• The Korean Journal of Physiology and Pharmacology
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• 제28권1호
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• pp.59-72
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• 2024

To investigate the mechanism of Wenshen Xuanbi Decoction (WSXB) in treating osteoarthritis (OA) via network pharmacology, bioinformatics analysis, and experimental verification. The active components and prediction targets of WSXB were obtained from the TCMSP database and Swiss Target Prediction website, respectively. OA-related genes were retrieved from GeneCards and OMIM databases. Protein-protein interaction and functional enrichment analyses were performed, resulting in the construction of the Herb-Component-Target network. In addition, differential genes of OA were obtained from the GEO database to verify the potential mechanism of WSXB in OA treatment. Subsequently, potential active components were subjected to molecular verification with the hub targets. Finally, we selected the most crucial hub targets and pathways for experimental verification in vitro. The active components in the study included quercetin, linolenic acid, methyl linoleate, isobergapten, and beta-sitosterol. AKT1, tumor necrosis factor (TNF), interleukin (IL)-6, GAPDH, and CTNNB1 were identified as the most crucial hub targets. Molecular docking revealed that the active components and hub targets exhibited strong binding energy. Experimental verification demonstrated that the mRNA and protein expression levels of IL-6, IL-17, and TNF in the WSXB group were lower than those in the KOA group (p < 0.05). WSXB exhibits a chondroprotective effect on OA and delays disease progression. The mechanism is potentially related to the suppression of IL-17 and TNF signaling pathways and the down-regulation of IL-6.

• Korean Circulation Journal
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• 제54권8호
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• pp.468-481
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• 2024

Background and Objectives: Although the clinical consequences of advanced heart failure (HF) may be similar across different etiologies of cardiomyopathies, their proteomic expression may show substantial differences in relation to underlying pathophysiology. We aimed to identify myocardial tissue-based proteomic characteristics and the underlying molecular pathophysiology in non-ischemic cardiomyopathy with different etiologies. Methods: Comparative extensive proteomic analysis of the myocardium was performed in nine patients with biopsy-proven non-ischemic cardiomyopathies (3 dilated cardiomyopathy [DCM], 2 hypertrophic cardiomyopathy [HCM], and 4 myocarditis) as well as five controls using tandem mass tags combined with liquid chromatography-mass spectrometry. Differential protein expression analysis, Gene Ontology (GO) analysis, and Ingenuity Pathway Analysis (IPA) were performed to identify proteomic differences and molecular mechanisms in each cardiomyopathy type compared to the control. Proteomic characteristics were further evaluated in accordance with clinical and pathological findings. Results: The principal component analysis score plot showed that the controls, DCM, and HCM clustered well. However, myocarditis samples exhibited scattered distribution. IPA revealed the downregulation of oxidative phosphorylation and upregulation of the sirtuin signaling pathway in both DCM and HCM. Various inflammatory pathways were upregulated in myocarditis with the downregulation of Rho GDP dissociation inhibitors. The molecular pathophysiology identified by extensive proteomic analysis represented the clinical and pathological properties of each cardiomyopathy with abundant proteomes. Conclusions: Different etiologies of non-ischemic cardiomyopathies in advanced HF exhibit distinct proteomic expression despite shared pathologic findings. The benefit of tailored management strategies considering the different proteomic expressions in non-ischemic advanced HF requires further investigation.

• Applied Microscopy
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• 제35권3호
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• pp.121-128
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• 2005

Ras 신호전달체계는 세포내 다양한 결합 분자들과 더불어 세포의 분열과 세포의 이동에 관여한다. Dynamin 단백질은 endocytosis와 분비과정에서 vesicle를 분리하는데 관여하는 것으로 알려져 있으며, 3가지 아형으로 구분된다. Dynamin I은 신경조직에서 만 발현되고, dynamin II는 모든 조직에서 발현되지만 dynamin III는 정소를 포함한 생식기계에서만 발현된다. 선행된 연구에서 NIH3T3 세포를 이용하여 ras과발현 세포주를 만들었으며, dynamin II와 ras의 신호전달체계에 있는 Grb2가 결합한다는 것을 보고하였다. 따라서, 본 연구는 ras 단백질이 과발현되는 세포 (NIH3T3 (ras))와 대조세포인 NIH3T3의 형태학적인 차이점을 분석하고, 이 두 세포들에서 dynamin II 단백질의 발현의 차이를 비교하고자 하였다. Dynamin II의 발현차이를 분석하기 위해 형광염색을 하여 공초점 레이저현미경으로 세포내 분석을 하였으며, western blot을 시행하여 생화학적인 발현차이를 보았다. 또한, 두 세포의 미세구조적인 분석을 위하여 SEM과 TEM을 사용하였다. Dynamin II는 NIH3T3 (ras) 세포에서 발현이 증가 하였으며, NIH3T3 세포에 비하여 좀더 방추형이 었으며, 작은 세포질 돌기가 세포막을 따라 다수 신장되어있음이 관찰되었다. 또한, NIH3T3 (ras) 세포의 endocytotic vesicle이 형성되는 부위에서 dynamin II의 발현이 증가하였다. 이러한 결과로 dynamin II는 ras신호전달체계의한 신호전달분자로서 작용을 할 것으로 사료된다.

• The Plant Pathology Journal
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• 제30권4호
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• pp.343-354
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• 2014

Rice blast disease caused by Magnaporthe oryzae is one of the most serious diseases of cultivated rice (Oryza sativa L.) in most rice-growing regions of the world. In order to investigate early response genes in rice, we utilized the transcriptome analysis approach using a 300 K tilling microarray to rice leaves infected with compatible and incompatible M. oryzae strains. Prior to the microarray experiment, total RNA was validated by measuring the differential expression of rice defense-related marker genes (chitinase 2, barwin, PBZ1, and PR-10) by RT-PCR, and phytoalexins (sakuranetin and momilactone A) with HPLC. Microarray analysis revealed that 231 genes were up-regulated (>2 fold change, p < 0.05) in the incompatible interaction compared to the compatible one. Highly expressed genes were functionally characterized into metabolic processes and oxidation-reduction categories. The oxidative stress response was induced in both early and later infection stages. Biotic stress overview from MapMan analysis revealed that the phytohormone ethylene as well as signaling molecules jasmonic acid and salicylic acid is important for defense gene regulation. WRKY and Myb transcription factors were also involved in signal transduction processes. Additionally, receptor-like kinases were more likely associated with the defense response, and their expression patterns were validated by RT-PCR. Our results suggest that candidate genes, including receptor-like protein kinases, may play a key role in disease resistance against M. oryzae attack.

• Journal of Plant Biotechnology
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• 제2권2호
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• pp.61-71
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• 2000

By screening a cDNA library of auxin-treated mung bean (Vigna radiata L.) hypocotyls, we have isolated two full-length cDNA clones, pVR-ACS6 and pVR-ACS7, for 1-aminocyclopropane-1-carboxylate (ACC) synthase, the rate-limiting enzyme in the ethylene biosynthetic pathway. While PVR-ACS6 corresponds to the previously identified PCR fragment pMBA1, pVR-ACS7 is a new cDNA clone. A comparison of deduced amino acid sequences among auxin-induced ACC synthases reveal that these enzymes share a high degree of homology (65-75%) to VR-ACS6 and VR-ACS7 polypeptides, but only about 50% to VR-ACS1 polypeptide. ACS6 and ACS7 are specifically induced by auxin, while ACS1 is induced by cycloheximide, and to lesser extent by excision and auxin treatment. Results from nuclear run-on transcription assay and RNA gel blot studies revealed that all three genes were transcriptionally active displaying unique patterns of induction by IAA and various hormones in etiolated hypocotyls. Particularly, 24-epibrassinolide (BR), an active brassinosteroid, specifically enhanced the expression of VR-ACS7 by distinct temporal induction mechanism compared to that of IAA. In addition, BR synergistically increased the IAA-induced VR-ACS6 and VR-ACS7 transcript levels, while it effectively abolished both the IAA- and kinetin-induced accumulation of VR-ACS1 mRNA. In light-grown plants, VR-ACS1 was induced by IAA in roots, whereas W-ACS6 in epicotyls. IAA- and BR-treatments were not able to increase the VR-ACS7 transcript in the light-grown tissues. These results indicate that the expression of ACC synthase multigene family is regulated by complex hormonal and developmental networks in a gene- and tissue-specific manner in mung bean plants. The VR-ACS7 gene was isolated, and chimeric fusion between the 2.4 kb 5'-upstream region and the $\beta$-glucuronidase (GUS) reporter gene was constructed and introduced into Nicotiana tobacum. Analysis of transgenic tobacco plants revealed the VR-ACS7 promoter-driven GUS activity at a highly localized region of the hypocotyl-root junction of control seedlings, while a marked induction of GUS activity was detected only in the hypocotyl region of the IAA-treated transgenic seedlings where rapid cell elongation occurs. Although there was a modest synergistic effect of BR on the IAA-induced GUS activity, BR alone failed to increase the GUS activity, suggesting that induction of VR-ACS7 occurs via separate signaling pathways in response to IAA and BR.

• Molecules and Cells
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• 제37권7호
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• pp.547-553
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• 2014

Glioblastoma multiforme (GBM) is one of the most common brain malignancies and has a very poor prognosis. Recent evidence suggests that the presence of cancer stem cells (CSC) in GBM and the rare CSC subpopulation that is resistant to chemotherapy may be responsible for the treatment failure and unfavorable prognosis of GBM. A garlic-derived compound, Z-ajoene, has shown a range of biological activities, including anti-proliferative effects on several cancers. Here, we demonstrated for the first time that Z-ajoene specifically inhibits the growth of the GBM CSC population. CSC sphere-forming inhibition was achieved at a concentration that did not exhibit a cytotoxic effect in regular cell culture conditions. The specificity of this inhibitory effect on the CSC population was confirmed by detecting CSC cell surface marker CD133 expression and biochemical marker ALDH activity. In addition, stem cell-related mRNA profiling and real-time PCR revealed the differential expression of CSC-specific genes, including Notch, Wnt, and Hedgehog, upon treatment with Z-ajoene. A proteomic approach, i.e., reverse-phase protein array (RPPA) and Western blot analysis, showed decreased SMAD4, p-AKT, 14.3.3 and FOXO3A expression. The protein interaction map (http://string-db.org/) of the identified molecules suggested that the AKT, ERK/p38 and $TGF{\beta}$ signaling pathways are key mediators of Z-ajoene's action, which affects the transcriptional network that includes FOXO3A. These biological and bioinformatic analyses collectively demonstrate that Z-ajoene is a potential candidate for the treatment of GBM by specifically targeting GBM CSCs. We also show how this systemic approach strengthens the identification of new therapeutic agents that target CSCs.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제32권1호
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• pp.36-41
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• 2006

Objective. The objective of this study was to examine the hypothesis that inflammatory synovial fluid from TMJ internal derangement initiates a transient increase in intracellular calcium concentration ([$Ca^{2+}$]i) in chondrocytes and the induced Ca2+ signaling affects iNOS/COX-2 gene expression patterns following exposure to inflamed synovial fluid. Materials and Methods. Two female adult patients with symptoms of TMD who agreed to participate in the study were selected for this study. Immortalized human juvenile costal chondrocyte C-28/I2 was grown to 80% confluency and synovial fluids from two patients were added respectively to culture media for 24 hours at the concentration of 100ng/10ml. Confocal laser scanning microscope (CLSM) was used to examine changes of intracellular calcium concentration ([$Ca^{2+}$]i). RT-PCR was performed to identify the expression profile of IL-1${\alpha}$, iNOS, COX-2. Results. Increased [$Ca^{2+}$]i was observed in chondrocytes subjected to inflamed synovial fluid compared to control cultures and in respective cultures exposed to inflamed synovial fluids from each patient, IL-1${\beta}$, COX-2 mRNA were detected. However, in neither case iNOS mRNA was expressed. IL-1${\alpha}$, COX-2, and iNOS mRNA were expressed in control culture. Conclusion. Our results show that immortalized chondrocytes cultured with inflamed synovial fluids from patients diagnosed as disc displacement without reduction and limitation in mouth opening showed increased calcium concentration and expression of COX-2 while inhibiting the production of iNOS, which in turn could adversely affect the chondrocytes in at least short term by hindering physiologic role of NO against inflammatory cascades. These findings suggest that inflamed synovial fluid may differentially regulate the transcriptomes of relevant inflammatory mediators, especially iNOS/COX-2 axis in chondrocytes through adjusting calcium transients.

• Journal of Plant Biotechnology
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• 제33권2호
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• pp.93-97
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• 2006

농작물 생산에 있어서 병원균 침입에 의한 피해를 줄이는 것은 아주 중요한 과제이다. 식물은 스스로도 생체 방어 신호 전달 기작을 가지고 있지만 그 피해를 줄이는데 있어서 한계가 있다. 본 연구는 콩과 식물에서 분리한 식물생체방어 신호전달 유전자인 SCaM-5를 토마토에 형질전환하여 형질전환 식물체를 작성하고 병 저항성에 관한 실험을 수행한 것이다. SCaM-5 유전자가 형질전환 된 토마토에서는 pathogen-related(PR-5) 유전자를 항상 발현시킴으로서 계속적으로 식물 생체방어 신호기작을 활성 시킨다는 사실을 확인하였다. 또한, SCaM-5를 형질전환 시킨 토마토는 식물에 막대한 피해를 주는 중요한 곰팡이 (P. capsici와 F. oxysporm)와 bacteria (Pst DC3000)에 대하여 병 저항성을 가진다는 것을 검증하였다.

• 환경정책연구
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• 제8권4호
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• pp.75-94
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• 2009

제3자재원조달 계약의 과정이 1단계 신호발생 게임과 2단계 주인-대리인 게임 등 2단계 게임 모형을 통해 분석된다. 2단계 게임의 해는 역진귀납법을 통해 구해진다. 2단계 게임에서, 에너지절약기업의 최적 노력수준, 에너지 사용자의 최적 보상 체계, 그리고 두 경기자의 보수는 각각의 부분게임에서 도출된다. 이렇게 해서 도출된 각각의 부분게임의 최적해는 서로 비교된다. 그 결과 우리는 만약 에너지절약기업의 수입을 감소하는 비율로 증가시키는 누진적인 판매세와 같은 에너지절약기업의 수입에 대한 제약이 존재한다면, 최적 분배 비율은 선형 보상 체계에서 'I'보다 작은 수준에서 유일하게 결정된다는 것을 알게 되었다. 즉 유일한 균형이 존재한다는 것이다. 부분게임 각각의 경우에 대한 자기충족적인 유일한 균형은 분리균형인 바, 이 균형에서 에너지사용자는 높은 기술수준을 보유하고 있는 에너지절약기업(H형 ESCO)의 에너지진단 제안은 받아들이되, 낮은 기술수준을 보유한 에너지절약기업(L형 ESCO)의 진단제안은 거절한다. L형 ESCO는 제3자 재원조달 시장에서 수익을 창출할 수 없게 된다. 반면, H형 ESCO는 L형과 H형의 진단수수료의 차이만큼 수익을 얻게 된다. 따라서 H형 ESCO의 균형에서의 수익은 자신의 기술수준뿐만 아니라 L형보다 더 빠르게 진보된 기술수준을 통해서 증가하게 된다. 에너지사용자는 어떠한 추가비용을 지출하지도 않으면서 기존 자신의 에너지시스템에서 ESCO가 3자재원조달 임무를 하도록 허용함으로써 일정분의 수익을 얻게 된다.

• Molecules and Cells
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• 제27권4호
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• pp.467-473
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• 2009

Our previous study suggested that OsBWMK1, a gene which encodes a member of the rice MAP kinase family, generates transcript variants which show distinct expression patterns in response to environmental stresses. The transcript variants are generated by alternative splicing and by use of alternative promoters. To test whether the two alternative promoters, pOsBWMK1L (promoter for the OsBWMK1L splice variant) and pOsBWMK1S (promoter for the OsBWMK1S splice variant), are biologically functional, we analyzed transgenic plants expressing GUS fusion constructs for each promoter. Both pOsBWMK1L and pOsBWMK1S are biologically active, although the activity of pOsBWMK1S is lower than that of pOsBWMK1L. Histochemical analysis revealed that pOsBWMK1L is constitutively active in most tissues at various developmental stages in rice and Arabidopsis, whereas pOsBWMK1S activity is spatially and temporally restricted. Furthermore, the expression of pOsBWMK1S::GUS was upregulated in response to hydrogen peroxide, a plant defense signaling molecule, in both plant species. These results suggest that the differential expression of OsBWMK1 splice variants is the result of alternative promoter usage and, moreover, that the mechanisms controlling OsBWMK1 gene expression are conserved in both monocot and dicot plants.