葛根素在体外对成骨细胞增殖和分化的影响

背景：成骨细胞是骨代谢平衡过程中的关键功能细胞,植物雌激素对成骨细胞的增殖和分化有重要影响,葛根素作为植物雌激素的一种,在体外以较大范围浓度对成骨细胞功能的影响仍少见报道。 目的：观察葛根素在体外对大鼠成骨细胞增殖和分化功能的影响。 方法：取新生Wistar大鼠的颅盖骨,对成骨细胞进行分离、培养、纯化及鉴定。将培养的成骨细胞随机分为对照组、10-3~10-10 mol/L不同浓度葛根素组,观察不同浓度葛根素对体外培养的成骨细胞增殖和碱性磷酸酶活性表达的影响。 结果与结论：细胞经葛根素处理后10-5~10-9 mol/L组成骨细胞增殖活性较对照组明显增加(P < 0.05),第3天增殖最快(P < 0.01),第4天开始下降;诱导第4天,各组碱性磷酸酶活性与对照组相比,差异均有显著性意义(P < 0.01),其中以10-6 mol/L组最显著(P < 0.01)。然而葛根素10-3 mol/L组成骨细胞增殖活性、碱性磷酸酶活性表达较对照组均减少(P < 0.05)。提示葛根素对成骨细胞的影响存在剂量依赖性,并且具有双向性,即在低浓度(10-5~10-8 mol/L)下刺激骨形成;在高浓度(10-3~10-4 mol/L)下抑制骨形成。     

中药促成骨细胞增殖和分化的机制与作用

背景：中药促进成骨细胞增殖和分化的效果已经得到肯定,其在细胞分子水平的药理机制研究取得了很大进展。 目的：对国内外应用中药诱导成骨细胞增殖和分化的药理机制研究进展作一综述。 方法：应用计算机检索CNKI,Pubmed和sciencedirect数据库中2001-01/2010-12关于中药促进成骨细胞增殖和分化机制研究的文章,在标题和摘要中以“中药,成骨细胞,增殖,分化,机制”或“Chinese herbs,osteoblast,proliferation,differentiation, mechanism”为检索词进行检索。初检得到176篇文献,最终选择30篇文献进行综述。 结果与结论：中药通过对成骨细胞基因、信号通路、细胞因子和OPG/RANK/RANKL系统的调控,以及类雌激素样作用促进成骨细胞增殖和分化,其作用机制明确,将中药引入骨组织工程领域具有良好的发展前景。 关键词：中药;成骨细胞;增殖;分化;机制;组织工程 doi:10.3969/j.issn.1673-8225.2012.07.037     

基于Toll样受体4通路研究黄芪甲苷促进成骨细胞增殖分化的作用

目的研究黄芪甲苷促进成骨细胞增殖分化的作用,并初步探讨作用机制。方法将成骨细胞MC3T3-E1分为对照组和实验组,对照组正常培养成骨细胞,实验组采用不同浓度黄芪甲苷干预成骨细胞,并孵育不同时间,MTT法观察对细胞增殖的影响,检测碱性磷酸酶(ALP)活性观察对细胞分化的作用,Western blot检测成骨细胞Toll样受体-4(TLR-4)蛋白表达。结果与对照组比较,随着黄芪甲苷浓度的增加,成骨细胞活性或ALP活性逐渐上升,并趋于稳定。延长黄芪甲苷作用时间,低浓度组细胞活性显著升高,而各组细胞ALP活性均显著上升。与对照组比较,实验组细胞中TLR-4表达显著下降。结论黄芪甲苷能够促进成骨细胞增殖和分化,其作用机制可能与TLR-4通路有关。     

三种波形磁场对大鼠成骨细胞增殖与分化的影响

目的：研究不同波形磁场辐照，对离体大鼠成骨细胞增殖与分化的影响。方法：用矩形、三角形和正弦三种典型波形磁场辐照离体成骨细胞。结果：频率15Hz，幅值5mT，矩形磁场促进细胞增殖（P〈0．01），抑制细胞分化（P〈0．01）;正弦磁场抑制细胞增殖（P〈0．01），促进细胞分化（P〈0．01）；三角彤磁场对细胞增殖和分化的影响尤显著性差异．结论：除强度和频率窗外，还需考虑波形的影响。     

葛根素通过下调miR-23a促进小鼠前成骨细胞增殖和分化的研究

目的 探讨葛根素是否通过下调miR-23a促进小鼠前成骨细胞增殖和分化。方法 将小鼠前成骨细胞MC3T3-E1分为对照组、葛根素组、葛根素+miR-NC组和葛根素+miR-23a过表达组,RT-qPCR检测细胞中miR-23a的表达,CCK-8法检测细胞增殖活性,碱性磷酸酶活性检测葛根素对细胞活力的影响,Western blot检测细胞中Runx2蛋白的表达。应用生物信息学和双荧光素酶报告基因实验验证miR-23a和Runx2的靶向调控关系。结果 与对照组相比,葛根素组MC3T3-E1细胞增殖活性增强,细胞中miR-23a表达降低,Runx2蛋白表达水平升高(P<0.05);与葛根素+miR-NC组相比,葛根素+miR-23a过表达组MC3T3-E1细胞增殖活性降低,细胞中Runx2蛋白表达降低(P<0.05);双荧光素酶报告基因实验证实miR-23a靶向负调控Runx2的表达。结论 葛根素促进小鼠前成骨细胞增殖和分化,机制可能与下调miR-23a进一步调控Runx2表达有关。     

低氧条件下成骨细胞的增殖与分化

背景：低氧可通过多种途径作用于成骨细胞影响骨代谢,对骨生成、骨愈合等产生负面影响。 目的：观察低氧对体外培养大鼠成骨细胞增殖、分化的影响,并探讨其分子机制。 方法：分离培养新生Wistar大鼠颅盖骨成骨细胞,取第2代细胞分别在常氧(体积分数20%O₂)与低氧(体积分数3%O₂)条件下培养。 结果与结论：低氧组成骨细胞增殖、碱性磷酸酶活性、骨钙素水平及茜素红结节形成数量均明显低于常氧组(P < 0.05或  P < 0.01),说明缺氧条件对成骨细胞的增殖、分化及功能有抑制作用;低氧组骨形成发生蛋白2及Runx2表达低于常氧组(P < 0.05或P < 0.01),说明低氧条件下大鼠成骨细胞Runx2、骨形成发生蛋白2的表达受抑制。结果表明低氧可通过抑制大鼠成骨细胞的Runx2、骨形成发生蛋白2的表达进一步抑制成骨细胞的增殖与分化。     

中药含药血清对体外成骨细胞增殖和分化影响的实验研究进展

近年来不少学者从细胞分子生物学角度开展了中医药对成骨细胞(OB)影响的实验研究,为中医药防治骨质疏松提供了分子生物学理论依据,其中运用体外成骨细胞培养和中药血清干预实验,研究骨质疏松的中药防治是常用的方法之一,也是中药防治骨质疏松症研究热点之一,本文主要回顾了含药血清对成骨细胞的增殖与分化方面的相关文献,这些研究对中药防治骨质疏松症奠定了基础。     

XW630对大鼠成骨细胞雌激素受体基因表达的影响

探讨抗骨质疏松新化合物XW630促进成骨作用的机理。从SD大鼠颅骨中分离培养成骨细胞，首次采用具有高灵敏度的逆转录—聚合酶链反应(RT—PCR)技术直接从大鼠成骨细胞中检测雌激素受体mRNA的表达。结果表明，XW630具有促进成骨细胞ERmRNA表达的作用，且呈时间依赖性。在相同浓度(10^-4mol／L)条件下，XW630作用强于四环素加哌嗪雌酮及单纯雌酚酮。XW630可能通过对ER基因表达的影响而在骨代谢中起作用。     

转铁蛋白受体基因对神经干细胞体外增殖和分化的影响研究

目的研究转铁蛋白受体基因(TfR)对神经干细胞(NSCs)体外增殖和分化的影响。方法分别将转铁蛋白受体基因神经干细胞(TfR-NSCs)和正常神经干细胞加入神经干细胞分化液分化7 d后,免疫荧光观察细胞分化及分别计算两种细胞胶质细胞和神经元细胞的分化率;CCK-8法检测其细胞在1 d、2 d、3 d时的A值来观察细胞增殖能力。结果转铁蛋白受体基因神经干细胞和正常神经干细胞均可分化成神经元和胶质细胞,且二者分化率未见明显差异(P0.05);转基因神经干细胞的增殖能力未受抑制(P0.05)。结论转铁蛋白受体基因神经干细胞的增殖分化未受明显影响,为下一步转铁蛋白受体转基因神经干细胞的应用提供支持。     

雌激素对成骨细胞的作用及作用机制

雌激素缺乏是绝经后骨质疏松(PMOP)的首要原因,雌激素替代治疗(ERT)是PMOP的首选方案.雌激素对骨代谢有重要调节作用,雌激素在成骨细胞增殖、分化、功能表达、凋亡等过程中起调节作用.雌激素对成骨细胞的作用与ER亚型、细胞因子/生长因子的旁分泌和自分泌作用、成骨细胞的凋亡关系密切.雌激素对成骨细胞的作用机制是ERT的重要理论依据.     

透骨消痛胶囊促进成骨细胞的增殖和分化

背景：透骨消痛胶囊是福建中医药大学治疗骨关节炎的临床验方,以往的研究多集中于其对软骨的作用。 目的：观察透骨消痛胶囊对成骨细胞增殖、分化及骨重塑相关因子表达的影响。 方法：以透骨消痛胶囊干预大鼠成骨细胞ROS17/2.8,分空白对照组和各浓度透骨消痛胶囊组,MTT法测定细胞增殖,比色法检测成骨细胞分化的生物标志物碱性磷酸酶活性,ELISA法检测骨钙素,茜素红染色观察骨矿化结节,实时定量PCR和ELISA法检测骨重塑相关因子骨保护素/核因子κB受体活化因子配体的表达。 结果与结论：与对照组相比,质量浓度为0.25-2 g/L透骨消痛胶囊能显著促进ROS17/2.8细胞增殖(P < 0.05),上调碱性磷酸酶活性(P < 0.05)、骨钙素的表达水平(P < 0.05)和矿化结节面积(P < 0.05),增加骨保护素/核因子κB受体活化因子配体的比例(P < 0.05)。提示透骨消痛胶囊防治骨关节炎的作用机制可能与其促进成骨细胞的增殖、分化以及调节骨重塑平衡的作用有关。     

Estrogen receptor beta expression in invasive breast cancer

The aim of this work was to determine the extent of estrogen receptor beta (ER-beta) expression in invasive breast cancer (BrCA) and whether ER-beta expression is correlated with response to adjuvant hormonal therapy with tamoxifen (AHTT). Immunohistochemical staining (IHC) for estrogen receptor alpha (ER-alpha) and ER-beta was performed on sections of formalin-fixed and paraffin-embedded tissue from 47 unselected invasive breast carcinomas (BrCA). IHC for ER-beta was also performed on sections of BrCA from 118 women who were treated with mastectomy and AHTT. Survival analysis was performed using the Kaplan-Meier method and the log-rank test. Of the 47 unselected BrCA, 17 (36%) were negative for ER-alpha and of these, 8 (47% of ER-alpha negative cases and 17% of all 47 patients) were ER-beta positive. Five of the 8 ER-alpha negative and ER-beta positive cases were positive for ER biochemically. There was no correlation between ER-beta positivity and overall survival in the unselected group. By contrast, in the group of women treated with AHTT, expression of ER-beta in more than 10% of cancer cells was associated with better survival (P = .0077), even in women with node-negative BrCA (P = .0069). In conclusion, our results show that a significant number of women with BrCA are positive for ER-beta only, and may be determined to be ER-negative when currently available IHC is used. ER-beta status is a significant predictor of response to AHTT in women with BrCA. Larger studies with multivariate analysis are needed to confirm these findings.     

Estrogen receptor beta is expressed in human colorectal adenocarcinoma

Estrogen receptor beta (ER-beta) has recently been detected in a human colon cancer cell line. The aim of this work was to determine whether ER-beta is expressed in human colorectal carcinoma (CRC) tissue and the extent of this expression. ER-beta expression in CRC was investigated by immunohistochemical staining of sections of formalin-fixed, paraffin-embedded tissue from 55 CRC. The percent of positive cells was recorded. ER-beta immunoreactivity was always present in normal epithelium and adenomas in the same sections of some CRC and was always nuclear. In CRC, nuclear ER-beta immunoreactivity was detected in >10% of the cancer cells in 67% of the cases and was almost always associated with cytoplasmic immunoreactivity. There were no statistically significant differences between the ER-beta-positive and -negative groups in regard to depth of invasion, nodal metastases, or survival, regardless of the cut-off value used. We conclude that (1) a significant number of CRCs are positive for ER-beta. (2) estrogen may play an important role in the proliferation of normal colonic epithelium, and (3) there is differential localization of ER-beta immunoreactivity between normal colon, adenomas, and CRCs. Whether different ER-beta isoforms are differentially expressed in CRCs, and whether human CRCs respond to treatment with antiestrogens, is the subject of studies currently in progress.     

Effects of apatite cements on proliferation and differentiation of human osteoblasts in vitro

Although apatite cement (AC) and sintered hydroxyapatite (s-HAP) are known to show good osteoconductivity, it is not clear whether or not the degree of their osteoconductivity is the same. In addition, it has not been clarified whether or not it is dependent on the type of AC; conventional AC (c-AC), fast-setting AC (fs-AC) or anti-washout AC (aw-AC). The aim of this study was to investigate the effects of ACs on cultured human osteoblasts, as they may provide a useful index of osteoconductivity. Human osteoblasts were cultured on the surfaces of ACs and s-HAP, and were evaluated with respect to cell attachment, proliferation, and differentiation. We found that ACs and s-HAP showed similar cell attachment. No significant difference between ACs and s-HAP was found with respect to the proliferation of osteoblasts. In contrast, we found that the differentiation of osteoblasts was enhanced on the surface of ACs compared with that of s-HAP. However, there was no difference among the types of AC. We therefore concluded that AC may show better osteoconductivity than s-HAP, and that osteoconductivity of AC may be similar, regardless of the type of AC.     

Estrogen receptor modulators and estrogen receptor beta immunolabelling in human umbilical vein endothelial cells

Human umbilical vein endothelial cells (HUVEC) exposed to the female sex hormone estradiol show different kinds of effects including increased elasticity, activation of plasma membrane Na⁺/H⁺ exchange, prostacyclin production, prevention of apoptosis and many others. The aim of this study was the systematic analysis of the immunolabelling of estrogen receptors (ERs), ERα and ERβ, in HUVEC after stimulation with different commercially available ER modulators and ER agonists or antagonists. HUVEC response to these substances was shown to be regulated via ERβ. ERα immunolabelling or up-regulation was abrogated after application of estrogen derivatives, selective estrogen receptor modulators (SERM) and ER agonists or antagonists. Immunolabelling of ERβ was significantly increased by estradiol, estrone, ethinylestradiol and tumour necrosis factor alpha (TNFα). SERM, such as Tamoxifen, and pure antagonists, such as ICI 182.780, stimulated ERβ in HUVEC at low concentrations, whereas higher concentrations inhibited ERβ immunolabelling. The pure estrogen receptor agonist 2,3-bis (4-hydroxyphenyl) proprionitrile (DPN) exhibited its activating potential at low concentrations. In contrast, higher concentrations resulted in a down-regulation of ERβ. Estrogenic effects in HUVEC, independent of stimulation or inhibition, are mediated via the ERβ. SERM such as Tamoxifen and ER antagonists such as ICI 182.780 act as ER activators in low concentrations, whereas higher concentrations lead to inhibitory effects.