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Expression of neurotransmitter receptors in oral keratinocytes and their response to agonists 神经递质受体在口腔角质形成细胞中的表达及其对激动剂的反应
Pub Date : 2021-03-31 DOI: 10.11620/IJOB.2021.46.1.39
Eun Ji Choi, Sung-Ho Chang, Se-Young Choi, Youngnim Choi
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引用次数: 0
The antibacterial effect of Endoseal TCS mixed with water-soluble mangostin derivatives of Garcinia mangostana L. ethanol extract against Enterococcus faecalis and Staphylococcus aureus Endoseal TCS与山竹果水溶性山竹苷衍生物乙醇提取物混合对粪肠球菌和金黄色葡萄球菌的抑菌效果
Pub Date : 2021-03-31 DOI: 10.11620/IJOB.2021.46.1.45
Tae-Young Park, Y. Lim, Jin Hee Kim, Dae Sung Lee, J. Kook
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引用次数: 0
Anticancer effects of D-pinitol in human oral squamous carcinoma cells d -蒎醇对人口腔鳞状癌细胞的抗癌作用
Pub Date : 2020-12-30 DOI: 10.11620/IJOB.2020.45.4.152
Hyunjin Shin, T. Bang, Kang haemi, Park,Bong Soo, Kim Inryoung
D-pinitol is an analog of 3-methoxy-D-chiro-inositol found in beans and plants. D-pinitol has anti-inflammatory, antidiabetic, and anticancer effects. Additionally, D-pinitol induces apoptosis and inhibits metastasis in breast and prostate cancers. However, to date, no study has investigated the anticancer effects of D-pinitol in oral cancer. Therefore, in this study, whether the anticancer effects of D-pinitol induce apoptosis, inhibit the epithelialto- mesenchymal transition (EMT), and arrest cell cycle was investigated in squamous epithelial cells. D-pinitol decreased the survival and cell proliferation rates of CAL-27 and Ca9-22 oral squamous carcinoma cells in a concentration- and time-dependent manner. Evidence of apoptosis, including nuclear condensation, poly (ADP-ribose) polymerase, and caspase-3 fragmentation, was also observed. D-pinitol inhibited the migration and invasion of both cell lines. In terms of EMT-related proteins, E-cadherin was increased, whereas N-cadherin, Snail, and Slug were decreased. D-pinitol also decreased the expression of cyclin D1, a protein involved in the cell cycle, but increased the expression of p21, a cyclin-dependent kinase inhibitor. Hence, D-pinitol induces apoptosis and cell cycle arrest in CAL-27 and Ca9-22 cells, demonstrating an anticancer effect by decreasing the EMT.
d -蒎醇是在豆类和植物中发现的3-甲氧基- d -氨基肌醇的类似物。d -蒎醇具有抗炎、抗糖尿病和抗癌作用。此外,d -蒎醇还能诱导乳腺癌和前列腺癌的细胞凋亡和抑制转移。然而,到目前为止,还没有研究调查d -蒎醇在口腔癌中的抗癌作用。因此,本研究探讨了D-pinitol在鳞状上皮细胞中是否具有诱导细胞凋亡、抑制上皮间质转化(epithelial - alto- mesenchymal transition, EMT)和阻滞细胞周期的抗癌作用。d -蒎醇降低CAL-27和Ca9-22口腔鳞癌细胞的存活率和细胞增殖率呈浓度依赖性和时间依赖性。还观察到细胞凋亡的证据,包括核凝聚、聚(adp -核糖)聚合酶和caspase-3断裂。d -蒎醇抑制两种细胞系的迁移和侵袭。emt相关蛋白方面,E-cadherin升高,N-cadherin、Snail和Slug降低。D-pinitol还降低了参与细胞周期的蛋白cyclin D1的表达,但增加了周期蛋白依赖性激酶抑制剂p21的表达。因此,D-pinitol诱导CAL-27和Ca9-22细胞凋亡和细胞周期阻滞,通过降低EMT显示出抗癌作用。
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引用次数: 2
Inhibitory effects of Coptis chinensis extract on the growth and biofilm formation of Streptococcus mutans and Streptococcus sobrinus 黄连提取物对变形链球菌和斜纹链球菌生长和生物膜形成的抑制作用
Pub Date : 2020-12-30 DOI: 10.11620/IJOB.2020.45.4.143
S. Kim, Y. Song, H. A. Lee, H. Na, C. Jung, Gyung Yun Bek, Jin Chung
Streptococcus mutans and Streptococcus sobrinus play important roles in dental caries. Coptis chinensis is a natural product with antimicrobial activity against enterobacteria; however, its effects on oral streptococci are still unknown. Therefore, the effects of C. chinensis on the growth and biofilm formation of the representative cariogenic bacteria S. mutans and S. sobrinus were investigated for the possible use of C. chinensis as an anticaries agent. The C. chinensis extract was diluted with sterile distilled water, and 0.1–2.5% of the extract was used in the experiment. The effects of the C. chinensis extract on the growth and glucan formation of S. mutans and S. sobrinus were measured by viable cell counting and spectrophotometry at 650 nm absorbance, respectively. Crystal violet staining was also carried out to confirm the C. chinensis extract’s inhibitory effect on biofilm formation. The C. chinensis extract significantly inhibited the growth of S. mutans and S. sobrinus at concentrations of ≥ 0.3% as compared with the control group. The viable cell count of colonies decreased by 1.7-fold and 1.2-fold at 2.5% and 1.25%, respectively, compared with the control group. The biofilm formation of S. mutans and S. sobrinus was inhibited by > 20-fold at C. chinensis extract concentrations of ≥ 1.25% as compared with the control group. In summary, the C. chinensis extract inhibited the growth and biofilm and glucan formation of S. mutans and S. sobrinus . Therefore, C. chinensis might be a potential candidate for controlling dental caries.
变形链球菌和隐球菌在龋齿中起重要作用。黄连是一种对肠杆菌具有抗菌活性的天然产物;然而,其对口腔链球菌的影响尚不清楚。因此,本研究旨在探讨中华葡萄球菌对具有代表性的蛀牙细菌变形链球菌和sobrinus的生长和生物膜形成的影响,为中华葡萄球菌作为一种抗蛀牙剂的可能性进行研究。用无菌蒸馏水稀释五味子提取液,提取液用量为0.1-2.5%。采用活细胞计数法和650 nm光度法分别测定了五味子提取物对变形链球菌和sobrinus生长和葡聚糖形成的影响。结晶紫染色也证实了三叶草提取物对生物膜形成的抑制作用。当浓度≥0.3%时,与对照组相比,紫荆提取物显著抑制了变形链球菌和sobrinus的生长。2.5%和1.25%时,菌落活细胞数分别比对照组减少1.7倍和1.2倍。≥1.25%的紫荆提取物对变形链球菌和sobrinus生物膜的形成抑制作用是对照组的20倍以上。综上所述,五味子提取物对变形链球菌和sobrinus的生长、生物膜和葡聚糖的形成具有抑制作用。因此,中华月牙可能是防治龋病的潜在候选者。
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引用次数: 0
Induction of anti-aquaporin 5 autoantibodies by molecular mimicry in mice 分子模拟诱导小鼠抗水通道蛋白5自身抗体
Pub Date : 2020-12-30 DOI: 10.11620/IJOB.2020.45.4.211
Ahreum Lee, Youngnim Choi
Molecular mimicry is the most common mechanism that breaches self-tolerance. We previously identified autoantibodies to aquaporin-5 (AQP5) in the sera of patients with Sjogren’s syndrome and found that the aquaporin of Prevotella melaninogenica (PmAqp), an oral commensal, is highly homologous to human AQP5. This study aimed to test whether PmAqp can induce anti-AQP5 autoantibodies via molecular mimicry. From the amino acid sequence of PmAqp, an immunizing peptide; i.e., PmE-L, was designed, which contained both the B cell epitope “E” and T cell epitope. C57BL/6 and BALB/c mice were subcutaneously immunized with linear or cyclic forms of PmE-L emulsified in incomplete Freund’s adjuvant. The concentrations of the antibodies in sera were measured using enzymelinked immunosorbent assays. Both linear and cyclic PmE-L induced high levels of antibodies against not only the immunized peptides but also autoantibodies against AQP5E and antibodies against PmE, a Pm homolog of AQP5E. In C57BL/6 mice; however, the cyclic form of PmE-L was more efficient than the linear form in inducing autoantibodies against AQP5E that contained a cyclic epitope. The levels of anti-PmE antibodies and anti-AQP5E autoantibodies showed a strong positive correlation (r = 0.95, p < 0.0005), suggesting molecular mimicry. Collectively, the mice produced anti-AQP5E autoantibodies in response to a PmAqp-derived peptide. This model proved to be useful for studying the mechanisms of autoantibody production by molecular mimicry.
分子模仿是最常见的破坏自我容忍的机制。我们之前在干燥综合征患者的血清中发现了水通道蛋白-5 (AQP5)的自身抗体,并发现黑色素生成普雷沃特菌(PmAqp)的水通道蛋白与人类AQP5高度同源。本研究旨在检测PmAqp是否可以通过分子模拟诱导抗aqp5自身抗体。免疫肽PmAqp的氨基酸序列;即PmE-L,它同时包含B细胞表位“E”和T细胞表位。C57BL/6和BALB/c小鼠皮下接种不完全弗氏佐剂乳化的线性或环状PmE-L。采用酶联免疫吸附法测定血清中抗体的浓度。线性PmE- l和环状PmE- l都诱导了高水平的抗体,不仅针对免疫肽,而且针对AQP5E的自身抗体和针对PmE (AQP5E的Pm同源物)的抗体。C57BL/6小鼠;然而,环状形式的PmE-L在诱导针对含有环状表位的AQP5E的自身抗体方面比线性形式的更有效。抗pme抗体水平与抗aqp5e自身抗体水平呈强正相关(r = 0.95, p < 0.0005),提示分子模拟。总的来说,小鼠产生抗aqp5e自身抗体,以响应pmaqp衍生的肽。该模型可用于研究分子模拟产生自身抗体的机制。
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引用次数: 1
Alteration of cellular events in tooth development by chemical chaperon, Tauroursodeoxycholic acid treatment 化学伴侣子、金牛座脱氧胆酸处理对牙齿发育中细胞事件的改变
Pub Date : 2020-12-30 DOI: 10.11620/IJOB.2020.45.4.190
Eui-Seon Lee, Y. P. Aryal, Tae-Young Kim, Elina Pokharel, Harim Kim, S. Sung, Wern-Joo Sohn, Lee Youngkyun, Chang-Hyeon An, K. J. Young
Several factors, including genetic and environmental insults, impede protein folding and secretion in the endoplasmic reticulum (ER). Accumulation of unfolded or mis-folded protein in the ER manifests as ER stress. To cope with this morbid condition of the ER, recent data has suggested that the intracellular event of an unfolded protein response plays a critical role in managing the secretory load and maintaining proteostasis in the ER. Tauroursodeoxycholic acid (TUDCA) is a chemical chaperone and hydrophilic bile acid that is known to inhibit apoptosis by attenuating ER stress. Numerous studies have revealed that TUDCA affects hepatic diseases, obesity, and inflammatory illnesses. Recently, molecular regulation of ER stress in tooth development, especially during the secretory stage, has been studied. Therefore, in this study, we examined the developmental role of ER stress regulation in tooth morphogenesis using in vitro organ cultivation methods with a chemical chaperone treatment, TUDCA. Altered cellular events including proliferation, apoptosis, and dentinogenesis were examined using immunostaining and terminal deoxynucleotidyl transferase dUTP nick end labeling assay. In addition, altered localization patterns of the formation of hard tissue matrices related to molecules, including amelogenin and nestin, were examined to assess their morphological changes. Based on our findings, modulating the role of the chemical chaperone TUDCA in tooth morphogenesis, especially through the modulation of cellular proliferation and apoptosis, could be applied as a supporting data for tooth regeneration for future studies.
一些因素,包括遗传和环境的损害,阻碍蛋白质折叠和内质网(ER)的分泌。未折叠或错误折叠的蛋白在内质网中的积累表现为内质网应激。为了应对内质网的这种病态状况,最近的数据表明,未折叠蛋白反应的细胞内事件在管理内质网的分泌负荷和维持蛋白质平衡方面起着关键作用。牛磺酸去氧胆酸(TUDCA)是一种化学伴侣和亲水性胆汁酸,已知通过减轻内质膜应激来抑制细胞凋亡。大量研究表明,TUDCA影响肝脏疾病、肥胖和炎症性疾病。近年来,人们对内质网应激在牙齿发育特别是分泌阶段的分子调控进行了研究。因此,在本研究中,我们采用化学伴侣处理(TUDCA)的体外器官培养方法,研究内质网应激调节在牙齿形态发生中的发育作用。使用免疫染色和末端脱氧核苷酸转移酶dUTP缺口末端标记法检测细胞增生、凋亡和牙本质发生等改变的细胞事件。此外,研究人员还检测了与淀粉原蛋白和巢蛋白等分子相关的硬组织基质形成的定位模式的改变,以评估其形态学变化。基于我们的研究结果,调节化学伴侣蛋白TUDCA在牙齿形态发生中的作用,特别是通过调节细胞增殖和凋亡,可以作为未来牙齿再生研究的支持数据。
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引用次数: 0
Antitumor effects of octyl gallate on hypopharyngeal carcinoma cells 没食子酸辛酯对下咽癌细胞的抗肿瘤作用
Pub Date : 2020-12-30 DOI: 10.11620/IJOB.2020.45.4.218
Trang Ntk, H. Yoo
The antitumor effects of octyl gallate (OG) were investigated on FaDu human hypopharyngeal squamous carcinoma cells. At various concentrations, OG inhibited the proliferation of FaDu cells by suppressing cell cycle regulators and induced apoptosis by activating caspase 3 and its downstream poly (ADP-ribose) polymerase, thereby damaging DNA. Immunoblotting demonstrated that OG significantly suppressed the expression of integrin family proteins (integrin α4, αv, β3, β4), hindering cell adhesion. The reduced expression of integrins subsequently mediated the mitogenactivated protein kinase signaling pathway to stimulate the activation of extracellular signal-regulated kinases and c-jun N-terminal kinases, leading to apoptosis. Thus, OG demonstrated antitumor activity on hypopharyngeal squamous carcinoma cells by suppressing cell proliferation and inducing apoptosis.
研究没食子酸辛酯(OG)对FaDu人下咽鳞癌细胞的抗肿瘤作用。在不同浓度下,OG通过抑制细胞周期调节因子抑制FaDu细胞的增殖,并通过激活caspase 3及其下游多聚核糖(adp -核糖)聚合酶诱导细胞凋亡,从而损伤DNA。免疫印迹显示,OG显著抑制整合素家族蛋白(整合素α4、αv、β3、β4)的表达,阻碍细胞粘附。整合素的表达减少随后介导有丝分裂原激活的蛋白激酶信号通路,刺激细胞外信号调节激酶和c-jun n端激酶的激活,导致细胞凋亡。由此可见,OG通过抑制下咽鳞癌细胞增殖和诱导细胞凋亡,表现出抗肿瘤活性。
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引用次数: 0
Role of microglial activation on neuronal excitability in rat substantia gelatinosa 小胶质细胞活化对大鼠明胶质神经元兴奋性的影响
Pub Date : 2020-12-30 DOI: 10.11620/IJOB.2020.45.4.225
A. Park, S. Chun
Glial cells, including astrocytes and microglia, interact closely with neurons and modulate pain transmission, particularly under pathological conditions. In this study, we examined the excitability of substantia gelatinosa (SG) neurons of the spinal dorsal horn using a patch clamp recording to investigate the roles of microglial activation in the nociceptive processes of rats. We used xanthine/xanthine oxidase (X/XO), a generator of superoxide anion (O2∙–), to induce a pathological pain condition. X/XO treatment induced an inward current and membrane depolarization. The inward current was significantly inhibited by minocycline, a microglial inhibitor, and fluorocitrate, an astrocyte inhibitor. To examine whether toll-like receptor 4 (TLR4) in microglia was involved in the inward current, we used lipopolysaccharide (LPS), a highly specific TLR4 agonist. The LPS induced inward current, which was decreased by pretreatment with Tak-242, a TLR4-specific inhibitor, and phenyl N-t-butylnitrone, a reactive oxygen species scavenger. The X/XO-induced inward current was also inhibited by pretreatment with Tak-242. These results indicate that the X/XO-induced inward current of SG neurons occurs through activation of TLR4 in microglial cells, suggesting that neuroglial cells modulate the nociceptive process through central sensitization.
胶质细胞,包括星形胶质细胞和小胶质细胞,与神经元密切相互作用并调节疼痛传递,特别是在病理条件下。本研究通过膜片钳记录大鼠脊髓背角明胶质(SG)神经元的兴奋性,探讨了小胶质细胞的激活在大鼠伤害感受过程中的作用。我们使用产生超氧阴离子(O2∙-)的黄嘌呤/黄嘌呤氧化酶(X/XO)诱导病理性疼痛。X/XO处理诱导向内电流和膜去极化。向内电流被米诺环素(一种小胶质细胞抑制剂)和氟柠檬酸盐(一种星形胶质细胞抑制剂)显著抑制。为了研究小胶质细胞中的toll样受体4 (TLR4)是否参与了内向电流,我们使用了脂多糖(LPS),一种高度特异性的TLR4激动剂。LPS诱导向内电流,通过tlr4特异性抑制剂Tak-242和活性氧清除剂苯基n -t-丁基硝基酮预处理降低电流。Tak-242预处理也能抑制X/ xo诱导的内向电流。这些结果表明,X/ xo诱导的SG神经元向内电流是通过激活小胶质细胞中的TLR4发生的,表明神经胶质细胞通过中枢致敏调节伤害感受过程。
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引用次数: 0
Green tea polyphenol (–)-epigallocatechin-3-gallate prevents ultraviolet-induced apoptosis in PC12 cells 绿茶多酚(-)-表没食子儿茶素-3-没食子酸酯防止紫外线诱导的PC12细胞凋亡
Pub Date : 2020-12-30 DOI: 10.11620/IJOB.2020.45.4.179
S. Woo, Kim Yoon-Jung, Bangrong Cai, S. Park, Young Ho Kim, Ok-Joon Kim, I. Kang, Won-Jae Kim, Ji-Yeon Jung
Green tea polyphenol (–)-epigallocatechin-3-gallate (EGCG) is a potent antioxidant with protective effects against neurotoxicity. However, it is currently unclear whether EGCG protects neuronal cells against radiation-induced damage. Therefore, the objective of this study was to investigate the effects of EGCG on ultraviolet (UV)-induced oxidative stress and apoptosis in PC12 cells. The effects of UV irradiation included apoptotic cell death, which was associated with DNA fragmentation, reactive oxygen species (ROS) production, enhanced caspase-3 and caspase-9 activity, and poly (ADP-ribose) polymerase cleavage. UV irradiation also increased the Bax/Bcl-2 ratio and mitochondrial pathway-associated cytochrome c expression. However, pretreatment with EGCG before UV exposure markedly decreased UV-induced DNA fragmentation and ROS production. Furthermore, the UV irradiationinduced increase in Bax/Bcl-2 ratio, cytochrome c upregulation, and caspase-3 and caspase-9 activation were each ameliorated by EGCG pretreatment. Additionally, EGCG suppressed UV-induced phosphorylation of p38 and rescued UV-downregulated phosphorylation of ERK. Taken together, these results suggest that EGCG prevents UV irradiationinduced apoptosis in PC12 cells by scavenging ROS and inhibiting the mitochondrial pathways known to play a crucial role in apoptosis. In addition, EGCG inhibits UV-induced apoptosis via JNK inactivation and ERK activation in PC12 cells. Thus, EGCG represents a potential neuroprotective agent that could be applied to prevent neuronal cell death induced by UV irradiation.
绿茶多酚(-)-表没食子儿茶素-3-没食子酸酯(EGCG)是一种有效的抗氧化剂,对神经毒性具有保护作用。然而,目前还不清楚EGCG是否能保护神经细胞免受辐射引起的损伤。因此,本研究旨在探讨EGCG对紫外线(UV)诱导的PC12细胞氧化应激和凋亡的影响。紫外线照射的影响包括细胞凋亡,这与DNA断裂、活性氧(ROS)的产生、caspase-3和caspase-9活性的增强以及聚(adp -核糖)聚合酶的裂解有关。紫外线照射也增加了Bax/Bcl-2比值和线粒体途径相关的细胞色素c表达。然而,在紫外线照射前用EGCG预处理可显著降低紫外线诱导的DNA断裂和ROS的产生。此外,紫外线照射诱导的Bax/Bcl-2比值升高、细胞色素c上调、caspase-3和caspase-9活化均可通过EGCG预处理得到改善。此外,EGCG抑制了紫外线诱导的p38磷酸化,并挽救了紫外线下调的ERK磷酸化。综上所述,这些结果表明EGCG通过清除活性氧和抑制已知在凋亡中起关键作用的线粒体途径来阻止紫外线照射诱导的PC12细胞凋亡。此外,EGCG通过JNK失活和ERK活化抑制uv诱导的PC12细胞凋亡。因此,EGCG是一种潜在的神经保护剂,可用于预防紫外线照射诱导的神经元细胞死亡。
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引用次数: 0
Increased SOX2 expression in three-dimensional sphere culture of dental pulp stem cells 牙髓干细胞三维球培养中SOX2表达增加
Pub Date : 2020-12-30 DOI: 10.11620/IJOB.2020.45.4.197
Eun Jin Seo, I. Jang
Mesenchymal stem cells in the dental pulp exhibit a tendency for differentiation into various dental lineages and hold great potential as a major conduit for regenerative treatment in dentistry. Although they can be readily isolated from teeth, the exact characteristics of these stem cells have not been fully understood so far. When compared to twodimensional (2D) cultures, three-dimensional (3D) cultures have the advantage of enriching the stem cell population. Hence, 3D-organoid culture and 3D-sphere culture were applied to dental pulp cells in the current study. Although the establishment of the organoid culture proved unsuccessful, the 3D-sphere culture readily initiated the stable generation of cell aggregates, which continued to grow and could be passaged to the second round. Interestingly, a significant increase in SOX2 expression was detected in the 3D-spheroid culture compared to the 2D culture. These results indicate the enrichment of the stemness-high population in the 3D-sphere culture. Thus, 3D-sphere culture may act as a link between the conventional and 3D-organoid cultures and aid in understanding the characteristics of dental pulp stem cells.
牙髓间充质干细胞具有向不同牙系分化的趋势,在牙科再生治疗中具有巨大的潜力。尽管这些干细胞可以很容易地从牙齿中分离出来,但到目前为止,人们还没有完全了解这些干细胞的确切特征。与二维(2D)培养相比,三维(3D)培养具有丰富干细胞群的优势。因此,本研究将3d类器官培养和3d球培养应用于牙髓细胞。虽然类器官培养的建立失败,但3d球培养很容易启动细胞聚集体的稳定产生,细胞聚集体继续生长并可以传代到第二轮。有趣的是,与2D培养相比,在3d球体培养中检测到SOX2的表达显著增加。这些结果表明,在3d球培养中,高干性群体丰富。因此,3d球培养可以作为传统和3d类器官培养之间的联系,并有助于理解牙髓干细胞的特性。
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引用次数: 0
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International Journal of Oral Biology
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