1,25(OH)_2D_3对去卵巢大鼠骨组织成骨细胞、骨细胞凋亡的影响

目的观察1,25(OH)_2D_3对去卵巢大鼠骨组织中成骨细胞、骨细胞凋亡的影响。方法 36只雌性SD大鼠随机分成四组,对尼尔雌醇组和1,25(OH)_2D_3组分别给予尼尔雌醇(0.1mg/kg)和1,25(OH)_2D_3(0.05μg/kg)治疗12周。12周后,以DXA法测定大鼠全身的骨密度;放射免疫法测定各组大鼠血清中骨钙素及雌二醇的水平;处死各组大鼠,采用3′-OH末端DNA原位标记技术和透射电镜检测骨细胞、成骨细胞凋亡。结果 12周后,去卵巢组大鼠的骨密度和血清雌二醇水平明显降低,骨钙素含量升高,与假手术组相比,差异有显著性(P0.05)。1,25(OH)_2D_3可以增加去卵巢大鼠的全身骨密度和血清骨钙素含量(P0.05),但是不增加血清雌二醇的水平(P0.05)。1,25(OH)_2D_3可以抑制骨细胞、成骨细胞凋亡,与去卵巢组相比差异有显著性(P0.05)。结论 1,25(OH)_2D_3对去卵巢大鼠骨质疏松症具有防治作用,其部分机制可能为1,25(0H)_2D_3抑制了骨细胞、成骨细胞凋亡,从而调节骨重建。     

维生素D受体作用的分子基础

维生素D受体 (VDR)是由 6个功能区组成 ,每个功能区分工不同但又相互协调。配体结合区介导与 1 ,2 5(OH) 2 D3结合并与视黄酸X受体形成异二聚体 ,DNA结合区则通过两个锌指结构选择性地识别靶基因上DR3型维生素D反应元件 (VDRE)并由此改变局部DNA的构象 ,在其他协同转录激活 /抑制因子和AF 1、AF 2两个亚区的共同参与下激活转录过程 ,进行基因表达。在这一过程中 ,磷酸化对VDR转录活性的意义目前仍不清楚。VDR的异常与多种疾病有关。     

H13D联合Alen对去卵巢骨质疏松大鼠骨密度及骨生物力学指标的影响

目的探讨骨肽(OGP)衍生物H13D联合阿仑磷酸钠(Alen)对去卵巢(OVX)骨质疏松大鼠骨密度及骨生物力学指标的影响。方法健康雌性SD大鼠40只,随机分为假手术组、H13D组、Alen组、联合组、OVX组。其中假手术组手术取出卵巢再送回,余四组行双侧卵巢切除术,制作OVX大鼠模型。H13D组建模后皮下注射H13D10 nmol/100 g;Alen组皮下注射Alen 10μg/100 g;联合组同时给予HBD、Alen;OVX组、假手术组注射与上述药物等体积的PBS,均1次/d。12周后处死大鼠,检测L1～4椎骨与左侧股骨骨密度、血清钙、磷、碱性磷酸酶(ALP)水平,取右侧股骨行股骨3点弯曲试验,L5椎骨行腰椎压缩试验。结果骨密度从高到低依次为联合组、假手术组、Alen组、H13D组、OVX组(P均<0.05)。假手术组血清ALP水平低于余四组,联合组、H13D组、Alen组ALP低于OVX组(P均<0.05);L1～4椎骨能量吸收H13D组、联合组优于假手术组及OVX组(P均<0.05);股骨最大载荷、弹性载荷H13D组优于OVX组、Alen组、假手术组(P均<0.05);股骨弯曲能量联合组高于Alen组(P<0.05)。结论 H13D联合Alen可明显增加OVX骨质疏松大鼠的骨密度,改善骨生物力学指标,两药具有明显的协同作用。     

广西壮、汉族绝经后妇女维生素D 受体基因型与骨密度、骨代谢的关系

目的探讨维生素D受体基因(VDR)型在壮、汉族绝经后妇女中的分布及其与骨密度、骨代谢的关系.方法在广西居住20年以上的绝经后汉族妇女116名,壮族妇女82名.记录年龄、绝经年龄,测量身高、体重.采用双能X线吸收法测定骨密度(BMD);用聚合酶链反应-限制性片段长度多态性(PCR -RFLP)法测定受试者的VDR基因型;测定血清骨钙素(osteocalcin,OC)、尿脱氧吡啶啉(deoxypyridinoline,DPD)和尿肌酐(creatinine,Cr).结果壮、汉族妇女年龄、绝经年限、体重、体重指数、BMD、VDR基因型频率分布无显著性差异(P>0.05);BB、Bb、bb基因型检出率分别为6.57%、66.16%和27.27%;BB基因型组第2腰椎(L2)BMD比bb基因型组低10.03%,第4腰椎(L4)BMD分别较bb、Bb基因型组低9.63%和12.44%(P<0.05);BB基因型组骨质疏松发生率最高(46.15%),Bb基因型组次之(19.85%),bb基因型组最低(14.81%)(P<0.05);BB基因型组OC最低,与Bb、bb 基因型组比较也有显著性差异(P<0.05);三组间尿DPD排泄率(DPD/Cr)差异无统计学意义.结论 VDR基因型可作为预测广西壮、汉族绝经后妇女骨质疏松危险性的遗传学标志.     

老年危重症患者血清1,25(OH)2D3水平与APACHE Ⅱ评分及预后的关系

目的探讨老年危重症患者血清1,25-羟维生素D3(1,25(OH)2D3)水平与急性生理与慢性健康状况评分(APACHE)Ⅱ评分及预后的关系。方法选择2012年6月至2014年6月该院收治的老年(≥60岁)危重症患者共180例,依据APACHEⅡ评分由低到高分为高、中、低三组,并依据患者死亡存活状况,分为存活组和死亡组。采集患者空腹静脉血,采用酶联免疫吸附试验(ELISA)检测血清1,25(OH)2D3水平,记录患者院内死亡率和器官衰竭数量,计算患者APACHEⅡ评分,分析血清1,25(OH)2D3水平与APACHEⅡ评分、院内死亡率的相关性。结果三组患者血清1,25(OH)2D3水平逐渐降低,器官衰竭数量和院内死亡率逐渐升高(P0.05)。存活组血清1,25(OH)2D3水平高于死亡组,而器官衰竭数量和APACHEⅡ评分均低于死亡组(P0.05)。患者血清1,25(OH)2D3水平与APACHEⅡ评分呈负相关性(r=-0.378,P0.05),与院内死亡率呈负相关关系(r=-0.294,P0.05)。结论老年危重患者血清1,25(OH)2D3水平与患者APACHEⅡ评分、预后均相关,其与APACHEⅡ评分系统相结合,可评价患者的危重病情和预后,有重要的临床价值。     

1,25-(OH)2D3对UUO模型大鼠ILK及TGF-β1表达的影响

目的 探讨1,25-(OH)2D3减轻单侧输尿管梗阻(UUO)大鼠肾间质纤维化的作用及其机制.方法 将96只雄性SD大鼠随机分为对照组、UUO组、假手术组、干预组各24只,其中UUO组和干预组均于左肾下极处游离并结扎左输尿管建立UUO模型,假手术组开腹后仅游离左输尿管,对照组不行特殊处理.其后干预组予1,25-(OH)2D3 0.5 μg(溶于1 mL花生油中)灌胃、余三组均予1 mL花生油灌胃;各组分别于术后第1、3、7、14天每天各处死6只大鼠,留取左侧肾脏标本,光镜观察肾脏组织形态学改变及肾脏病理损害,免疫组化法观察整合来连接激酶( ILK)、转化生长因子(TGF) -β1蛋白表达,RT-PCR法检测ILK mRNA表达.结果 ①与对照组相比,UUO组肾皮质变薄、肾间质炎细胞浸润明显增多、纤维化随时间延长而加重(P＜0.05);与UUO组相比,干预组肾组织炎细胞浸润减少、纤维化程度减轻、病变积分明显降低(P＜0.05).②与对照组相比,UUO组ILK蛋白阳性表达随时间延长显著增加(P＜0.01);与UUO组比较,干预组ILK蛋白表达减少(P＜0.01);与对照组相比,UUO组TGF-β1蛋白随时间延长阳性表达显著增加(P＜0.01);与UUO组比较,干预组TGF-β1蛋白表达减少;ILK蛋白表达与TGF-β1蛋白表达呈显著正相关(r =0.872,P＜0.05).③与对照组比较,UUO组和干预组ILK mRNA表达均随时间延长明显增强(P＜0.01),其中干预组表达在7、14 d无显著差异;干预组各期均显著低于UUO组(P＜0.01).结论 1,25-(OH) 2D3可减轻UUO大鼠肾间质纤维化、保护肾脏功能,可能机制为间接或直接抑制肾间质ILK、TGF-β1表达;此为防治肾间质纤维化提出了新的方向.     

甲亢患者骨密度、骨代谢指标及维生素D受体的变化

本研究发现甲状腺功能亢进症 (甲亢 )患者腰椎、股骨颈、Ward三角和大转子区骨密度降低 ,血清钙、血清磷、血清碱性磷酸酶及血清骨钙素升高 ,血淋巴细胞内维生素D受体 (VDR)含量降低。FT3与FT4与骨密度呈负相关。提示甲亢导致高转换型骨丢失 ,它可引起骨密度降低 ,骨代谢指标及VDR含量异常。     

1,25(OH)2D3对应用糖皮质激素治疗的肾病患者骨代谢的影响

目的观察1,25(OH)2D3对应用糖皮质激素治疗的原发性肾脏病患者骨代谢的影响。方法将16例原发性肾脏病患者随机分为治疗组和对照组。治疗组在应用糖皮质激素治疗的同时给予0.25μg的1,25(OH)2D3,每日1次口服,碳酸钙0.75,每日3次口服。对照组单纯给予碳酸钙0.75,每日3次口服。所有患者在治疗前及治疗12周时应用双能X线吸收法骨密度仪测定腰椎和股骨颈的骨密度,同时测定有关骨代谢指标,包括血全段甲状旁腺素,骨钙素,尿脱氧吡啶啉,血钙、磷以及血清白蛋白,24小时尿蛋白定量、尿钙和尿磷。结果治疗12周时两组患者的腰椎和股骨颈骨密度均下降,两组间比较无显著性差异(P>0.05);治疗12周时对照组患者的骨钙素较治疗前明显下降(P<0.05),而治疗组无明显差异;尿脱氧吡啶啉在两组中均下降,但治疗组下降明显(P<0.05);两组患者血全段甲状旁腺素、24小时尿钙和尿磷治疗前后及组间比较均无显著性差异(P>0.05);应用1,25(OH)2D3治疗者无1例出现高钙血症。结论应用糖皮质激素治疗12周的原发性肾脏病患者其腰椎及股骨颈骨密度下降明显。1,25(OH)2D3可促进骨形成,抑制骨的吸收。     

1,25(OH)2D3对体外培养成骨细胞骨保护素mRNA表达的影响

维生素Ｄ体内活性形式 1,2 5 (ＯＨ) 2 Ｄ3具有促进骨吸收及骨形成作用。其骨吸收作用通过成骨细胞介导 ,然而机制尚未完全清楚。骨保护素 (ｏｓｔｅｏｐｒｏｔｅｇｅｒｉｎ ,ＯＰＧ)是近年分离出的一种因子 ,由成骨细胞合成和分泌 ,在破骨细胞的分化形成中起信号传导作用〔1〕。我们于 2 0 0 0年 6～ 12月实验观察 1,2 5 (ＯＨ) 2 Ｄ3对体外培养乳鼠成骨细胞ＯＰＧ基因表达的影响 ,以探讨其对骨吸收功能的调节机制。　　一、材料与方法　　 1.细胞培养 :取新生 2 4ｈＳＤ大鼠颅骨 ,Ⅰ型胶原酶分阶段消化收集细胞 ,5 %ＣＯ2 、3 7℃培养 ,每…     

不同糖耐量人群血清25(OH)D3水平及与胰岛β细胞功能的关系

目的 比较不同糖耐量人群血清25(OH)D3水平,分析其与胰岛β细胞功能的关系.方法 共纳入131例受试者,包括新诊断2型糖尿病患者(T2DM组)50例,糖调节受损者(IGR组)45名和糖耐量正常者(NGT组)36名.收集临床资料并检测相关生化指标,采用酶联免疫吸附法测定空腹血清25(OH)D3水平.结果 NGT组、IGR组和T2DM组血清25(OH)D3水平依次下降(F=25.984,P＜0.05).血清25 (OH)D3与体重指数、腰围、臀围、腰臀比、空腹血糖、口服葡萄糖耐量试验2 h血糖(2 hPG)呈负相关(r=-0.600 ～-0.175,P均＜0.05),与空腹胰岛素(FINS)、胰岛素曲线下面积、稳态模型评估-胰岛β细胞功能指数、早相胰岛素分泌指数(△INS30/△G30)呈直线正相关(r=0.296～0.693,P均＜0.05).多元逐步回归分析显示,2hPG、△INS30/△G30是血清25(OH)D3水平的独立相关因素(β=0.204,-0.178,P均＜0.05).结论 IGR者与T2DM患者25(OH)D3水平降低.血清25(OH)D3与胰岛素分泌功能呈正相关,与肥胖、血糖水平呈负相关.     

The vitamin D receptor (VDR) start codon polymorphism in primary hyperparathyroidism and parathyroid VDR messenger ribonucleic acid levels.

Vitamin D regulates parathyroid cell proliferation and secretion of PTH. Increased prevalence of the polymorphic vitamin D receptor (VDR) alleles b, a, and T has been reported in sporadic primary hyperparathyroidism (PHPT), suggesting that these genetic variants may predispose to the disease. Recently, another polymorphism in the VDR gene was related to bone mineral density, and this VDR-FokI polymorphism causes different lengths of the VDR, implying possible functional consequences. The VDR-FokI polymorphism was studied in 182 postmenopausal women with sporadic PHPT and in matched controls. No significant differences in distribution of the VDR-FokI genotypes could be detected between the groups, although there was a tendency toward overrepresentation of the F allele in the PHPT patients (P = 0.05). There were no significant associations with age, serum calcium, serum PTH, bone mineral density, or parathyroid tumor weight. The VDR genotypes were unrelated to VDR and PTH messenger ribonucleic acid levels in the parathyroid adenomas of 42 PHPT patients. In 23 PHPT patients, the Ca2+-PTH set-points were determined in vivo and were unrelated to the VDR alleles. We suggest that the VDR-FokI polymorphism has at most a minor pathogenic importance in the development of PHPT.     

慢性肾功能衰竭患者维生素D受体基因多态性与肾性骨病的关系

目的 探讨慢性肾功能衰竭（CRF）患者维生素D受体（VDR）基因多态性与肾性骨病（RO）发病的关系。方法 用RFLPs法检测87例CRF患者VDR基因型,用超声骨密度检测法检测患者跟骨骨密度（骨超声振幅衰减BUA）,用放免法检测患者血全段甲状旁腺素（iPTH）。然后分析相应数据,考察VDR与BUA及甲状旁腺素的关系。结果 VDR基因型主要分布在Bb和bb两型（97.68%）。Bb与bb二型患者BUA均低于正常参考值,且三组数据比较差异有非常显著性（P＜0.01）。三型间血全段甲状旁腺素水平亦有差异（P＜0.01）。结论 VDR基因型对慢性肾衰患者骨代谢有决定性影响,为筛选患者中RO和继发甲状旁腺机能亢进症（SHPT）高发人群提供参考。     

Functional vitamin D receptor (VDR) in the t-tubules of cardiac myocytes: VDR knockout cardiomyocyte contractility

We have previously shown that the active form of vitamin D, 1,25 dihydroxyvitamin D3 [1,25(OH)(2)D(3)], has both genomic and rapid nongenomic effects in heart cells; however, the subcellular localization of the vitamin D receptor (VDR) in heart has not been studied. Here we show that in adult rat cardiac myocytes the VDR is primarily localized to the t-tubule. Using immunofluorescence and Western blot analysis, we show that the VDR is closely associated with known t-tubule proteins. Radioligand binding assays using (3)H-labeled 1,25(OH)(2)D(3) demonstrate that a t-tubule membrane fraction isolated from homogenized rat ventricles contains a 1,25(OH)(2)D(3)-binding activity similar to the classic VDR. For the first time, we show that cardiac myocytes isolated from VDR knockout mice show accelerated rates of contraction and relaxation as compared with wild type and that 1,25(OH)(2)D(3) directly affects contractility in the wild-type but not the knockout cardiac myocyte. Moreover, we observed that acute (5 min) exposure to 1,25(OH)(2)D(3) altered the rate of relaxation. A receptor localized to t-tubules in the heart is ideally positioned to exert an immediate effect on signal transduction mediators and ion channels. This novel discovery is fundamentally important in understanding 1,25(OH)(2)D(3) signal transduction in heart cells and provides further evidence that the VDR plays a role in heart structure and function.     

Serum levels of vitamin D metabolites and bone remodelling in hyperthyroidism

Serum levels of 25-hydroxyvitamin D, 24,25-dihydroxyvitamin D and 1,25-dihydroxyvitamin D were measured in 25 untreated hyperthyroid patients in whom histomorphometric evaluations of iliac crest bone biopsies were performed after in vivo tetracycline doublelabeling. The serum concentration of 25-hydroxyvitamin D was normal. The serum concentration of 1,25-dihydroxyvitamin D was reduced (p < 0.02) compared to normal whereas the serum concentration of 24,25-dihydroxyvitamin D was increased (p < 0.02). The bone changes were characterized by an enhanced turn-over in trabecular and cortical bone leading to an increased porosity of cortical bone and mobilisation of bone mineral. The observed changes in vitamin D metabolism could be explained by a reduced renal 1-alpha-hydroxylase activity secondary to hypercalcaemia with suppressed parathyroid secretion and hyperphosphataemia. The bone changes were unrelated to the serum levels of vitamin D metabolites. In trabecular bone the appositional rate and mineralization rate of osteoid were increased and the mineralization lag time was shortened showing that the mineralization and formation of osteoid in the hyperthyroid state can progress with an enhanced rate in spite of a reduced mean serum level of the active vitamin D metabolite, 1,25-dihydroxyvitamin D.     

Effect of vitamin D receptor (VDR) genotypes on the risk for osteoporosis in type 1 Gaucher disease

The almost universal finding of osteopenia in Gaucher disease and the unpredictable advent of avascular necrosis and/or pathological fractures are clinically relevant issues that are not necessarily reversed by specific enzyme replacement therapy (ERT). However, genetic/epigenetic factors may explain variance for bone mineral density (BMD) in the normal population. Among the candidate genes, vitamin D receptor (VDR) polymorphisms have been correlated with BMD at the lumbar spine (LS) and femoral neck (FN) and/or fracture risk in various populations. Helsinki Committee approval was received. DNA samples from 15 healthy individuals and 83 non-neuronopathic adult patients with recent BMD data at LS and FN were tested for VDR polymorphisms FokI, ApaI, TaqI, and BsmI. Chi-square, Fisher’s exact, analysis of variance (ANOVA), Scheffe post hoc, and non-parametric Kruskal-Wallis ANOVA were applied as appropriate. P values <0.05 was considered statistically significant. There were no significant differences between patients and controls in frequency of any of polymorphic genotypes. The ApaI and BsmI genotypes were correlated with the N370S/N370S Gaucher genotype (p = 0.029, and p = 0.061, respectively). There was a correlation between the BsmI genotype and skeletal involvement (p = 0.018) and a trend to significance with Z scores at LS (p = 0.084). There was a statistically significant correlation between the Taq1 variants and BMD at the FN (p = 0.030) with the TT variant associated with lower BMD. As in other populations, VDR polymorphic genotype may be an independently sorting modifier in the prediction of BMD and bone involvement in Gaucher disease.     

维生素D受体基因多态性对尿毒症伴继发性甲状旁腺功能亢进的影响

目的　评价维生素D受体 (VDR)基因多态性对尿毒症伴继发性甲状旁腺功能亢进( 2°HPT)严重程度的影响。方法　对 1 0 6例尿毒症维持性血液透析患者采用聚合酶链反应———限制性片段长度 (PCR RFLP)方法分析VDR基因Apa I位点多态性 ,有ApaI酶切位点的等位基因规定为a ,没有者规定为A。结果　 1 0 6例患者VDR基因ApaI多态性频率分别为AA 1 7%、Aa54 .7%、aa 2 8.3%。aa基因组患者血清iPTH浓度 [( 339.8± 357.4) pg/ml]明显高于AA基因组[( 1 51 .9± 1 6 7.3) pg/ml]和Aa基因组患者 [( 1 91 .5± 1 6 9.8) pg/ml],两组比较差异有显著性 ( P <0 .0 5)。aa基因组患者骨钙蛋白浓度 [( 36 .2± 56 .0 )ng/ml]明显高于AA基因组 [( 9.2± 6 .1 )ng/ml]和Aa基因组患者 [( 1 5.3± 2 1 .6 )ng/ml],两组比较差异有显著性 ( P <0 .0 5)。三组间血钙、磷、镁及一般情况均无明显差别 ,但aa基因组降钙素浓度明显低于Aa基因组 [( 30 .1 6± 1 7.59)vs( 73.2 2± 6 8.2 6 ) pg/ml,P <0 .0 1 ]。结论　a等位基因患者较非a等位基因患者易产生严重的 2°HPT。