淫羊藿对骨的作用研究进展

淫羊藿 (HerbaEpimdii)为小檗科植物 ,又名仙灵脾、三枝九叶草等 ,具有“补肾壮阳 ,益精健骨”之功效。淫羊藿多糖、淫羊藿苷、淫羊藿甙和黄酮类为其主要成份[1 ] 。淫羊藿的作用广泛 ,如对心血管系统、中枢神经系统、血液系统、免疫系统、抗炎、抗骨质疏松、抗衰老、抗肿瘤等均有作用 ,本文就其对骨的作用的研究现状作一综述。一、淫羊藿对成骨细胞增殖与分化的影响成骨细胞是蛋白分泌型细胞 ,能产生Ⅰ型胶原 ,合成分泌骨基质 ,具有高碱性磷酸酶 (ALP)活性 ,并能吸收和转运钙离子 ,是骨形成和骨再建的重要功能细胞。目前认为 ,ALP活性…     

力学刺激与淫羊藿苷耦合作用抑制疲劳损伤诱导破骨细胞分化的分子机制研究

目的 探讨力学刺激与淫羊藿苷(ICA)耦合作用对疲劳载荷下破骨细胞分化的影响及可能存在的分子机制。方法 体外培养小鼠单核巨噬细胞系RAW264.7,空白对照组为α-MEM完全培养基;疲劳载荷下破骨细胞模型组更换为破骨细胞培养液(含质量浓度为40 ng/ml巨噬细胞集落刺激因子和40 ng/ml破骨细胞分化因子的α-MEM完全培养基),并对RAW264.7细胞施加5 000 με基底拉伸应变;力学刺激+ICA组经上述细胞因子和高应变(5 000 με)刺激后,更换为含有1×10^-5 mol/L ICA的α-MEM完全培养基,并施加1 000 με基底拉伸应变。采用抗酒石酸酸性磷酸酶(TRAP)检测试剂盒检测破骨细胞中TRAP的活性变化;实时定量PCR检测破骨细胞标志性基因TRAP、组织蛋白酶K(CTSK)和基质金属蛋白酶9(MMP-9)的mRNA表达;蛋白质印迹法(Western Blot)分析核因子κB(NF-κB)异位情况。结果 与模型组相比,力学刺激(1 000 με的基底拉伸)和ICA(1×10^-5 mol/L)耦合作用能显著抑制破骨细胞中TRAP的活性(P<0.01),减少破骨细胞的形成;显著下调标志性基因TRAP、CTSK和MMP-9的mRNA表达,差异均具有统计学意义(均P<0.01);通过抑制NF-κB信号通路中P65、P50、NF-κB抑制蛋白-α(IκB-α)的磷酸化来抑制破骨细胞的生成。结论 力学刺激与ICA耦合作用可有效抑制疲劳载荷下破骨细胞的分化及其骨吸收功能,其作用机制可能是通过调节NF-κB信号通路来实现。     

淫羊藿苷治疗骨不连的分子机制

文题释义：骨不连：骨组织具有强大的自我修复能力,给予适当治疗后大多数骨折都会很快很好地愈合。然而,仍有一小部分骨折难以愈合,当骨折超过3个月仍未愈合,称为延迟愈合,当骨折超过9个月仍未达到骨性连接,则称为骨不连。淫羊藿苷：为淫羊藿干燥茎叶的提取物,呈淡黄色针状结晶粉末,相对分子质量为676.65,属黄酮类化合物,现代药理学研究发现其具有很强的生物活性,对骨组织、免疫系统、肿瘤组织、神经系统、生殖系统、内分泌系统和心血管系统等具有显著作用。背景：骨不连是骨科临床上常见的并发症,严重影响患者的身心健康和生活质量。近年来大量的研究发现淫羊藿苷在促进骨折愈合和治疗骨缺损方面有着显著作用;骨不连和骨折愈合是对立共存的,有关骨折愈合机制的研究实际上就是治疗骨不连的研究。目的：综述淫羊藿苷在骨不连治疗过程中的分子机制研究进展。方法：第一作者应用计算机以“icariin,bone nonunion,bone marrow mesenchymal stem cells,periosteal cell,osteoblasts,osteoclast”及“淫羊藿苷、骨不连、骨髓间充质干细胞、骨膜细胞、成骨细胞、破骨细胞”作为主题词检索PubMed、中国知网、万方、维普数据库,共检索到542篇文献,按入选标准及排除标准进行筛选,最终纳入44篇文献。结果与结论：淫羊藿苷通过促进骨髓间充质干细胞和骨膜细胞增殖及向成骨细胞分化、促进成骨细胞增殖与成熟、抑制破骨细胞的破骨作用,有效促进骨折愈合和治疗骨不连。目前绝大部分研究仍处于基础实验阶段,尚需要大量临床研究,对于相关蛋白及基因方面也有待于进一步研究,希望为骨不连的中药治疗提供新思路。ORCID: 0000-0002-2013-743X(余绍涌)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     

淫羊藿苷对糖皮质激素诱导的骨微血管内皮细胞损伤的保护作用

目的体外建立糖皮质激素诱导骨微血管内皮细胞（Human bone microvascular endothelial cell,HBMEC）损害的模型,研究糖皮质激素对HBMEC的损害情况;探讨淫羊藿苷（Icariin,ICA）对糖皮质激素所致HBMEC损伤的保护作用。方法HBMEC分离与体外培养,建立体外糖皮质激素杀伤细胞模型,用MTF法观察ICA对激素损伤的HBMEC活性的影响,测定细胞生存率,并对细胞进行台盼蓝染色以观察ICA对激素诱导的HBMEC坏死的影响。结果糖皮质激素诱导HBMEC损害,台盼蓝染色显示细胞损害为坏死而非凋亡,并呈现一定的浓度依赖和时间依赖性;ICA能保护激素诱导的HB—MEC损害,使细胞生存率明显提高,ICA能显著抑制激素诱导的内皮细胞坏死。结论ICA具有保护激素诱导的HBMEC损伤的作用,这为早期干预治疗激素性骨坏死提供了实验药理依据。     

淫羊藿苷促进骨折愈合作用机制的研究进展

近年来 , 国内外学者对中药淫羊藿主要有效成分淫羊藿苷加速骨折愈合的机制做了一系列研究,目前研究结果显示, 淫羊藿苷加速骨折愈合的机制主要是通过激活 Wnts、BMP、MAPK、ER 等相关信号通路促进 BMSCs、ASCs 的迁移归巢,进行成骨性分化为成骨细胞,同时促进成骨细胞的成熟、矿化以形成骨痂,并抑制破骨细胞的骨吸收活性。本文从淫羊藿苷对相关细胞以及对血管重建的作用方面对淫羊藿苷加速骨折愈合的作用机制做一回顾综述,以期为后续的研究提供参考。     

淫羊藿苷调节细胞增殖作用的研究进展

淫羊藿苷是传统补益中药淫羊藿的有效活性成分之一,研究发现可影响多种细胞的增殖。淫羊藿苷主要促进骨组织相关细胞增殖,包括成骨细胞、软骨细胞、骨膜细胞、间充质干细胞,淫羊藿苷抑制多种肿瘤细胞增殖,包括骨肉瘤细胞、乳腺癌细胞和卵巢癌细胞等,淫羊藿苷亦可调节其他细胞的增殖,包括神经干细胞、平滑肌细胞等。但淫羊藿苷对细胞增殖促进和抑制的双重调节机制需要进一步研究,方能为其在细胞增殖紊乱相关疾病中的应用提供理论依据。     

淫羊藿苷调节酸性微环境减轻绝经后老年骨质疏松性疼痛

背景：前期研究已证明,淫羊藿苷在促进骨形成和抑制骨吸收方面具有重要作用,但其对骨质疏松介导的骨痛产生的影响尚未见报道。目的：探讨淫羊藿苷减轻绝经后老年性骨质疏松性骨痛的可能机制。方法：(1)动物实验：将200只C57BL/6小鼠随机分为4组：假手术组、模型组、模型+淫羊藿苷组,模型+碳酸酐酶Ⅱ抑制剂组。除假手术组外,其余各组摘除小鼠卵巢建立绝经后骨质疏松模型。模型+淫羊藿苷组在造模后第2天灌胃淫羊藿苷,每2周进行1次疼痛行为学实验并取材,持续20周。microCT检测股骨骨量、苏木精-伊红染色及TRAP染色检测破骨细胞活性、免疫荧光染色检测神经元形态及相关离子通道表达。(2)细胞实验：提取小鼠骨髓来源的破骨细胞前体细胞,在体外使用RANKL/M-CSF体系诱导分化为破骨细胞并添加不同浓度淫羊藿苷(1,10μmol/L)干预。采用抗酒石酸酸性磷酸酶染色检测破骨细胞分化,采用鬼笔环肽染色检测破骨细胞肌动蛋白环,采用骨板吸收实验检测破骨细胞噬骨功能,采用pH计检测体系pH值,采用Western Blot检测破骨细胞分化相关蛋白表达。此外,提取小鼠背根神经节来源神经细胞并用淫羊藿苷处理,采用C...     

淫羊藿总黄酮的药理学研究

目的 研究淫羊藿总黄酮(total flavonoids of epimedium,TFE)对人脐静脉内皮细胞损伤的保护作用.方法 将人脐静脉内皮细胞体外培养、传代后随机分为六组:空白对照组、TFE模型组、溶血性磷脂酰胆碱(LPC)损伤组、LPC+TFE(50 mg/L)组、LPC+TFE(100 mg/L) 、LPC+TFE(200mg/L)组,比较淫羊藿总黄酮对内皮细胞损伤的作用.结果 TFE模型组与空白组比较,各组数据无明显差异;LPC损伤组中MDA值、LDH值和细胞内KOS水平较空白组显著增加,而NO含量降低;在加入LPC+TFE的三组中,随着TFE的浓度增加,细胞上清液中NO含量升高,MDA和LDH含量以及ROS水平降低.结论 淫羊藿总黄酮对内皮细胞损伤具有保护作用.     

淫羊藿苷防治血管性痴呆的实验研究

目的：观察淫羊藿苷(Icariin，ICA)对血管性痴呆(vascular dementia。VD)大鼠学习记忆的保护作用．探讨ICA防治VD的作用机制。方法：采用永久性结扎双侧颈总动脉的方法(2-VO法)以及脑缺血再灌注方法(I10-R10-I10)制作VD大鼠模型，利用Morris水迷宫检测ICA对大鼠空间辨别学习记忆能力的影响；用光化学法检测大脑皮层及海马组织中SOD活性、MDA和AChE含量．并检测大鼠血清中AChE的含量；用免疫组化方法检测海马内AChE及ChAT的水平，并通过图象处理软件进行定量分析；     

中药淫羊藿苷抑制肝癌HepG2.2.15细胞增殖和免疫逃逸作用研究

目的:观察中药淫羊藿苷(ICA)对HepG2.2.15细胞增殖及对CD3AK细胞杀伤活性的影响,探讨ICA对肝癌细胞Fas/FasL途径免疫逃逸的逆转作用,为ICA的开发应用提供新的理论和实验依据.方法:MTT法检测细胞增殖和细胞杀伤活性;流式细胞术检测细胞表面分子表达水平和细胞凋亡率.结果:50 μg/mlICA作用HepG2.2.15细胞48、72小时的增殖抑制作用明显,抑制率分别为22.04%、29.68%(P＜0.05),呈时间依赖效应.HepG2.2.15细胞经ICA处理后,FasL的表达率由16.22%显著下降至8.29%,Fas表达率由0.79%提高到1.70%(P＞0.05).ICA可明显抑制HepG2.2.15细胞诱导Jurkat细胞凋亡,凋亡率从46.66%下降为18.20%.ICA处理HepG2.2.15细胞后,不同效靶比的CD3AK细胞的杀伤活性,可分别由对照组的15.81%、35.04%、42.85%显著提高至42.58%、67.55%、88.93%(P＜0.05,P＜0.01),呈效靶比依赖效应.结论:ICA能有效地抑制HepG2.2.15细胞增殖,并有一定时间依赖效应;ICA可下调FasL的表达,上调细胞表面Fas的表达,对HepG2.2.15细胞诱导的T淋巴细胞凋亡作用有一定阻断,逆转肿瘤细胞的免疫逃逸作用;ICA显著增强HepG2.2.15细胞对CD3AK细胞杀伤的敏感性.     

X-Ray Microprobe Analysis of Articular Cartilage

An electron X-ray microprobe has been used to determine quantitatively in situ the concentration of negatively charged fixed groups as well as the sulphur content of human articular cartilage.

Operating conditions were investigated and procedures were developed for the study of both 'thick' (>20 mμ) and 'thin' (<20 mμ) sections. A resolution of the order of a few microns could be obtained.

Calibration experiments were carried out using calcium sulphate-geIatin standards.

By determining simultaneously the total fixed charge density and the sulphur content in any given spot it was possible t o deduce the proportions of chondroitin and keratan sulphate.

The results obtained so far indicate that the surface layer of articular cartilage contains mainly chondroitin sulphate and that there is no hyaluronic acid present.

Differences were also studied in fixed charge density and sulphur content between the pericellular regions and the interterritorial matrix in the different zones of cartilage.

It was found that whilst there were no differences in the superficial zone, in the middle and deep zones the pericellular region contained a slightly higher concentration of negatively charged groups and sulphur. Moreover the ratio of fixed charge density to sulphur content was a little higher in the pericellular region than in the inter- territorial matrix.     

力学刺激在破骨细胞分化中的作用

骨组织细胞主要有成骨细胞、破骨细胞、骨细胞3种类型,与细胞外基质共同维持骨的结构完整性和功能性。破骨细胞专门负责骨吸收,通过释放酸性物质和蛋白水解酶降解骨基质,发挥骨吸收作用;通过与成骨细胞相互协调,共同维持骨稳态。破骨细胞增多,导致骨吸收增加,从而引起骨质疏松症等骨骼疾病;破骨细胞生成过少,将导致骨吸收减弱等相关疾病,例如骨硬化症。因此,对于破骨细胞功能的精确调控,在维持骨稳态的平衡中发挥极为关键的作用。既往研究多从生物化学角度解释和阐述各种生物因素对破骨细胞的调控。然而越来越多的研究证实,力学刺激在破骨细胞分化过程中发挥着重要的作用。本文着眼于力学刺激对破骨细胞分化的影响,讨论力学刺激在其中可能发挥的作用,并对该领域的新发现以及未来的发展进行探讨。     

Parathyroid hormone induces bone resorption in human peripheral blood mononuclear cells

Osteoclasts are known to derive from a macrophage colony-stimulating factor (M-CSF)-dependent precursor shared with macrophages. Cells capable of forming osteoclasts are present in peripheral blood. We characterized this population by incubating human peripheral blood mononuclear cells (PBMCs) with osteoclast-inductive UMR 106 cells, human macrophage colony stimulating factor (hM-CSF) and parathyroid hormone (PTH) or 1,25(OH)₂vitamin D3 on slices of devitalised cortical bone. We found that PBMCs were capable of substantial bone resorption, to levels comparable to those of haemopoietic tissue. Cells plated at very low densities and screened for the presence or absence of excavations revealed a linear relationship (r = 0.994) between the number of cells plated and the number of excavations formed. The limiting dilution analysis suggested that 1 in every 300–600 plated cells (0.15–0.3% of the PBMC population) had the capacity to resorb bone. The precursor was found in the rapidly adherent fraction, and typically generated very small numbers of excavations, suggesting that it was a relatively mature cell type. Co-cultures of PBMCs with UMR 106 cells would not generate osteoclasts without PTH/1,25(OH)₂vitamin D3, even with M-CSF, indicating that osteoclast-induction by stromal cells is not attributable to hormonal induction of M-CSF in UMR 106 cells, but that PTH induces some other activity, necessary for osteoclast but not macrophage formation, in UMR 106 cells. Osteoclasts did not form if PTH was omitted in the first few days of the culture period. Thus, osteoclasts appear to form not from cells committed to macrophage differentiation, but from a discrete subpopulation of relatively mature bipotential or osteoclast-committed precursors which, in the absence of an osteoclast-inductive stimulus, become irreversibly lost to the osteoclast lineage.     

Rapid inactivation and apoptosis of osteoclasts in the maternal skeleton during the bone remodeling reversal at the end of lactation

There is a rapid reversal in maternal skeletal metabolism and bone remodeling from accelerated bone resorption during lactation to skeletal rebuilding after lactation. The purpose was to determine the changes that occur in maternal osteoclasts during the transition from lactation to postlactation. Skeletal samples were taken from female rats on days 10 and 19 of lactation and 1 and 7 days after lactation. The pups were weaned on day 20. There was a rapid change in the osteoclast population after weaning, resulting in less resorption surface. Osteoclasts detached from bone surfaces, lost their ruffled borders, and became fragmented with immunocytochemical evidence of apoptosis within 24 hr after lactation. Concomitant with the rapid regression in the osteoclast population was an over fivefold increase in maternal calcitonin (CT) levels at 24 hr after weaning. Serum calcium and estrogen (E2) increased, but prolactin (PRL) and PTH decreased after weaning. The hormone changes, particularly that of CT, are consistent with the rapid regression of the osteoclast population at the end of lactation. These changes are similar to a reversal phase of a bone remodeling cycle where bone formation commences when resorption ceases on bone surfaces and suggests that the fate of osteoclasts during bone remodeling is programmed cell death. These results also suggest that bone remodeling is well synchronized prior to, during, and after lactation to accommodate the mineral requirements of the offspring as well as the mother. Anat Rec, 290:65–73, 2007. © 2006 Wiley‐Liss, Inc.     

Osteoclast formation and function in pigmented villonodular synovitis

Pigmented villonodular synovitis (PVNS) is a synovial tumour-like lesion that frequently causes osteolysis. PVNS contains numerous macrophages and osteoclast-like giant cells. In this study, we have analysed the cytochemical and functional characteristics of mononuclear and multinucleated cells in PVNS and determined the cellular and humoral mechanisms underlying giant cell formation and resorption in PVNS. Giant cells and CD14(+) and CD14(-) mononuclear cell populations were isolated from PVNS synovial tissue and cultured alone or in the presence and absence of the osteoclastogenic factors, RANKL and M-CSF. Osteoclast formation and activity was assessed by expression of TRAP and evidence of lacunar resorption. Giant cells in PVNS expressed an osteoclast-phenotype (CD51(+) , TRAP(+) , CD14(-) , HLA-DR(-) ) and were formed only in cultures of mononuclear cells that expressed the macrophage marker CD14. Osteoclast formation required RANKL and occurred in both the presence and absence of exogenous M-CSF. CD14(-) cells in PVNS expressed RANKL. Lacunar resorption by PVNS-derived giant cells was abolished by the addition of the bisphosphonate, zoledronate. Our findings indicate that osteoclasts form by a RANKL-dependent mechanism from CD14(+) mononuclear phagocytes in PVNS. Osteoclast formation occurred even in the absence of exogenous M-CSF, a finding which is in keeping with over-expression of M-CSF playing a pathogenic role in this condition. Anti-osteoclast resorptive treatment may be useful to control osteolysis in PVNS.     

THE EFFECT ON CHICK OSTEOCLASTS OF INFECTION WITH PARAMYXOVIRUSES

The detection of virus in osteoclasts from Pagetic patients is now well known, but it has yet to be shown convincingly that the presence of virus in Pagetic osteoclasts influences their behaviour. In this study, osteoclasts from embryonic chick tibiae were infected with canine distemper virus or measles virus and compared with mock-infected controls. Infection was confirmed using virus-specific fluorescent antibodies. It was found that virus infection did not alter osteoclast morphology or tartrate-resistant acid phosphatase (TRAP) activity. Both infected and mock-infected osteoclasts produced resorption pits on bovine bone slices; these could be divided into two distinct size classes with a computer-based measuring system. Virus infection significantly increased the proportion of the larger size class of resorption pit. These results suggest that virus infection can increase bone resorption by osteoclasts, lending further support to the hypothesis that viruses play a role in Paget's disease of bone.     

骨水泥与非骨水泥型全膝关节假体置换效果的系统评价

背景：国外研究调查显示,在全膝关节置换中有95.2%采用骨水泥进行假体固定,但是也有专家学者认为使用骨水泥固定假体风险高。 目的：应用Cochrane系统评价的方法评价膝关节置换中骨水泥型与非骨水泥型假体置换的效果差异。 方法：检索Medline(1996年1月至2011年8月)、Embase(1980年1月至2011年8月)、Cochranelibrary (2011年8月)、中国生物医学文献数据库(CBM, 1990年1月至2011年8月)及相关参考文献,收集骨水泥型和非骨水泥型假体全膝关节置换的随机对照试验,采用Cochrane的方法学评价文献质量,应用RevMan5.1.2 进行Meta分析。比较骨水泥型和非骨水泥型假体在术后生存率、稳定性、相关并发症、翻修率、异位骨化的差异。并使用GRADEpro version3.2.2软件对纳入研究进行证据评级。 结果与结论：纳入8个随机对照试验,共1 381例患者,实验组(骨水泥组)676例,对照组(非骨水泥组)705例。4个研究比较了骨水泥组和非骨水泥组假体置换后≤5年组的膝关节生存率,两组差异有显著性意义,说明置换后≤5年骨水泥组生存率高。4个研究比较了置换后>5年组膝关节生存率,两组差异有显著性意义,说明置换后>5年组骨水泥假体组生存率高。3个研究在不同随访时间比较了置换后假体稳定性,结果显示两组间在假体稳定性上差异无显著性。4个研究在不同随访时间比较了置换后假体相关的并发症,两组间在假体相关并发症上比较差异无显著性意义。5个研究在不同随访时间比较了置换后翻修率,两组间在假体翻修率上差异无显著性意义。3个研究在不同随访时间比较了置换后假体异位骨化,两组间在置换后假体异位骨化上差异无显著性意义。两组置换后疗效均较术前明显提高,组间比较的Meta 分析结果显示,不论是≤5年组还是>5年组,均是骨水泥型假体生存率均高于非骨水泥型假体生存率,两组在稳定性、相关并发症、翻修率、异位骨化等方面差异均无显著性意义(P > 0.05)。     

月骨的临床解剖学及人工假体的研究进展*

目前临床上治疗月骨缺血性坏死的手术方法较多,不同手术方式各有优势,但尚无一种方法在改善患者的疼痛及恢复腕关节功能上明显优于其他治疗。因此,探索新的治疗月骨缺血性坏死的手法方式是手外科医师研究的趋势。近年来,随着对月骨解剖和生物力学分析的深入研究,人工月骨假体置换成为治疗月骨缺血坏死的重要手段。本文旨在结合相关文献资料,对月骨的临床解剖学研究和人工假体的临床应用进行综述。     

髋关节假体体外磨损试验的应用及研究进展

体外磨损试验(ISO 14242)基于良好的实验室控制条件和操作规范模拟人体行走步态的关节承受载荷和运动角度范围,广泛用于比较髋关节假体不同材料、假体设计的耐磨损性能.本文基于体外磨损试验和临床磨损评价的研究文献,介绍体外磨损标准试验方法及其在髋关节设计开发中的应用,探讨体外磨损标准方法的局限、研究进展以及未来发展方向...     

全髋翻修髋臼骨缺损中颗粒打压植骨的应用

背景：髋臼重建植骨分为颗粒植骨、结构植骨和混合植骨,而颗粒骨由于制作简便、愈合时间短等优点而被大量使用。 目的：探讨全髋人工关节翻修中髋臼AAOSIII型骨缺损颗粒打压植骨技术的应用要点和临床效果。 方法：16例髋关节翻修AAOSIII骨缺损患者采用单纯颗粒打压植骨技术、结合金属器械或结构植骨处理髋臼AAOSIII型骨缺损。对比治疗前及治疗后定期随访的髋关节Harris评分及影像学资料,评估颗粒打压植骨对该型髋臼骨缺损的治疗效果。 结果与结论：按照治疗后3,6,12个月及每半年周期随访中,16例患者髋关节疼痛症状均得到明显缓解,并恢复行走功能。16例患者治疗后Harris髋关节功能评分由治疗前平均48.00分提高到84.94分(P < 0.01),其中优5例,良8例,可2例,差1例,优良率为81%,患者满意率为94%。16例患者术后X射线片示髋臼旋转中心均恢复(接近)正常,臼杯骨性覆盖良好,植骨颗粒获得放射学骨整合,臼杯假体未发生移位,假体植入物未发生位置改变或断裂现象。结果提示颗粒打压植骨技术修复翻修髋臼AAOSIII型骨缺损,有效重建了髋臼的骨性结构,保留并恢复了髋臼的骨量,具有良好的技术优势和临床疗效。