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Characterization of immune cells in the rat intestinal mucosa and liver involved in inflammation caused by LPS and evaluation of the effects of N-acetylcysteine and disulfiram (well-known sulfur drugs) for this inflammation LPS引起的大鼠肠黏膜和肝脏炎症中免疫细胞的特征及n -乙酰半胱氨酸和双硫仑(众所周知的含硫药物)对这种炎症的作用的评价
IF 2.3 4区 生物学 Q4 CELL BIOLOGY Pub Date : 2025-06-06 DOI: 10.1016/j.acthis.2025.152272
Anthea Miller , Giorgia Pia Lombardo , Laura Spiccia , Valentina Natale , Alba Migliorato , Marek Bednarski , Małgorzata Iciek , Anna Bilska-Wilkosz , Mateusz Sablik , Eugenia Rita Lauriano , Magdalena Kotańska , Simona Pergolizzi
Lipopolysaccharide (LPS)-induced inflammation is an experimental rat model often used as a tool for testing new drugs as candidates for treating various diseases associated with inflammation. New methods now allow for precise imaging of tissues and changes induced by various factors. To increase knowledge about LPS-induced inflammation and promote strategies for investigating new therapies, this study aims to characterize immune cells involved in inflammation in the rat intestinal mucosa and liver and to evaluate the therapeutic effect of two well-known sulfur drugs N-acetylcysteine (NAC) and disulfiram (DSF) on this model LPS was administered intraperitoneally to rats once a day, for 10 days. NAC and DSF were administered 5 h after LPS. At the end of experiment, animals were euthanized, and the intestine and liver were collected. The immune cells of the intestinal mucosa and liver were characterized with the following antibodies: Toll-like receptors (TLR2 and TLR4), smooth muscle alpha-actin (α-SMA), major histocompatibility complex II (MHC-II), and serotonin (5-HT). In samples obtained from inflamed rat intestinal mucosa, it was possible to detect TLR2-positive and TLR4-positive cells, and numerous α-SMA-positive cells, indicating an inflammatory state. Furthermore, an increase in serotonin positive neuroendocrine cells compared to normal was demonstrated, which could be associated with intestinal inflammation. The number of these positive cells was much smaller in the samples derived from animals treated with NAC or DSF, suggesting anti-inflammatory action of these drugs. In the inflamed rat liver, several immune cells positive for these antibodies were observed and NAC or DSF decreased the amount of these positive cells. In conclusion, this study shows that bacterial LPS can activate various innate immune system cell populations, such as dendritic cells, neutrophils, Kupffer cells, myofibroblasts and enterocytes. Moreover, this study demonstrates the beneficial effects on NAC and DSF in alleviating inflammation and relieving tissue fibrosis in the LPS-induced inflammation in the rat intestinal mucosa and liver.
脂多糖(LPS)诱导的炎症是一种实验性大鼠模型,经常被用作测试新药作为治疗各种炎症相关疾病的候选药物的工具。现在的新方法允许对组织和由各种因素引起的变化进行精确成像。为了增加对脂多糖诱导炎症的认识,促进研究新疗法的策略,本研究旨在表征大鼠肠黏膜和肝脏中参与炎症的免疫细胞,并评估两种著名的硫药物n -乙酰半胱氨酸(NAC)和双硫仑(DSF)对该模型的治疗效果,LPS每天腹腔注射一次,持续10天。LPS后5 h给予NAC和DSF。实验结束时,对动物实施安乐死,取肠、肝。肠黏膜和肝脏免疫细胞的主要抗体为toll样受体(TLR2和TLR4)、平滑肌α-肌动蛋白(α-SMA)、主要组织相容性复合体II (MHC-II)和血清素(5-HT)。在炎症大鼠肠黏膜样品中,可以检测到tlr2阳性和tlr4阳性细胞,以及大量α- sma阳性细胞,表明炎症状态。此外,与正常相比,血清素阳性神经内分泌细胞增加,这可能与肠道炎症有关。在NAC或DSF处理的动物样本中,这些阳性细胞的数量要少得多,这表明这些药物具有抗炎作用。在炎症大鼠肝脏中,观察到一些免疫细胞对这些抗体呈阳性,NAC或DSF减少了这些阳性细胞的数量。综上所述,本研究表明细菌LPS可以激活多种先天免疫系统细胞群,如树突状细胞、中性粒细胞、库普弗细胞、肌成纤维细胞和肠细胞。此外,本研究还证实了NAC和DSF在lps诱导的大鼠肠黏膜和肝脏炎症中具有减轻炎症和缓解组织纤维化的有益作用。
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引用次数: 0
Exosomal delivery of AZD5582 to overcome TRAIL resistance as an optimal therapy against triple-negative breast cancer 外泌体递送AZD5582克服TRAIL耐药是治疗三阴性乳腺癌的最佳方法
IF 2.3 4区 生物学 Q4 CELL BIOLOGY Pub Date : 2025-05-24 DOI: 10.1016/j.acthis.2025.152270
Wanting Zhang , Quanjiang Li , Rui Tian , Zhujie Deng , Ronghui Sun , Xiubin Kuang , Jun Peng , Bin Xie , Chen Huang , Zhengqiang Yuan
Triple-negative breast cancer (TNBC) is a highly aggressive subtype of breast cancer that lacks effective targeted therapies mainly due to drug resistance. Therefore, there is an urgent need to develop highly effective therapeutic strategies for TNBC. Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) selectively induces apoptosis in transformed and cancerous cells, indicating its potential for anti-cancer therapy. Unfortunately, the clinical trials of recombinant TRAIL (rTRAIL) had actually failed due to the very common TRAIL resistance in cancers. Exosomal delivery of TRAIL (EV-T) has been shown to greatly improve the cytotoxicity of TRAIL. Moreover, the TRAIL resistance was closely correlated with the upregulation of inhibitors of apoptosis proteins (IAPs) in cancer cells. As a potent antagonist of IAPs, AZD5582 (AZD) was previously shown to drastically sensitize TRAIL response. Herein, we hypothesize that AZD may be loaded into EV-T for co-delivery of AZD and TRAIL to make a synergistic combination anticancer therapy against TNBC. First, TRAIL-expressing mesenchymal stem cells (MSCs) were used to prepare EV-Ts. Then, AZD was loaded into EV-T by ultrasound to prepare the composite nanodrug AZD@EV-T. EV encapsulation significantly improved AZD stability and cellular delivery efficiency, leading to the synergistically augmented cytotoxicity and apoptosis induction in both breast and kidney cancer cell lines, whilst sparing the normal MSCs. The potential mechanisms underlying the synergism appeared to be associated with the concomitant upregulation of death receptor 5 (DR5) and expressional suppression of various anti-apoptotic factors. Importantly, the AZD@EV-T therapy triggered strikingly enhanced growth inhibition and apoptosis in the subcutaneously established BT549 xenograft tumors, consequently leading to the complete tumor regression. It also demonstrated significant potential for treating kidney cancer in A498 kidney tumor organoids. Together, AZD@EV-T effectively overcomes TRAIL resistance and may represent a highly effective and innovative anticancer therapy for both TNBC and kidney cancers.
三阴性乳腺癌(TNBC)是一种高度侵袭性的乳腺癌亚型,主要由于耐药而缺乏有效的靶向治疗。因此,迫切需要开发高效的TNBC治疗策略。肿瘤坏死因子(Tumor necrosis factor, TNF)相关凋亡诱导配体(apoptosis-inducing ligand, TRAIL)在转化细胞和癌细胞中选择性诱导凋亡,表明其具有抗癌治疗的潜力。不幸的是,重组TRAIL (rTRAIL)的临床试验实际上失败了,因为TRAIL在癌症中非常常见。外泌体递送TRAIL (EV-T)已被证明可大大改善TRAIL的细胞毒性。此外,TRAIL耐药与癌细胞中凋亡蛋白抑制剂(IAPs)的上调密切相关。作为一种有效的IAPs拮抗剂,AZD5582 (AZD)先前被证明能显著致敏TRAIL反应。在此,我们假设AZD可能被装载到EV-T中,共同递送AZD和TRAIL,从而形成针对TNBC的协同联合抗癌治疗。首先,利用表达trail的间充质干细胞(MSCs)制备EV-Ts。然后,通过超声将AZD加载到EV-T中,制备复合纳米药物AZD@EV-T。EV包封显著提高了AZD的稳定性和细胞递送效率,从而协同增强了乳腺癌和肾癌细胞系的细胞毒性和凋亡诱导,同时保留了正常的MSCs。协同作用的潜在机制似乎与死亡受体5 (DR5)的上调和各种抗凋亡因子的表达抑制有关。重要的是,AZD@EV-T治疗在皮下建立的BT549异种移植肿瘤中引发了显著增强的生长抑制和细胞凋亡,从而导致肿瘤完全消退。在A498肾肿瘤类器官中也显示出治疗肾癌的显著潜力。总之,AZD@EV-T有效地克服了TRAIL耐药性,可能代表了TNBC和肾癌的一种高效和创新的抗癌疗法。
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引用次数: 0
Lysophosphatidic acid (LPA) receptor signaling enhances malignant potential in highly migratory lung cancer cells under hypoxic conditions 溶血磷脂酸(LPA)受体信号在缺氧条件下增强高迁移肺癌细胞的恶性潜能
IF 2.3 4区 生物学 Q4 CELL BIOLOGY Pub Date : 2025-05-23 DOI: 10.1016/j.acthis.2025.152268
Moemi Tamura, Miwa Takai, Mao Yamamoto, Narumi Yashiro, Anri Taniguchi, Yuka Kusumoto, Shion Nagano, Nanami Shimomura, Toshifumi Tsujiuchi
Hypoxia contributes to tumor progression, promoting cancer cell motility, invasion and metastasis. Lysophosphatidic acid (LPA) receptors are implicated in the pathogenesis of cancer. In this study, we investigated the role of LPA receptor signaling in modulating malignant behavior under hypoxic conditions (1 % O2) in lung cancer cells. We generated highly migratory A549-R12 cells from the parental lung cancer A549 cells for this purpose. LPAR1 and LPAR2 expression levels were lower in both A549 and A549-R12 cells cultured at 1 % O2 compared to those cultured at 21 % O2, while LPAR3 expression remained unchanged between the two cell lines. Cell motility increased in both A549 and A549-R12 cells cultured at 1 % O2. Notably, A549-R12 cells exhibited greater motility under 1 % O2 conditions than A549 cells. Treatment with AM966 (an LPA1 antagonist) and (2S)-OMPT (an LPA3 agonist) further increased the motility of A549-R12 cells, while GRI-977143 (an LPA2 agonist) decreased their motility. Moreover, the invasion activity of A549-R12 cells was higher than that of A549 cells, with 1 % O2 conditions significantly enhancing A549-R12 cell invasion. AM966 and (2S)-OMPT stimulated, whereas GRI-977143 inhibited, the invasion of A549-R12 cells. In the presence of LPA, cell viability to cisplatin (CDDP) was higher in A549-R12 cells cultured at both 21 % and 1 % O2 compared to A549 cells. These results suggest that LPA receptor signaling plays a key role in regulating malignant progression in highly migratory lung cancer cells under hypoxic conditions.
缺氧有助于肿瘤的进展,促进癌细胞的运动、侵袭和转移。溶血磷脂酸(LPA)受体与癌症的发病机制有关。在这项研究中,我们研究了LPA受体信号在低氧条件下(1 % O2)在肺癌细胞中调节恶性行为的作用。为此,我们从亲代肺癌A549细胞中产生了高度迁移的A549- r12细胞。在1 % O2下培养的A549和A549- r12细胞中,LPAR1和LPAR2的表达水平低于在21 % O2下培养的细胞,而LPAR3的表达在两种细胞系之间保持不变。在1 % O2条件下培养的A549和A549- r12细胞的细胞活力均增加。值得注意的是,A549- r12细胞在1 % O2条件下比A549细胞表现出更大的运动性。AM966 (LPA1拮抗剂)和(2S)-OMPT (LPA3激动剂)进一步增加了A549-R12细胞的运动性,而GRI-977143 (LPA2激动剂)降低了它们的运动性。此外,A549- r12细胞的侵袭活性高于A549细胞,1 % O2条件显著增强A549- r12细胞的侵袭活性。AM966和(2S)-OMPT刺激A549-R12细胞的侵袭,而GRI-977143抑制A549-R12细胞的侵袭。在LPA存在的情况下,与A549细胞相比,在21 %和1 % O2下培养的A549- r12细胞对顺铂(CDDP)的存活率更高。这些结果表明LPA受体信号在缺氧条件下调节高迁移性肺癌细胞的恶性进展中起关键作用。
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引用次数: 0
Non-specific background in immunoglobulin G staining can be effectively eliminated using heating or a catalase inhibitor 免疫球蛋白G染色中的非特异性背景可以通过加热或过氧化氢酶抑制剂有效地消除
IF 2.3 4区 生物学 Q4 CELL BIOLOGY Pub Date : 2025-05-19 DOI: 10.1016/j.acthis.2025.152269
Kairan Yang , Ting Xu , Chengkang Lin , Zuisu Yang , Haiyan Lyu , Falei Yuan
Immunoglobulin G (IgG) staining is a widely used method for assessing blood-brain barrier (BBB) integrity. However, significant non-specific binding is often observed in many studies, which can interfere with the accurate interpretation of results. In this study, the horseradish peroxidase (HRP)-based polymer method and the streptavidin-biotin complex (SABC) method were used to perform IgG staining. The effects of hydrogen peroxide, heating, and a catalase inhibitor on reducing background staining were evaluated in brain sections from untreated mice and those subjected to middle cerebral artery occlusion (MCAO). The results showed that immunohistochemistry without hydrogen peroxide pretreatment still produced minimal background in paraffin-embedded sections. However, IgG staining with hydrogen peroxide pretreatment led to substantial background in vibratome sections. Compared to the SABC method, a mixture of the catalase inhibitor 3-amino-1,2,4-triazole and hydrogen peroxide reduced background staining by 35.4 % ± 5.7 % in the cortex of untreated mouse brains and by 36.9 % ± 1.8 % in the contralateral cortex of MCAO mice when using the polymer method. Additionally, heating at 75°C was sufficient to eliminate non-specific binding in brain sections from both untreated and MCAO mice. Hydrogen peroxide pretreatment alone was ineffective in removing background staining in brain sections from either untreated or MCAO mice. In summary, this study demonstrates that hydrogen peroxide pretreatment is effective in reducing background only when combined with a catalase inhibitor but is unnecessary when the tissue is heated. Heating is a simple and effective method for removing the IgG staining background when detecting BBB leakage.
免疫球蛋白G (IgG)染色是一种广泛使用的评估血脑屏障(BBB)完整性的方法。然而,在许多研究中经常观察到显著的非特异性结合,这可能会干扰结果的准确解释。本研究采用辣根过氧化物酶(HRP)聚合物法和链亲和素-生物素复合物(SABC)法进行IgG染色。过氧化氢、加热和过氧化氢酶抑制剂对降低背景染色的影响在未治疗的小鼠和大脑中动脉闭塞(MCAO)小鼠的脑切片中进行了评估。结果表明,未经双氧水预处理的免疫组织化学方法在石蜡包埋切片中仍产生极小的背景。然而,过氧化氢预处理的IgG染色在振动体切片中导致大量背景。与SABC方法相比,过氧化氢酶抑制剂3-氨基-1,2,4-三唑和过氧化氢的混合物在使用聚合物方法时,未处理小鼠大脑皮层的背景染色减少35.4% % ± 5.7 %,在MCAO小鼠对侧皮层的背景染色减少36.9 % ± 1.8 %。此外,在75°C下加热足以消除未处理和MCAO小鼠脑切片中的非特异性结合。单独过氧化氢预处理对未处理或MCAO小鼠脑切片的背景染色均无效。总之,这项研究表明,过氧化氢预处理只有在与过氧化氢酶抑制剂联合使用时才能有效地降低背景,而当组织被加热时则没有必要。加热是检测血脑屏障渗漏时去除IgG染色背景的一种简单有效的方法。
{"title":"Non-specific background in immunoglobulin G staining can be effectively eliminated using heating or a catalase inhibitor","authors":"Kairan Yang ,&nbsp;Ting Xu ,&nbsp;Chengkang Lin ,&nbsp;Zuisu Yang ,&nbsp;Haiyan Lyu ,&nbsp;Falei Yuan","doi":"10.1016/j.acthis.2025.152269","DOIUrl":"10.1016/j.acthis.2025.152269","url":null,"abstract":"<div><div>Immunoglobulin G (IgG) staining is a widely used method for assessing blood-brain barrier (BBB) integrity. However, significant non-specific binding is often observed in many studies, which can interfere with the accurate interpretation of results. In this study, the horseradish peroxidase (HRP)-based polymer method and the streptavidin-biotin complex (SABC) method were used to perform IgG staining. The effects of hydrogen peroxide, heating, and a catalase inhibitor on reducing background staining were evaluated in brain sections from untreated mice and those subjected to middle cerebral artery occlusion (MCAO). The results showed that immunohistochemistry without hydrogen peroxide pretreatment still produced minimal background in paraffin-embedded sections. However, IgG staining with hydrogen peroxide pretreatment led to substantial background in vibratome sections. Compared to the SABC method, a mixture of the catalase inhibitor 3-amino-1,2,4-triazole and hydrogen peroxide reduced background staining by 35.4 % ± 5.7 % in the cortex of untreated mouse brains and by 36.9 % ± 1.8 % in the contralateral cortex of MCAO mice when using the polymer method. Additionally, heating at 75°C was sufficient to eliminate non-specific binding in brain sections from both untreated and MCAO mice. Hydrogen peroxide pretreatment alone was ineffective in removing background staining in brain sections from either untreated or MCAO mice. In summary, this study demonstrates that hydrogen peroxide pretreatment is effective in reducing background only when combined with a catalase inhibitor but is unnecessary when the tissue is heated. Heating is a simple and effective method for removing the IgG staining background when detecting BBB leakage.</div></div>","PeriodicalId":6961,"journal":{"name":"Acta histochemica","volume":"127 3","pages":"Article 152269"},"PeriodicalIF":2.3,"publicationDate":"2025-05-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144084107","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Retraction notice to “miR-103–3p attenuates liver injury with severe acute pancreatitis by inhibiting pyroptosis through miR-103-3p/NLRP1 axis” [Acta Histochem. 126 (2024) 152211] 关于“miR-103-3p通过miR-103-3p/NLRP1轴抑制焦亡减轻重症急性胰腺炎肝损伤”的撤回通知[journal of histochemistry . 126(2024) 152211]。
IF 2.4 4区 生物学 Q4 CELL BIOLOGY Pub Date : 2025-05-15 DOI: 10.1016/j.acthis.2025.152258
Wenquan Zhang , Min Du , Yingjian Jiang , Jiang Wang , Yue Yu , DianLiang Zhang
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引用次数: 0
Quantifying mechanical, proliferation, and migrational properties of bulk ovarian cancer cells and ovarian cancer stem cells 定量测定大块卵巢癌细胞和卵巢癌干细胞的力学、增殖和迁移特性
IF 2.3 4区 生物学 Q4 CELL BIOLOGY Pub Date : 2025-05-10 DOI: 10.1016/j.acthis.2025.152260
Crystal Brown , Emily Stidham , Elly Beck , Ali Eickhoff , Brianna Hill , Kalyani Nair , Craig Cady
There are a limited number of studies analyzing ovarian cancer stem cell properties. The goal of this study was to analyze the mechanical and migrational properties of ovarian cancer stem cells with the well-researched bulk ovarian cancer cells. Through the use of atomic force microscopy (AFM), the mechanical properties of both cell types were gathered. In preparation for AFM analysis, an optimal fixation method was developed and performed on the cells. AFM analysis provided mechanical properties for both cell types, including cellular stiffness, maximum adhesion force, surface area, and mean surface roughness. Cell proliferation and transwell migration assays were assessed to determine the aggressiveness of both cancer cell types. A clear trend between both cell types was expected for the mechanical and potential aggression properties. The data from both analyses were used to create a baseline for migration, proliferation, and mechanical properties of ovarian cancer stem cells and bulk ovarian cancer cells. Further studies will assess if these properties are impacted by chemotherapy exposure.
分析卵巢癌干细胞特性的研究数量有限。本研究的目的是分析卵巢癌干细胞的力学和迁移特性与大量研究的卵巢癌细胞。通过原子力显微镜(AFM),收集了两种细胞类型的力学性能。为了准备AFM分析,我们开发了一种最佳的固定方法并对细胞进行了固定。AFM分析提供了两种细胞类型的力学性能,包括细胞刚度、最大附着力、表面积和平均表面粗糙度。评估细胞增殖和跨井迁移试验以确定两种癌细胞类型的侵袭性。两种细胞类型在机械和潜在的攻击特性上有明显的趋势。这两项分析的数据被用来创建卵巢癌干细胞和散装卵巢癌细胞的迁移、增殖和力学特性的基线。进一步的研究将评估这些特性是否受到化疗暴露的影响。
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引用次数: 0
Corrigendum to “DRAM1 plays a tumor suppressor role in clear cell renal cell carcinoma through modulating Akt signaling” [Acta Histochem. 124 (2022) 151874] “DRAM1通过调节Akt信号通路在透明细胞肾细胞癌中发挥肿瘤抑制作用”的更正[j].组织化学学报,124(2022)151874。
IF 2.3 4区 生物学 Q4 CELL BIOLOGY Pub Date : 2025-05-09 DOI: 10.1016/j.acthis.2025.152259
Qingyan Feng, Meijuan Cheng, Jingjing Jin, Shenglei Zhang, Yaling Bai, Jinsheng Xu
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引用次数: 0
SNHG16 suppression enhances M2 macrophage polarization and inhibits VSMC migration in atherosclerosis 抑制SNHG16可增强动脉粥样硬化中M2巨噬细胞极化,抑制VSMC迁移
IF 2.3 4区 生物学 Q4 CELL BIOLOGY Pub Date : 2025-05-06 DOI: 10.1016/j.acthis.2025.152248
Bing Gao , Maogong Xi , Ying Cui , Kai Wang , Hui Zhang , Yiyong Wang
Atherosclerosis (AS) significantly impacts both cardiovascular and cerebrovascular health, making it an important area of research for potential therapeutic interventions. This study investigates the role of lncRNA SNHG16 in macrophage polarization and its effects on the progression of AS. We assessed the expression of SNHG16 in macrophages and vascular smooth muscle cells (VSMCs) treated with oxidized low-density lipoprotein (ox-LDL) using qPCR. Cell proliferation was evaluated via EdU assay and western blotting, while flow cytometry and immunofluorescence were employed to analyze the polarization of macrophages. Foam cell formation was examined using Oil Red O staining. In a co-culture system, VSMCs treated with ox-LDL were cultured alongside macrophages pretreated with sh-SNHG16, and VSMC viability, migration, and motility were assessed using CCK-8, migration, and scratch assays. The levels of inflammatory cytokines, including IL-6, IL-10, TGFβ, and TNFα, were quantified by ELISA. Our results show that SNHG16 expression is upregulated in ox-LDL-treated cells, which correlates with enhanced macrophage proliferation. Inhibition of SNHG16 promoted M1-to-M2 macrophage polarization, reducing foam cell formation and inflammation. Furthermore, SNHG16 knockdown limited VSMC viability and motility, while attenuating ox-LDL-induced inflammatory responses. In conclusion, suppression of SNHG16 favors M2 macrophage polarization and presents a potential therapeutic target for AS management.
动脉粥样硬化(AS)显著影响心脑血管健康,使其成为潜在治疗干预的重要研究领域。本研究探讨lncRNA SNHG16在巨噬细胞极化中的作用及其对AS进展的影响。我们使用qPCR方法评估了氧化低密度脂蛋白(ox-LDL)处理巨噬细胞和血管平滑肌细胞(VSMCs)中SNHG16的表达。EdU法、western blotting法检测细胞增殖,流式细胞术、免疫荧光法检测巨噬细胞极化情况。油红O染色检测泡沫细胞形成。在共培养系统中,用ox-LDL处理的VSMC与用sh-SNHG16预处理的巨噬细胞一起培养,使用CCK-8、迁移和划痕试验评估VSMC的活力、迁移和运动性。ELISA法测定各组炎症因子IL-6、IL-10、tgf - β、tnf - α水平。我们的研究结果表明,SNHG16在ox- ldl处理的细胞中表达上调,这与巨噬细胞增殖增强有关。抑制SNHG16促进巨噬细胞M1-to-M2极化,减少泡沫细胞形成和炎症。此外,SNHG16敲低限制了VSMC的活力和运动,同时减轻了ox- ldl诱导的炎症反应。综上所述,抑制SNHG16有利于M2巨噬细胞极化,是治疗AS的潜在靶点。
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引用次数: 0
Histological, histochemical, and morphometric analysis of epidermal Leydig cells and histochemical characterization of epidermal apical cells in juvenile and adult axolotls (Ambystoma mexicanum) 幼年和成年美西螈表皮间质细胞的组织学、组织化学和形态计量学分析及表皮顶端细胞的组织化学特征
IF 2.3 4区 生物学 Q4 CELL BIOLOGY Pub Date : 2025-04-24 DOI: 10.1016/j.acthis.2025.152255
Omar Betancourt-León , Verónica Rodríguez-Mata , Antonieta Martínez-Guerrero , Armando Pérez-Torres
Ambystoma mexicanum, also known as the axolotl, is a paedomorphic urodele. Metamorphosis can be induced experimentally, and the most significant changes occur in the skin. These include thinning of the epidermis, increased keratinization of the stratified squamous epithelium, and loss of Leydig cells (LCs). Similar epidermal changes are observed in other metamorphic urodeles. Epidermal cells are responsible for the secretory function of the skin in juvenile amphibians, whereas dermal glands perform this function in adults after metamorphosis. In the axolotl, this occurrence is still partially understood. The only recognized epidermal secretory cells in juvenile A. mexicanum are the LCs, whose specific secretion products have not yet been characterized from the histochemical standpoint. Additionally, the persistence of LCs in adulthood, when mucous and serous (granular-protein secretion) glands are abundant, remains a matter of debate. The present study aims to describe the morphological and histochemical changes in the epidermis of 10 cutaneous regions from juvenile (4 months old) and adult (24 and 48 months old) non-metamorphic A. mexicanum, with a particular focus on the amount and histochemical characteristics of LCs. Results indicate that the juvenile epidermis is a stratified cuboidal epithelium formed by three strata: basal, spinosum (containing the LCs), and apical. The most superficial layer contains cuboidal cells that lack the characteristics of a true stratum corneum. In adults, the stratum apical is also formed by squamous cells, suggesting a transition to a cornified and squamous layer as age increases. Histochemical methods demonstrated that LCs are most likely serous and not mucous cells. On the other hand, cuboidal cells of the juvenile apical stratum would be responsible for producing mucous secretion components. Morphometric analysis revealed a significant decrease in both LCs and the epidermal thickness in the 24-month-old adult axolotl compared to the juvenile. While LC count and epidermal thickness in the 48-month-old adult showed a slight increase compared to the 24-month-old adult, these differences were not statistically significant and far lower than those observed in the juvenile axolotl, which exhibited the highest number of LCs and a thicker epidermis. These natural axolotl epidermal changes indicate a gradual transition toward a morphology resembling metamorphic skin as age advances. The decreased number of LCs and the transition from cuboid cells to squamous cells in the stratum apical suggest that both cell types may naturally disappear entirely at some point during development.
墨西哥Ambystoma mexicanum,也被称为蝾螈,是一种幼童形的水生动物。变态可以通过实验诱导,最显著的变化发生在皮肤上。这些包括表皮变薄,层状鳞状上皮角化增加,间质细胞(LCs)丢失。类似的表皮变化在其他变态的尾虫中也可见。在幼年两栖动物中,表皮细胞负责皮肤的分泌功能,而在变态后的成年两栖动物中,真皮腺则执行这一功能。在美西螈中,这种现象仍然被部分理解。在墨西哥青霉幼崽中唯一被识别的表皮分泌细胞是LCs,其特异性分泌产物尚未从组织化学的角度进行表征。此外,成年期粘液腺和浆液腺(颗粒蛋白分泌腺)丰富时,LCs的持续性仍存在争议。本研究旨在描述幼年(4个月大)和成年(24个月和48个月大)非变质a . mexicanum皮肤10个区域表皮的形态学和组织化学变化,特别关注LCs的数量和组织化学特征。结果表明,幼代表皮是由基层、棘层(含LCs)和顶层三层组成的层状立方上皮。最浅层含有立方体细胞,缺乏真正角质层的特征。在成人中,根尖层也由鳞状细胞形成,表明随着年龄的增长,向锥形和鳞状层过渡。组织化学方法表明,lc很可能是浆液细胞,而不是黏液细胞。另一方面,幼体顶端层的立方细胞可能负责产生粘液分泌成分。形态计量学分析显示,与幼体相比,24月龄成年美西螈的LCs和表皮厚度均显著降低。虽然与24月龄的成虫相比,48月龄的成虫LC数量和表皮厚度略有增加,但差异无统计学意义,且远低于幼鱼,幼鱼的LC数量最多,表皮更厚。这些自然的美西螈表皮变化表明,随着年龄的增长,它们逐渐向类似于变质皮肤的形态过渡。细胞数量的减少和根尖层中长方体细胞向鳞状细胞的转变表明,这两种细胞类型可能在发育过程中的某一时刻自然完全消失。
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引用次数: 0
Proposal for a simple and easy-to-implement protocol for three-dimensional tissue imaging that is compatible with observation using a confocal microscope 建议一个简单和易于实施的协议,三维组织成像,是兼容的观察使用共聚焦显微镜
IF 2.3 4区 生物学 Q4 CELL BIOLOGY Pub Date : 2025-04-23 DOI: 10.1016/j.acthis.2025.152257
Takuto Matano , Kiyotada Naitou , Jannatul Ferdous , Takahiko Shiina , Mitsuya Shiraishi
Tissue observation has traditionally been limited to obtaining two-dimensional information from thinly sliced tissues due to issues with light transmission and antibody penetration. In recent years, three-dimensional tissue observation methods combining tissue clearing and deep immunostaining methods have been reported. However, due to the significantly different procedures in these methods from conventional immunostaining methods and the requirement for an expensive and specialized light-sheet microscope for tissue observation, the widespread adoption of these methods has been limited. To promote the shift from the current two-dimensional tissue observation to three-dimensional tissue observation using a combination of tissue clearing and immunostaining, it is essential to establish a simple and easy-to-implement protocol that is compatible with observation using a confocal microscope, which is available in many facilities. In this study, we first examined the effects of tissue clearing and staining conditions of immunostaining with thin tissue slices. We showed that CUBIC-L enhances immunolabeling without diminishing the immunoreactivity of antigens. We also showed that high detergent concentrations enhance the intensity of immunoreactivity and that a two-step staining procedure is suitable for our proposed protocol. Based on the results, we propose a simple protocol that can be easily adapted from conventional methods and is compatible with confocal microscopes. The results of this study are expected to facilitate a shift from traditional methods to three-dimensional tissue observation techniques that combine tissue clearing and immunostaining, contributing to the broader adoption of three-dimensional tissue observation.
由于光透射和抗体穿透的问题,组织观察传统上仅限于从薄片组织中获取二维信息。近年来,有报道称三维组织观察方法结合了组织清除和深度免疫染色方法。然而,由于这些方法的操作步骤与传统的免疫染色方法有很大不同,而且需要使用昂贵的专业光片显微镜进行组织观察,因此这些方法的广泛应用受到了限制。为了促进从目前的二维组织观察向结合使用组织清除和免疫染色的三维组织观察的转变,有必要建立一个简单易行的方案,该方案应与使用共聚焦显微镜进行的观察相兼容,而共聚焦显微镜在许多设备中都可以使用。在本研究中,我们首先研究了组织清理和染色条件对薄组织切片免疫染色的影响。我们发现,CUBIC-L 可增强免疫标记,而不会降低抗原的免疫活性。我们还发现,高浓度去污剂能增强免疫反应的强度,而且两步染色程序适合我们提出的方案。基于这些结果,我们提出了一种简单的方案,它可以很容易地从传统方法中进行调整,并与共聚焦显微镜兼容。本研究的结果有望推动三维组织观察技术从传统方法向结合组织清除和免疫染色的方法转变,从而促进三维组织观察技术的广泛应用。
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引用次数: 0
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Acta histochemica
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