EHE 30 ㎍/ml did not show any cytotoxicity on LX-2 cells. Pre-treatment of EHE (30 ㎍/mL) significantly inhibited α-smooth muscle actin expression induced by TGF-β1. Additionally, EHE significantly decreased Smad2 and Smad3 phosphorylations, Smad binding element-driven luciferase activity and plasminogen activator inhibitor type 1 expression by TGF-β1. Furthermore, increases of matrix metalloproteinases 2 genes by TGF-β1 was also attenuated by EHE treatment. Conclusion : These results suggest that EHE has an ability to suppress fibrogenic process in activated HSC via inhibition of TGF-β1-TGFBR mediated canonical (Smad dependent) pathway.

• Clinical and Experimental Pediatrics
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• v.49 no.5
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• pp.558-564
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• 2006

Purpose : Transcranial electromagnetic stimulation(TMS) is a noninvasive method which stimulates the central nervous system through pulsed magnetic fields without direct effect on the neurons. Although the neurobiologic mechanisms of magnetic stimulation are unknown, the effects on the brain are variable according to the diverse stimulation protocols. This study aims to observe the effect of the magnetic stimulation with two different stimulation methods on the cultured hippocampal slices. Methods : We obtained brains from 8-days-old Spague-Dawley rats and dissected the hippocampal tissue under the microscope. Then we chopped the tissue into 450 µm thickness slices and cultured the hippocampal tissue by Stoppini's method. We divided the inserts, which contained five healthy cultured hippocampal slices respectively, into magnetic stimulation groups and a control group. To compare the different effects according to the frequency of magnetic stimulation, stimulation was done every three days from five days in vitro at 0.67 Hz in the low stimulation group and at 50 Hz in the high stimulation group. After N-methyl-D-aspartate exposure to the hippocampal slices at 14 days in vitro, magnetic stimulation was done every three days in one and was not done in another group. To evaluate the neuronal activity after magnetic stimulation, the $NeuN/{\beta}$-actin ratio was calculated after western blotting in each group. Results : The expression of NeuN in the magnetic stimulation group was stronger than that of the control group, especially in the high frequency stimulation group. After N-methyl-D-aspartate exposure to hippocampal slices, the expression of NeuN in the magnetic stimulation group was similar to that of the control group, whereas the expression in the magnetic non-stimulation group was lower than that of the control group. Conclusion : We suggest that magnetic stimulation increases the neuronal activity in cultured hippocamal slices, in proportion to the stimulating frequency, and has a neuroprotective effect on neuronal damage.

• Microbiology and Biotechnology Letters
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• v.44 no.4
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• pp.571-576
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• 2016

In this study, the anti-inflammatory activities of the 80% ethanol extract of Dystaenia takeshimana (DT) were investigated using Raw 264.7 cells treated with lipopolysaccharide (LPS). The effect of DT extract on the production of pro-inflammatory factors (iNOS, COX-2) in LPS-stimulated Raw 264.7 macrophages was examined. The cytotoxic effect of DT extract on macrophage cells (Raw 264.7) was examined by the 3-[4, 5-dimethyl-thiazol-2-yl]-2, 5-diphenyl-tetrazoliumbromide (MTT) assay. Treatment with DT extract showed 100% or more cell viability at the concentration $1,000{\mu}g/ml$. The inhibitory effect of DT extract on protein expression of inducible NOS (iNOS) and cyclooxygenase-2 (COX-2) was measured by western blotting using the concentrations 50, 100, and $500{\mu}g/ml$, with ${\beta}-actin$ used as the positive control. Consequently, the protein expression of iNOS, and COX-2 as observed by western blotting, was decreased by 56%, 61.6%, respectively with $500{\mu}g/ml$ DT extract. Inhibition of iNOS and COX-2 mRNA expression was measured by reverse transcription- polymerase chain reaction (PCR) using DT extract concentrations 50, 100, and $500{\mu}g/ml$, with GAPDH used as a positive control. Consequently, the mRNA expression of iNOS and COX-2 as observed by reverse-transcription-PCR was decreased by 77.9% and 83.3%, respectively at $500{\mu}g/ml$ concentration of DT extract. In conclusion, DT extract may affect inflammatory factors as a potential anti-inflammatory agent.

• Microbiology and Biotechnology Letters
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• v.45 no.2
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• pp.178-183
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• 2017

This study examined a new functional cosmetic material possessing application possibility of Sambucus sieboldiana var. pendula (Nakai) (SS) extract. For this, we analyzed the toxic effect of the SS extract on macrophages (RAW 264.7 cells) by performing MTT assay. Results of the MTT assay showed ${\geq}100%$ cell viability after treatment with $500{\mu}g/ml$ SS extract. To determine the anti-inflammatory activity of the SS extract, we examined its inhibitory effect on lipopolysaccharide (LPS)-induced NO production in RAW 264.7 cells by performing Griess assay. Result of the Griess assay showed that the SS extract inhibited LPS-induced NO production in a concentration-dependent manner. Next, we examined the effect of the SS extract on the production of proinflammatory factors inducible NOS (iNOS) and cyclooxygenase-2 (COX-2) in LPS-stimulated RAW 264.7 cells. First, we determined the inhibitory effect of 50, 100, and $500{\mu}g/ml$ SS extract on iNOS and COX-2 protein expression by performing western blot analysis, with ${\beta}$-actin as a positive control. Results of western blotting showed that treatment with $500{\mu}g/ml$ SS extract decreased iNOS and COX-2 protein expression by 31.2% and 54.7%, respectively. Next, we determined the inhibitory effect of 50, 100, and $500{\mu}g/ml$ SS extract on iNOS and COX-2 mRNA expression by performing reverse transcription-polymerase chain reaction (PCR), with GAPDH as a positive control. Results of reverse transcription-PCR showed that treatment with $500{\mu}g/ml$ SS extract decreased the mRNA expression of iNOS and COX-2 by 72.2% and 89%, respectively. These results suggest that the SS extract is a highly valuable natural compound because of its functional components and anti-inflammatory activity.

• Tuberculosis and Respiratory Diseases
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• v.49 no.4
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• pp.453-465
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• 2000

Background : Lung carcinogenesis is a multistage process involving alterations in multiple genes and diverse pathway. Mutational activation of oncogenes and inactivation of tumor suppressor genes, and subsequent increased genetic instability are the major genetic events. The p53 gene and FHIT gene as tumor suppressor genes contribute to the pathogenesis of lung cancer, evidenced by mutation, microsatellite instability(MI) and loss of heterozygosity(LOH). Methods : We analysed genetic mutations of p53 and FHIT gene in 29 surgical specimens of nonsmall cell lung cancer using PCR-single strand conformation polymorphism, DNA sequencing and RT-PCR. MI and LOH were analyzed in loci of D3S1285, D9S171, and TP53. Results : In 2 cases, point mutation of p53 gene was observed on exon 5. MI of 3 times and LOH of 14 times were observed in at least one locus. In terms of the location of microsatellite, D3S1285 as a marker of FH1T was observed in 5 cases out of 26 specimens; D9S171 as a marker of p16 in 5 out of 17; and TP53 as a marker of p53 in 7 out of 27. In view of histologic type, squamous cell carcinoma presented higher frequency of microsatellite alteration, compared to others. Mutation of FHIT gene was observed in 11 cases and 6 cases of those were point mutation as a silent substitution on exon 8. FHIT mRNA expression exhibited deletion on exon 6 to 9 in 4 cases among 15 specimens, presenting beta-actin normally. Conclusion : Our results show comparable frequency of genetic alteration in nonsmall cell lung cancer to previous studies of Western countries. Microsatellite analysis might have a role as a tumor marker especially in squamous cell carcinoma. Understanding molecular abnormalities involved in the pathogenesis could potentially lead to prevention, earlier diagnosis and the development of novel investigational approaches to the treatment of lung cancer.