Qingyi decoction attenuates intestinal epithelial cell injury via the calcineurin/nuclear factor of activated T-cells pathway

BACKGROUND Recent studies have demonstrated that dysfunction of the intestinal barrier is a significant contributing factor to the development of severe acute pancreatitis(SAP).A stable intestinal mucosa barrier functions as a major anatomic and functional barrier,owing to the balance between intestinal epithelial cell(IEC)proliferation and apoptosis.There is some evidence that calcium overload may trigger IEC apoptosis and that calcineurin(CaN)/nuclear factor of activated Tcells(NFAT)signaling might play an important role in calcium-mediated apoptosis.AIM To investigate the potential mechanisms underlying the therapeutic effect of Qingyi decoction(QYD)in SAP.METHODS A rat model of SAP was created via retrograde infusion of sodium deoxycholate.Serum levels of amylase,tumor necrosis factor(TNF-α),interleukin(IL)-6,D-lactic acid,and diamine oxidase(DAO);histological changes;and apoptosis of IECs were examined in rats with or without QYD treatment.The expression of the two subunits of CaN and NFAT in intestinal tissue was measured via quantitative realtime polymerase chain reaction and western blotting.For in vitro studies,Caco-2 cells were treated with lipopolysaccharide(LPS)and QYD serum,and then cell viability and intracellular calcium levels were detected.RESULTS Retrograde infusion of sodium deoxycholate increased the severity of pancreatic and intestinal pathology and the levels of serum amylase,TNF-α,and IL-6.Both the indicators of intestinal mucosa damage(D-lactic acid and DAO)and the levels of IEC apoptosis were elevated in the SAP group.QYD treatment reduced the serum levels of amylase,TNF-α,IL-6,D-lactic acid,and DAO and attenuated the histological findings.IEC apoptosis associated with SAP was ameliorated under QYD treatment.In addition,the protein expression levels of the two subunits of CaN were remarkably elevated in the SAP group,and the NFATc3 gene was significantly upregulated at both the transcript and protein levels in the SAP group compared with the control group.QYD significantly restrained CaN and NFATc3 gene expression in the intestine,which was upregulated in the SAP group.Furthermore,QYD serum significantly decreased the LPS-induced elevation in intracellular free Ca^(2+)levels and inhibited cell death.CONCLUSION QYD can exert protective effects against intestinal mucosa damage caused by SAP and the protective effects are mediated,at least partially,by restraining IEC apoptosis via the CaN/NFATc3 pathway.

Calcineurin/Nuclear Factor of Activated T-Cell Pathway in Cutaneous Squamous Cell Carcinoma

Introduction Cutaneous squamous cell carcinoma (CSCC),a keratinocyte-derived skin neoplasm with malignant potential,1 represents 20%-50% of skin cancers and currently has an increasing incidence in the United States.2 Ultraviolet (UV)solar radiation is the primary risk factor for the development of CSCC,and the cumulative exposure received over a lifetime plays a major role in this development.3 Mutations in the p53 gene are the most common genetic abnormalities,causing nonfunctional p53 protein production and cells with damaged DNA replicate in CSCC.

转录因子NFATc在钙神经素介导的脑缺血再灌注损伤中的作用(英文)

目的 研究转录因子NFATc及NF κB在钙神经素介导的脑缺血再灌注损伤中的作用。方法 Westernblotting和EMSA分子生物学技术。结果 与对照组相比较,CsA明显减低I/R组Fas配体和NFATc的蛋白表达;对照组、I/R组和CsA处理组I κB α蛋白表达无显著区别;未观察到对照组、I/R组和CsA处理组有phospho I κB α蛋白表达;与对照组相比较,CsA明显减低I/R组Fas配体启动子远端和Fas配体启动子近端NFAT结合位点的NFAT DNA结合活性(P<0 01)。结论 转录因子NFATc参与钙神经素介导的脑缺血再灌注损伤,促进CD95配体分子的转录表达;NF κB可能未参与钙神经素介导的脑缺血再灌注损伤的作用机制。

Inhibiting 5-hydroxytryptamine receptor 3 alleviates pathological changes of a mouse model of Alzheimer's disease

Extracellular amyloid beta(Aβ) plaques are main pathological feature of Alzheimer’s disease.However,the specific type of neuro ns that produce Aβ peptides in the initial stage of Alzheimer’s disease are unknown.In this study,we found that 5-hydroxytryptamin receptor 3A subunit(HTR3A) was highly expressed in the brain tissue of transgenic amyloid precursor protein and presenilin-1 mice(an Alzheimer’s disease model) and patients with Alzheimer’s disease.To investigate whether HTR3A-positive interneurons are associated with the production of Aβ plaques,we performed double immunostaining and found that HTR3A-positive interneurons were clustered around Aβ plaques in the mouse model.Some amyloid precursor protein-positive or β-site amyloid precursor protein cleaving enzyme-1-positive neurites near Aβ plaques were co-localized with HTR3A interneurons.These results suggest that HTR3A-positive interneurons may partially contribute to the generation of Aβ peptides.We treated 5.0-5.5-month-old model mice with tro pisetron,a HTR3 antagonist,for 8 consecutive weeks.We found that the cognitive deficit of mice was partially reversed,Aβ plaques and neuroinflammation we re remarkably reduced,the expression of HTR3 was remarkably decreased and the calcineurin/nuclear factor of activated T-cell 4 signaling pathway was inhibited in treated model mice.These findings suggest that HTR3A interneurons partly contribute to generation of Aβ peptide at the initial stage of Alzheimer’s disease and inhibiting HTR3 partly reve rses the pathological changes of Alzheimer’s disease.

Molecular mechanisms of triggering,amplifying and targeting RANK signaling in osteoclasts

Osteoclast differentiation depends on receptor activator of nuclear factor-κB(RANK) signaling,which can be divided into triggering,amplifying and targeting phases based on how active the master regulator nuclear factor of activated T-cells cytoplasmic 1(NFATc1) is. The triggering phase is characterized by immediateearly RANK signaling induced by RANK ligand(RANKL) stimulation mediated by three adaptor proteins,tumor necrosis factor receptor-associated factor 6,Grb-2-associated binder-2 and phospholipase C(PLC)γ2,leading to activation of IκB kinase,mitogen-activated protein kinases and the transcription factors nuclear factor(NF)-κB and activator protein-1(AP-1). Mice lacking NF-κB p50/p52 or the AP-1 subunit c-Fos(encoded by Fos) exhibit severe osteopetrosis due to a differentiation block in the osteoclast lineage. The amplification phase occurs about 24 h later in a RANKLinduced osteoclastogenic culture when Ca2+ oscillation starts and the transcription factor NFATc1 is abundantly produced. In addition to Ca2+ oscillation-dependent nuclear translocation and transcriptional auto-induction of NFATc1,a Ca2+ oscillation-independent,osteoblastdependent mechanism stabilizes NFATc1 protein in dif-ferentiating osteoclasts. Osteoclast precursors lacking PLCγ2,inositol-1,4,5-trisphosphate receptors,regulator of G-protein signaling 10,or NFATc1 show an impaired transition from the triggering to amplifying phases. The final targeting phase is mediated by activation of numerous NFATc1 target genes responsible for cell-cell fusion and regulation of bone-resorptive function. This review focuses on molecular mechanisms for each of the three phases of RANK signaling during osteoclast differentiation.

凋亡相关蛋白3及活化T细胞核因子3在卵巢上皮性肿瘤组织中的表达及与临床病理特征相关性研究

目的 探讨凋亡相关蛋白3（Apoptosis associated protein 3,APR3）及活化T细胞核因子3（Nuclear factor of activated T-cell,NFAT3）在卵巢上皮性肿瘤组织中的表达及与临床病理特征的相关性.方法 收集2014年4月至2015年4月住院治疗的92例卵巢上皮性肿瘤患者,其中恶性肿瘤患者23例,交界性肿瘤患者24例,良性肿瘤患者45例,采用免疫组化方法检测所有人组患者肿瘤组织中APR3和NFAT3表达,比较不同类型卵巢上皮性肿瘤的APR3和CaN-NFAT3的表达差异,分析恶性卵巢上皮性肿瘤患者APR3和NFAT3的表达与临床病理因素的相关性,分析恶性卵巢上皮性肿瘤患者APR3和NFAT3的表达相关性.结果 恶性卵巢上皮性肿瘤APR3阳性表达率（78.26％）明显高于交界性肿瘤（41.67％）和良性肿瘤（22.22％）,差异均有统计学意义（χ^2=5.864、7.632,均P〈0.05）;恶性卵巢上皮性肿瘤APR3表达与组织分化程度、临床分期、腹腔脏器和淋巴转移及腹水均明显相关（χ^2=7.425、7.262、8.421、5.031,均P〈0.05）.恶性卵巢上皮性肿瘤NFAT3阳性表达率（56.52％）明显高于交界性肿瘤（29.17％）和良性肿瘤（17.78％）,差异均有统计学意义（χ^2=6.829、7.547,均P〈0.05）;恶性卵巢上皮性肿瘤NFAT3表达与组织分化程度、临床分期及腹腔脏器和淋巴结转移均明显相关（χ^2=5.253、6.367、8.021,均P〈0.05）.恶性卵巢上皮性肿瘤患者APR3的表达与NFAT3的表达呈正相关（r=0.032,P〈0.05）.结论 APR3和NFAT3在恶性卵巢上皮性肿瘤中的表达明显增加,与患者组织分化程度、临床分期、腹腔脏器和淋巴转移密切相关,且两者表达呈明显正相关,可能与恶性卵巢上皮性肿瘤发生和发展有关.