• Biomolecules & Therapeutics
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• v.27 no.1
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• pp.101-106
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• 2019

Most diabetic patients experience diabetic mellitus (DM) urinary bladder dysfunction. A number of studies evaluate bladder smooth muscle contraction in DM. In this study, we evaluated the change of bladder smooth muscle contraction between normal rats and DM rats. Furthermore, we used pharmacological inhibitors to determine the differences in the signaling pathways between normal and DM rats. Rats in the DM group received an intraperitoneal injection of 65 mg/kg streptozotocin and measured blood glucose level after 14 days to confirm DM. Bladder smooth muscle contraction was induced using acetylcholine (ACh, $10^{-4}M$). The materials such as, atropine (a muscarinic receptor antagonist), U73122 (a phospholipase C inhibitor), DPCPX (an adenosine $A_1$ receptor antagonist), udenafil (a PDE5 inhibitor), prazosin (an ${\alpha}_1$-receptor antagonist), papaverine (a smooth muscle relaxant), verapamil (a calcium channel blocker), and chelerythrine (a protein kinase C inhibitor) were pre-treated in bladder smooth muscle. We found that the DM rats had lower bladder smooth muscle contractility than normal rats. When prazosin, udenafil, verapamil, and U73122 were pre-treated, there were significant differences between normal and DM rats. Taken together, it was concluded that the change of intracellular $Ca^{2+}$ release mediated by PLC/IP3 and PDE5 activity were responsible for decreased bladder smooth muscle contractility in DM rats.

• Applied Chemistry for Engineering
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• v.29 no.4
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• pp.452-460
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• 2018

This study was conducted to investigate the physiological activities of Houttuynia cordata extracts and fractions. H. cordata extracts were extracted with 50% ethanol and the ethyl acetate fractions were obtained from the extracts. Minimum inhibitory concentration (MIC) values of the ethyl acetate fraction for S. aureus and B. subtilis were $78{\mu}g/mL$ and $312{\mu}g/mL$, respectively, indicating the high activity against gram-positive bacteria. The free radical scavenging activity ($FSC_{50}$) for 1,1-diphenyl-2-picrylhydrazyl (DPPH) was higher in the ethyl acetate fraction with $12.00{\mu}g/mL$ compared to that of $27.15{\mu}g/mL$ for 50% ethanol extract. The total antioxidant activity ($OSC_{50}$) values for reactive oxygen species (ROS) produced in $Fe^{3+}-EDTA/H_2O_2$ system by a luminol-dependent chemiluminescence method were 2.91 and $0.983{\mu}g/ml$ for the 50% ethanol extract and ethyl acetate fraction, respectively. To investigate cellular protective effects on the HaCaT cell, the intracellular ROS scavenging activity was measured after UVB irradiation and the ethyl acetate fraction of H. cordata showed the activity in a concentration-dependent from $1.6{\mu}g/mL$ and a reduction rate of 54.3% at a maximum concentration of $12.5{\mu}g/mL$. Also, HaCaT cell protective effect against $H_2O_2$-mediated decreased the cell viability of the ethyl acetate fraction of H. cordata which significantly increased the cell viability from $0.8{\mu}g/mL$ and the maximum cell viability showed 86.9%. The ethyl acetate fraction of the H. cordata extracts was analyzed by TLC and HPLC. As a result, quercitrin, isoquercitrin, hyperoside, chlorogenic acid, neochlorogenic acid, cryptochlorogenic acid, rutin and afzelin were identified. From the above results, it was suggested that the extracts and fractions of H. cordata have a potential to be applied in the field of cosmetics as a natural antioxidant/preservative capable of protecting the cell membrane from the oxidative stress by eliminating ROS and exhibiting the antimicrobial effect.

• The Journal of Internal Korean Medicine
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• v.21 no.2
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• pp.259-266
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• 2000

Objectives : The effects of aqueous extracts of Cortex ulmi pumilae (a traditional medicine for cancer treatment in oriental medicine) on the induction of apoptotic cell death were investigated in human liver origm hepatoma cell lines, HepG2. Methods : The death of HepG2 cells was markedly induced by the addition of extracts of Cortex ulmi pumilae in a dose-dependent manner. The apoptotic characteristic ladder pattern of DNA strand break was not observed in cell death of HepG2. In addition, it was not shown nucleus chromatin condensation and fragmentation under hoechst staining. However, by the using annexin V staining assay, externalizations of phosphatidylserine in HepG2 cell which were treated with Cortex ulmi pumilae extracts were detected in the early time (at 9 hr after extract treatment). Furthermore, LDH release was not detected in this early stage. Therefore, Cortex ulmi pumilae extracts-induced cell death of HepG2 cells is mediated by apoptotic death signal processes. Result : The activity of caspase 3-like proteases remained in a basal level in HepG2 cells which treated with the extract of Cordyceps sinensis. However, it was markedly increased in HepG2 cells which treated with two extracts of Cortex ulmi pumilae (C.U.P.-C, C.U.P.-K) which were differently extracted (respectively, 2.3 and 3.3 fold). On a while, the phosphotransferase activities of JNK1 was markedly induced in HepG2 cells which were treated with two extracts of Cortex ulmi pumilae. On the contrary, the activation of transcriptional activator, activating protein1(AP-1) and NF-kB were severely decreased by these two extracts of Cortex ulmi pumilae (C.U.P.-C, C.U.P.-K). In addition, antioxidants (GSH and NAC) and intracellular $Ca2^+$ level regulator (Bapta/AM and Thapsigargin) did not affect Cortex ulmi pumilae extracts-induced apoptotic death of HepG2 cells. Conclusions : In conclusion, our results suggest that two extracts of Cortex ulmi pumilae (C.U.P.-C, C.U.P.-K) induces the apoptotic death of human liver origin hepatoma HepG2 cells via activation of caspase 3-like proteases as well as JNK1, and inhibition of transcriptional activators, AP-1 and $NK-{\kappa}B$.

• Current Research on Agriculture and Life Sciences
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• v.14
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• pp.111-122
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• 1996

S. chibaensis J-59 produced an extracellular xylanase in a CSL medium composed of 1.5% com steep liquor, 0.1% $MgSO_4{\cdot}7H_2O$, 0.012% $CoCl_2{\cdot}6H_2O$, and 0.15% glucose containing xylan. but it did not produce in the culture medium containing xylose. The production of enzyme reached to a maximum level (0.83 uints/ml) when bacteria were cultured in 2.5 l jar fermentor for 48hrs at $30^{\circ}C$ and pH 7.0. Furthermore, S. chibaensis J-59 produced an intracellular glucose isomerase in a medium containing xylan and/or xylose. Xylanase was purified 29-fold over the culture supernatants of S. chibaensis J-59 by ammonium sulfate fractionation, chromatography on DEAE-Sephadex A-50, and gel filtration on Sephadex G-200. The purified enzyme is a monomeric enzyme with a native molecular mass of 25 kDa and a subunit molecular mass of 25 kDa. The purified enzyme requires $Mg^{2+}$ for activity, $Ca^{2+}$, $Co^{2+}$ is not an inhibitor but inhibit by $Fe^{3+}$, $Hg^{2+}$, and $Cu^{2+}$, sodium dodecyl sulfate, N-bromosuccinide. Pattern of hydrolysis demonstrated that the xylanase was an endo-splitting enzyme able to break down birchwood xylan at random giving xylobiose, xylotriose and xylotetrose as the main end products.

• Journal of Life Science
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• v.21 no.2
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• pp.183-193
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• 2011

Migration and differentiation of mesenchymal stem cells are crucial for tissue regeneration in response to injury. Sphingosine-1-phosphate (S1P) is a bioactive lipid that regulates a variety of biological processes, including proliferation, survival, differentiation and motility. In the present study, we determined the role of S1P in migration and differentiation of human bone marrow-derived mesenchymal stem cells (BMSCs). S1P stimulated migration of BMSCs in a dose- and time-dependent manner, and pre-incubation of the cells with pertussis toxin completely abrogated S1P-induced migration, suggesting involvement of Gi-coupled receptors in S1P-induced cell migration. S1P elicited elevation of intracellular concentration of $Ca^{2+}$ ($[Ca^{2+}]_i$) and pretreatment with VPC23019, an antagonist of $S1P_1/S1P_3$, blocked S1P-induced migration and increase of $[Ca^{2+}]_i$. Small interfering RNA-mediated knockdown of endogenous $S1P_1$ attenuated S1P-induced migration of BMSCs. Furthermore, S1P treatment induced expression of $\alpha$-smooth muscle actin ($\alpha$-SMA), a smooth muscle marker, and pretreatment with VPC23019 abrogated S1P-induced $\alpha$-SMA expression. S1P induced phosphorylation of p38 mitogen-activated protein kinase (MAPK), and pretreatment of cells with SB202190, an inhibitor of p38 MAPK, or adenoviral overexpression of a dominant-negative mutant of the p38 MAPK blocked S1P-induced cell migration and $\alpha$-SMA expression. Taken together, these results suggest that S1P stimulates migration and smooth muscle differentiation of BMSCs through an $S1P_1$-p38 MAPK-dependent mechanism.

• Journal of Life Science
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• v.34 no.7
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• pp.500-508
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• 2024

Recently, there has been increasing interest in enhancing the indoor air quality, particularly in response to the growing utilization of public facilities. The focus of this study was on assessing the efficacy and safety of an air sterilizer equipped with electrolytic salt catalysts. To that end, we evaluated the antimicrobial activity of the vapor spraying from the air sterilizer and its cytotoxicity in condensed form on human cell lines (HaCaT, BEAS-2B, and THP-1). Against the test organisms, which comprised five bacterial strains (Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Pseudomonas aeruginosa, Salmonella typhimurium) and one fungal strain (Candida albicans), the air sterilizer exhibited relatively high antimicrobial activities ranging from 10.89 to 73.98% following 1 and 3 hr of vapor spraying, which were notably time-dependent. Importantly, cytotoxicity assessments on human cells indicated no significant harmful effect even at a 1.0% concentration. Comprehensive safety evaluations included morphological observations, gene expression (Bcl-2, Bax) tests, and FACS analysis of intracellular ROS levels. Consistent with previous cytotoxicity findings, these estimates demonstrated no significant changes, highlighting the air sterilizer's safety and antimicrobial activities. In a simulated 20-hr operation within an indoor environment, the air sterilizer not only showed an 89.4% removal of total bacteria but also a 100.0% removal of Escherichia sp. and fungi. This research outlines the potential of the developed electrolytic salt catalyst air sterilizer to effectively remove indoor microbial pollutants without compromising human safety, underscoring the solution that it offers for improving indoor air quality.

• Microbiology and Biotechnology Letters
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• v.46 no.4
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• pp.360-371
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• 2018

Extracts and fractions of Anemarrhena asphodeloides Bunge were prepared and their physiological activities and components were analyzed. Antimicrobial activities of the ethyl acetate and aglycone fractions were $78{\mu}g/ml$ and $31{\mu}g/ml$, respectively, for Staphylococcus aureus and $156{\mu}g/ml$ and $125{\mu}g/ml$, respectively, for Pseudomonas aeruginosa. 1,1-Diphenyl-2-picrylhydrazyl free radical scavenging activities ($FSC_{50}$) of 50% ethanol extract, ethyl acetate fraction, and aglycone fraction of A. asphodeloides extracts were $146.2{\mu}g/ml$, $23.19{\mu}g/ml$, and $71.06{\mu}g/ml$, respectively. The total antioxidant capacity ($OSC_{50}$) in an $Fe^{3+}$-EDTA/hydrogen peroxide ($H_2O_2$) system were $17.5{\mu}g/ml$, $1.5{\mu}g/ml$, and $1.4{\mu}g/ml$, respectively. The cytoprotective effect (${\tau}_{50}$) in $^1O_2$-induced erythrocyte hemolysis was 181 min with $4{\mu}g/ml$ of the aglycone fraction. The ${\tau}_{50}$ of the aglycone fraction was approximately 4-times higher than that of (+)-${\alpha}$-tocopherol (${\tau}_{50}$, 41 min). Analysis of $H_2O_2$-induced damage of HaCaT cells revealed that the maximum cell viabilities for the 50% ethanol extract, ethyl acetate fraction, and aglycone fraction were 86.23%, 86.59%, and 89.70%, respectively. The aglycone fraction increased cell viability up to 11.53% at $1{\mu}g/ml$ compared to the positive control treated with $H_2O_2$. Analysis of ultraviolet B radiation-induced HaCaT cell damage revealed up to 41.77% decreased intracellular reactive oxygen species in the $2{\mu}g/ml$ aglycone fraction compared with the positive control treated with ultraviolet B radiation. The findings suggest that the extracts and fractions of A. asphodeloides Bunge have potential applications in the field of cosmetics as natural preservatives and antioxidants.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.20 no.12
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• pp.298-305
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• 2019

Cudrania tricuspidata has been reported to have many biological activities, including anti-inflammatory, anti-cancer, and antioxidant properties. However, the effects of C. tricuspidata root extract (CTE) on human platelet aggregation induced by collagen as well as the signaling pathways involved remain unknown. In the present study, we investigated the effect of CTE on human platelets. CTE inhibited platelet aggregation via down-regulation of thromboxane A₂ (TXA₂) by blocking cyclooxygenase-1 (COX-1) activity and intracellular Ca²⁺ mobilization in collagen-induced platelets. CTE also reduced the phosphorylation of phospholipase C (PLC) γ2 and syk. CTE regulated platelet aggregation via cyclic guanosine monophosphate (cGMP)-dependent phosphorylation of vasodilator-stimulated phosphoprotein (VASP) Ser²³⁹. In addition, administration of CTE (50 and 100 mg/kg) significantly reduced hyper-aggregated platelet aggregation by collagen (5 ㎍/mL) without hepatotoxicity in HFD (high fat diet)-fed rats. Taken together, these results suggest that CTE has anti-platelet effects both in vitro and in vivo. Therefore, CTE may be an effective therapeutic and preventive agent for cardiovascular disease, and is a safe and natural product.

• Journal of Yeungnam Medical Science
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• v.13 no.1
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• pp.46-58
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• 1996

Inositol 1,4,5-triphosphate($InsP_3$) is a second messenger for mobilizing intracellular $Ca^{2+}$. It can be dephosphorylated by soluble and particulate forms on $InsP_3$ 5-phosphatase, or phosphorylated to produce inositol 1,3,4,5-tetrakisphosphate($InsP_3$) by $InsP_3$ 3-kinase. These enzymes represent possible targets for the regulation of the $InsP_3/InsP_4$ signal. $InsP_3$ 3-kinase which catalyses th ATP-dependent phosphorylation of $InsP_3$ was purified from bovine brain tissue. All operation were carried out at $4^{\circ}C$. Fresh tissure was homogenized and centrifuged. The supernatant was pooled. Proteins were precipitated from 10% polyethylene glycol, and suspended solution was applied to DEAE cellulose column for chromatography. As the result of above procedure, two isozymes of $InsP_3$ 3-kinase, I and II were obtained. Each isozyme was applied to Matriz green gel, Calmodulin-Affigel 15 column and subsequent phenyl-TSK HPLC column. Specific activites(SA) and fold of puriety were observed at each purification step of chromatography. At DEAE cellulose chromatography, SA were I, 0.6 and II, 4.8 nM/min/mg, and folds were I, 17.2 and II, 16.6. At Matrix green gel chromatography, SA were I, 18 and II, 11 nM/min/mg, folds were I, 62.1 and II, 38.0. At calmodulin-Affigel 15 column chromatography, SA were I, 19 and II, 13 nM/min/mg, folds were I, 65.5 and II, 44.8. Finally $InsP_3$ kinase I and II were purified 3,103-fold and 2,310-fold, and SA were I, 900 and II, 670 nM/min/mg, respectively. SDS-polyacrylamide gel electrophoresis elucidated 3 distinct fractions of Mr of 145,000, 85,000 and 69,500 from isozyme I, and 2 distinct fractions of Mr of 79,000 and 57,000 from isozyme II.

• Korean Journal of Food Science and Technology
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• v.46 no.3
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• pp.369-374
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• 2014

The physiological characteristics of kiwi (Actinidia delicosa) fruit were analyzed, which inclued its nutritional composition, in vitro-antioxidant activities, and neuronal cell protective effects. The most abundant components of mineral, amino acid, and fatty acid were found to be potassium (K), glutamic acid, and a-linolenic acid, respectively. The major free sugars were fructose, glucose, and sucrose. In addition, ${\beta}$-carotene and vitamin C contents were $1.35{\mu}g/100mL$ and 29.21 mg/100 g, respectively. The 2,2'-azinobis-(3-ethylbenothiazline-6-sulfonic acid) (ABTS) radical-scavenging activity of the n-hexane fraction obtained from the kiwi extract was 10.52% at a concentration of $1000{\mu}g/mL$. The malondialdehyde (MDA) inhibition of the n-hexane fraction was found to increased in a dose-dependent manner. The intracellular reactive oxygen species (ROS) accumulation after hydrogen peroxide ($H_2O_2$) treatment of PC12 cells was significantly reduced in the presence of the n-hexane fraction compared to PC12 cells treated with $H_2O_2$ only. Moreover, in the a MTT assay, the n-hexane fraction showed in vitro-protective effects against $H_2O_2$-induced neurotoxicity.