• Microbiology and Biotechnology Letters
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• v.49 no.3
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• pp.283-288
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• 2021

This study investigated the anticancer activity of methanol extract from Platycarya strobilacea leaf in AGS human gastric cancer cells. We determined the cell viability effect of P. strobilacea using MTS assay. Apoptosis induction and cell cycle arrest were confirmed by fluorescein isothiocyanate and propidium iodide staining using cellometer K2. The mRNA expression levels of the Bcl-2 family were confirmed by reverse transcription-polymerase chain reaction. The cell viability was decreased in a dose-dependent manner treated with different concentrations of P. strobilacea. Total, early, and late apoptotic cells were dramatically increased, and the cell cycle was arrested at the sub-G1 phase. The mRNA expressions of Bcl-2 and Bcl-xL were reduced, whereas pro-apoptotic factors, Bax and Bak, were increased in a dose-dependent manner. These results suggested that P. strobilacea leaf extract induced significant apoptotic activity through an intrinsic mitochondria pathway.

• Laboraroty Animal Research
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• v.34 no.4
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• pp.195-202
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• 2018

Hyperglycemia is one of the major risk factors for stroke. Hyperglycemia can lead to a more extensive infarct volume, aggravate neuronal damage after cerebral ischemia. ${\alpha}$-Synuclein is especially abundant in neuronal tissue, where it underlies the etiopathology of several neurodegenerative diseases. This study investigated whether hyperglycemic conditions regulate the expression of ${\alpha}$-synuclein in middle cerebral artery occlusion (MCAO)-induced cerebral ischemic injury. Male Sprague-Dawley rats were treated with streptozotocin (40 mg/kg) via intraperitoneal injection to induce hyperglycemic conditions. MCAO were performed four weeks after streptozotocin injection to induce focal cerebral ischemia, and cerebral cortex tissues were obtained 24 hours after MCAO. We confirmed that MCAO induced neurological functional deficits and cerebral infarction, and these changes were more extensive in diabetic animals compared to non-diabetic animals. Moreover, we identified a decrease in ${\alpha}$-synuclein after MCAO injury. Diabetic animals showed a more serious decrease in ${\alpha}$-synuclein than non-diabetic animals. Western blot and reverse-transcription PCR analyses confirmed more extensive decreases in ${\alpha}$-synuclein expression in MCAO-injured animals with diabetic condition than these of non-diabetic animals. It is accepted that ${\alpha}$-synuclein modulates neuronal cell death and exerts a neuroprotective effect. Thus, the results of this study suggest that hyperglycemic conditions cause more serious brain damage in ischemic brain injuries by decreasing ${\alpha}$-synuclein expression.

• Korean Journal of Agricultural Science
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• v.46 no.3
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• pp.653-660
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• 2019

Ganoderma lucidum, an oriental polypore fungus and medicinal mushroom, has a long history of use for promoting health and longevity in Korea, China, and other Asian countries. This study was aimed at determining the anti-inflammatory activity and mechanism of action of Ganoderma lucidum in murine macrophage RAW 264.7 cells. Ganoderma lucidum was extracted with ethanol and freeze-dried. The anti-inflammatory effect (nitrite production) of Ganoderma lucidum extracts was tested using a nitric oxide (NO) colorimetric assay. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) was performed to quantify the mRNA expression of cytokines including tumor necrosis factor-${\alpha}$ ($TNF-{\alpha}$), interleukin $(IL)-1{\beta}$, and IL-6. Western blotting was performed to measure the expression levels of inflammation-related proteins, such as inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), nuclear factor kappa B ($NF-{\kappa}B$) p65, and phosphorylated $NF-{\kappa}B$ p65. The NO colorimetric assay showed that NO production increased with the treatment of lipopolysaccharide in (LPS)-activated RAW 264.7 macrophages and decreased with the cotreatment of Ganoderma lucidum extracts and LPS. Ganoderma lucidum extracts repressed the mRNA expressions of cytokines, which were increased after the LPS treatment. In addition, Ganoderma lucidum extracts inhibited the LPS-induced expression of iNOS and COX-2 and the LPS-induced phosphorylation of $NF-{\kappa}B$ p65. These results suggest that the Ganoderma lucidum extracts exert an anti-inflammatory activity by inhibiting $NF-{\kappa}B$ related proteins and cytokines.

• Journal of Physiology & Pathology in Korean Medicine
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• v.33 no.2
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• pp.109-115
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• 2019

Aster Yomena (AY) has been used in traditional medicine to treat diseases such as obesity, hyperlipidemia, atherosclerosis, diabetes and osteoarthritis. However, protective effect of AY on acute pancreatitis (AP) has not been reported. The present study examined the anti-inflammatory effects of an ethanol extract of AY on cerulein-induced AP. AP was induced in mice by intraperitoneally injecting cerulein ($50{\mu}g/kg$) hourly for 6 times. 70% ethanol extract of AY (0.1, 0.2, and 0.5 g/kg) was orally administered for 1 week before acute pancreatitis induction. The mouse was killed at 6 hours after the final cerulein injection. The pancreas and lung were rapidly removed for histological examination and myeloperoxidase (MPO) assay. Blood samples were taken to determine serum amylase and lipase activity. In addition real-time reverse transcription-polymerase chain reaction (RT-PCR) was also performed to investigate mRNA expression of proinflammatory cytokines such as $TNF-{\alpha}$. $IL-1{\beta}$, and IL-6. Administration of AY significantly ameliorated pancreatic weight to body weight ratio, histological damages and MPO activity during AP. In addition, AY inhibited the serum amylase and lipase activity during AP. Also, mRNA expression of $TNF-{\alpha}$, $IL-1{\beta}$ and IL-6 were inhibited by AY against AP. Our results revealed that pre-treatment of AY reduces the severity of cerulein-induced AP. Therefore, AY may have a protective effect drug against AP.

• Journal of Ginseng Research
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• v.43 no.2
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• pp.261-271
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• 2019

Background: Protopanaxatriol (PPT) is an aglycone of ginsenosides, which has high medicinal values. Production of PPT from natural ginseng plants requires artificial deglycosylation procedures of ginsenosides via enzymatic or physicochemical treatments. Metabolic engineering could be an efficient technology for production of ginsenoside sapogenin. For PPT biosynthesis in Panax ginseng, damarenediol-II synthase (PgDDS) and two cytochrome P450 enzymes (CYP716A47 and CYP716A53v2) are essentially required. Methods: Transgenic tobacco co-overexpressing P. ginseng PgDDS, CYP716A47, and CYP716A53v2 was constructed via Agrobacterium-mediated transformation. Results: Expression of the three introduced genes in transgenic tobacco lines was confirmed by Reverse transcription-polymerase chain reaction (RT-PCR). Analysis of liquid chromatography showed three new peaks, dammarenediol-II (DD), protopanaxadiol (PPD), and PPT, in leaves of transgenic tobacco. Transgenic tobacco (line 6) contained $2.8{\mu}g/g$ dry weight (DW), $7.3{\mu}g/g$ DW, and $11.6{\mu}g/g$ DW of PPT, PPD, and DD in leaves, respectively. Production of PPT was achieved via cell suspension culture and was highly affected by auxin treatment. The content of PPT in cell suspension was increased 37.25-fold compared with that of leaves of the transgenic tobacco. Transgenic tobacco was not able to set seeds because of microspore degeneration in anthers. Transmission electron microscopy analysis revealed that cells of phloem tissue situated in the center of the anther showed an abnormally condensed nuclei and degenerated mitochondria. Conclusion: We successfully achieved the production of PPT in transgenic tobacco. The possible factors deriving male sterility in transgenic tobacco are discussed.

• The Journal of Internal Korean Medicine
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• v.40 no.3
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• pp.356-368
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• 2019

Objectives: This study was designed to evaluate the efficacy of Pharbitidis Semen (PS) on obesity using 3T3-L1 cells and high-fat-diet rats. Methods: After the administration of PS at a concentration of 0.125, 0.25, and 0.50 mg/ml in 3T3-L1 cells, the degree of differentiation of adipocytes was measured by Oil Red O staining. The experimental animals were divided into five groups in vivo. Normal group: diet-fed normal group (Normal), Control group (Control): high-fat-diet (HFD)-fed, Positive control group (PC): HFD+Contrave 200 mg/kg, PS_Low group: HFD+PS 250 mg/kg group, PS_High group: HFD+PS 500 mg/kg. Results: The total cholesterol, low density lipoprotein-cholesterol (LDL-cholesterol), and triglycerides (TG) content of the PS medication groups were significantly lower than those of the control group. In addition, high density lipoprotein cholesterol (HDL-cholesterol) tended to increase, and free fatty acid, total lipid, and phospholipid tended to decrease, but the decrease was not statistically significant. The administration of PS inhibited the differentiation of 3T3-L1 adipocytes, and a statistically significant difference was found in the group administered with 0.125 mg/ml. The mRNA expression of $PPAR-{\gamma}$ and leptin, which are involved in the differentiation of adipocytes, was observed using reverse transcription polymerase chain reaction (RT-PCR). The expressions of $PPAR-{\gamma}$ and leptin significantly decreased in comparison with the control and the positive control.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.32 no.3
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• pp.13-22
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• 2019

Objectives : Armeniacae semen is the seed of Prunus armenica L. var. ansu MAXIM, and this is classified into Rosaceae. Armeniacae semen has been used for centuries in traditional oriental medicine for the treatment of pain and inflammatory diseases. Amygdalin is the major compound of Armeniacae semen, and it is now being used for the treatment of pain and cancer. Methods : In the present study, we compared the effects of an aqueous extract of Armeniacae semen and a solution of amygdalin extracted from Armeniacae semen on lipopolysaccharide(LPS)-stimulated prostaglandin E2 synthesis and nitric oxide production in mouse BV-2 microglial cells. For this study, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay, reverse transcription-polymerase chain reaction(RT-PCR), prostaglandin E2 immunoassay and nitric oxide detection were performed on mouse BV-2 microglial cells. Results : In the present study, an aqueous extract of Armeniacae semen and an amygdalin solution extracted from Armeniacae semen suppressed prostaglandin E2 synthesis and nitric oxide production by inhibiting the LPS-induced enhancement of cyclooxygenase-2(COX-2) mRNA and the inducible nitric oxide synthase mRNA in mouse BV-2 cells. For the cyclooxygenase-1(COX-1) expression, an aqueous extract of Armeniacae semen showed a more potent suppression effect compared to the amygdalin solution. However, the amygdalin solution more potently suppressed the LPS-induced COX-2 mRNA expression compared to the aqueous extract of Armeniacae semen. Conclusions : As a result, aqueous extract of Armeniacae semen and amygdalin exert anti-inflammatory and analgesic effects.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.54 no.5
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• pp.698-702
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• 2021

Yellow head virus (YHV) is a rod-shaped positive-sense single-stranded RNA virus, classified into the genus Okavirus, family Roniviridae, and order Nidovirales. In this study, 200 fleshy prawns (Fenneropenaeus chinensis) collected from the vicinity of Narodo in Goheung-gun, Korea, were screened for the presence of yellow head complex viruses and related genotype such as YHV genotype 8. The detection rate of YHV genotype 8 among the 200 fleshy prawns, determined using nested RT-PCR (reverse transcription polymerase chain reation), was 39.0%. Phylogenetic analysis of the ORF1b gene of YHV showed that eight distinct genetic lineages were detected. The four strains of YHV genotype 8 obtained in this study formed a robust clade with the YHV genotype 8 group that was first isolated from fleshy prawns in China suspected to have acute hepatopancreatic necrosis disease (AHPND).

• Journal of Microbiology and Biotechnology
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• v.31 no.8
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• pp.1069-1078
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• 2021

Sevoflurane postconditioning (SPostC) has been proved effective in cardioprotection against myocardial ischemia/reperfusion injury. It was also reported that heat shock protein 70 (HSP70) could be induced by sevoflurane, which played a crucial role in hypoxic/reoxygenation (HR) injury of cardiomyocytes. However, the mechanism by which sevoflurane protects cardiomyocytes via HSP70 is still not understood. Here, we aimed to investigate the related mechanisms of SPostC inducing HSP70 expression to reduce the HR injury of cardiomyocytes. After the HR cardiomyocytes model was established, the cells transfected with siRNA for HSP70 (siHSP70) or not were treated with sevoflurane during reoxygenation. The lactate dehydrogenase (LDH) level was detected by colorimetry while cell viability and apoptosis were detected by MTT and flow cytometry. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and Western blotting were used to detect HSP70, apoptosis-, cell cycle-associated factors, iNOS, and Cox-2 expressions. Enzyme-linked immuno sorbent assay (ELISA) was used to measure malondialdehyde (MDA) and superoxide dismutase (SOD). SPostC decreased apoptosis, cell injury, oxidative stress and inflammation and increased viability of HR-induced cardiomyocytes. In addition, SPostC downregulated Bax and cleaved caspase-3 levels, while SPostC upregulated Bcl-2, CDK-4, Cyclin D1, and HSP70 levels. SiHSP70 had the opposite effect that SPostC had on HR-induced cardiomyocytes. Moreover, siHSP70 further reversed the effect of SPostC on apoptosis, cell injury, oxidative stress, inflammation, viability and the expressions of HSP70, apoptosis-, and cell cycle-associated factors in HR-induced cardiomyocytes. In conclusion, this study demonstrates that SPostC can reduce the HR injury of cardiomyocytes by inducing HSP70 expression.

• Journal of Microbiology and Biotechnology
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• v.31 no.8
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• pp.1115-1122
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• 2021

Rotavirus (RV), as the main cause of diarrhea in children under 5 years, contributes to various childhood diseases. Valeriana jatamansi Jones is a traditional Chinese herb and possesses antiviral effects. In this study we investigated the potential mechanisms of V. jatamansi Jones in RV-induced diarrhea. MTT assay was performed to evaluate cell proliferation and the diarrhea mice model was constructed using SA11 infection. Mice were administered V. jatamansi Jones and ribavirin. Diarrhea score was used to evaluate the treatment effect. The enzyme-linked immunosorbent assay was performed to detect the level of cytokines. Western blot and quantitative reverse transcription-PCR were used to determine protein and mRNA levels, respectively. Hematoxylin-eosin staining was applied to detect the pathological change of the small intestine. TdT-mediated dUTP nick-end labeling was conducted to determine the apoptosis rate. The results showed V. jatamansi Jones promoted MA104 proliferation. V. jatamansi Jones downregulated phosphatidylinositol 3-kinase (PI3K) and protein kinase B (AKT) in protein level, which was consistent with the immunohistochemistry results. Moreover, V. jatamansi Jones combined with ribavirin regulated interleukin-1β (IL-1β), interferon γ, IL-6, tumor necrosis factor α, and IL-10, and suppressed secretory immunoglobulin A secretion to remove viruses and inhibit dehydration. V. jatamansi Jones + ribavirin facilitated the apoptosis of small intestine cells. In conclusion, V. jatamansi Jones may inhibit RV-induced diarrhea through PI3K/AKT signaling pathway, and could therefore be a potential therapy for diarrhea.