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Regulation of the plasminogen activator activity and inflammatory environment via transforming growth factor-beta regulation of sperm in porcine uterine epithelial cells

  • Kim, Su-jin;Cheong, Hee-Tae;Park, Choon-keun
    • Journal of Animal Reproduction and Biotechnology
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    • v.35 no.4
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    • pp.297-306
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    • 2020
  • The aims of the present study were to confirm that regulation of the PA and environment via TGF-β regulation of sperm by Percoll-separated in porcine uterine epithelial cells. And, it was performed to identify the cytokines (TGF-β1, 2 and 3, TGF-β receptor1 and 2; interleukin, IL-6, IL-8) and PA-related genes (urokinase-PA, uPA; tissue-PA, tPA; PA inhibitor, PAI; uPA-receptor, uPAR) by spermatozoa. The experiment used porcine uterus epithelial cells (pUECs) and uterine tissue epithelial cells, Boar sperm were separated by discontinuous Percoll density gradient (45/90%), and tissues were co-incubated with spermatozoa, followed by real-time PCR. PA activity was measured of sperm by discontinuous Percoll density gradient (45/90%) for 24 hours. To measure viability and acrosome damage of sperm double stained propidium iodide (PI) and SYBR-14 or FITC-PNA were used. In results, binding ratio of Percoll-separated sperm was found no differences, but sperms isolated from 90% Percoll layer reduced PA activity (p < 0.05). when co-cultured sperm selected Percoll in porcine uterus tissues epithelial cells, 90% layer sperm increased TGF-β R1, contrastively tPA and PAI-1 in comparison with control (p < 0.05). 45% sperm was decreased the expression of uPA (p < 0.05). TGF-β decreased PA activity in the supernatant collected from pUECs (p < 0.05). Especially, The group including uPA, PAI-1 were induce sperm intact, while it was reduced in sperm damage when compared to control (p < 0.05). Also, there was no significant difference group of tPA and tPA+I in the dead sperm and acrosome damage compared to control. The expression of tPA and PAI showed a common response. Percoll-separated spermatozoa in 90% layer reduced tPA and IL-related gene mRNA expression. Thus, Percoll-sparated sperm in 90% layer show that it can suppress inflammation through increased expression of TGF-β and downregulation of PA and IL in epithelial cells compared to 45% layer Percoll.

Transforming Growth Factor β Inhibits MUC5AC Expression by Smad3/HDAC2 Complex Formation and NF-κB Deacetylation at K310 in NCI-H292 Cells

  • Lee, Su Ui;Kim, Mun-Ock;Kang, Myung-Ji;Oh, Eun Sol;Ro, Hyunju;Lee, Ro Woon;Song, Yu Na;Jung, Sunin;Lee, Jae-Won;Lee, Soo Yun;Bae, Taeyeol;Hong, Sung-Tae;Kim, Tae-Don
    • Molecules and Cells
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    • v.44 no.1
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    • pp.38-49
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    • 2021
  • Airway mucus secretion is an essential innate immune response for host protection. However, overproduction and hypersecretion of mucus, mainly composed of the gel-forming MUC5AC protein, are significant risk factors for patients with asthma and chronic obstructive pulmonary disease (COPD). The transforming growth factor β (TGFβ) signaling pathway negatively regulates MUC5AC expression; however, the underlying molecular mechanism is not fully understood. Here, we showed that TGFβ significantly reduces the expression of MUC5AC mRNA and its protein in NCI-H292 cells, a human mucoepidermoid carcinoma cell line. This reduced MUC5AC expression was restored by a TGFβ receptor inhibitor (SB431542), but not by the inhibition of NF-κB (BAY11-7082 or Triptolide) or PI3K (LY294002) activities. TGFβ-activated Smad3 dose-dependently bound to MUC5AC promoter. Notably, TGFβ-activated Smad3 recruited HDAC2 and facilitated nuclear translocation of HDAC2, thereby inducing the deacetylation of NF-κB at K310, which is essential for a reduction in NF-κB transcriptional activity. Both TGFβ-induced nuclear translocation of Smad3/HDAC2 and deacetylation of NF-κB at K310 were suppressed by a Smad3 inhibitor (SIS3). These results suggest that the TGFβ-activated Smad3/HDAC2 complex is an essential negative regulator for MUC5AC expression and an epigenetic regulator for NF-κB acetylation. Therefore, these results collectively suggest that modulation of the TGFβ1/Smad3/HDAC2/NF-κB pathway axis can be a promising way to improve lung function as a treatment strategy for asthma and COPD.

Study on the Sasang Type Diagnosis Using Objective Biopsychological Measures (객관적인 생리심리 지표를 사용한 사상체질 진단 연구)

  • Chae, Han;Hwang, Yosun;Kim, Min Seong;Baek, Younghwa;Jeong, Kyungsik;Lee, Jeongyun;Lee, Siwoo;Lee, Soo Jin
    • Journal of Sasang Constitutional Medicine
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    • v.33 no.2
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    • pp.1-13
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    • 2021
  • Introduction The previous study showed that the psychological Sasang Personality Questionnaire (SPQ) and physical Body Mass Index (BMI) were clinically useful for predicting Sasang types diagnosed with certified clinical specialist and QSCCII. The purpose of current study was to examine its clinical usefulness using large sized clinical sample. Methods The age, sex, height, weight, circumference of waist and hip, Sasang Personality Questionnaire (SPQ), and Sasang type of 2,049 participants listed on the Korean Medicine Data Center were acquired. The anthropometrics of BMI, Ponderal Index (PI), Waist-Hip Ratio (WHR), and Basal Metabolic Rate (BMR) were calculated. The discriminant analysis using SPQ subscales, BMI, age and sex were performed to predict Sasang types. And, the ANCOVA with age and sex as covariate was also used to examine differences among diagnosed and predicted Sasang type groups in biopsychological features. Results The discriminant analysis showed 82.0% of percentage correctly predicted. And, the biopsychological characteristics of predicted Sasang type groups were significantly similar to those of diagnosed Sasang type groups. In both predicted and diagnosed Sasang type groups, the anthropometric measures were in increasing order of So-Eum, So-Yang, and Tae-Eum types, and the psychological measures were in increasing order of So-Eum, Tae-Eum, and So-Yang types as shown in previous studies. Discussion and Conclusion The well validated biopsychological variables of SPQ and BMI showed theoretical and clinical usefulness, and were found to be useful for diagnosing Sasang types in clinical setting. The current study might be useful for the East-West integrative medicine and evidence-based teaching.

Inhibition of miR-128 Abates Aβ-Mediated Cytotoxicity by Targeting PPAR-γ via NF-κB Inactivation in Primary Mouse Cortical Neurons and Neuro2a Cells

  • Geng, Lijiao;Zhang, Tao;Liu, Wei;Chen, Yong
    • Yonsei Medical Journal
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    • v.59 no.9
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    • pp.1096-1106
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    • 2018
  • Purpose: Alzheimer's disease (AD) is the sixth most common cause of death in the United States. MicroRNAs have been identified as vital players in neurodegenerative diseases, including AD. microRNA-128 (miR-128) has been shown to be dysregulated in AD. This study aimed to explore the roles and molecular mechanisms of miR-128 in AD progression. Materials and Methods: Expression patterns of miR-128 and peroxisome proliferator-activated receptor gamma ($PPAR-{\gamma}$) messenger RNA in clinical samples and cells were measured using RT-qPCR assay. $PPAR-{\gamma}$ protein levels were determined by Western blot assay. Cell viability was determined by MTT assay. Cell apoptotic rate was detected by flow cytometry via double-staining of Annexin V-FITC/PI. Caspase 3 and $NF-{\kappa}B$ activity was determined by a Caspase 3 Activity Assay Kit or $NF-{\kappa}B$ p65 Transcription Factor Assay Kit, respectively. Bioinformatics prediction and luciferase reporter assay were used to investigate interactions between miR-128 and $PPAR-{\gamma}$ 3'UTR. Results: MiR-128 expression was upregulated and $PPAR-{\gamma}$ expression was downregulated in plasma from AD patients and $amyloid-{\beta}$ $(A{\beta})-treated$ primary mouse cortical neurons (MCN) and Neuro2a (N2a) cells. Inhibition of miR-128 decreased $A{\beta}-mediated$ cytotoxicity through inactivation of $NF-{\kappa}B$ in MCN and N2a cells. Moreover, $PPAR-{\gamma}$ was a target of miR-128. $PPAR-{\gamma}$ upregulation attenuated $A{\beta}-mediated$ cytotoxicity by inactivating $NF-{\kappa}B$ in MCN and N2a cells. Furthermore, $PPAR-{\gamma}$ downregulation was able to abolish the effect of anti-miR-128 on cytotoxicity and $NF-{\kappa}B$ activity in MCN and N2a cells. Conclusion: MiR-128 inhibitor decreased $A{\beta}-mediated$ cytotoxicity by upregulating $PPAR-{\gamma}$ via inactivation of $NF-{\kappa}B$ in MCN and N2a cells, providing a new potential target in AD treatment.

Improvement of the Beam-Wave Interaction Efficiency Based on the Coupling-Slot Configuration in an Extended Interaction Oscillator

  • Zhu, Sairong;Yin, Yong;Bi, Liangjie;Chang, Zhiwei;Xu, Che;Zeng, Fanbo;Peng, Ruibin;Zhou, Wen;Wang, Bin;Li, Hailong;Meng, Lin
    • Journal of the Korean Physical Society
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    • v.73 no.9
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    • pp.1362-1369
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    • 2018
  • A method aimed at improving the beam-wave interaction efficiency by changing the coupling slot configuration has been proposed in the study of extended interaction oscillators (EIOs). The dispersion characteristics, coupling coefficient and interaction impedance of the high-frequency structure based on different types of coupling slots have been investigated. Four types of coupled cavity structures with different layouts of the coupling slots have been compared to improve the beam-wave interaction efficiency, so as to analyze the beam-wave interaction and practical applications. In order to determine the improvement of the coupling slot to a coupled cavity circuit in an EIO, we designed four nine-gap EIOs based on the coupled cavity structure with different coupling slot configurations. With different operating frequencies and voltages takes into consideration, beam voltages from 27 to 33 kV have been simulated to achieve the best beam-wave interaction efficiency so that the EIOs are able to work in the $2{\pi}$ mode. The influence of the Rb and the ds on the output power is also taken into consideration. The Rb is the radius of the electron beam, and the ds is the width of the coupling slot. The simulation results indicate that a single-slot-type EIO has the best beam-wave interaction efficiency. Its maximum output power is 2.8 kW and the efficiency is 18% when the operating voltage is 31 kV and electric current is 0.5 A. The output powers of these four EIOs that were designed for comparison are not less than 1.7 kW. The improved coupling-slot configurations enables the extended interaction oscillator to meet the different engineering requirements better.

Role of stearyl-coenzyme A desaturase 1 in mediating the effects of palmitic acid on endoplasmic reticulum stress, inflammation, and apoptosis in goose primary hepatocytes

  • Tang, Bincheng;Qiu, Jiamin;Hu, Shenqiang;Li, Liang;Wang, Jiwen
    • Animal Bioscience
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    • v.34 no.7
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    • pp.1210-1220
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    • 2021
  • Objective: Unlike mammals, goose fatty liver shows a strong tolerance to fatty acids without obvious injury. Stearyl-coenzyme A desaturase 1 (SCD1) serves crucial role in desaturation of saturated fatty acids (SAFs), but its role in the SAFs tolerance of goose hepatocytes has not been reported. This study was conducted to explore the role of SCD1 in regulating palmitic acid (PA) tolerance of goose primary hepatocytes. Methods: 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide was examined to reflect the effect of PA on hepatocytes viability, and quantitative polymerase chain reaction was used to detect the mRNA levels of several genes related to endoplasmic reticulum (ER) stress, inflammation, and apoptosis, and the role of SCD1 in PA tolerance of goose hepatocytes was explored using RNA interfere. Results: Our results indicated that goose hepatocytes exhibited a higher tolerant capacity to PA than human hepatic cell line (LO2 cells). In goose primary hepatocytes, the mRNA levels of fatty acid desaturation-related genes (SCD1 and fatty acid desaturase 2) and fatty acid elongate enzyme-related gene (elongase of very long chain fatty acids 6) were significantly upregulated with 0.6 mM PA treatment. However, in LO2 cells, expression of ER stress-related genes (x box-binding protein, binding immunoglobulin protein, and activating transcription factor 6), inflammatory response-related genes (interleukin-6 [IL-6], interleukin-1β [IL-1β], and interferon-γ) and apoptosis-related genes (bcl-2-associated X protein, b-cell lymphoma 2, Caspase-3, and Caspase-9) was significantly enhanced with 0.6 mM PA treatment. Additionally, small interfering RNA (siRNA) mediated downregulation of SCD1 significantly reduced the PA tolerance of goose primary hepatocytes under the treatment of 0.6 mM PA; meanwhile, the mRNA levels of inflammatory-related genes (IL-6 and IL-1β) and several key genes involved in the phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT), forkhead box O1 (FoxO1), mammalian target of rapamycin and AMPK pathways (AKT1, AKT2, FoxO1, and sirtuin 1), as well as the protein expression of cytochrome C and the apoptosis rate were upregulated. Conclusion: In conclusion, our data suggested that SCD1 was involved in enhancing the PA tolerance of goose primary hepatocytes by regulating inflammation- and apoptosis-related genes expression.

Improvement of blood glucose homeostasis in mice fed with Capsosiphon fulvescens extract-added whole wheat cookie (매생이 추출물 첨가 통밀 쿠키의 마우스 혈당 항상성 개선 효과)

  • Lim, Jae-Min;Chun, Su-Hyun;Jeong, Yu-Jin;Lee, Kwang-Won
    • Korean Journal of Food Science and Technology
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    • v.53 no.3
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    • pp.313-320
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    • 2021
  • The present study aimed to investigate the effect of whole wheat cookie supplemented with Capsosiphon fulvescens (CF) extract on serum glucose homeostasis in C57BL/6 mice. This study examined whether the same effect was demonstrated for whole wheat cookie in comparison to previous research documenting the glucose-lowering effect of food products combined with CF extract. Mice were divided into three groups depending on the diet administered: normal cookie (NC), whole wheat cookie (WC), and WC blended with CF extract (WCFE). After 4 weeks of administering the experimental diet, the blood glucose level, serum insulin level, and homeostatic model assessment for insulin resistance index were found to be significantly lower in the WCFE group than in the NC and WC groups. These results suggest that whole wheat cookie containing CF extract is effective in preventing insulin resistance and maintaining blood glucose homeostasis.

Design of Processor Lever Controller for Electric Propulsion System of Naval Ship (전기추진 함정용 프로세서 레버 제어기 설계)

  • Shim, Jaesoon;Lee, Hunseok;Jung, Sung-Young;Oh, Jin-Seok
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.25 no.1
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    • pp.134-145
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    • 2021
  • It is common to optimize the propulsion control system through a so-called tuning process that modifies the parameter values of the propulsion control software during a ship commissioning. However, during this process, if the error of the initial setting value is large, the tuning time may take too long, or the propulsion equipment can be seriously damaged. Therefore, we conducted research on the design of a propulsion controller that applied a Processor lever controller even for inexperienced people with relatively little experience in tuning propulsion control software to be able to reduce the tuning time while protecting the propulsion system. Through simulation, by comparing the execution result of propulsion control lever commands through the PI controller without applying the Processor lever controller. We analyzed the improvement of the Overshoot and propulsion performance. The simulation results showed that the safety of the propulsion system increased because Overshoot of approximately 9.74%, which occurred when the Processor lever function was not applied.

Association between soluble forms of the receptor for advanced glycation end products and periodontal disease: a retrospective study

  • Kim, Keun-Suh;Lee, Yun Jong;Ahn, Soyeon;Chang, Yoon-Seok;Choi, Yonghoon;Lee, Hyo-Jung
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.47 no.6
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    • pp.445-453
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    • 2021
  • Objectives: Periodontitis is the most common chronic disease that causes tooth loss and is related to systemic diseases such as cardiovascular disease and diabetes. An objective indicator of the current activity of periodontitis is necessary. Soluble forms of the receptor for advanced glycation end products (sRAGE) are markers that reflect the status of inflammatory diseases. In this study, the relationship between sRAGE and periodontitis was analyzed to determine whether it can be used to diagnose the current state of periodontitis. Patients and Methods: Eighty-four patients without any systemic diseases were diagnosed with periodontitis using three classifications of periodontitis. Demographics and oral examination data such as plaque index (PI), bleeding on probing (BOP) index, and probing pocket depth (PPD) were analyzed according to each classification. In addition, correlation and partial correlation between sRAGE and the values indicating periodontitis were analyzed. Results: In each classification, the level of sRAGE tended to decrease if periodontitis was present or severe, but this change was not statistically significant. sRAGE and periodontitis-related variables exhibited a weak correlation, among which the BOP index showed a relatively strong negative correlation (ρ=-0.20). Based on this, on analyzing the correlation between the BOP index and sRAGE in the group with more severe periodontitis (PPD≥5 mm group, severe group of AAP/CDC [American Academy of Periodontology/Centers for Disease Control and Prevention], periodontitis group of López), the correlation further increased (ρ=-0.23, -0.40, -0.50). Partial correlation analysis of the sRAGE and BOP index showed a stronger negative correlation (ρ=-0.36, -0.55, -0.45). Conclusion: sRAGE demonstrated a tendency to decrease upon increased severity of periodontitis according to the classifications used. Above all, the correlation with the BOP index, which reflects the current state of periodontitis, was higher in the group with severe periodontitis. This indicates that the current status of periodontitis can be diagnosed through sRAGE.

Ginsenoside Rg3 in combination with artesunate overcomes sorafenib resistance in hepatoma cell and mouse models

  • Chen, Ying-Jie;Wu, Jia-Ying;Deng, Yu-Yi;Wu, Ying;Wang, Xiao-Qi;Li, Amy Sze-man;Wong, Lut Yi;Fu, Xiu-Qiong;Yu, Zhi-Ling;Liang, Chun
    • Journal of Ginseng Research
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    • v.46 no.3
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    • pp.418-425
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    • 2022
  • Background: Sorafenib is effective in treating hepatoma, but most patients develop resistance to it. STAT3 signaling has been implicated in sorafenib resistance. Artesunate (ART) and 20(R)-ginsenoside Rg3 (Rg3) have anti-hepatoma effects and can inhibit STAT3 signaling in cancer cells. This study aimed to evaluate the effects of Rg3 in combination with ART (Rg3-plus-ART) in overcoming sorafenib resistance, and to examine the involvement of STAT3 signaling in these effects. Methods: Sorafenib-resistant HepG2 cells (HepG2-SR) were used to evaluate the in vitro anti-hepatoma effects of Rg3-plus-ART. A HepG2-SR hepatoma-bearing BALB/c-nu/nu mouse model was used to assess the in vivo anti-hepatoma effects of Rg3-plus-ART. CCK-8 assays and Annexin V-FITC/PI double staining were used to examine cell proliferation and apoptosis, respectively. Immunoblotting was employed to examine protein levels. ROS generation was examined by measuring DCF-DA fluorescence. Results: Rg3-plus-ART synergistically reduced viability of, and evoked apoptosis in HepG2-SR cells, and suppressed HepG2-SR tumor growth in mice. Mechanistic studies revealed that Rg3-plus-ART inhibited activation/phosphorylation of Src and STAT3 in HepG2-SR cultures and tumors. The combination also decreased the STAT3 nuclear level and induced ROS production in HepG2-SR cultures. Furthermore, overactivation of STAT3 or removal of ROS diminished the anti-proliferative effects of Rg3-plus-ART, and removal of ROS diminished Rg3-plus-ART's inhibitory effects on STAT3 activation in HepG2-SR cells. Conclusions: Rg3-plus-ART overcomes sorafenib resistance in experimental models, and inhibition of Src/STAT3 signaling and modulation of ROS/STAT3 signaling contribute to the underlying mechanisms. This study provides a pharmacological basis for developing Rg3-plus-ART into a novel modality for treating sorafenib-resistant hepatoma.