1,25-二羟维生素D3对支气管哮喘大鼠气道炎症和肺内诱导型一氧化氮合酶的影响

目的本文探讨1,25-二羟维生素D3[1,25-dihydroxy vitmin D3,1,25-(OH)2D3]对于支气管哮喘(简称哮喘)大鼠气道炎症和肺内诱导型一氧化氮合酶(induced nitric oxide synthase,iNOS)的影响。方法50只Wistar大鼠随机分为四组:正常对照组,哮喘组,预防处理组和治疗组。用卵蛋白作为致敏原制备哮喘大鼠模型,第一组预防处理组在致敏前三天给予口服1,25-(OH)2D3,共给药3周,第二组预防组则于致敏第1天和第7天给予肌注维生素D3。治疗组则于支气管激发开始前给药处理。应用动物肺功能仪测定大鼠对于乙酰甲胆碱刺激的气道反应性,测定支气管肺泡灌洗液(BALF)细胞总数及嗜酸粒细胞计数和肺组织病理变化。检测肺组织中一氧化氮(NO)含量,iNOS活性和iNOS mRNA表达水平,用免疫组织化学染色方法检测肺组织iNOS的表达并观察iNOS在气道组织分布的改变。结果1,25-(OH)2D3预防及治疗组可减轻BALF中细胞总数及嗜酸粒细胞计数,同时降低肺组织中NO含量、iNOS活性及大鼠肺组织中iNOS mRNA的表达水平。免疫组织化学结果显示,干预组可降低哮喘大鼠气道组织中上皮细胞和巨嗜细胞中iNOS阳性。结论1,25-(OH)2D3可降低哮喘大鼠气道炎症及iNOS表达水平,提示1,25-(OH)2D3对于哮喘可能有潜在的治疗作用。     

基质金属蛋白酶与支气管哮喘气道重塑

气道炎症和重塑是支气管哮喘（简称哮喘）的主要特征。细胞外基质（extracellular matrix,ECM）的降解和沉积失衡是导致呼吸道壁结构异常构建的重要原因,而基质金属蛋白酶（matrix metalloproteinases,MMP）则是调节ECM代谢的主要限速酶。各种MMP及其抑制剂之间可以互相调节,也可作用于细胞因子等多种炎症因子,以增强或减弱其生物学效应。气道重塑的过程也是MMP在气道壁沉积的过程,因此对MMP及其抑制剂的深入研究,对哮喘炎症及气道重塑机制的进一步明确有着重要意义。本文着重就MMP的生物学特性以及它们在哮喘气道重塑中所起的作用进行综述。     

基质金属蛋白酶与支气管哮喘气道重塑

气道炎症和重塑是支气管哮喘(简称哮喘)的主要特征.细胞外基质(extracellular matrix,ECM)的降解和沉积失衡是导致呼吸道壁结构异常构建的重要原因,而基质金属蛋白酶(matrixmetailoproteinases,MMP)则是调节ECM代谢的主要限速酶.各种MMP及其抑制剂之间可以互相调节.也可作用于细胞因子等多种炎症因子.以增强或减弱其生物学效应.气道重塑的过程也是MMP在气道壁沉积的过程,因此对MMP及其抑制剂的深入研究,对哮喘炎症及气道重塑机制的进一步明确有着重要意义.本文着重就MMP的生物学特性以及它们在哮喘气道重塑中所起的作用进行综述.     

1,25-二羟维生素D3水平与新生儿呼吸道感染分析研究

目的 探讨1,25-二羟维生素D₃[1,25-(OH)₂-D₃]水平对新生儿呼吸道感染的影响,为新生儿呼吸道感染预防与治疗提供依据。方法 收集2020年1月至2020年6月在首都医科大学附属北京世纪坛医院新生儿重症监护病房诊断为急性呼吸道感染的新生儿120例作为呼吸道感染组,选择同期住院出生的116例健康足月新生儿作为对照组,同时测定各组新生儿血清1,25-(OH)₂-D₃水平。依据1,25-(OH)₂-D₃含量为四组,血清1,25-(OH)₂-D₃含量>250nmol/L为过量,血清1,25-(OH)₂-D₃含量75～250nmol/L为适宜,血清1,25-(OH)₂-D₃含量50～74nmol/L为不足,血清1,25-(OH)₂-D₃含量<50n...     

基质金属蛋白酶-9与支气管哮喘

基质金属蛋白酶 9在哮喘患者的支气管肺泡灌洗液、痰、血中都增加。MMP 9mRNA的表达和免疫活性在哮喘患者的支气管镜检物中增加。本文就MMP 9的生物学特征、调节以及与气道炎症、气道重塑、气流阻塞、气道高反应性及激素治疗的关系作一综述     

孟鲁司特钠、布地奈德对哮喘小鼠气道重塑及肺组织MMP-9、TIMP-1表达的影响

目的观察孟鲁司特钠、布地奈德对哮喘小鼠气道重塑及肺组织基质金属蛋白酶-9(MMP-9)、基质金属蛋白酶抑制物-1(TIMP-1)表达的影响。方法将40只雌性小鼠随机分为4组各10只,模型组、孟鲁司特钠组、布地奈德组采用卵蛋白(OVA)致敏及OVA连续滴鼻激发建立哮喘模型,对照组予同等剂量生理盐水;在每次激发前30 min,孟鲁司特钠组予孟鲁司特钠0.25 mL/只(0.1 mg/mL)灌服,布地奈德组用1 mg布地奈德+8 mL生理盐水雾化吸入20 min,对照组以生理盐水进行腹腔注射及雾化吸入。HE染色观察小鼠肺组织病理变化,Image-Pro Plus6.0彩色图像分析系统测定支气管壁及平滑肌厚度,免疫组化法测定肺组织MMP-9和TIMP-1。结果对照组无支气管收缩表现,支气管组织结构正常;较对照组,模型组出现支气管收缩、黏膜充血水肿及炎症细胞浸润等情况较重,孟鲁司特钠组和布地奈德组较模型组明显减轻。与对照组比较,模型组、孟鲁司特钠组、布地奈德组支气管壁、平滑肌厚度增加,MMP-9、TIMP-1阳性率增高(P<0.05或0.01);与模型组比较,孟鲁司特钠组、布地奈德组支气管壁、平滑肌厚度减小,MMP-9、TIMP-1阳性率降低(P<0.05或0.01)。结论孟鲁司特钠和布地奈德可部分改善哮喘小鼠气道重塑并降低肺组织MMP-9和TIMP-1的表达。     

1,25-二羟维生素D3的免疫调节机理及其应用进展

1,25-二羟维生素D3[1,25(OH)2D3]不仅具有调节钙磷代谢的作用,还可影响多种细胞的生长和分化,具有调节免疫系统的作用.该文综述1,25(OH)2D3的免疫调节机制,为进一步开发更安全有效的类似物,并作为新兴的免疫调节剂应用于临床提供理论依据.     

1,25-二羟基维生素D3对糖尿病小鼠周围神经功能影响的研究

目的 探讨1,25-二羟基维生素D₃[1,25(OH)₂D₃]对DM小鼠周围神经功能的影响。方法C57BL/6J小鼠腹腔注射STZ构建DM小鼠模型,随机分为DM组(DM)、DM第1周1,25(OH)₂D₃治疗组(1w-DM)、DM第2周1,25(OH)₂D₃治疗组(2w-DM)、DM周围神经病变1,25(OH)₂D₃治疗组(DPN)以及DM第1周溶剂对照组(1w-DMC)、DM第2周溶剂对照组(2w-DMC)、DPN溶剂对照组(DPNC),每组各15只,并设置正常对照组(NC)10只。建模后第1、2、4周分别对1w-DM、2w-DM、DPN组予1,25(OH)₂D₃腹腔注射,连续5 d;监测各组血糖、体重、触觉、热痛觉及协调能力,8周后收集血清检测25(OH)D₃水平。结果 与DM组比较,1w-DM、2w-DM组第8周体重升高...     

MMP-9、TIMP-1分别对哮喘小鼠和气道重塑小鼠作用的比较研究

目的研究基质金属蛋白酶-9（matrix metalloproteinase-9,MMP-9）和基质金属蛋白酶抑制剂-1（tissue inhibitor matrix metalloproteinase-1,TIMP-1）在小鼠哮喘模型和气道重塑模型中表达作用的区别。方法30只BALB／c雌性小鼠按随机数字表法分为哮喘组（n=10）、气道重塑组（n=10）和生理盐水对照组（n=10）。通过鸡卵清白蛋白（ovabumin,OVA）滴鼻和腹腔注射的的方法分别建立小鼠哮喘模型和气道重塑模型。通过病理学观察气道炎症和气道重塑。用免疫组化法测定MMP-9和TIMP-1蛋白的表达;RT-PCR测定MMP-9和TIMP-1的基因水平表达。结果病理学显示哮喘组气道痉挛,大量炎性细胞浸润;气道重塑组呈现气道上皮指状增生,管腔内容物增多,上皮下纤维化。免疫组化结果显示哮喘组MMP-9为8868.8±3544.5,TIMP-1为4783±1508.1,二者比值为1.85;气道重塑组MMP-9为4383.1±2498.6,TIMP-1为6542.3±3026.6,气道重塑组MMP-9／TIMP-1的比值为0.67。二者有统计学意义（P〈0.01）。RT-PCR结果显示气道重塑组MMP-9 mRNA的表达低于哮喘组,而气道重塑组TIMP-1 mRNA的表达高于哮喘组。结论MMP-9主要在哮喘气道炎症中表达增高,而TIMP-1则在气道重塑过程中表达升高,二者的比例失调可能是哮喘的发病机制之一。     

咳喘宁对支气管哮喘大鼠气道重塑及MMP-9、TIMP-1的影响

目的 观察咳喘宁对支气管哮喘大鼠气道形态学和基质金属蛋白酶-9（MMP-9）、基质金属蛋白酶组织抑制剂-1（TIMP-1）的影响.方法 40只SD大鼠随机分为正常组、模型组、咳喘宁高、低剂量组（27 和13.5 g生药/kg体重）、桂龙咳喘宁胶囊对照组（0.41 g/kg体重）,每组8只.除正常组外以卵蛋白致敏并吸入激发法制备大鼠支气管哮喘模型,各治疗组均从第1次哮喘激发开始（造模第3周）至处死前每天灌胃给药,激发并给药4 w后处死大鼠,取肺组织HE染色,彩色图像分析仪测量支气管管壁厚度、平滑肌厚度,采用ELISA双抗体夹心法测定肺组织MMP-9、TIMP-1含量.结果 与正常组比较,模型组大鼠支气管管壁和平滑肌厚度、肺组织MMP-9、TIMP-1含量及二者比值明显增加（P＜0.01）;与模型组比较,各治疗组均可显著降低支气管管壁和平滑肌厚度（P＜0.01）,降低肺组织MMP-9、TIMP-1含量以及二者比值（P＜0.05或P＜0.01）;且咳喘宁高、低剂量组优于桂龙咳喘宁胶囊组（P＜0.05或P＜0.01）.结论咳喘宁可通过降低肺组织MMP-9和TIMP-1含量,调节二者比值,抑制支气管哮喘大鼠气道壁增厚和平滑肌增生肥大,从而抑制支气管哮喘大鼠气道重塑.     

H2型松弛素对慢性支气管哮喘小鼠气道重塑与肺组织细胞周期蛋白D1表达的影响

目的 研究H2型松弛素(H2Relaxin)对慢性支气管哮喘(简称哮喘)模型小鼠气道重塑及肺组织细胞周期蛋白D1 (cyclin D1)表达的影响,探素松弛素在哮喘治疗中的可能作用.方法 将40只BALB/c小鼠按随机数字表法分为健康对照组、哮喘组、阴性对照组(vehicle)、松弛素组4组,每组10只;卵清白蛋白(OVA)致敏、激发建立慢性哮喘小鼠模型;vehicle组和松弛素组每日分别给予生理盐水和松弛素(0.25 mg·kg-1 ·d-1)皮下注射.HE和马森(Masson)染色观察各组小鼠气道炎症及胶原沉积情况;采用免疫组织化学半定量法测定气道壁α-平滑肌肌动蛋白( α-SMA)的表达;酶联免疫吸附测定( ELISA)法检测肺组织羟脯氨酸含量;用逆转录-PCR(RT-PCR)及免疫印迹法(Western blot)分别测定各组小鼠肺组织中cyclin D1mRNA和蛋白表达水平.结果 哮喘组、vehicle 组与健康对照组相比,嗜酸粒细胞浸润增多,气道管腔狭窄,平滑肌层增厚,胶原纤维增生,而松弛素组上述改变较此2组轻.与健康对照组相比,哮喘组、vehicle组小鼠支气管α-SMA阳性染色面积/支气管基底膜周径显著增加(均P ＜0.05),松弛素组小鼠支气管α-SMA阳性染色面积/支气管基底膜周径低于哮喘组和vehicle组(均P＜0.05);哮喘组和vehicle组小鼠羟脯氨酸含量[每克肺组织中分别为(0.68±0.10)mg、(0.67±0.10) mg]高于健康对照组的(0.26±0.05)mg(q值分别为16.61和16.01,均P＜0.01),松弛素组小鼠羟脯氨酸含量为(0.40±0.06)mg,低于哮喘组和vehicle组(q值分别为10.88和10.26,均P＜0.05);Western blot检测结果显示,cyclin D1在哮喘组、vehicle组和松弛素组的表达(分别为1.38±0.18、1.50±0.10和0.72±0.13)高于健康对照组的0.38±0.10(q值分别为13.00、14.65和4.49,均P＜0.05),而松弛素组表达量低于哮喘组与vehicle组(q值分别为8.51和10.16,均P＜0.05).哮喘组与vehicle组各项检测指标间差异无统计学意义(P＞0.05).结论 松弛素可以显著减轻慢性哮喘小鼠气道炎症反应和平滑肌层的增厚,延缓哮喘小鼠气道重塑进程,该作用可能部分与其下调cyclin D1表达有关.     

Effect of 24,25-dihydroxyvitamin D3 on 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] metabolism in vitamin D-deficient rats infused with 1,25-(OH)2D3

Previous studies revealed that administration of 24,25-dihydroxyvitamin D3 [24,25-(OH)2D3] to calcium (Ca)-deficient rats causes a dose-dependent reduction in markedly elevated serum 1,25-(OH)2D3 level. Although the results suggested that the metabolism of 1,25-(OH)2D3 was accelerated by 24,25-(OH)2D3, those experiments could not define whether the enhanced metabolism of 1,25-(OH)2D3 played a role in the reduction in the serum 1,25-(OH)2D3 level. In the present study, in order to address this issue more specifically, serum 1,25-(OH)2D3 was maintained solely by exogenous administration through miniosmotic pumps of 1,25-(OH)2D3 into vitamin D-deficient rats. Thus, by measuring the serum 1,25-(OH)2D3 concentration, the effect of 24,25-(OH)2D3 on the MCR of 1,25-(OH)2D3 could be examined. Administration of 24,25-(OH)2D3 caused a dose-dependent enhancement in the MCR of 1,25-(OH)2D3, and 1 microgram/100 g rat.day 24,25-(OH)2D3, which elevated serum 24,25-(OH)2D3 to 8.6 +/- 1.3 ng/ml, significantly increased MCR and suppressed serum levels of 1,25-(OH)2D3. The effect of 24,25-(OH)2D3 on 1,25-(OH)2D3 metabolism developed with a rapid time course, and the recovery of iv injected [1 beta-3H]1,25-(OH)2D3 in blood was significantly reduced within 1 h. In addition, there was an increase in radioactivity in the water-soluble fraction of serum as well as in urine, suggesting that 1,25-(OH)2D3 is rapidly degraded to a water-soluble metabolite(s). Furthermore, the reduction in serum 1,25-(OH)2D3 was associated with a reduction in both serum and urinary Ca levels. Because the conversion of [3H]24,25-(OH)2D3 to [3H]1,24,25-(OH)2D3 or other metabolites was minimal in these rats, 24,25-(OH)2D3 appears to act without being converted into other metabolites. These results demonstrate that 24,25-(OH)2D3 rapidly stimulates the metabolism of 1,25-(OH)2D3 and reduces its serum level. It is suggested that 24,25-(OH)2D3 plays a role in modifying serum 1,25-(OH)2D3 concentrations by affecting the metabolism of 1,25-(OH)2D3 and may have a therapeutic values in the treatment of hypercalcemia or hypercalciuria caused by 1,25-(OH)2D3 excess.     

1,25-二羟维生素D3对被动致敏人气道平滑肌细胞的抑制作用

目的 检测1,25-二羟维生素D3[1,25-(OH)2D3]对被动致敏的人气道平滑肌细胞(HASMC)的增殖及其表达基质金属蛋白酶-9(MMP-9)和解整合素-金属蛋白酶33(ADAM33)的影响,探讨其调节支气管哮喘(简称哮喘)患者气道重塑的可能机制.方法用10%哮喘患者血清被动致敏HASMC,以10%非哮喘患者血清为对照.四甲基偶氮唑盐(MTT)法检测不同浓度1,25-(OH)2D3对HASMC细胞增殖活力的变化并确定其有效作用浓度.然后以有效作用浓度的1,25-(OH)2D3预处理HASMC,MTT法测定细胞增殖活力,流式细胞仪测定细胞周期,实时荧光定量PCR及蛋白免疫印迹法分别检测细胞中MMP-9及ADAM33的表达情况.结果 (1)1,25-(OH)2D3在(10-9～10-7)mol/L浓度下能浓度依赖性地抑制被动致敏的HASMC增殖(P<0.05);(2)10-7 mol/L的1,25-(OH)2D3能时间依赖性地抑制被动致敏的HASMC增殖并特异性抑制细胞周期中G1/S的转化;(3)VD组MMP-9及ADAM33蛋白表达较哮喘组分别下降了(63.4±3.6)%和(50.9±2.9)%,但仍显著高于对照组(P<0.01);(4)VD组MMP-9及ADAM33 mRNA表达较哮喘组分别下降了(52.2±2.5)%和(67.8±3.2)%,但仍显著高于对照组(P<0.01).结论 1,25-(OH)2D3能从多个层面抑制被动致敏的HASMC的功能,这可能是其调节哮喘气道重塑的作用机制之一.

Abstract：

Objective To investigate the effects of 1,25-(OH)2D3 on the proliferation of passively sensitized human airway smooth muscle cells(HASMCs) and their expressions of MMP-9 and a disintegrin and metalloprotease 33(ADAM33). Methods HASMCs were passively sensitized with 10% serum from asthmatic patients. MTT colorimetri assay was used to examine the effect of 1,25-(OH)2D3 on cell proliferation at different concentrations(10-10 mol/L, 10-9 mol/L, 10-8 mol/L, 10-7 mol/L).By this way, its optimal inhibitory concentration was determined. And then the effects of 1,25-(OH)2D3 at the optimal concentration on cell proliferation was examined by the same MTT assay and cell cycle analysis by flow cytometry. The expressions of MMP-9 and ADAM33 in HASMCs were studied by real-time quantitative RT-PCR and Western blotting analysis. Results (1)Inhibition of cell proliferation by 1,25-(OH)2D3 was barely detectable at 10-10 mol/L. But with the increasing concentration ranging from 10-9 mol/L to 10-7 mol/L, 1,25-(OH)2D3 markedly inhibited the cell proliferation concentration-dependently and reached the maximum effect at the concentration of 10-7 mol/L.Accordingly,10-7 mol/L was chosen as the optimal concentration of 1,25-(OH)2D3 for the following study. (2)At the concentration of 10-7 mol/L,1,25-(OH)2D3 inhibited the cell proliferation of passively sensitized HASMCs in a time-dependent manner and hampered the G1/S transition. (3)1,25-(OH)2D3 pretreatment attenuated the MMP-9 and ADAM33 protein levels in passively sensitized HASMCs by (63.4±3.6)% and (50.9±2.9)%,respectively (P<0.01). (4)1,25-(OH)2D3 significantly inhibited the MMP-9 and ADAM33 mRNA levels in passively sensitized HASMCs by (52.2±2.5)% and (67.8±3.2) %, respectively (P<0.01). Conclusion 1,25-(OH)2D3 has a direct inhibitory effect on passively sensitized HASMCs in vitro,including the inhibition of cell proliferation and the expressions of MMP-9 and ADAM33,which maybe associated with the beneficial role of 1,25-(OH)2D3 in the prevention and therapy of asthmatic airway remodeling.     

1,25-Dihydroxyvitamin D3 [1,25-(OH)2D3] increases insulin-like growth factor I (IGF-I) receptors in clonal osteoblastic cells. Study on interaction of IGF-I and 1,25-(OH)2D3

We previously reported a cooperative effect between insulin-like growth factor I (IGF-I) and 1,25-dihydroxy-vitamin D3 [1,25-(OH)2D3] in murine clonal osteoblastic cells, MCT3T3-E1. In the present study, the possible mechanism of interaction between these hormones was investigated. The effect of IGF-I on 1,25-(OH)2D3 receptors in MC3T3-E1 cells was examined. The affinity and hormone binding capacity of 1,25-(OH)2D3 receptors were not altered by IGF-I. Immunoblot analysis showed about 54 kilodaltons (kDa) 1,25-(OH)2D3 receptors, similar to that observed for mouse fibroblasts. The synthesis of IGF-I by the cells under a serum-free condition was determined by RIA. The assay revealed immunoreactive IGF-I secreted by MC3T3-E1 cells (1.79 +/- 0.04 x 10(-9) M, mean +/- SE, n = 5). Rat GH significantly increased the concentration of IGF-I, but 1,25-(OH)2D3 did not. IGF-I radioligand-receptor assay revealed specific binding of IGF-I to MC3T3-E1 cells. The relative potency of IGF-I-related peptides to bind with the cells was in the order of IGF-I much greater than multiplication-stimulating activity (the rat homologue of IGF-II) greater than insulin, and the receptor protein migrated as a 130-kDa band in autoradiography. Scatchard analysis showed a significant increase in IGF-I binding sites by 50% after 3-day treatment with 5 x 10(-11) M 1,25-(OH)2D3, without any change in affinity. These results indicate that the interaction of IGF-I and 1,25-(OH)2D3 in the culture of MC3T3-E1 cells may be mediated by the effect of 1,25-(OH)2D3 on IGF-I receptors.     

Effect of 1,25-(OH)2D3 (a vitamin D analogue) on passively sensitized human airway smooth muscle cells

BACKGROUND AND OBJECTIVES: In asthma, airway smooth muscle cell (ASMC) hyperplasia plays an important role in airway remodelling. Increased expression of matrix metalloproteinases-9 (MMP-9), a disintegrin and metalloprotease 33 (ADAM33) in ASMCs are also relevant to asthmatic airway remodelling. 1,25-dihydroxyvitamin D(3) (1,25-(OH)(2)D(3)) has potent antiproliferative properties in vitro in various cell types; however, its role in ASMCs is not well understood. This study investigated the effect of 1,25-(OH)(2)D(3) on passively sensitized human bronchial (airway) smooth muscle cell (HASMC) proliferation and MMP-9 and ADAM33 expressions. METHODS: The effect of 1,25-(OH)(2)D(3) on cell proliferation was examined by 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium-bromide colorimetry assay; cell cycle analysis by flow cytometry; and immunocytochemical staining for proliferating cell nuclear antigen (PCNA). The expression of MMP-9 and ADAM33 in HASMCs was investigated by real-time quantitative PCR and Western Blot analysis. RESULTS: 1,25-(OH)(2)D(3) effectively suppressed passively sensitized HASMC proliferation, proliferating cell nuclear antigen expression and G(1)/S transition in HASMCs passively sensitized with asthmatic serum. Further analysis showed that 1,25-(OH)(2)D(3) significantly down-regulated the expressions of protein for MMP-9 and ADAM33, as well as their mRNA levels in passively sensitized HASMCs. CONCLUSIONS: 1,25-(OH)(2)D(3) has direct inhibitory effects on passively sensitized HASMCs in vitro, including inhibition of cell proliferation and expression of MMP-9 and ADAM33, suggesting a possible beneficial role for 1,25-(OH)(2)D(3) in preventing and treating asthmatic airway remodelling.     

Effect of vitamin D2 and vitamin D3 on the serum concentrations of 1,25(OH)2D2, and 1,25(OH)2D3 in normal subjects

Serum concentrations of vitamin D2 and vitamin D3 metabolites were measured in 19 normal subjects before and during treatment with either vitamin D2 or vitamin D3, 4000 IU per day for 8 weeks. Vitamin D2 treatment increased the serum concentration of 1,25(OH)2D2, but a corresponding decrease in 1,25(OH)2D3 resulted in an unchanged serum concentration of total 1,25(OH)2D. During treatment with vitamin D3, the serum concentration of 1,25(OH)2D metabolites was unchanged. We conclude that the production of 1,25(OH)2D is tightly regulated and that 1 alpha-hydroxylase does not discriminate between D2 and D3 metabolites in normal subjects.     

血清25羟维生素D3或1,25二羟维生素D3水平与慢性阻塞性肺疾病关系的Meta分析

目的：系统评价血清25(OH)D3或1,25(OH)2 D3与 COPD 相关病例对照研究,进一步明确血清维生素 D 水平与 COPD 之间的关系。方法计算机检索 PubMed、EMBASE、The Cochrane Library、中国生物医学文献数据库(CBM)、中国知网(CNKI)数据库、万方数据库、维普数据库,并辅以文献追溯的方法,收集国内外发表的相关病例对照研究,检索时间均从建库至2015年11月。2位研究者按纳入排除标准筛选文献并评价纳入研究质量,应用 RevMan 5.2软件进行 Meta 分析。结果共纳入15篇病例对照研究,包含1948例 COPD患者及1549例健康对照者。25(OH)D3浓度水平结果：Meta分析结果表明无论是急性加重期 COPD还是稳定期 COPD,与健康对照组相比,其25(OH)D3浓度水平差异均有统计学意义,COPD组25(OH)D3浓度低于健康对照组。急性加重期组vs对照组,WMD=-14.28,95%CI =(-23.82~-4.73),Z=2.93,P <0.01;稳定期组vs对照组 WMD=-4.46,95%CI =(-7.36~-1.57),Z=3.02,P <0.01;急性加重期与稳定期相比,25(OH)D3浓度水平差异亦有统计学意义[WMD=-2.66,95%CI =(-4.19~-1.12), Z=3.38,P<0.01,见图2];1,25(OH)2 D3浓度水平结果：Meta分析结果表明无论是急性加重期COPD还是稳定期 COPD,与健康对照组相比,其1,25(OH)2 D3浓度水平差异均有统计学意义, COPD组1,25(OH)2 D3浓度低于健康对照组。急性加重期组vs对照组,WMD=-15.09,95%CI =(-17.97~-12.22),Z =10.3,P <0.01;稳定期组 vs 对照组,WMD=-9.62,95%CI =(-12.55~-6.70),Z=3.02,P<0.01。结论 COPD患者体内25(OH)D3与1,25(OH)2 D3浓度水平均显著降低,维生素D缺乏可能参与COPD的发生、发展,并影响COPD患者的预后。     

Contrasting effects of exogenous 1,25-dihydroxyvitamin D [1,25-(OH)2D] versus endogenous 1,25-(OH)2D, induced by dietary calcium restriction, on vitamin D receptors

The biological actions of 1,25-dihydroxyvitamin D [1,25-(OH)2D] are mediated by specific binding of the hormone with an intracellular vitamin D receptor, which ultimately regulates expression of genes within the target tissues. The quantity of vitamin D receptors varies between target tissues and within target tissues, depending on the physiological state of the animal. One factor that can modulate tissue vitamin D receptor content is 1,25-(OH)2D. In the present study performed in male rats, exogenous administration of 36 ng 1,25-(OH)2D3/day for 7 days increased plasma 1,25-(OH)2D concentrations 5-fold above those in control rats (to 261 +/- 17 pg/ml). Compared with those in control rats, 1,25-(OH)2D3 treatment resulted in a 1.5-fold increase in duodenal vitamin D receptor content (351 +/- 16 vs. 520 +/- 21 fmol/mg protein) and a 3-fold increase in renal vitamin D receptor content (60.3 +/- 5.2 vs. 193.8 +/- 22.7 fmol/mg protein). The effects of endogenously produced 1,25-(OH)2D on tissue vitamin D receptor content were studied by feeding rats either a 0.02% or 1% calcium diet for 2, 7, 14, or 21 days. Rats fed the low calcium diet exhibited plasma 1,25-(OH)2D concentrations similar to (day 7) or exceeding (days 14 and 21) those achieved by exogenous administration of 1,25-(OH)2D3, yet duodenal vitamin D receptor content was not up-regulated by dietary calcium restriction at any time point. The renal vitamin D receptor content of calcium restricted rats was 20-38% lower (P less than 0.05) than that in rats fed a calcium-replete diet 7, 14, and 21 days after initiation of the dietary treatments. These data suggest that under physiological conditions, increased plasma concentrations of 1,25-(OH)2D do not result in up-regulation of tissue vitamin D receptor concentrations, and that dietary calcium restriction must induce some factor(s) that results in down-regulation of vitamin D receptors in the kidney.     

细胞周期蛋白D1在支气管哮喘小鼠肺组织中的表达及其与气道重塑的关系

目的 研究细胞周期蛋白D1(Cyclin D1)在支气管哮喘(简称哮喘)小鼠肺组织中的表达,探讨Cyclin D1在哮喘及气道重塑中的作用.方法 将40只SPF级BALB/c小鼠按随机数字表法分为正常对照组(A组)、哮喘雾化2周组(B组)、哮喘雾化4周组(C组)、哮喘雾化8周组(D组)4组,每组10只.用10%鸡卵白蛋白(OVA)致敏和1%OVA激发小鼠建立哮喘模型.分析支气管肺泡灌洗液(BALF)中嗜酸粒细胞(EOS)计数及分类;用动物肺功能仪检测各组小鼠肺功能状况;用苏木精-伊红(HE)染色观察气道炎症及细胞浸润情况;用图像分析软件观察气道壁及平滑肌层变化情况;用逆转录-聚合酶链反应(RT-PCR)及实时定量(Real-time)PCR测定肺组织中Cyclin D1 mRNA水平表达变化;用Western blot法观察肺组织中Cyclin D1的蛋白表达变化.结果 BALF分析结果提示,B、C、D组EOS计数分别为(42.6±0.9)×104/L、(54.7±1.4)×104/L、(44.8±2.4)×104/L,与A组[(3.4±0.5)×104/L]比较差异有统计学意义(q值分别为79.75、91.42、84.82,P均＜0.01);对小鼠呼气阻力检测发现,乙酰胆碱浓度为45 μg/kg时B、C、D组分别为(5.27±0.16)cm·L-1·min-1、(6.68士0.20)cm·L-1·min-1、(7.14±0.41)cm·L-1·min-1,与A组[(4.11±0.15)cm·L-1·min-1]比较差异有统计学意义(q值分别为5.58、6.39、7.11,P均＜0.05);支气管平滑肌面积/管腔内周长(Wam/Pi)B组为2.8±0.6,C组为4.8±0.6,D组为6.4±0.7,与A组(2.4±0.4)比较差异有统计学意义(q值分别为6.40、8.28、9.27,P＜0.05);管壁面积/管腔内周长(Wat/Pi)B组为6.4±0.8,C组为8.3±1.2,D组为9.3±1.0,与A组(5.6±1.0)比较差异有统计学意义(q值分别为2.80、4.83、6.37,P均＜0.05);Western blot检测发现Cyclin D1在B、C、D组表达量分别为0.587±0.015、0.808±0.029、0.826±0.022,与A组(0.404±0.016)比较差异有统计学意义(q值分别为5.87、8.08、8.26,P均＜0.01);相关性分析结果提示呼气阻力和Cyclin D1水平表达呈正相关(r=0.83,P＜0.05).结论 Cyclin D1在哮喘小鼠肺组织中表达增加,其表达与气道反应性呈正相关,Cyclin D1可能通过细胞外信号调节激酶(ERK)信号通路参与气道重塑过程.