1,25(OH)2D3抑制嘌呤霉素氨基核苷酸肾病大鼠足细胞凋亡

目的 观察1，25（OH）2D3对嘌呤霉素氨基核苷酸（PAN）肾病大鼠足细胞凋亡的影响。 方法 72只雄性SD大鼠随机分为健康对照组（NC）、PAN组和1，25（OH）2D3治疗组 [1，25（OH）2D3 0.2 μg·kg-1·d-1灌胃]。一次性尾静脉注射PAN 100 mg/kg体质量建立足细胞损伤的PAN肾病动物模型。于3、7、14、21 d分批处死动物，分别检测不同时间点尿蛋白量（24 h）和肾功能。光镜和透射电镜观察肾组织学改变。TUNEL法检测足细胞凋亡。RT-PCR、免疫荧光、免疫组化分别检测nephrin、TGF-β1 mRNA和蛋白的表达。Western印迹检测磷酸化（p）-Smad2/3的表达。 结果 （1）PAN组各时间点BUN、Scr、尿蛋白量（24 h）[7 d时，（20.26±4.87） mg比（1.01±0.41） mg，P < 0.01]均高于同期的NC组，而肾小球足细胞显著减少[14 d时，(10.9±4.2) 个/肾小球切面比(31.9±6.2)个/肾小球切面，P ＜ 0.01]，且足突增宽融合。1，25（OH）2D3治疗组各时间点尿蛋白量（24 h）[7 d时（9.95±3.82） mg]和BUN、Scr显著低于PAN组（P < 0.05），且肾脏病理改变减轻。（2）PAN组7 d时nephrin mRNA和蛋白的表达显著降低，nephrin由正常的沿毛细血管襻线状分布向颗粒状、团快状改变，足细胞凋亡数显著增加[14 d时，（37.4±7.9）个/肾小球切面]。与PAN组相比，1，25（OH）2D3治疗组各时间段nephrin mRNA和蛋白的表达显著增加，且保持着正常的沿毛细血管襻线状分布，足细胞凋亡数显著减少[14 d时，(21.9±6.2) 个/肾小球切面，P < 0.01]。（3）PAN组TGF-β1 mRNA和蛋白的表达以及p-Smad2/3蛋白的表达均高于NC组（P < 0.01），1，25（OH）2D3治疗组TGF-β1 mRNA和蛋白的表达以及p-Smad2/3蛋白的表达低于PAN组（P < 0.01）。 结论 1，25（OH）2D3能有效地抑制PAN诱导的足细胞凋亡，减少尿蛋白，其对足细胞损伤的保护作用可能与抑制TGF-β1信号通路有关。     

骨化三醇联合仙灵骨葆治疗女性骨质疏松症的临床研究

目的通过骨化三醇联合仙灵骨葆、阿仑膦酸钠分别治疗女性症状型骨质疏松症患者,对比骨密度、疼痛感觉、碱性磷酸酶指标改善情况观察其疗效,考察骨化三醇联合仙灵骨葆对骨质疏松症具体临床效果,提供临床参考。方法选取河北省徐水县原发性骨质疏松症女性患者90人,随机分为两组,每组45人,分别使用骨化三醇胶丸＋仙灵骨葆胶囊（A组）,阿仑膦酸钠（B组）治疗6个月,对比骨密度、疼痛感觉、碱性磷酸酶指标改善情况。结果A组完成治疗38人,B组完成治疗36人;两组治疗6个月后骨密度、疼痛感觉、碱性磷酸酶指标均明显改善（P〈0．05）,两组比较差异无统计学意义（P〉0．05）,两组个别患者出现药物不良反应,均程度较轻,停药后症状可逆,但是联合用药组有更好的患者依从性。结论骨化三醇胶丸联合仙灵骨葆胶囊治疗女性骨质疏松症有良好疗效。     

钙三醇干预甲状旁腺素诱导的心肌细胞内钙超载和细胞肥大

目的探讨甲状旁腺素（PTH）对鼠心肌细胞内游离钙（[Ca^2＋]i）和心肌肥大的影响及其机制,以及钙三醇的干预作用。方法培养的新生大鼠心肌细胞以Fluo-3／AM负载,通过激光共聚焦显微术（LSCM）测定细胞内[Ca^2＋]i;以细胞面积和细胞蛋白含量作为心肌细胞肥大指标,体外实验观察PTH瞬时和持续刺激对鼠心肌细胞[Ca^2＋]i和肥大的影响,以及钙三醇的干预作用。结果①在细胞外Ca^2＋为2．5mmol／L时,PTH1-34在0．1和1μmol浓度下的刺激可促使心肌细胞静息[Ca^2＋]i荧光强度（FI）快速上升;1μmol浓度PTH1-34刺激,在细胞外液无钙,或应用10μmol硝苯地平预处理时,心肌细胞静息钙则无明显上升。②培养的心肌细胞应用PTH1-34 0．01和0．1μmol刺激7天后,心肌细胞内钙荧光强度、心肌细胞面积和蛋白含量均较对照组显著增加。若以PTH1-34 0．1μmol刺激,并同时加入0．001μmol的钙三醇干预,上述指标明显降低（P〈0．01）;但应用0．1μmol高浓度钙三醇干预,上述指标则未见显著改善。结论PTH1-34刺激可显著增加培养心肌细胞[Ca^2＋]i,诱导细胞肥大;适宜浓度钙三醇具有保护作用,电压依赖型钙通道的开放引起的细胞外钙内流增加是PTH1-34诱导上述变化的重要机制之一。     

血液透析滤过串联血液灌流联合骨化三醇冲击治疗对血透患者继发性甲状旁腺功能亢进的临床疗效

目的探讨血液透析滤过串联血液灌流联合骨化三醇冲击治疗对维持性血液透析患者并发继发性甲状旁腺功能亢进的临床疗效。 方法选取2019年01月至2019年06月60例维持性血液透析并发继发性甲状旁腺功能亢进的患者作为研究对象，其中男性42例，女性18例；年龄20～68岁，平均(52±17.2)岁。对照组30例，每周常规血液透析3次，联合骨化三醇冲击治疗；研究组30例，每周常规血液透析1次，血液透析滤过1次，血液透析滤过＋血液灌流1次，联合骨化三醇冲击治疗。观察两组患者血钙、血磷、血甲状旁腺素、碱性磷酸酶(ALP)等各项指标变化，并对两组患者的临床症状进行比较分析。 结果两组患者血甲状旁腺素均降低(P<0.05)；研究组患者的血甲状旁腺素、血磷、碱性磷酸酶明显低于对照组(P<0.05)；对照组治疗后血钙水平上升，较治疗前差异有统计学意义(P<0.05)；研究组治疗前后血钙水平差异无统计学意义(P>0.05)。 结论对维持性血液透析继发甲状旁腺功能亢进患者应用血液透析滤过串联血液灌流联合骨化三醇冲击治疗不仅能有效降低血甲状旁腺素、血磷、碱性磷酸酶水平，不影响血钙水平，且更能改善患者临床症状，提高生活质量。     

Hypocalcemia reduces insulin turnover but not insulin-mediated glucose metabolism in piglets

Disturbances of glucose metabolism similar to type II diabetes are often accompanied by low extra cellular calcium concentrations, as shown in periparturient ruminants. Hypocalcemia reduces insulin secretion and possibly endogenous glucose production, but whether hypocalcemia also affects peripheral actions of insulin is not clear. The problem was investigated with piglets in this study. Insulinmediated turnover of glucose and the clearance of insulin were measured in 7 control piglets (+D) and in 10 piglets with inherited calcitriol deficiency (−D). The measurements involved continuous intravenous infusions of insulin (1, 2, 5, 7.5 and 10 mU·kg⁻¹·min⁻¹) and glucose to maintain a glucose concentration of 7 mmol · l⁻¹ (combined insulin and glucose clamps). Both groups of piglets were studied under normo- and hypocalcemic conditions. During hypocalcemia and with equal insulin infusion rates, −D and +D piglets attained significantly higher (28%–133%) steady state insulin concentrations in plasma than during normocalcemia. This showed that the clearance rates of insulin during hypocalcemia were 48% lower in +D and 20% lower in −D piglets than during normocalcemia. With equivalent insulin infusion rates +D piglets developed significantly higher (15%–48%) steady state insulin concentrations in plasma than −D piglets, indicating that the clearance rate of insulin was higher in the calcitriol-deficient state. The relationship between insulin concentration in plasma and glucose turnover (measured with [3-³H]glucose) was not influenced by the calcemic and vitamin D status of the piglets. This indicated that the peripheral action of insulin on glucose metabolism was not influenced by the extracellular calcium concentration and vitamin D status. Received: March 2001 / Accepted in revised form: July 2002 Correspondence to C. Schlumbohm     

Interferon Gamma,but not Calcitriol Improves the Osteopetrotic Phenotypes in ADO2 Mice

ADO2 is a heritable osteosclerotic disorder that usually results from heterozygous missense dominant negative mutations in the chloride channel 7 gene (CLCN7). ADO2 is characterized by a wide range of features and severity, including multiple fractures, impaired vision due to secondary bony overgrowth and/or the lack of the optical canal enlargement with growth, and osteonecrosis/osteomyelitis. The disease is presently incurable, although anecdotal evidence suggests that calcitriol and interferon gamma‐1b (IFN‐G) may have some beneficial effects. To identify the role of these drugs for the treatment of ADO2, we utilized a knock‐in (G213R mutation in Clcn7) ADO2 mouse model that resembles the human disease. Six‐week‐old ADO2 heterozygous mice were administered vehicle (PBS) or calcitriol or IFN‐G 5 times per week for 8 weeks. We determined bone phenotypes using DXA and μCT, and analyzed serum biochemistry and bone resorption markers. ADO2 mice treated with all doses of IFN‐G significantly (p<0.05) attenuated the increase of whole body aBMD and distal femur BV/TV gain in both male and female compared to the vehicle group. In contrast, mice treated with low and medium doses of calcitriol showed a trend of higher aBMD and BV/TV whereas high dose calcitriol significantly (p<0.05) increased bone mass compared to the vehicle group. The calcium and phosphorus levels did not differ between vehicle and IFN‐G or calcitriol treated mice; however, we detected significantly (p<0.05) elevated levels of CTX/TRAP5b ratio in IFN‐G treated mice. Our findings indicate that while IFN‐G at all doses substantially improved the osteopetrotic phenotypes in ADO2 heterozygous mice, calcitriol treatment at any dose did not improve the phenotype and at high dose further increased bone mass. Thus, use of high dose calcitriol therapy in ADO2 patients merits serious reconsideration. Importantly, our data support the prospect of a clinical trial of IFN‐G in ADO2 patients.     

Effect of Rocaltrol on Bone Mass in Patients with Pulmonary Disease Treated with Corticosteroids

The aim of this study was to evaluate the effect of calcitriol on bone mass in patients with corticosteroid induced osteoporosis. Thirty-seven patients (26 females, 11 males, mean age 66.4 years) with pulmonary disease under long-term treatment with corticosteroids (5–10 mg prednisolone daily) and osteopenia/osteoporosis verified by dual-energy x-ray absorptiometry (DEXA) measurement were enrolled into the study. Rocaltrol was prescribed to 30/37 of the patients, the rest of the patients (6 females, 1 male) served as controls. In the treatment group, there was a slight increase of bone mass in the hip and lumbar vertebrae (L1–L4), whereas the control group showed a decrease of bone mass (change rate of bone mass in patients +0.8% and +1.0%, respectively, vs. –1.9% and –0.3%, respectively, in the control group). The preliminary results of our study suggest a beneficial role for the treatment of corticosteroid induced osteoporosis with Rocaltrol, which is well-tolerated by patients and cost-efficient in patient management.     

绝经后妇女骨密度与血清1，25-（OH）2D3，25-OHD3浓度关系的研究

目的：研究绝经后妇女无优势手臂骨密度与血清1,25-（OH）2D3,25-OHD3含量的关系。方法：采用定量超声骨量分析系统（QUS）测量82例绝经后妇女无优势前臂远端骨密度（BMD）以诊断骨质疏松,采用放射免疫法（RIA）和ELISA检测骨质疏松组和骨量正常组妇女血清中1,25-（OH）2D3,25-OHD3水平。结果：绝经后骨质疏松组妇女血清1,25-（OH）2D3,25-OHD3含量分别为（26.97±6.78）pg/ml和（34.75±12.62）nmol/L,骨量正常组分别为（34.75±12.62）pg/ml和（42.03±12.63）nmol/L,骨质疏松妇女血清1,25-（OH）2D3、25-OHD3含量明显低于骨量正常组（P〈0.01）。绝经后妇女无优势前臂骨密度与血清1,25-（OH）2D3含量存在明显相关性（r=0.387,P=0.001）,25-OHD3含量与前臂骨密度也存在相关性。结论：血清1,25-（OH）2D3水平与无优势前臂骨密度明显相关,是绝经后妇女骨量降低的重要原因之一。     

Monocyte fructose 1,6-bisphosphatase and cytidine deaminase enzyme activities: potential pharmacodynamic measures of calcitriol effects in cancer patients

Purpose: To determine, in peripheral blood monocytes (PBM), whether the enzymatic activities of fructose 1,6-bisphosphatase (FBPase), cytidine deaminase (CDDase) and 24-hydroxylase (CYP24), enzymes regulated by calcitriol are useful pharmacodynamic (PD) measures of calcitriol effects in cancer patients. Methods: Cancer patients enrolled in a phase I clinical trial of calcitriol and carboplatin were studied. Baseline and calcitriol-induced changes in FBPase, CDDase and CYP24 activities were measured in PBM collected before, 6, 24, and 48 h after administration of calcitriol, prior to carboplatin, in doses ranging from 4 to 11 μg daily for 3 consecutive days (QD×3). Normal FBPase, CYP24 and CDDase activities were measured in PBM from untreated healthy volunteers. Results: Baseline activities in PBM from cancer patients and healthy volunteers were (median and range): 1.0 (0.0–43.5) and 4.4 (3.1– 8.2) nmol/min/mg protein for FBPase (P = 0.002); 2.5 (0.9–9.3) and 0.8 (0.4–2.0) fmol/h/10⁶ cells for CYP24 (P = 0.016), and 5.6 (2.5–22.3) and 6.6 (1.1–47.4) nmol/min/mg protein for CDDase (P > 0.05), respectively. All calcitriol doses achieved peak serum calcitriol levels > ×3 the physiological levels, increased cancer patient PBM FBPase activity to normal levels and decreased CDDase activity to undetectable levels within 48 h, with no significant change in CYP24 activity. These enzyme activity changes were not associated with hypercalcemia. Conclusions: Calcitriol treatment-induced increase in FBPase and decrease in CDDase activities in cancer patient PBM are potential early and sensitive non-hypercalcemia PD measures of calcitriol effects.