头低位卧床对恒河猴骨代谢、糖脂代谢的抑制效应及其相关性分析

空间失重引起的骨代谢调节失衡是航天员遭受的最严重的危害之一.骨代谢失衡还有可能影响机体的糖脂代谢平衡.本研究利用恒河猴头低位卧床模拟失重效应实验方法,分析头低位卧床过程中恒河猴血清中骨代谢、糖脂代谢指标变化情况及其相关性.卧床组恒河猴血清中BAP在头低位卧床实验开始7天便出现了显著下降（P<0.05）,血清胰岛素、高密度脂肪酸含量在7天显著下降并一直维持在较低水平,血糖含量在7天时显著下降,但在21天时明显回升.分析发现,骨钙素与血糖、皮质醇、高密度脂肪酸含量间均存在相关性,这表明头低位卧床模拟失重效应实验中骨与糖脂代谢之间存在相互调控.      

模拟失重对心血管自主神经调节功能的影响

载人航天或模拟失重后，航天员会出现运动能力与立位耐力降低，其发生机理与多种因素的改变有关．为进一步验证这种变化与中枢神经系统调节功能障碍的可能相关，本文研究模拟失重过程中心脏与外周血管自主神经调节功能的动态变化及与卧床后立位耐力降低的关系．结果表明，6名被试者的HRV谱的总功率（TP）及低频（LF）、高频（HF）成份均减少，而LF：HF比值在卧床后期有增大趋势．卧床后HUT初始6min所有被试者心率明显快于卧床前的相应值．说明模拟失重后心脏迷走神经反应与外周血管交感神经活动水平降低，心脏交感神经活动水平逐渐升高．但卧床后HUT时心血管自主神经调节反应基本正常．     

模拟微重力对人骨髓间充质干胞向成骨细胞分化中细胞信号通路影响的分析

通过模拟来研究微重力对hMSC向成骨细胞分化的影响,并利用相关信号通路的激活剂或抑制剂来调节这一分化过程．研究结果表明,在成骨细胞分化诱导条件下,微重力降低了hMSC向成骨细胞定向分化的能力,并且成骨细胞标记性基因的表达明显降低,Runt相关转录因子2（Runx2）的表达受到抑制．相反,过氧化物酶体增殖激活受体γ（PPARγ2）的表达则增加．同时,微重力也降低了ERK的磷酸化水平,而增加了p38MAPK的磷酸化水平．使用p38MAPK的抑制剂SB203580能够抑制p38MAPK的磷酸化,但不能降低PPARγ2的表达水平．骨形态发生蛋白（BMP）能增加Runx2的表达水平．成纤维细胞生长因子2（FGF2）增加了ERK的磷酸化水平,但也不能显著增加成骨细胞标记性基因的表达水平．采用BMP,FGF2和SB203580三种因子组合来调控微重力下的成骨细胞分化能力,结果表明三者的协同作用能显著逆转微重力对成骨细胞定向分化的生物学效应．研究结果还说明,模拟微重力应该是通过不同的细胞信号通路来抑制成骨细胞分化和提升脂肪细胞分化的．     

模拟微重力对人骨髓间充质干细胞向成骨细胞分化中细胞信号通路影响的分析

通过模拟来研究微重力对hMSC向成骨细胞分化的影响,并利用相关信号通路的激活剂或抑制剂来调节这一分化过程.研究结果表明,在成骨细胞分化诱导条件下,微重力降低了hMSC向成骨细胞定向分化的能力,并且成骨细胞标记性基因的表达明显降低, Runt相关转录因子2(Ruax2)的表达受到抑制.相反,过氧化物酶体增殖激活受体γ(PPARγ2)的表达则增加.同时,微重力也降低了ERK的磷酸化水平,而增加了p38MAPK的磷酸化水平.使用p38MAPK的抑制剂SB203580能够抑制p38MAPK的磷酸化,但不能降低PPARγ2的表达水平.骨形态发生蛋白(BMP)能增加Runx2的表达水平.成纤维细胞生长因子2(FGF2)增加了ERK的磷酸化水平,但也不能显著增加成骨细胞标记性基因的表达水平.采用BMP,FGF2和SB203580三种因子组合来调控微重力下的成骨细胞分化能力,结果表明三者的协同作用能显著逆转微重力对成骨细胞定向分化的生物学效应.研究结果还说明,模拟微重力应该是通过不同的细胞信号通路来抑制成骨细胞分化和提升脂肪细胞分化的.      

基板和流速对羟基磷灰石生长的影响

通过共混制备聚二甲基硅氧烷（PDMS）/羟基磷灰石（HAP）和PDMS/生物玻璃复合材料作为生长基板，研究了静止条件下不同基板上HAP的生长.设计了一套流速加载装置，观察模拟体液流速对生物玻璃基板上HAP晶体生长的影响.研究发现在静止条件下，HAP晶体在PDMS复合材料上比在生物玻璃上生长更快，尺寸更大；随着流速的增加，HAP晶体尺寸更大而且无定形沉淀数量减少.      

模拟失重(HDT—30°)和噪声复合因素对大鼠神经-内分泌-免疫功能的影响

本研究结果表明，大鼠在声级为８５ｄＢ（Ａ）噪声环境下悬吊（ＨＤＴ－３０°）１０昼夜后，淋巴细胞转化率及白细胞介素２（ＩＬ２）明显降低，血浆皮质酮含量显著升高，下丘脑内多巴胺（ＤＡ）和去甲肾上腺素（ＮＡ）含量呈升高趋势，表明机体处于应激状态．单纯８５ｄＢ（Ａ）噪声组除多巴胺及Ｔ淋巴细胞增殖反应外上述指标也有同样的变化规律，但没有复台因素组明显．本文出现的神经－内分泌－免疫系统功能失调可能是航天员易患感染性疾病的原因之一．     

模拟空间失重状态下流体桥振动实验研究

就微重力环境下粘滞流体桥振动特性问题，采用ＰＴＦ（平台流体浴槽）装置，在模拟空间失重状态下进行实验研究。实验就不同桥长（不同细长度）、不同位置在不同外界挠动频率下的振动特性针对不同粘滞系数反复进行实验，测试并详细记录了各参数下桥界面振幅随频率变化的数据和谱图（计数据２０００余，谱图８１张）。实验数据和谱图的分析结果表明：①谐频随桥细长度增加向低频方向移动；②谐频和细长度关系是双曲反比例关系。     

骨特异性转录因子Runx2对抗空间骨丢失效应的初步研究

为了构建稳定过表达Runx2 (骨特异性转录因子)的C2C12 (小鼠成肌细胞)和MG63 (前成骨细胞)细胞株, 并用于研究Runx2在对抗空间骨丢失效应中的作用. 利用实时定量聚合酶链式反应鉴定Runx2下游基因I型胶原、碱性磷酸酶表达情况, 在二维回转器中培养稳定细胞株, 通过定量聚合酶链式反应,观察在模拟失重效应下Runx2基因对其下游基因表达的影响. 结果表明, 通过筛选获得稳定转染的C2C12-Runx2和MG63-Runx2细胞株, 经鉴定都能过表达Runx2. 转染后的细胞 I 型胶原和碱性磷酸酶mRNA表达增高. 回转组与对照组相比, MG63, C2C12-Runx2, MG63-Runx2细胞的I型胶原和碱性磷酸酶mRNA表达降低, 但在模拟失重效应下, 转染细胞中I型胶原和碱性磷酸酶的mRNA表达下降程度明显低于未转染细胞株. 所构建的C2C12-Runx2和MG63-Runx2细胞株比较稳定, 并证实Runx2能部分对抗失重引起的成骨特异性分子的降低.      

基于认知过程的飞行员脑力负荷动态预测

现代军用飞机座舱驾驶系统信息高度密集、任务复杂多变。为探讨信息加工类型与多任务协同对飞行员脑力负荷的影响，依据ACT-R认知模块与脑力负荷决定性因素的关联性，将飞行员的脑力负荷划分为感知负荷与认知负荷，并基于四维多资源干扰理论，考虑多任务协同作用时的资源干扰对脑力负荷的影响，提出了基于认知过程的飞行员脑力负荷动态预测模型。为校验所提模型，选取16名被试完成4种模拟飞行任务的脑力负荷评价实验，结果显示：不同飞行任务在飞行绩效、NASA-TLX主观评价、平均扫视时间与扫视频率下的主效应显著（P<0.05），总脑力负荷预测值与NASA-TLX主观评价、扫视频率和心率呈显著正相关，平均脑力负荷预测值与飞行绩效、瞳孔直径和平均扫视时间呈显著正相关。所提模型对飞行员脑力负荷的动态预测与评价具有应用价值。     

太阳风中航天器带电与尾迹效应的模拟

航天器充电和尾迹效应会对周围等离子体造成扰动,影响测量装置结果的准确性.利用SPIS （Spacecraft Plasma Interaction Software）分别模拟了航天器与太阳风的相互作用,考察了光电效应以及航天器尺度对表面充电情况和尾迹效应的影响.结果表明:太阳风环境下,等离子体密度稀薄,电子电流比光电子电流小得多,航天器表面为正电势,航天器后部有清晰的尾迹结构,尾迹带负电;光电效应可改变尾迹结构,与无光电效应相比,光电效应使得航天器尾迹尺度变大;由于太阳风定向运动动能大于航天器表面势能,航天器的尾迹结构与其几何尺寸有关,航天器尺寸越大,尾迹尺度越大.      

The declined phosphorylation of Heat shock protein 27 in rat cardiac muscle after hindlimb unloading

Hindlimb unloading can induce the cardiac atrophy and diminished cardiac function, however, the mechanisms responsible for which remain elusive. The chronic volume unloading of heart, which decreases the local mechanical stress, may lead to cardiac atrophy after hindlimb unloading. Many studies showed that integrin signaling, p38 MAPK, Heat shock protein 27 and cytoskeleton involved in the hypertrophic growth induced by mechanical stress. However, the mechanisms responsible for cardiac atrophy after hindlimb unloading are still unclear. In this study, we used the tail-suspended, hindlimb unloading rat model to simulate the effects of microgravity. Western blot analysis was used to detect the protein expression of Heat shock protein 27, focal adhesion kinase, p38 MAPK and their phosphorylation levels in rat cardiac muscle after 14d hindlimb unloading. The results showed that the phosphorylation levels of both Heat shock protein 27 and p38 MAPK were decreased significantly in rat cardiac muscle after hindlimb unloading. However, the phosphorylation level of focal adhesion kinase was not decreased significantly. The results suggested that Heat shock protein 27, the downstream of p38 MAPK, might play a critical role in the cardiac atrophy in response to simulated microgravity induced by hindlimb unloading.     

Effects of microgravity on bone and calcium homeostasis.

Mechanical function is known to be of crucial importance for the maintenance of bone tissue. Gravity on one hand and muscular effort on the other hand are required for normal skeletal structure. It has been shown by numerous experimental studies that loss of total-body calcium, and marked skeletal changes occur in people who have flown in space. However, most of the pertinent investigations have been conducted on animal models, including rats and non-human primates, and a reasonably clear picture of bone response to spaceflight has emerged during the past few years. Osteopenia induced by microgravity was found to be associated with reduction in both cortical and trabecular bone formation, alteration in mineralization patterns and disorganization of collagen, and non-collagenous protein metabolism. Recently, cell-culture techniques have offered a direct approach of altered gravity effects at the osteoblastic-cell level. But the fundamental mechanisms by which bone and calcium are lost during spaceflight are not yet fully known. Infrequency and high financial cost of flights have created the necessity to develop on-Earth models designed to mimic weightlessness effects. Antiorthostatic suspension devices are now commonly used to obtain hindlimb unloading in rats, with skeletal effects similar to those observed after spaceflight. Therefore, actual and "simulated" spaceflights, with investigations conducted at whole body and cellular levels, are needed to elucidate pathogeny of bone loss in space, to develop effective countermeasures, and to study recovery processes of bone changes after return to Earth.     

Effects of microgravity on muscle and cerebral cortex: a suggested interaction.

The "slow" antigravity muscle adductor longus was studied in rats after 14 days of spaceflight (SF). The techniques employed included standard methods for light microscopy, neural cell adhesion molecule (N-CAM) immunocytochemistry and electron microscopy. Light and electron microscopy revealed myofiber atrophy, segmental necrosis and regenerative myofibers. Regenerative myofibers were N-CAM immunoreactive (N-CAM-IR). The neuromuscular junctions showed axon terminals with a decrease or absence of synaptic vesicles, degenerative changes, vacant axonal spaces and changes suggestive of axonal sprouting. No alterations of muscle spindles was seen either by light or electron microscopy. These observations suggest that muscle regeneration and denervation and synaptic remodeling at the level of the neuromuscular junction may take place during spaceflight. In a separate study, GABA immunoreactivity (GABA-IR) was evaluated at the level of the hindlimb representation of the rat somatosensory cortex after 14 days of hindlimb unloading by tail suspension ("simulated" microgravity). A reduction in number of GABA-immunoreactive cells with respect to the control animals was observed in layer Va and Vb. GABA-IR terminals were also reduced in the same layers, particularly those terminals surrounding the soma and apical dendrites of pyramidal cells in layer Vb. On the basis of previous morphological and behavioral studies of the neuromuscular system after spaceflight and hindlimb suspension it is suggested that after limb unloading there are alterations of afferent signaling and feedback information from intramuscular receptors to the cerebral cortex due to modifications in the reflex organization of hindlimb muscle groups. We propose that the changes observed in GABA immunoreactivity of cells and terminals is an expression of changes in their modulatory activity to compensate for the alterations in the afferent information.     

Inhibitory effect of simulated microgravity on differentiating preosteoblasts

The bone loss induced by microgravity is partly due to the decrease of mature osteoblasts. In the present study, we employed the random positioning machine (RPM) to simulate microgravity and investigated the acute effects of simulated microgravity on the differentiation of 2T3 preosteoblasts. Following 7 days’ culture under normal (1 g) condition, cells were exposed to simulated microgravity for 24 h. The results showed that 24 h treatment of simulated microgravity significantly decreased alkaline phosphatase (ALP) activity without changing the cell morphology. In addition, the mRNA expressions of osteogenic genes, including runt-related gene 2 (Runx2), osterix, osteocalcin (OC), type I collagen (Col I) and bone morphogenetic protein (BMP), were dramatically downregulated. Moreover, western blot analysis of total extracellular signal-regulated kinase (Erk) and phosphorylated Erk (p-Erk) indicated that p-Erk level, which represents the Erk activation status, was increased. Taken together, our results suggested that acute exposure to simulated microgravity inhibited osteoblast differentiation through modulating the expression of osteogenic genes and the Erk activity. These findings provide new insight for bone loss due to microgravity and unloading.     

Alterations in glucose and protein metabolism in animals subjected to simulated microgravity.

Reduction of physical activity due to disease or environmental restraints, such as total bed rest or exposure to spaceflight, leads to atrophy of skeletal muscle and is frequently accompanied by alterations in food intake and the concentration of metabolic regulatory hormones such as insulin. Hindlimb suspension of laboratory rats, as a model for microgravity, also shows marked atrophy of gravity dependent muscles along with a reduced gain in body weight. Suspended rats exhibit enhanced sensitivity to insulin-induced glucose uptake when compared with normal control rats and resistance to insulin action when compared with control rats matched similarly for reduced body weight gain. These changes are accompanied by decreased insulin binding and tyrosine kinase activity in soleus but not plantaris muscle, unchanged glucose uptake by perfused hindlimb and decreased sensitivity but not responsiveness to insulin-induced suppression of net proteolysis in hindlimb skeletal muscle. These findings suggest that loss of insulin sensitivity during muscle atrophy is associated with decreased insulin binding and tyrosine kinase activity in atrophied soleus muscle along with decreased sensitivity to the effects of insulin on suppressing net protein breakdown but not on enhancing glucose uptake by perfused hindlimb.     

Comparison of hindlimb unloading and partial weight suspension models for spaceflight-type condition induced effects on white blood cells

Animal models are frequently used to assist in the determination of the long- and short-term effects of space flight. The space environment, including microgravity, can impact many physiological and immunological system parameters. It has been found that ground based models of microgravity produce changes in white blood cell counts, which negatively affects immunologic function. As part of the Center of Acute Radiation Research (CARR), we compared the acute effects on white blood cell parameters induced by the more traditionally used animal model of hindlimb unloading (HU) with a recently developed reduced weightbearing analog known as partial weight suspension (PWS). Female ICR mice were either hindlimb unloaded or placed in the PWS system at 16% quadrupedal weightbearing for 4 h, 1, 2, 7 or 10 days, at which point complete blood counts were obtained. Control animals (jacketed and non-jacketed) were exposed to identical conditions without reduced weightbearing. Results indicate that significant changes in total white blood cell (WBC), neutrophil, lymphocyte, monocyte and eosinophil counts were observed within the first 2 days of exposure to each system. These differences in blood cell counts normalized by day 7 in both systems. The results of these studies indicate that there are some statistically significant changes observed in the blood cell counts for animals exposed to both the PWS and HU simulated microgravity systems.     

Personality,social support and affective states during simulated microgravity in healthy women

This study investigated the time-course of stress and recovery states and their relations to social support and personality traits in healthy women during a long-term head-down tilt bed rest. Personality, social support and affective states were assessed in 16 women exposed to simulated microgravity for a 60-day duration involving three stages: a 20-day baseline control period (BDC), a 60-day head-down tilt bed rest (HDT) and a 20-day post-HDT ambulatory recovery period (R+). Participants were divided into two groups: an exercise (Exe, n = 8) and a control group (Ctl, n = 8). All the participants experienced significantly more stress during the HDT period. But exercise did not improve the impaired effects of simulated microgravity. The Exe group perceived more stress and less recovery than the Ctl group during the HDT period. Among the five major personality factors, only Neuroticism was related to both social and affective variables. Neuroticism was positively associated with stress and negatively associated with recovery and social support (S-SSQ). Practical implications in psychological countermeasures for better dealing with the key human factor in spaceflights are discussed.     

Parathyroid hormone-related protein is a gravisensor in lung and bone cell biology.

Parathyroid Hormone-related Protein (PTHrP) has been shown to be essential for the development and homeostatic regulation of lung and bone. Since both lung and bone structure and function are affected by microgravity, we hypothesized that 0 x g down-regulates PTHrP signaling. To test this hypothesis, we suspended lung and bone cells in the simulated microgravity environment of a Rotating Wall Vessel Bioreactor, which simulates microgravity, for up to 72 hours. During the first 8 hours of exposure to simulated 0 x g, PTHrP expression fell precipitously, decreasing by 80-90%; during the subsequent 64 hours, PTHrP expression remained at this newly established level of expression. PTHrP production decreased from 12 pg/ml/hour to 1 pg/ml/hour in culture medium from microgravity-exposed cells. The cells were then recultured at unit gravity for 24 hours, and PTHrP expression and production returned to normal levels. Based on these findings, we have obtained bones from rats flown in space for 2 weeks (Mission STS-58, SL-2). Analysis of PTHrP expression by femurs and tibias from these animals (n=5) revealed that PTHrP expression was 60% lower than in bones from control ground-based rats. Interestingly, there were no differences in PTHrP expression by parietal bone from space-exposed versus ground-based animals, indicating that the effect of weightlessness on PTHrP expression is due to the unweighting of weight-bearing bones. This finding is consistent with other studies of microgravity-induced osteoporosis. The loss of the PTHrP signaling mechanism may be corrected using chemical agents that up-regulate this pathway. In conclusion, PTHrP represents a stretch-sensitive paracrine signaling mechanism that may sense gravity.     

微重力下胶原蛋白纤维化及羟基磷灰石结晶的初步研究

在长期空间飞行过程中, 骨质丢失是一个严重问题. 羟基磷灰石(HAP)晶体是骨骼的主要成分, 骨骼中的胶原蛋白纤维在HAP生长结晶过程中起到关键作用. 研究了胶原蛋白纤维化过程在模拟微重力和常重力条件下的变化, 对以胶原 蛋白纤维作为模板生长出的HAP晶体形貌进行了观察. 结果表明, 不同浓度胶原蛋白溶液中形成的胶原蛋白纤维, 其内部孔隙数量和尺寸在模拟微重力条件下要明显大于常重力条件下, 胶原蛋白纤维内部孔隙的分布也不同于常重力条 件下的结果. 以模拟微重力条件下形成的胶原蛋白纤维为模板生长出的HAP 晶体主要为立方体状, 而以常重力条件下形成的胶原蛋白纤维为模板生长出的 HAP晶体形貌主要为板状. 该结果有助于未来进一步阐明空间骨质丢失的机理.      

Evaluation of upper body muscle activity during cardiopulmonary resuscitation performance in simulated microgravity

Performance of efficient single-person cardiopulmonary resuscitation (CPR) is vital to maintain cardiac and cerebral perfusion during the 2–4 min it takes for deployment of advanced life support during a space mission. The aim of the present study was to investigate potential differences in upper body muscle activity during CPR performance at terrestrial gravity (+1Gz) and in simulated microgravity (μG). Muscle activity of the triceps brachii, erector spinae, rectus abdominis and pectoralis major was measured via superficial electromyography in 20 healthy male volunteers. Four sets of 30 external chest compressions (ECCs) were performed on a mannequin. Microgravity was simulated using a body suspension device and harness; the Evetts–Russomano (ER) method was adopted for CPR performance in simulated microgravity. Heart rate and perceived exertion via Borg scores were also measured. While a significantly lower depth of ECCs was observed in simulated microgravity, compared with +1Gz, it was still within the target range of 40–50 mm. There was a 7.7% decrease of the mean (±SEM) ECC depth from 48 ± 0.3 mm at +1Gz, to 44.3 ± 0.5 mm during microgravity simulation (p < 0.001). No significant difference in number or rate of compressions was found between the two conditions. Heart rate displayed a significantly larger increase during CPR in simulated microgravity than at +1Gz, the former presenting a mean (±SEM) of 23.6 ± 2.91 bpm and the latter, 76.6 ± 3.8 bpm (p < 0.001). Borg scores were 70% higher post-microgravity compressions (17 ± 1) than post +1Gz compressions (10 ± 1) (p < 0.001). Intermuscular comparisons showed the triceps brachii to have significantly lower muscle activity than each of the other three tested muscles, in both +1Gz and microgravity. As shown by greater Borg scores and heart rate increases, CPR performance in simulated microgravity is more fatiguing than at +1Gz. Nevertheless, no significant difference in muscle activity between conditions was found, a result that is favourable for astronauts, given the inevitable muscular and cardiovascular deconditioning that occurs during space travel.