1α,25-二羟基维生素D3抑制人结肠癌细胞系SW480中β-catenin转录活性

目的探讨1α,25-二羟基维生素D_3[1,25(OH)_2D_3]对人结肠癌细胞系SW480中β-catenin转录活性的影响及作用机制。方法 1,25(OH)_2D_3(100 nmol/L)干预和溶剂对照处理SW480细胞48 h;用双荧光素酶报告系统检测β-catenin的转录活性;Western blot检测1,25(OH)_2D_3干预后β-catenin在细胞核/质内的定位变化;用RT-qPCR和Western blot检测β-catenin和FOXM1 mRNA及蛋白水平。采用siRNA敲降SW480细胞中FOXM1后,检测1,25(OH)_2D_3干预后β-catenin转录活性的变化。结果 1)在SW480细胞中,1,25(OH)_2D_3能显著降低β-catenin的转录活性(P0.05),但不影响β-catenin mRNA和蛋白表达水平; 2)1,25(OH)_2D_3上调SW480细胞中FOXM1蛋白水平的表达(P0.05); 3)敲降SW480细胞中FOXM1降低1,25(OH)_2D_3对β-catenin转录活性的抑制作用(P0.05)。结论 1,25(OH)_2D_3通过上调FOXM1的表达,发挥抑制β-catenin转录活性的作用。     

25-羟基维生素D化学发光定量免疫分析方法的建立

目的 建立一种竞争法定量检测人血清血浆中25-羟基维生素D[25-(OH)D]的免疫分析方法.方法 本研究以磁颗粒-链酶亲和素-生物素为固相分离系统,样本中加入解离剂使维生素D结合蛋白(vitamin D binding protein,VDBP)变性而使25-(OH)D游离出来,用生物素标记25-(OH)D,用吖碇酯...     

1, 25二羟基胆钙化醇诱导大鼠骨髓单核细胞向破骨细胞转化的机制

目的:探讨1,25二羟基胆钙化醇(1,25(OH)2D3)诱导大鼠骨髓单核细胞向破骨细胞转化时明胶酶表达及其参与骨陷窝形成机制。方法:分离乳鼠骨髓内细胞,诱导生成破骨样细胞。姬姆莎、抗酒石酸酸性磷酸酶(TRAP)染色鉴定。扫描电镜观察诱导出的细胞贴附于骨片上形成的骨陷窝,明胶酶谱检测细胞培养液中明胶酶表达水平。结果:单核细胞经1,25(OH)2D3诱导,第9日生成大量的破骨样细胞。姬姆莎染色显示出多核(≥3个),TRAP染色阳性,扫描电镜观察破骨细胞在骨片上培养时产生骨陷窝。1,25(OH)2D3组基质金属蛋白酶2(MMP-2)表达水平增加显著。结论:1,25(OH)2D3诱导单核细胞产生大量破骨细胞。参与骨陷窝形成的明胶酶是MMP-2。这可能是破骨细胞通过某些机制,促进其他非破骨细胞分泌有活性的MMP-2参与噬骨。     

常住北京市城区少儿血清维生素A和25-羟基维生素D水平分析

目的:检测北京市城区少儿冬季血清维生素A(VA)和25-羟基维生素D(25-OH VD)水平,分析营养状况。方法:2012年12月至2013年2月本院儿童保健门诊体检的0-14岁少儿292例,按年龄分为0-1岁、2-3岁、4-6岁和7-14岁四组,完成问卷调查和体格检查后,采用高效液相色谱法测定每例少儿血清VA水平,采用高效液相质谱串联法测定每例少儿血清25-OH VD水平,分析各年龄组VA、25-OH VD水平差异及每组低水平VA、25-OH VD检出率。结果:(1)292例少儿血清VA平均水平为(1.09±0.29)μmol/L,不同年龄组VA水平有显著性差异(F=10.96,P0.01),以0-1岁组最低(0.9±0.31)μmol/L;所有受检少儿低水平VA检出率为43.84%(128/292),不同年龄组低水平VA检出率有显著性差异(χ2=34.74,P0.01),0-1岁组检出率最高(76.32%)。(2)292例少儿血清25-OH VD平均水平(22.12±8.51)ng/ml,不同年龄组水平有显著性差异(F=7.56,P0.01),7-14岁组最低(18.34±6.34)ng/ml;所有受检者中低水平25-OH VD检出率为80.17%(233/292),不同年龄组低水平25-OH VD检出率有显著性差异(χ2=13.79,P0.05),4-6岁组最高(87.05%)。结论:冬季,常住北京市城区0-1岁幼儿的VA最低,7-14岁的25-OH VD最低,值得关注。     

1α，25-二羟基维生素D3通过TLR4信号通路抑制川崎病血清诱导的THP-1细胞炎症因子表达

目的:研究1α,25-二羟基维生素D3[1,25-(OH)2D3]是否通过调控巨噬细胞Toll样受体4(TLR4)信号通路的活化影响川崎病炎症反应的发生。方法:收集襄阳市第一人民医院收治的73例川崎病患儿血清,另收集急性上呼吸道感染患儿(感染组)和儿保科体检儿童(对照组)的血清各30例,利用试剂盒检测各组25-羟基维生素D3[25-(OH)D3]水平的变化,并绘制受试者工作特征(ROC)曲线,确定诊断截断点。体外细胞实验,利用采集的血清诱导THP-1巨噬细胞构建川崎病固有免疫模型,实验分组为:15%正常血清组、15%川崎病血清组及15%川崎病血清+1,25-(OH)2D3(1 nmol/L、10 nmol/L和100 nmol/L)组[1,25-(OH)2D3均于血清刺激前加入]。加入血清刺激24h后,利用RT-qPCR检测TLR4、白细胞介素1β(IL-1β)和肿瘤坏死因子α(TNF-α)的mRNA表达水平,ELISA检测细胞培养上清液中TNF-α的含量;Western blot检测TLR4和p-P65的蛋白水平;免疫荧光技术检测TLR4蛋白的表达。结果:川崎病患儿体内白细胞数、中性...     

1，25双羟维生素D3对成骨样细胞骨架和基因表达的影响

用荧光免疫法研究了１，２５双羟基维生素Ｄ３对大鼠成骨样细胞（ＲＯＳ１７／２．８）细胞骨架的影响和用斑点杂交技术研究了１，２５（ＯＨ）２Ｄ３对人成骨样细胞（ＯＳ－７３２）基因表达的影响。结果表明：在１０＾－７ｍｏｌ／Ｌ１，２５（ＯＨ）２Ｄ３连续作用４ｄ后，大鼠ＲＯＳ１７／２．８细胞的微管蛋白和波形纤维蛋白明显改善，纤粘蛋白荧光明显增强。而在１，２５（ＯＨ）２Ｄ３作用前后ＯＳ－７３２细胞ｃ－ｍｙｃ和ｍ     

HBeAg阴性慢性乙型肝炎患者血清25-羟基维生素D3的检测及临床意义

目的 探究HBeAg阴性慢性乙型肝炎患者血清25-羟基维生素D3的水平变化及临床意义。方法 选取2015年3月~2018年5月81例HBeAg阴性且非肝硬化的慢性乙型肝炎患者作为观察组,依据血清谷丙转氨酶（ALT）水平分为观察组A 41例（ALT处于正常水平且HBV DNA水平＜2000 IU/ml,持续时间高于6个月）和观察组B 40例（ALT处于升高水平且HBV DNA水平≥2000 IU/ml,持续时间高于6个月）。同时选取同期健康体检人群40例作为对照组。分别检测三组血清25-羟基维生素D3的浓度。结果 观察组血清25-羟基维生素D3低于健康人群,统计学意义显著（P＜0.001）,三组间比较,统计学意义显著（P＜0.001）。观察组患者血清25-羟基维生素D3与年龄、BMI、ALT、AST、Hb、WBC、PLT及AFP均无明显相关性（P＞0.05）。而与HBV DNA含量间存在显著负相关（r=-3.981,P＜0.05）。结论 HBeAg阴性慢性乙型肝炎患者血清25-羟基维生素D3存在异常,并与乙肝病毒复制相关,可能参与机体和乙肝病毒的免疫反应过程。     

葛根素通过Notch1信号通路抑制Raw264.7细胞向破骨细胞的分化

背景：前期研究表明葛根素干预后破骨细胞的分化被抑制,Notch1、HES1、Jagged1等Notch信号通路相关蛋白表达量下降,但Notch1信号通路对于葛根素抑制破骨细胞分化的作用机制尚不明确。目的：探究Notch信号通路对葛根素抑制小鼠巨噬细胞Raw264.7分化为破骨细胞的影响。方法：将Raw264.7细胞分为7组干预培养,空白对照组采用DMEM高糖完全培养基培养,破骨细胞诱导组采用破骨诱导培养基培养,葛根素干预组在破骨诱导的同时加入50μmol/L葛根素培养,葛根素+Notch1 si RNA对照组、葛根素+Notch1 si RNA组、葛根素+Notch1过表达对照组、葛根素+Notch1过表达组分别采用Notch1 si RNA对照序列、Notch1 si RNA序列、Notch1过表达对照质粒、Notch1过表达质粒转染Raw264.7细胞后,加入破骨诱导培养基和葛根素进行培养。培养7 d后,采用抗酒石酸酸性磷酸酶染色观察破骨细胞的数量和大小,F-actin染色观察破骨细胞骨架形成情况,RT-PCR检测破骨细胞形成标志物的基因表达水平。结果与结论：(1)抗酒石酸酸性磷酸...     

晚期糖基化终末产物可影响破骨细胞的骨吸收功能

BACKGROUND: The effects of advanced glycation end products (AGEs) on osteoclast-induced bone resorption is controversial and the underlying mechanisms remain unclear. Most of the studies indicate that AGEs can enhance bone resorption, while some others show the opposite effects. OBJECTIVE:To investigate the effects of AGEs on osteoclast-induced inorganic matrix dissolution and organic component degradation and the underlying mechanisms. METHODS: RAW 264.7 cells were induced to generate osteoclasts, and AGEs (50-400 µg/mL) or control-bovine serum albumin (100 µg/mL) was added since the beginning of the induction. The effect of AGEs on bone resorption was evaluated by analyzing the area of resorption pits on the Osteo Assay Surface plates and the expression of cathepsin K. Furthermore, the number of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells, nuclei per osteoclasts and the expression of integrin ανβ3 were detected. RESULTS AND CONCLUSION: The area of resorption pits and expression of cathepsin K in AGEs groups were significantly decreased compared with the control group, and this inhibiting effect became more obvious with the increase of AGEs concentration. TRAP staining also showed that number of TRAP-positive multinucleated cells and nuclei per osteoclast were significantly reduced in an AGE dose-dependent manner. Quantitative PCR revealed that the expression of integrin ανβ3 decreased significantly with the extension of AGEs incubation time. These data indicate that AGEs can exert inhibitory effects on organic and inorganic matrix degradation induced by osteoclasts. The underlying mechanism may be involved in the inhibitory effects of AGEs on directed differentiation and cell fusion of osteoclast precursor cells, and migration and adhension of osteoclasts. 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

Effect of 1,25-dihydroxyvitamin D3 on interferon gamma production in vitro

In recent years, receptors for calcitriol (the active form of vitamin D3) have been identified in monocytes and activated, but not resting, human B and T lymphocytes suggesting that it may be involved in immune regulation. Because lymphokines are central in the regulation and modulation of immune or inflammatory responses and since the calcium translocation is involved in the mitogen-induced activation of lymphocytes, we thought it interesting to study the role of calcitriol on interferon gamma (IFN-gamma) production in vitro. In this study, we report that calcitriol inhibits the IFN-gamma production by staphylococcal enterotoxin A-stimulated peripheral blood mononuclear cells (PBMC) in a dose-dependent fashion. The inhibitory effect was less potent in calcium ionophore A23187-stimulated PBMC and was absent in resting PBMC.     

1,25-二羟维生素D3对1型糖尿病预防效果观察

目的:1,25-二羟维生素D3[1,25-(OH)2D3]对连续多次小剂量链脲菌素(the mu ltip le low dose streptozotoc in,MLDS)诱导的自身免疫性糖尿病小鼠的预防作用及其机制初探。方法:小鼠分为三组。正常对照组:连续5天腹腔注射与糖尿病组等容量柠檬酸盐缓冲液;糖尿病组:连续5天腹腔注射STZ(40mg.kg-1),以血糖水平持续高于16.7mmol/L为成模标准;预防组:先隔日腹腔注射1,25-(OH)2D3(5μg.kg-1),共15次,然后再连续5天腹腔注射STZ(40mg.kg-1)。实验结束后各组动物处死收集血清并采集胰腺检测诱导型一氧化氮合酶(iNOS)活性及血清胰岛素水平。结果:MLDS诱发的自身免疫性糖尿病模型在第四周基本建成。MLDS使小鼠血糖、血清及胰腺iNOS活性升高,血清胰岛素水平下调;预防组小鼠注射STZ前给予1,25-(OH)2D3有明显降血糖和上调血清胰岛素水平作用,同时抑制血清及胰腺iNOS活性,与糖尿病组比较有显著性差异(P<0.05)。结论:1,25-(OH)2D3可有效预防MLDS诱导的自身免疫性糖尿病的发生。该效应可能与1,25-(OH)2D3抑制iNOS活性有关。     

Inhibition of tropoelastin expression by 1,25-dihydroxyvitamin D3.

Elastin production is modulated by steroid hormones and is dependent on calcium. Because vitamin D3 is involved in the regulation of calcium metabolism and influences the expression of various extracellular matrix proteins, we investigated whether vitamin D3 influences tropoelastin expression. Three elastin-producing, bovine cell types, auricular chondroblasts, nuchal ligament fibroblasts and arterial smooth muscle cells, were treated with the principal active metabolite of vitamin D3, 1,25-dihydroxyvitamin D3 (1,25[OH]2D3), and with 24,25 dihydroxyvitamin D3 (24,25[OH]2D3). Tropoelastin levels in culture media and cell layers, as measured by an enzyme-linked immunoassay, decreased in a dose and exposure dependent manner after treatment with 1,25(OH)2D3; 24,25(OH)2D3 had no effect on tropoelastin production relative to solvent-treated controls. The maximal effective dose of 1,25(OH)2D3 was 10(-7) M for 48 hr, which resulted in a severalfold reduction of tropoelastin production, and decreased tropoelastin levels were detected at 8 hr after treatment. Reduction of tropoelastin protein production was paralleled by a decrease of equal magnitude in the steady-state levels of tropoelastin mRNA. Vitamin D3 metabolites had no effect on DNA or total protein synthesis. These results suggest that vitamin D3 may be an important modulator of elastin expression.     

1,25-dihydroxyvitamin D3 and development of tuberculosis in cattle

This report describes the presence and activity of 1,25-dihydroxyvitamin D3 (1,25-D3) in experimental bovine tuberculosis. Animals that went on to develop tuberculous lesions exhibited a rapid transient increase in serum 1,25-D3 within the first 2 weeks following infection with Mycobacterium bovis. 1,25-D3-positive mononuclear cells were later identified in all tuberculous granulomas by immunohistochemical staining of postmortem lymph node tissue. These results suggest a role for 1,25-D3 both at the onset of infection and in the development of the granuloma in these infected animals. Using a monoclonal antibody to the vitamin D receptor (VDR) as a VDR agonist, we confirmed that activation of the vitamin D pathway profoundly depresses antigen-specific, but not mitogenic, bovine peripheral blood T-cell responses (proliferation and gamma interferon production). Investigation of the mechanism of this suppression showed that the VDR antibody modified the expression of CD80 by accessory cells, such that a significant positive correlation between T-cell proliferation and accessory cell CD80 emerged.     

VDR-dependent regulation of mast cell maturation mediated by 1,25-dihydroxyvitamin D3

1,25-Dihydroxyvitamin D3 [1,25(OH)2D3] is a secosteroid hormone that regulates bone metabolism, controls calcium homeostasis, and possesses immunomodulatory properties. We show here that 1,25(OH)2D3 contributes to the regulation of development and function of mast cells, which play a critical role in several inflammatory disorders. 1,25(OH)2D3 promotes apoptosis and inhibits maturation of mouse bone marrow-derived mast cell precursors. Dose-dependent inhibition of mast cell differentiation by 1,25(OH)2D3 is observed at discrete, intermediate stages of mast cell development, identified by expression of c-kit, FcepsilonRI, and IL-3 receptor-alpha chain, and depends on the expression of the vitamin D receptor (VDR). It is important that mast cell progenitors obtained from VDR-ablated mice undergo an accelerated maturation in vitro and give rise to more responsive mast cells than wild-type. Furthermore, histological analysis of mast cell density in peripheral tissues reveals a moderate increase in the number of mast cells in the skin of VDR-deficient mice compared with wild-type animals. These data support the hypothesis of a physiological role of 1,25(OH)2D3 in mast cell development and suggest novel, therapeutic uses of 1,25(OH)2D3 analogs.     

Modulation of cell-cell and cell-antigen interactions by 1,25-dihydroxyvitamin D3 and vitamin D3 sulfate in vitro: a study on pregnancy lymphocytes and hybridoma cells

The effect of 1,25-dihydroxyvitamin D₃ (1,25(OH)₂D₃ was investigated in single cell cytotoxicity assays, using K-562 target cells. The action of vitamin D₃ sulfate (VD₃S) in natural cytotoxicity assays as well as its effect on the antigen-specific adherence of hybridoma cells has also been studied. In the single cell cytotoxicity assay 1,25(OH₂)D₃ dose-dependently and significantly increased the binding of PBMC to target, the number of lysed target cells and NK activity, RU486, a compound known as a potent blocker of progesterone and glucocorticoid receptors, suppressed the effect of 1,25(OH₂)D₃ in all systems. VD₃S dose-dependently decreased the natural cytotoxicity of PBMC and the binding of hybridoma cells to antigen immobilized on plastic surfaces. The results suggest that both 1,25(OH₂)D₃ and VD₃S are potent     

Vitamin D endocrine system and bone tissue mineral metabolism in rats with adjuvant arthritis: Effect of 1,25-dihydroxyvitamin D3

Institute of Nutrition, Russian Academy of Medical Sciences. Vitaminy Research and Production Combine, Moscow. (Presented by Academician of the Russian Academy of Medical Sciences T. T. Berezov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 114, No. 11, pp. 510–512, November, 1992.     

Inhibition of production and function of interleukin-6 by 1,25-dihydroxyvitamin D3

1,25-Dihydroxyvitamin D3 (1,25-(OH)2D3) has been shown to interfere with immunoglobulin production and lymphocyte proliferation in vitro. These lymphocyte functions are influenced by interleukin (IL)-6 produced by antigen presenting cells. Hence, the ability of 1,25-(OH)2D3 to interfere with the production and function of IL-6 was investigated. 1,25-(OH)2D3 and the analogue MC 903 inhibited IL-6 production by LPS-stimulated human mononuclear cells. The precursor 25-OH D3 was ineffective. Likewise, 1,25-(OH)2D3 but not 25-OH D3 inhibited rIL-6-driven as well as rIL-1 alpha/beta-driven proliferation of murine thymocytes. This effect of 1,25-(OH)2D3 was partially or totally overcome by larger concentrations of rIL-6 as well as by rIL-2 and ionomycin. Consistently, the production of IL-6 and IL-2 in rIL-1 driven thymocyte cultures were found to be reduced by 1,25-(OH)2D3. Inhibition of production and function of IL-6 may therefore be involved in 1,25-(OH)2D3-mediated regulation of lymphocyte functions in vitro.