• Korean Journal of Microbiology
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• v.28 no.2
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• pp.114-119
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• 1990

A yeast chromosomal superkiller gene (SK13) was cloned and expressed in $ski3^{-}$ Saccharomyces cerevisiae strains. The gene was fused to the structural region of E. coli lacZ gene at its C-terminus in a yeast-E. coli shuttle vector, pSR605. The fused gene complemented $ski3^{-}$ strains with SK13 activity and the quantitative level of expression was measured as determined by assaying $\beta$-galactosidase activity. The SDS-polyacrylamide gel electrophoresis and the Western blot analysis of this fused protein showed the immuno-reacted bands with a protein of the estimated molecular size (ca.250Kd).

• Journal of Ginseng Research
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• v.45 no.6
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• pp.683-694
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• 2021

Background: Ginsenoside Rg1 (Rg1) has been well documented to be effective against various cardiovascular disease. The aim of this study is to evaluate the effect of Rg1 on mechanical stress-induced cardiac injury and its possible mechanism with a focus on the calcium sensing receptor (CaSR) signaling pathway. Methods: Mechanical stress was implemented on rats through abdominal aortic constriction (AAC) procedure and on cardiomyocytes and cardiac fibroblasts by mechanical stretching with Bioflex Collagen I plates. The effects of Rg1 on cell hypertrophy, fibrosis, cardiac function, [Ca²⁺]_i, and the expression of CaSR and calcineurin (CaN) were assayed both on rat and cellular level. Results: Rg1 alleviated cardiac hypertrophy and fibrosis, and improved cardiac decompensation induced by AAC in rat myocardial tissue and cultured cardiomyocytes and cardiac fibroblasts. Importantly, Rg1 treatment inhibited CaSR expression and increase of [Ca²⁺]_i, which similar to the CaSR inhibitor NPS2143. In addition, Rg1 treatment inhibited CaN and TGF-b1 pathways activation. Mechanistic analysis showed that the CaSR agonist GdCl₃ could not further increase the [Ca²⁺]_i and CaN pathway related protein expression induced by mechanical stretching in cultured cardiomyocytes. CsA, an inhibitor of CaN, inhibited cardiac hypertrophy, cardiac fibrosis, [Ca²⁺]_i and CaN signaling but had no effect on CaSR expression. Conclusion: The activation of CaN pathway and the increase of [Ca²⁺]_i mediated by CaSR are involved in cardiac hypertrophy and fibrosis, that may be the target of cardioprotection of Rg1 against myocardial injury.

• Herbal Formula Science
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• v.31 no.3
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• pp.171-182
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• 2023

Objectives : Saposhnikoviae Radix (SR) and Glehniae Radix (GR) have been frequently used in traditional medicine to treat diseases related to 'wind' syndrome, but there have been cases where it has been mixed in a state where the plant of origin is not clear. In this study, to select materials for conducting preclinical cerebral infarction research, the network pharmacology analysis method was used to select suitable medicinal materials for the study. Methods : In this study, a Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) based network pharmacology analysis method was used, and oral bioavailability (OB), drug likeness (DL), Caco-2 and BBB permeability were utilized to select compounds with potential activity. For the values of each variable used in this study, OB ≥ 20%, DL ≥ 0.18, Caco-2 ≥ 0, and BBB ≥ -0.3 were applied, then networks of bioactive compounds, target proteins, and target diseases was constructed. STRING database was used to construct a protein-protein interaction network. Results : It was confirmed that SR rather than GR has various target proteins and target diseases based on network pharmacological analysis using TCMSP database. And it was analyzed that the bioactive compounds only in SR act more on neurovascular diseases, and both drugs are expected to be effectively used for cardiovascular diseases. Conclusions : In our future study, SR will be used in an ischemic stroke mouse model, and the mechanism of action will be explored focusing on apoptosis and cell proliferation.

• Journal of Physiology & Pathology in Korean Medicine
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• v.26 no.6
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• pp.874-885
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• 2012

In the present study, the possibility of whether the pharmacological effects of Scutellariae Radix Aqueous Extracts(SR) were favorably changed by report that lipopolysaccharide(LPS)-induced rat acute lung injury was treated with Red-Koji(Monascus purpureus 12002) fermentation. Three different dosages of Red-Koji fermented SR extract(fSR), 125, 250 and 500 mg/kg were orally administered once a day for 28 days before LPS(Escherichia coli 0111:B4) treatments, and then 5 hours after LPS treatment(500 ${\mu}g$/head, intra trachea instillation), all rats were sacrificed. Changes in the body weights, lung weights, pulmonary transcapillary albumin transit, arterial gas parameters(pH, $PaO_2$ and $PaCO_2$) bronchoalveolar lavage fluid(BALF) protein, lactate dehydrogenase(LDH) and proinflammatory cytokine tumor necrosis factor-${\alpha}$(TNF-${\alpha}$), interleukin-$1{\beta}$(IL-$1{\beta}$) contents, total cell numbers, neutrophil and alveolar macrophage ratios, lung malondialdehyde(MDA), myeloperoxidase(MPO), proinflammatory cytokine TNF-${\alpha}$ and IL-$1{\beta}$ contents were observed with histopathology of the lung, changes on luminal surface of alveolus(LSA), thickness of alveolar septum, number of polymorphonuclear neutrophils(PMNs). As results of LPS-injection, dramatical increases in lung weights, pulmonary transcapillary albumin transit increases in $PaCO_2$, decreases in pH of arterial blood and $PaO_2$, increases of BALF protein, LDH, TNF-${\alpha}$ and IL-$1{\beta}$ contents, total cells, neutrophil and alveolar macrophage ratios, lung MDA, MPO, TNF-${\alpha}$ and IL-$1{\beta}$ contents increases were detected with decreases in LSA and increases of alveolar septum and PMNs numbers, respectively as compared with intact control. Especially fSR 125 mg/kg showed quite similar favorable effects on the LPS-induced acute lung injuries as compared with 60 mg/kg of ${\alpha}$-lipoic acid and 250 mg/kg of SR. The results suggest that over 125 mg/kg of fSR extracts showed favorable effects on the LPS-induced acute lung injury mediated by their antioxidant and anti-inflammatory effects. Moreover, increases of the pharmacological effects of SR on LPS-induced acute lung injury were observed by Red-Koji fermentation in this study, at least 2-fold higher.

• Molecules and Cells
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• v.39 no.2
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• pp.149-155
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• 2016

In the heart, excitation-contraction (E-C) coupling is mediated by $Ca^{2+}$ release from sarcoplasmic reticulum (SR) through the interactions of proteins forming the $Ca^{2+}$ release unit (CRU). Among them, calsequestrin (CSQ) and histidine-rich $Ca^{2+}$ binding protein (HRC) are known to bind the charged luminal region of triadin (TRN) and thus directly or indirectly regulate ryanodine receptor 2 (RyR2) activity. However, the mechanisms of CSQ and HRC mediated regulation of RyR2 activity through TRN have remained unclear. We first examined the minimal KEKE motif of TRN involved in the interactions with CSQ2, HRC and RyR2 using TRN deletion mutants and in vitro binding assays. The results showed that CSQ2, HRC and RyR2 share the same KEKE motif region on the distal part of TRN (aa 202-231). Second, in vitro binding assays were conducted to examine the $Ca^{2+}$ dependence of protein-protein interactions (PPI). The results showed that TRN-HRC interaction had a bell-shaped $Ca^{2+}$ dependence, which peaked at pCa4, whereas TRN-CSQ2 or TRN-RyR2 interaction did not show such $Ca^{2+}$ dependence pattern. Third, competitive binding was conducted to examine whether CSQ2, HRC, or RyR2 affects the TRN-HRC or TRN-CSQ2 binding at pCa4. Among them, only CSQ2 or RyR2 competitively inhibited TRN-HRC binding, suggesting that HRC can confer functional refractoriness to CRU, which could be beneficial for reloading of $Ca^{2+}$ into SR at intermediate $Ca^{2+}$ concentrations.

• Proceedings of the Korean Biophysical Society Conference
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• 2001.06a
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• pp.66-66
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• 2001

Diabetic cardiomyopathy has been suggested to be caused by abnormal intracellular $Ca^{2＋}$ homeostasis in the myocardium, which is partly due to a defect in calcium transport by the cardiac sarcoplasmic reticulum (SR). In the present study, the underlying mechanism for this functional derangement was investigated with respect to SR $Ca^{2＋}$-ATPase and phospholamban (PLB, the inhibitor of SR $Ca^{2+}$-ATPase).(omitted)d)

• Journal of Korean Medicine Rehabilitation
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• v.21 no.3
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• pp.1-11
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• 2011

Objectives : This study was performed to evaluate the effects of root of Scutellariae Radix(SR) water extract against inflammatory response in the spinal cord injury(SCI). Methods : SCI was induced by mechanical contusion following laminectomy of 10th thoracic vertebra in Sprague-Dawley rat. SR was orally given once a day for 7days after SCI. Myeloperoxidase(MPO) positive neutrophils infiltration was examined. Inducible nitric oxide synthase(iNOS) and tumor necrosis factor-${\alpha}$(TNF-${\alpha}$) expressions were observed with immunohistochemistry. Glial fibrillary acidic protein(GFAP) positive astrocytes were examined using immuno-fluorescence. Results : 1. SR reduced MPO-positive neutrophils infiltration in peri-damage regions of the contusive SCI-induced rats. 2. SR reduced iNOS positive cells in the white matter of the contusive SCI-induced rats. 3. SR reduced TNF-${\alpha}$ positive cells in the gray and white matter of the contusive SCI-induced rats. 4. SR reduced cell number and size of astrocytes in peri-damage regions of the contusive SCI-induced rats. Conclusions : These results suggest that SR plays an inhibitory role against inflammatory response in the SCI.

• Korean journal of food and cookery science
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• v.28 no.1
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• pp.41-49
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• 2012

This purpose of this study was to develop a tea with small red bean which have been known to have effect regarding anti-obesity, fatigue recovery, edema recovery, blood circulation, etc. In order to provide baseline data for small red bean tea we investigated the general components, antioxidative effect and proanthocyanidin analysis in small red beans[Phaseolus angularis W.F. Wight.]. Physicochemical analysis(pH, sugar content, salinity, turbidity), color, anti oralmicrobial activity, content of saponin and sensory test of small red bean with different boiling time in 8 minutes(SR1), 16 minutes(SR2), 24 minutes(SR3), 32 minutes(SR4), 40 minutes(SR5) was also measured. It was shown that the crude fat, carbohydrate, moisture, crude protein, crude ash content of small red bean were 1.0%, 63.9%, 12.8%, 18.7%, 3.6%. DPPH free radical scavenging activity, the total phenolic compounds content and flavonoid content increased significantly (p<0.001). The results of analyzed proanthocyanidin was distinguished by characteristic UV-visible spectra with absorption maximum at 320 nm($t_R$ 7.589 min). As the boiling time(8 minutes:SR1, 16 minutes:SR2, 24 minutes:SR3, 32 minutes:SR4, 40 minutes:SR5) of small red beans increaseds, the pH significantly decreased(p<0.001). The sugar content, salinity and turbidity significantly increased(p<0.001). Moreover, Hunter L, a and b values, crude saponin also increased(p<0.001). The results of analyzed activity against oral bacteria, S. mutans, S. sobrinus, P. intermedia and P. gingivalis showed a higher antibacterial activities than E. coli and S. aureus. MIC was measured that S. mutans, S. sobrinus, P. intermedia and P. gingivalis showed a lower MICs than E. coli and S. aureus. The results regarding sensory test measures, In case of color, refreshing taste and overall quality, SR3 had the highest preference overall among tested samples. In cases conceming odor and taste, SR5 had the highest preference and with regards to sweetness and saltyness, SR4 had the highest preference.

• Molecules and Cells
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• v.40 no.9
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• pp.632-642
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• 2017

The DevSR (DosSR) two-component system, which is a major regulatory system involved in oxygen sensing in mycobacteria, plays an important role in hypoxic induction of many genes in mycobacteria. We demonstrated that overexpression of the kinase domain of Mycobacterium tuberculosis (Mtb) PknB inhibited transcriptional activity of the DevR response regulator in Mycobacterium smegmatis and that this inhibitory effect was exerted through phosphorylation of DevR on Thr180 within its DNA-binding domain. Moreover, the purified kinase domain of Mtb PknB significantly phosphorylated RegX3, NarL, KdpE, TrcR, DosR, and MtrA response regulators of Mtb that contain the Thr residues corresponding to Thr180 of DevR in their DNA-binding domains, implying that transcriptional activities of these response regulators might also be inhibited when the kinase domain of PknB is overexpressed.

• The Journal of Korean Physical Therapy
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• v.19 no.4
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• pp.1-14
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• 2007

Background: It is generally accepted that skeletal muscle contraction is triggered by nerve impulse and intracellular $Ca^{2+}\;([Ca^{2+}]_i)$ released from intracellular $Ca^{2+}$ stores such as sarcoplasmic reticulum (SR). Specifically, this process, called excitation-contraction (E-C) coupling, takes place at intracellular junctions between the plasma membrane, the transverse (T) tubule L-type $Ca^{2+}$ channel (dihydropyridine-sensitive L-rype $Ca^{2+}$ channel, DHPR, also called tetrads), and the SR $Ca^{2+}$ release channel (ryanodine-sensitive $Ca^{2+}$ release channel, RyR, also called feet) of internal $Ca^{2+}$ stores in skeletal muscle cells. Furthermore, it has been reported that the $Ca^{2+-}$ dependent and -independent contraction determine the expression of skeletal muscle genes, thus providing a mechanism for tightly coupling the extent of muscle contraction to regulation of muscle plasticity-related excitation-transcription (E-T) coupling. Purpose: Expression and activity of plasticity-associated enzymes in gastrocnemius muscle strips have not been well studied, however. Methods: Therefore, in this study the expression and phosphorylation of E-C and E-T coupling-related mediators such as protein kinases, ROS(reactive oxygen species)- and apoptosis-related substances, and others in gastrocnemius muscles from rats was examined. Results: I found that expression and activity of MAPKs (mitogen-activated protein kinases, ERK1/2, p38MAPK, and SAPK/JNK), apoptotic proteins (cleaved caspase-3, cytochrome c, Ref-1, Bad), small GTP-binding proteins (RhoA and Cdc42), actin-binding protein (cofilin), PKC (protein kinase C) and $Ca^{2+}$ channel (transient receptor potential channel 6, TRPC6) was observed in rat gastrocnemius muscle strips. Conclusion: These results suggest that MAPKs, ROS- and apoptosis-related enzymes, cytoskeleton-regulated proteins, and $Ca^{2+}$ channel may in part functionally import in E-C and E-T coupling from rat skeletal muscles.