目的 研究木香烃内酯(CT)对生长期大鼠骨质量的影响.方法 1 月龄雌性 SD 大鼠 30 只,体质量(99±3)g,随机分为 3 组:对照组(Control),9 mg/(kg·d)的 CT处理组(CT-9)和 18 mg/(kg·d)的 CT处理组(CT-18),每组 10 只.适应性饲养 3 d后,灌胃给药,Control组每天灌服等体积蒸馏水,每周称一次体质量,每 4 周检测一次全身骨密度(BMD).8 周后安乐死所有大鼠,检测股骨和椎骨离体 BMD、进行 micro CT 扫描和骨生物力学实验.结果 各组大鼠实验过程中体质量未出现统计学意义,8 周时 CT-9 组和 CT-18 组的全身 BMD 及离体股骨、椎骨BMD均显著高于 Control组,CT-18 组的股骨弹性模量和最大载荷显著高于 Control组,CT-9 组和 CT-18 组的相对骨体积、骨小梁数和体积骨密度均显著高于 Control组,骨小梁分离度显著低于 Control 组.结论 CT 能提高生长期大鼠的骨质量,具有开发预防和治疗骨质疏松药物的潜力.

目的 研究仙灵骨葆胶囊对生长期大鼠骨强度的影响.方法 将 20 只 1 月龄 SPF级 SD雌性大鼠按随机数字表法分为对照组和仙灵骨葆组,每组 10 只.仙灵骨葆组每天灌服 378 mg/kg仙灵骨葆胶囊,对照组每天给予等体积蒸馏水.实验期间每两周测一次全身骨密度,待第 6 周仙灵骨葆组显著高于对照组后安乐死取材.取脏器、股骨、胫骨和椎骨进行病理学分析、骨密度检测、生物力学检测、micro CT 分析以及血清生化指标检测.结果 实验期间各组大鼠体质量呈上升趋势,两组间比较并无统计学意义,脏器无明显病理学改变;给药 6 周后仙灵骨葆组全身骨密度、股骨骨密度和椎骨骨密度都显著高于对照组(P<0.05);仙灵骨葆组股骨最大载荷和屈服强度显著高于对照组(P<0.05),椎骨最大载荷也显著高于对照组(P<0.05);仙灵骨葆组胫骨骨体积分数、骨小梁厚度和骨小梁数显著高于对照组(P<0.05),骨小梁分离度显著低于对照组(P<0.05);与对照组相比,仙灵骨葆组血清 OCN 含量上升、TRACP-5b含量下降,差异具有统计学意义(P<0.05).结论 仙灵骨葆胶囊可能通过抑制骨吸收与促进骨形成来提高生长期大鼠骨强度,具有通过提高生长期大鼠峰值骨量来预防后期骨质疏松的潜力.

骨质疏松症是目前老龄化社会急需解决的慢性骨科疾病,是一种以骨量丧失、骨微结构恶化为特征的全身性骨骼疾病,因此如何有效地预防和治疗骨质疏松症越来越受到人们的关注.槲皮素是自然界广泛存在天然黄酮醇类化合物,具有雌激素样作用,可抑制骨吸收和促进骨形成.槲皮素通过抑制破骨细胞、调控骨髓间充质干细胞的细胞机制,促进骨形成、抑制骨吸收的信号通路机制,以及雌激素信号途径改善骨质疏松.综述了槲皮素改善骨质疏松的作用机制,希望为槲皮素的临床应用提供参考.

It is known that low-frequency pulsed electromagnetic fields (PEMFs) can promote the differentiation and maturation of rat calvarial osteoblasts (ROBs) cultured in vitro. However, the mechanism that how ROBs perceive the physical signals of PEMFs and initiate osteogenic differentiation remains unknown. In this study, we investigated the relationship between the promotion of osteogenic differentiation of ROBs by 0.6 mT 50 Hz PEMFs and the presence of polycystin2 (PC2) located on the primary cilia on the surface of ROBs. First, immunofluorescence staining was used to study whether PC2 is located in the primary cilia of ROBs, and then the changes of PC2 protein expression in ROBs upon treatment with PEMFs for different time were detected by Western blotting. Subsequently, we detected the expression of PC2 protein by Western blotting and the effect of PEMFs on the activity of alkaline phosphatase (ALP), as well as the expression of Runx-2, Bmp-2, Col-1 and Osx proteins and genes related to bone formation after pretreating ROBs with amiloride HCl (AMI), a PC2 blocker. Moreover, we detected the expression of genes related to bone formation after inhibiting the expression of PC2 in ROBs using RNA interference. The results showed that PC2 was localized on the primary cilia of ROBs, and PEMFs treatment increased the expression of PC2 protein. When PC2 was blocked by AMI, PEMFs could no longer increase PC2 protein expression and ALP activity, and the promotion effect of PEMFs on osteogenic related protein and gene expression was also offset. After inhibiting the expression of PC2 using RNA interference, PEMFs can no longer increase the expression of genes related to bone formation. The results showed that PC2, located on the surface of primary cilia of osteoblasts, plays an indispensable role in perceiving and transmitting the physical signals from PEMFs, and the promotion of osteogenic differentiation of ROBs by PEMFs depends on the existence of PC2. This study may help to elucidate the mechanism underlying the promotion of bone formation and osteoporosis treatment in low-frequency PEMFs.

中老年人骨质疏松症( OP)不仅极大降低患者的生活质量,还严重威胁其生命安全,给社会带来了巨大的负担[1].因此寻找安全可靠的治疗方法尤为重要.中草药因不良反应小得到了国内外学者的重视及广大患者的肯定[2].杜仲被认为是防治OP的经典用药,《本草纲目》记载其具有"治腰膝痛,补中益精气,坚筋骨,强志"的功效[3].

Objective:To study the effects of a simulated plateau environment on fracture healing in rats.Methods:A rat model of mid-femoral fracture was established by hacksaw truncation and intramedullary fixation with Kirschner wires in 60 male Wistar rats which were divide into 2 groups ( n=30) by the random number table method. The rats in the control group were raised in the animal experiment center of The 940 Hospital of Joint Logistic Support Force of Chinese PLA at an altitude of 1,400 m, while the rats in the plateau group were placed in an animal experimental cabin in a simulated plateau environment at a simulated altitude of 5,000 m. The body weight was weighed once a week and X-ray films were taken every 2 weeks. Blood samples were collected after 4 weeks for detection of biochemical indicators of bone metabolism. After 8 weeks, the femurs of the surgical side were taken for bone biomechanical detection and the bone mineral density of the healthy side was detected. After 4 and 8 weeks, the femurs of the surgical side were taken for in vitro Micro-CT scanning and angiography detection. After 1, 2, 4 and 8 weeks, the femurs of the surgical side were taken for bone histopathologic detection. Results:During the entire experiment, no rats in the control group died while the mortality rate of the rats in the plateau group was as high as 26.7% (8/30). In the plateau group, some organs were pathologically damaged in the rats, fracture union was delayed, and the callus differentiated and matured slowly with the chondrocytes still dominant at the 8th week. The bone mineral density and the maximum load of the femur in the plateau group were significantly lower than those in the control group ( P< 0.05). Angiography showed that the rats in the plateau group had microvascular proliferation which did not penetrate the fracture end at the 8th week. The bone formation indexes like osteocalcin, procollagen type Ⅰ N-terminal propeptide (PⅠNP), and osteoprotegerin of the rats in the plateau group were significantly lower than those in the control group at the 4th week ( P<0.05). The bone resorption indexes like tartrate resistant acid phosphatase 5b (TRACP-5b) and receptor activator for nuclear factor-κB ligand (RANKL) in the plateau group were significantly higher than those in the control group ( P<0.05). Conclusion:A simulated plateau environment at an altitude of 5,000 m may lead to delayed fracture healing in rats.

Objective To determine the effect of pinoresinol diglucoside (PDG) on bone quality in growing rats and evaluate its potential as an anti-osteoporosis drug. Methods A total of 30 1-month-old female SD rats (body weight 107±7 g) were randomly divided into 3 groups: control group (Control), 25 mg/(kg·d) PDG treatment group (PDG-25) and 50 mg/(kg·d) PDG treatment group (PDG-50), with 10 rats in each group. The rats in the PDG-25 group and PDG-50 group were given corresponding doses of PDG intragastrically every day, and those of the control group was given equal volume distilled water. Their body mineral density (BMD) of the whole body was measured in 2, 4 and 6 weeks after treatment. In the end of 6 weeks, all rats were sacrificed, and organ coefficients of important organs were calculated. Micro-CT scanning, bone biomechanical test and serum biochemical tests of bone metabolism were performed. Results There were no significant differences in body weight and main organ coefficients among the groups. At 6 weeks, the BMD value was significantly higher in the PDG-25 group and PDG-50 group than the Control group (P < 0.05). The PDG-50 group had significantly higher relative bone volume, trabecular number and volumetric bone mineral density (P < 0.05), lower trabecular separation (P < 0.05) than the Control group, and the bone microstructure of PDG-25 group and PDG-50 group was significantly better than that of Control group. The PDG-50 group had elevated elastic modulus of femur (P < 0.05), larger maximum load of vertebrae (P < 0.05). The PDG-50 group had increased content of serum osteocalcin (P < 0.01), but obviously lower content of serum tartrate resistant acid phosphatase 5b (P < 0.01), when compared with the Control group. The content of serum osteocalcin in the PDG-25 group was significantly higher than that in the Control group (P < 0.05). Conclusion PDG can effectively improve bone mineral density, optimize bone microstructure, enhance bone strength, promote bone formation, inhibit bone resorption and improve bone quality.

目的:探究瞬时受体电位阳离子通道V亚家族成员4(transient receptor potential cation channel sub?family V member 4,TRPV4)在H9C2细胞中缺氧环境下的作用机制.方法:采用H9C2细胞建立缺氧损伤模型,检测缺氧不同时间细胞内乳酸脱氢酶(lactate dehydrogenase,LDH)释放率、丙二醛(malondialdehyde,MDA)含量、活性氧(reactive oxygen species,ROS)水平及过氧化氢酶(catalase,CAT)和超氧化物歧化酶(superoxide dismutase,SOD)活性.将细胞分为常氧组(0 h)、HC-067047(TRPV4抑制剂)预处理+常氧(0 h)组、缺氧(24 h)组和HC-067047预处理+缺氧(24 h)组,使用Fluo-4 AM检测细胞内Ca2+含量,JC-1检测线粒体膜电位,Western blot检测TRPV4蛋白水平,RT-qPCR检测TRPV4的mRNA水平,DCFH-DA检测ROS含量,酶标仪检测CAT和SOD活性,CCK-8法检测细胞活力.结果:与0 h组相比,随着缺氧时间的延长,LDH释放率提高(P<0.05),细胞内ROS和MDA含量上升(P<0.05),CAT和SOD活性增强(P<0.05),氧化损伤逐渐加重并在24 h达到顶峰.此外,缺氧后细胞出现钙超载现象,线粒体膜电位下降,TRPV4被激活.使用TRPV4抑制剂HC-067047预处理后,H9C2的氧化损伤明显被抑制,细胞活力上升(P<0.05),并且钙超载现象和线粒体膜电位下降均有所逆转.结论:在心肌细胞缺氧后,TRPV4被激活,Ca2+内流,线粒体膜电位受损,ROS增多,氧化损伤加重,抗氧化系统被激活,CAT和SOD活性上升,进而调控氧化与抗氧化平衡.

OBJECTIVETo explore whether the effect of low-frequency pulsed electromagnetic fields (PEMFs) in promoting osteoblast mineralization and maturation is related to the primary cilia, polycystin2 (PC2) and sAC/PKA/CREB signaling pathway.METHODSWe detected the expression levels of PC2, sAC, PKA, CREB and their phosphorylated proteins in primary rat calvarial osteoblasts exposed to 50 Hz 0.6 mT PEMFs for 0, 5, 15, 30, 60, 90, and 120 min. We blocked PC2 function with amiloride hydrochloride and detected the changes in the activity of sAC/PKA/CREB signal pathway and the mineralization and maturation of the osteoblasts. These examinations were repeated in the osteoblasts after specific knockdown of PC2 via RNA interference and were the co-localization of PC2, sAC, PKA, CREB and their phosphorylated proteins with the primary cilia were using immunofluorescence staining. The expressions of PC2 and the signaling proteins of sAC/PKA/CREB pathway were detected after inhibition of primary ciliation by RNA interference.RESULTSThe expression levels of PC2, sAC, p-PKA and p- CREB were significantly increased in the osteoblasts after exposure to PEMFs for different time lengths (P < 0.01). Blocking PC2 function or PC2 knockdown in the osteoblasts resulted in failure of sAC/PKA/CREB signaling pathway activation and arrest of osteoblast mineralization and maturation. PC2, sAC, p-PKA and p-CREB were localized to the entire primary cilia or its roots, but PKA and CREB were not detected in the primary cilia. After interference of the primary cilia, PEMFs exposure no longer caused increase of PC2 expression and failed to activate the sAC/PKA/CREB signaling pathway or promote osteoblast mineralization and maturation.CONCLUSIONPC2, located on the surface of the primary cilia of osteoblasts, can perceive and transmit the physical signals from PEMFs and promote the mineralization and maturation of osteoblasts by activating the PC2/ sAC/PKA/CREB signaling pathway.

目的 研究松脂醇二葡萄糖苷(pinoresinol diglucoside,PDG)对青年大鼠骨代谢的影响.方法 将实验大鼠按随机数字表达法分为Control组、PDG-25组和PDG-50组.PDG-25组和PDG-50组每天分别按25 mg·kg-1和50 mg·kg-1的剂量灌服PDG,Control组每天灌服等体积蒸馏水.6周后处死所有大鼠,取脏器、血清、股骨、胫骨以及椎骨,用于组织病理学检测、离体骨密度检测、Micro CT成像分析、血清生化指标检测、双荧光标记观察以及股骨骨组织蛋白表达分析.结果 各组大鼠主要脏器的组织病理学结果均未见明显异常;与Control组相比,PDG-25组和PDG-50组的离体骨密度均明显增高;Micro CT结果显示,PDG-50组的骨体积分数、骨小梁数以及骨小梁厚度均明显升高,但骨小梁分离度明显下降;PDG-25组和PDG-50组荧光标记间距明显增大;PDG-25组和PDG-50组血清OPG含量明显提高,但RANKL含量明显降低;PDG-25组和PDG-50组BMP-2、Runx-2、OSX和OPG表达量明显增多,RANKL表达量明显减少.结论 25 mg·kg-1和50 mg·kg-1的PDG可能通过促进骨形成和抑制骨吸收来提高青年大鼠骨质量与骨强度.

Osteoporosis is defined as a systemic skeletal disease characterised by low bone mass and microarchitectural deterioration of bone tissue, with a consequent increase in bone fragility and susceptibility to fracture. Nowadays more than 200 million individuals are suffering from osteoporosis and still the number of affected people is dramatically increasing due to an aging population and longer life, representing a major public health problem. At present, great progress has been made in the development of drugs for the treatment of osteoporosis, but drug treatment can not reverse bone loss, and will bring a series of toxic and side effects to patients. A large number of studies have shown that homing disorders, osteogenic differentiation and cytokine effect of bone marrow mesenchymal stem cells play an important role in the pathogenesis of osteoporosis. As a new method, transplantation of bone marrow mesenchymal stem cells can not only avoid the side effects of drug therapy, but also fundamentally treat osteoporosis. It has great potential and application value, but there are also many problems to be solved.