目的 探究维生素D(vitamin D,VD)缺乏经Hippo-YAP信号通路抑制胎盘的发育,进而导致胎儿宫内生长受限的发生机制.方法 按照体质量将4w龄SD雌性大鼠随机分为VD缺乏组(vitamin D deficiency,VDD)与对照组(CTRL),VDD组采用VD缺乏饲料喂养,CTRL组采用标准饲料喂养,喂养8周后雌雄合笼.结果 妊娠13d(GD13)与 GD18 时,VDD 组孕鼠的血清 25(OH)D 浓度分别为(2.82±0.18)ng/ml 和(14.67±7.92)ng/ml,对照组孕鼠的血清25(OH)D浓度分别为(36.58±10.58)ng/ml和(19.92±6.91)ng/ml,组间比较在GD13时有差异(P＜0.0001).GD13 时,VDD 组孕鼠的胚胎着床数(12.83±1.17)显著低于 CTRL 组的(15.83±1.47)(P＜0.01);GD18 时,VDD组孕鼠胚胎着床数(12.83±2.23),显著低于CTRL组的(15.67±1.03)(P＜0.05),VDD 组孕鼠的胎儿流产率也明显上升.GD18时,VDD组孕鼠的胎盘重量平均为(2.93±0.42)g,显著低于CTRL组的(6.38±0.48)g(P＜0.0001);VDD组孕鼠的胎盘海绵体滋养层细胞面积为(11.63±0.03)％,显著小于CTRL组的(22.63±0.01)％(P＜0.05);VDD组的活胎数,孕鼠孕期增重显著降低(P＜0.01或P＜0.0001),吸收胎数显著增加(P＜0.05).GD13时,与CTRL组相比,VDD组孕鼠胎盘组织中YAP1蛋白表达水平显著降低(P＜0.01),P-YAP蛋白表达水平显著增高(P＜0.001).结论 母体VD缺乏是导致妊娠后胚胎发育迟缓的重要原因,其机制可能涉及Hippo-YAP信号通路的激活,在孕前及孕期合理补充VD对胎儿健康具有重要意义.

ObjectiveTo investigate the effect of iron on serum 25-hydroxyvitamin D3 (25-(OH)D3), 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) and the expression of vitamin D receptor (VDR) in rat kidney. Methods With 7 days′ adaptive feeding, thirty newly weaned male Sprague-Dawley (SD) rats were randomly divided into a control group (n = 6) and a model group (n = 24) according to body weight. The rats in the control group were fed with normal diet and those in the model group were fed with iron deficiency diet for 6 weeks. After successful modeling, the rats in model group were randomly divided into four group (6 rats in each group) with iron deficiency, and low, moderate and high iron according to hemoglobin (Hb) content. The rats in the control group and the iron deficiency group were given normal saline, and the rats in the low, moderate and high iron groups were given iron dextran at dosages of 11, 33 and 99 mg/kg by gastric gavage, respectively. After 4 weeks, the rats were anesthetized with 8% chloral hydrate. Blood samples of the rats were collected from the abdominal aorta for serum isolation and kidney tissue specimens were collected and stored at – 80 ℃ for later detections. Serum 25-(OH)D3 and 1,25-(OH)2D3 were detected with enzyme-linked immunosorbent assay (ELISA) kit method. Renal tissue iron content of the rats was measured with biochemical kit method. Protein expression of vitamin D receptor (VDR) in the rats′ kidney was determined with Western blot and immunohistochemistry method. Results The model rats′ Hb was significantly lower than that of the control rats (P < 0.05); the Hb of the rats in low, moderate and high iron groups were significantly increased compared to that of the rats in the iron deficiency group (all P < 0.05). Significantly decreased serum 25-(OH)D3/1,25-(OH)2D3 and VDR in kidney were detected in the rats of iron deficiency group contrasting to those of the rats in control group (P < 0.05 for all). In the rats of low, moderate and high iron groups, serum 25-(OH)D3 and 1,25-(OH)2D3 increased significantly (P < 0.05 for all) and with the increases of serum 25-(OH)D3 and 1,25-(OH)2D3, the VDR in kidney was gradually up-regulated in comparison with those in the rats of iron deficiency group. ConclusionIn rats, iron content affects the activation of VD3 and the expression of VDR protein in kidney.

目的 探讨维生素D(vitamin D,VD)与铁联合对脂多糖(lipopolysaccharide,LPS)所致的结肠炎性细胞的影响及作用机制.方法 将SW480细胞随机分为对照组(control),模型组(model,1μg/ml LPS),25(OH)D3干预组[1 p g/ml LPS+500 ng/ml 25(OH)D3],铁干预组[1 μ g/ml LPS+1 mmol/L 柠檬酸铁铵(ferric ammonium citrate,FAC)],联合干预组[1μg/ml LPS+500 ng/ml25(OH)D3+1 mmol/L FAC],干预24h后收集细胞,免疫荧光检测各组细胞活性氧(reactive oxygen species,ROS)水平;酶联免疫吸附试剂 盒(enzyme linked immunosorbent assay,ELISA)检测细胞IL-6、TNF-α、1,25(OH)2D3水平;western blots法检测各组细胞p38、p-p38、CYP27B1和CYP24A1蛋白表达水平.结果 与对照组相比,模型组的ROS、IL-6和TNF-α 水平明显增加(P＜0.05),p38蛋白磷酸化水平也明显增加(P＜0.05),CYP27B1蛋白表达水平增加,CYP24A1表达降低,25(OH)D3+FAC联合干预组的1,25(OH)2D3水平增加(P＜0.05);与模型组比较,25(OH)D3+FAC联合干预组的ROS、IL-6和TNF-α 水平明显降低(P＜0.05),p38蛋白磷酸化水平与CYP27B1蛋白表达明显降低(P＜0.05),CYP24A1表达增加.结论 维生素D与铁联合能够有效降低LPS所致的结肠炎性细胞的炎症水平,该过程可能是由p38丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)信号通路介导的.

目的 探讨铁和维生素D(VD)对孕期SD大鼠的贫血是否存在交互作用.方法 24只雌性SD大鼠适应性喂养7d后,按体质量随机分为正常组(+Fe+VD:Fe=181.6 mg/kg,VD3=2250 IU/kg,)、缺维生素D组(+Fe-VD:Fe=181.6 mg/kg,VD3=0 IU/kg,)、缺铁组(-Fe+VD:Fe=24.2 mg/kg,VD3=2250 IU/kg,)、缺铁缺维生素D组(-Fe-VD:Fe=24.2 mg/kg,VD3=0 IU/kg)4组,每组6只.8w后,雌雄鼠1:1合笼.孕鼠GD18d时,用2％戊巴比妥钠溶液进行腹腔注射麻醉,腹主动脉取血.结果 血清铁(serum iron,SI)和血清铁蛋白(serum ferritin,SF)在+Fe+VD、+Fe-VD、-Fe+VD、-Fe-VD组依次降低;血清可溶性转铁蛋白受体水平(serum soluble transferrin receptor,sTfR)在各组依次升高;与+Fe+VD组相比,-Fe-VD组的血清25-(OH)D3水平较低(P<0.01);与+Fe+VD组相比,其他各组血红蛋白(hemoglobin,Hb)、红细胞数目(number of red blood cells,RBC)、红细胞压积(hematocrit,HCT)、平均红细胞体积(mean red blood cell volume,MCV)、平均红细胞血红蛋白含量(average red blood cell hemoglobin content,MCH)等的水平均较低(P<0.05);与+Fe+VD组相比,其他各组的红细胞分布宽度变异系数(coefficient of variation of red blood cell distribution width,RDW)水平均较高(P<0.001).铁和维生素D3交互作用在Hb、RBC、HCT、RDW、MCV、MCH、MCHC水平差异均有统计学意义(P<0.05).结论 在妊娠期贫血发展进程中,铁与维生素D3可能存在交互作用.

Introduction: Maternal vitamin D deficiency (VDD) is associated with intrauterine growth restriction (IUGR), but the exact mechanism remains unclear. Here we explored the mechanism through which VDD induced IUGR.Methods: Female SD rats were fed a control normal diet (VD > 800 IU/Kg) or VDD diet (VD: 0 IU/Kg) for 8 weeks. Then, females were mated with 12-week-old male SD rats, and fetal and placental tissue were collected on the gestational day 13 (GD13) or 18 (GD18) to analyze the effects of VDD on pregnancy outcome and embryonic development. In vitro, the VDR gene of HTR-8/SVneo cells was knocked down to establish VDD model. Then, HTR-8/SVneo cells were treated with the MST1/2 inhibitor XMU-MP-1 or 0.1 mu M/L calcitriol for 24 h (h). The mechanism of Hippo-YAP signaling pathway in VDD-induced placental dysplasia was further investigated by western blot, invasion assay, wound healing assay and Hoechst/PI staining. Results: The IUGR of the pregnant rats in the VDD group was significant, the placental structure and function were damaged, and there was an obvious inflammatory response, accompanied by a significant increase in the level of the transcription co-activator YAP phosphorylation. In vitro, VDD significantly inhibited the migratory and invasive abilities of HTR-8/SVneo cells, accompanied by decreased EMT capacity and increased apoptosis. When intervening with XMU-MP-1 in advance, we found that the effects of VDD were neutralized by Hippo-YAP signaling blocker.Discussion: Maternal VDD causes placental dysplasia and IUGR, and these abnormal changes may be associated with the activation of Hippo-YAP signaling pathway.

目的 探讨血清维生素C(vitamin C,VC)和维生素D(vitamin D,VD)水平不同的溃疡性结肠炎(ulcerative colitis,UC)患者中肠道菌群、炎症因子及氧化应激的差异.方法 收集2016年12月至2019年1月在山西医科大学第一附属医院及同煤集团总医院确诊的50例UC患者,根据血清25(OH)D3水平将其分为非VD缺乏组(≥20ng/ml)和VD缺乏组(<20ng/ml),比较两组间VC、肠道菌群、炎症因子以及氧化应激指标的差异.结果 非VD缺乏组VC水平、抗氧化物质SOD均高于VD缺乏组;粪便变形菌门、血清促炎因子TNF-α和IL-1β水平低于VD缺乏组.结论 UC患者血清中VC的水平可能影响VD的水平,充足的VD和VC可维持肠道菌群稳定性、抑制促炎因子产生和增加UC患者的抗氧化能力.

Abstract Background Iron and vitamin D (VD) is essential to health. Previous studies have shown that iron homeostasis has a potential effect on VD metabolism, but the mechanism is not fully understood. Objectives To explore the relationship between VD metabolism and iron metabolism, as well as the regulatory mechanism of iron on VD metabolism. Methods 40 male rats were fed adaptively for 7 days and randomly divided into control (C, n = 6 normal diet) group and model (M, n = 24 iron deficient diet) by simple randomization, the latter was used to establish iron deficiency anemia (IDA) model. After 6 weeks of feeding, the M group was randomly divided into: iron deficiency group (DFe), low iron group (LFe), medium iron group (MFe) and high iron group (HFe) by block randomization. Different doses of iron dextran (based on iron content (100 g·bw·d)): 0, 1.1, 3.3 and 9.9 mg) were given respectively. After 4 weeks, the rats were anesthetized with 8% chloral hydrate, Blood (collected from the abdominal aorta), liver and kidney tissues were collected. The serum and tissues were separately packed and frozen at -80℃ for testing. Results The results showed that the levels of hemoglobin (Hb), red blood cell (RBC), serum iron (SI), liver iron, and kidney iron in DFe group were lower than those in the other four groups, while the levels of total iron-binding capacity (TIBC), transferrin (TF) and transferrin receptor (Tfr) in DFe group were higher than those in other groups; The serum levels of 25-(OH)D3 and 1,25-(OH)2D3 in DFe group were significantly lower than those in C group (P < 0.05). The correlation analysis showed that the levels of 25-(OH)D3 and 1,25-(OH)2D3 were negatively correlated with TIBC, TF and Tfr no correlation with SI. Western blotting, immunofluorescence, and q-PCR results showed that compared with C group, the protein and gene expressions of CYP2R1, CYP27A1, and CYP24A1 in DFe group were down-regulated, and the expression of CYP27B1 protein and gene was up-regulated in DFe group. Conclusion Iron may be involved in the metabolism of VD3 by regulating the expression of VD3 hydroxylase, suggesting that appropriate iron supplementation might promote the activation of VD3.