细胞不同代谢类型的热化学研究Ⅰ.生长、非生长及内源代谢水平的量热测定

细胞是生命的基本单位．对其代谢过程能量输出水平的直接测定，对细胞生理学来说是重要的实验基础数据，对于生物生理和研究能量代谢方面也有一定的理论意义．当然细胞的多样性及其代谢过程的复杂性使这一研究工作相当困难·作为开展研究的第一步我们选择便于培养处理的单细胞生物大肠杆菌作为研究对象，并对其所进行时三种基本代谢类型29Llxi：L谢；非生长依队和内派代谢分别进行研究，本文分别测定了＊烟三种不同类型代谢的热功率输出水平，得到了单个细胞在上述代谢过程热功率输出的定量数据．实验结果表明，不同类型代谢的能量输出水平…     

不同条件下离体大鼠肝脏线粒体能量代谢微量热分析

为了探究线粒体的能量代谢过程,本文以离体大鼠肝脏线粒体为模型,利用多通道、高灵敏度的热活性检测仪TAM Ⅲ,实时监测了不同线粒体浓度、不同底物、不同缓冲液、几种呼吸抑制剂以及Ca²⁺和线粒体渗透转换孔抑制剂CsA存在时线粒体的能量代谢,获得了完整的热功率―时间曲线,并通过计算得到了线粒体能量代谢的热动力学参数。通过分析发现：（1）线粒体浓度越大,代谢越快;（2）直接底物琥珀酸钠使线粒体代谢更快;（3）高浓度Ca²⁺能够刺激线粒体快速产热,且在长期代谢进程中,线粒体渗透转换孔抑制剂CsA并不能改变Ca²⁺造成的影响;（4）不同缓冲液对线粒体代谢的影响基于其组分的不同,缓冲液中含有呼吸底物;（5）呼吸抑制剂都能抑制线粒体的能量代谢,尤其是复合物IV的抑制剂NaN₃,高浓度下使代谢停止。     

等温微量热法在生命科学研究中的应用

简要介绍了等温微量热法的原理、典型的仪器及其在生命科学研究中应用所具有的特点。通过它可获得完整的细胞代谢过程产热曲线及其热动力学方程；可以研究细胞器的代谢规律；可以获取生物大分子与小分子相互作用的热动力学特征。     

微量热法表征邻香草醛氨基葡萄糖Schiff碱锌配合物的抗菌活性

用微量热法测定了大肠杆菌（细菌）及其在Ｚｎ－ｏ－ＶＧ（ｏ－ＶＧ：邻香草醛氨基葡萄糖Ｓｃｈｉｆｆ碱）作用下不同温度下的热谱；计算了它的生长速率常数ｋ、传找时间Ｇ、抑制率Ｉ、总产热量Ｑ、单个细菌的平均产热Ｑ０的单位时间平均每个细菌的产热量＾－量Ｑ０；建立了细菌生长代谢各种参量之间的函数关系，探讨了在不同温度和药物作用下细菌的生长代谢；发现可用ｔｇ、ｔｒ和ｔ表征细菌的生长代谢和药物的抗菌活性。     

量热法研究线粒体代谢的热力学和动力学行为

线粒体是细胞中极为重要的细胞器，是产生细胞所必需的“富能”物质ATP的重要场所，为细胞活动提供所需化学能．在生命体能量代谢过程中除一部分能量用于合成ATP外，其余则以热的形式释出．用精密热量计测出线粒体代谢过程中的热量输出对了解线粒体的功能和代谢机制具有十分重要的意义．用微量热法研究线粒体体外代谢已有一些报导［‘，’］．本工作用精密热量计和差式扫描量热仪侧定了水稻线粒体体外代谢热谱和DSC曲线，计算了水稻线粒体活性增长速率常数，比较了不同保藏时间的水稻线粒体体外代谢的差异，并初步探讨了水稻线粒体在变…     

微量量热法研究增溶性辅料吐温80对红细胞生理活性的影响

建立灵敏、专属性较好地表征增溶性辅料吐温80对红细胞生理活性影响的评价方法.采用微量量热法以生命体能量代谢特征参数衰亡速率常数(k)、最大热功率(Pmax)、总发热量(Q)、衰亡时间(t)为评价指标,考察增溶性辅料吐温80作用下红细胞的能量动态变化过程.结果表明,随着吐温80浓度(C)的增大,红细胞生长代谢特征参数总发热量(Q)降低、最大热功率(Pmax)增强、衰亡时间(t)减小、衰亡速率常数(k)减小,Q,k与ln C间量效关系良好(相关系数r>0.95);不同厂家吐温80作用于红细胞的总发热量(Q)存在显著性差异(P<0.05).与肉眼观察法、分光光度法、细胞计数等常规方法比较,微量量热法较能实时在线、全息地监测增溶性辅料对红细胞生理活性的动态过程,尤其是对亚致死效应细胞测定的分辨能力.该法可用于增溶性辅料吐温80质量标准的生物测定,亦为红细胞能量代谢等相关研究提供了实验参考.     

微量量热法研究増溶性辅料吐温80对红细胞生理活性的影响

建立灵敏、专属性较好地表征増溶性辅料吐温80对红细胞生理活性影响的评价方法.采用微量量热法以生命体能量代谢特征参数衰亡速率常数(k)、最大热功率(Pmax)、总发热量(Q)、衰亡时间(t)为评价指标,考察增溶性辅料吐温80作用下红细胞的能量动态变化过程.结果表明,随着吐温80浓度(C)的增大,红细胞生长代谢特征参数总发热量(Q)降低、最大热功率(Pmax)增强、衰亡时间(t)减小、衰亡速率常数(k)减小,Q,k与lnC间量效关系良好(相关系数r0.95);不同厂家吐温80作用于红细胞的总发热量(Q)存在显著性差异(P0.05).与肉眼观察法、分光光度法、细胞计数等常规方法比较,微量量热法较能实时在线、全息地监测增溶性辅料对红细胞生理活性的动态过程,尤其是对亚致死效应细胞测定的分辨能力.该法可用于增溶性辅料吐温80质量标准的生物测定,亦为红细胞能量代谢等相关研究提供了实验参考.     

肿瘤坏死因子α和γ-干扰素对白血病细胞作用的31PNMR分析

用^31P核磁共振观察了肿瘤坏死因子α（TNF－α）和γ－干扰素（IFN－γ）联合对人淋巴瘤白血病细胞系Molt-4作用后含磷化合物的变化；Molt-4细胞系的^31P谱主要由三磷酸腺苷（ATP）、无机磷（Pi)、磷酸单酯等的共振峰组成；^31P谱显示γ－干扰素、肿瘤坏死因子α及γ－干扰素和肿瘤坏死因子α联合对Molt-4细胞作用后，三磷酸腺式苷的β位磷的共振峰与无机磷的峰高比值（hATP/hPi)不同程度地降低；同时根据Pi的化学位移对Molt-4细胞胞内pH的变化进行了监测。     

细菌生长的热动力学研究进展

本文简单介绍了绝热式和热导式量热计及理论模型,细菌生长的动力学模型,着重介绍了近五年中热动力学在研究细菌生长代谢过程中的应用;通过测定不同条件下细菌生长代谢过程的产热曲线,求得细菌生长代谢的热力学和动力学参数,研究细菌的生长条件和药物的抑制作用。     

温度及药物诱导肿瘤细胞凋亡的实验观察

将微量热法和透射电镜技术相结合,研究了温度对人卵巢癌细胞代谢的影响,不同温度下癌细胞超微结构的变化和加抗肿瘤药物(嘧福绿)的协同作用.从宏观到微观,深入探讨了热疗与化疗协同作用对肿瘤细胞杀伤作用的机理.实验结果表明,在高温下细胞代谢降低,肿瘤细胞的形态发生一系列恶性变化,且高热可能诱导肿瘤细胞的凋亡;热疗加化疗的协同作用则能加速肿瘤细胞的凋亡,使细胞的能量代谢更低,随着作用时间的延长,最终导致细胞死亡.     

Microcalorimetric study of virus infection; The effects of hyperthermia and 1b recombinant homo interferon on the infection process of BHK-21 cells by foot and mouth disease virus

The metabolic process of BHK-21 cell line infected by foot and mouth disease virus (FMDV) was determined by using LKB-2277 Bioactivity Monitor. The aim of the present study is to investigate the metabolic thermal power of the virus infection process of BHK-21 cells, the effects of combinational treatments of hyperthermia and 1b recombinant homo interferon on this process. In contrast to the metabolic thermal power of uninfected BHK-21 cells, the thermogenetic curves show that the energy metabolism mechanisms of BHK-21 cells were significantly changed by the virus infection process. The maximum thermal power decreased and the time needed to reach the maximal thermal power increased with the increasing concentration of interferon. The results also show that the infection process was thermosensitive. But no apparent synergetic effect of the combinational treatments of hyperthermia and interferon was observed. The present microcalorimetric results are in accordance with the cytomorphology observations.     

Novel intramolecular coordination chemistry of some new metallocene complexes

The metabolic thermogenic curves of liver mitochondria isolated from the livers of Cyprinus Carpio vol and its parents were determined at 28°C by using an LKB‐2277 Bioactivity Monitor. The results indicated that their thermogenic curves are different The total heat output and total time of the metabolism of the liver mitochondria of the hybrid F₁ (Cyprinus Carpio val) are more than those of its parents, and its maximum heat power is between that of the female parent and male parent. The relationship between their metabolic thermogenic curves and character of mitochondrial metabolism, and thermokinetics and the heterosis were analyzed and discussed. The character of the mitochondrial thermogenic curves reflected the physiologic character of heterosis. The microcalorimetric method proved to be a probable and sensitive tool for the assessment of heterosis.     

Microcalorimetric studies on the mitochondria metabolism of Cyprinus Carpio val and its parents

The metabolic thermogenic curves of liver mitochondria isolated from the livers of Cyprinus Carpio vol and its parents were determined at 28°C by using an LKB‐2277 Bioactivity Monitor. The results indicated that their thermogenic curves are different The total heat output and total time of the metabolism of the liver mitochondria of the hybrid F₁ (Cyprinus Carpio val) are more than those of its parents, and its maximum heat power is between that of the female parent and male parent. The relationship between their metabolic thermogenic curves and character of mitochondrial metabolism, and thermokinetics and the heterosis were analyzed and discussed. The character of the mitochondrial thermogenic curves reflected the physiologic character of heterosis. The microcalorimetric method proved to be a probable and sensitive tool for the assessment of heterosis.     

一种新型希夫碱及其3d,4f配合物的抗菌活性

用微量热法研究一种新型希夫碱及其3d,4f配合物(2L, 2LZnYb)对大肠杆菌和金黄色葡萄球菌的抗菌活性, 得到了在它们作用下大肠杆菌和金黄色葡萄球菌生长代谢的产热曲线, 并且基于分析生长代谢和非生长代谢的产热曲线建立的热动力学方程, 获得了它们的抗菌活性. 结果表明, 两种化合物(ZL, 2LZnYb)对大肠杆菌的生长代谢有强的活性(IC50分别为6.1 和5.1 mg·L-1), 但对金黄色葡萄球菌的生长代谢的活性弱得多(IC50分别为310.1 和595.5 mg·L-1). Zn 和Yb的导入使化合物对大肠杆菌生长代谢的抑制作用稍微增加, 但大大降低了对金黄色葡萄球菌的抑制作用. 对于非生长代谢, 两种化合物的活性有很大的差别. 无论对大肠杆菌还是金黄色葡萄球菌, 由于配体2L的导入, 表现出显著的抑制作用, 2L的MSC50为6.4和209.7 mg·L-1. 配体2L可能成为新的抗菌先导化合物.     

A microcalorimetric study of the action of mercuric chloride on the metabolism of mitochondria isolated from Cyprinus carpio liver tissue

After the occurrence of 'Minamata disease' in 1950, mercury aroused much more attention, and lots of studies concerned have been made. The purpose of the present paper is to study the effect of mercuric chloride on the mitochondria suspension isolated from the liver tissue of Cyprinus carpio from the direct viewpoint of energy by using the microcalorimetric method. The metabolic thermogenic curves of the mitochondria suspension at 25°C were obtained, and the mitochondria metabolic thermokinetic equations were established, from which we obtained the thermodynamic and thermokinetic parameters: thermogenic rate constant (k), heat output (Q), average heat power (P_av), etc. Experimental results indicated that low concentration of mercuric chloride (5 nmol Hg²⁺/(mg protein)) stimulates the thermogenesis of mitochondria, suggesting a strong effect of uncoupling action, while high concentration of mercuric chloride (20 nmol Hg²⁺/(mg protein)) inhibits the metabolism of mitochondria completely, suggesting a fatal effect on the phosphorylation system. The effect of Hg²⁺ on mitochondria is concentration-depended, from which the probable reaction mechanism of Hg²⁺ to the mitochondria was proposed. So the microcalorimetric method can be used in the toxicology research.     

Exploring the in vitro anti-inflammatory activity of gross saponins of Tribulus terrestris L. fruit by using liquid chromatography-mass spectrometry-based cell metabolomics approach

Our previous studies confirmed the efficacy of gross saponins of Tribulus terrestris L. fruit in treating cerebral ischemia. This study aimed to investigate the related mechanisms in vitro. The lipopolysaccharide-induced BV2 cells model was constructed and treated with gross saponins at different concentrations to explore its anti-inflammatory activity. The cell metabolite changes were tracked by liquid chromatography-mass spectrometry (LC-MS)-based metabolomics, and the metabolic biomarkers and related metabolic pathways were analyzed. Molecular biochemistry analysis was further used to verify the relevant inflammatory pathways. The results showed that the saponins reduced nitric oxide release and the secretion of tumor necrosis factor-alpha, interleukin-1β, and interleukin-6 from lipopolysaccharide-induced BV2 cells. Metabolic perturbations occurred in lipopolysaccharide-treated BV2 cells, which could be reversed by drug treatment via mainly regulating glycerophospholipid metabolism, tryptophan metabolism, purine metabolism pathways, etc. The western blot analysis demonstrated that saponin could suppress the activation of the inflammatory-related signaling pathway. The present study explored the in vitro anti-inflammatory mechanism of gross saponins of Tribulus terrestris L. fruit using an LC-MS-based cell metabolomics approach, which confirms the great potential of LC-MS for drug efficacy evaluation and can be applied in other herbal medicine-related analyses.     

GC/MS-based metabolomics reveals fatty acid biosynthesis and cholesterol metabolism in cell lines infected with influenza A virus

Metabolomics is the downstream of systems biology and has drawn significant interest for studying the metabolic networks from cells to organisms. To profile the metabolites in two different cell lines (A549 and AGS) infected with influenza A virus, gas chromatography coupled with mass spectrometry (GC/MS) was employed. Some differentiating metabolites in the cell lines were tentatively identified using reference library, interpreted and visualized by applying principal components analysis (PCA) and cluster heat map. Consequently, metabolic flux profiling allowed the differentiation of fatty acid biosynthesis and cholesterol metabolism during viral replication in the cell lines. The change in fatty acid turnover was also observed. Metabolomics investigation also revealed the different responses between A549 and AGS cell lines to the virus infection. From the pattern recognition results, AGS cell line might be more susceptible to influenza A virus. Regarding the fact that AGS is a poorly differentiated gastric adenocarcinoma cell line whereas A549 is a relatively differentiated lung tumor one, it is speculated that viral replication might be associated with the cell differentiations.     

The application of heat conduction microcalorimetry to study the metabolism and pharmaceutical modulation of cultured mammalian cells

The heat produced by animal cells in culture can be used as the primary indicator of the kinetics of their metabolism because the scalar flux of it is a function of the metabolic flux. The validity of the relationship between heat and metabolism was demonstrated theoretically through the concept of thermal advancement and in experiments by the use of continuous cultures. This validation permitted the application of heat flux as a probe of the metabolic state of cells in culture. It consisted of an on-line heat conduction microcalorimeter that measures the instantaneous heat flow and dividing the smoothed signal with one obtained simultaneously using a dielectric spectrometer that records the change in capacitance as an estimate of the amount of viable biomass. In this mini-review, it is shown with Chinese hamster ovary cells (CHO320) genetically engineered to produce interferon-γ (IFN-γ) that heat flux is an early signal of deteriorating metabolism in cultures that produce considerable amounts of toxic lactate under fully aerobic conditions. The early detection favours the use of heat flux as the control variable in fed-batch cultures. This is a particularly useful finding in the context of the pharmaceutical industry because it will help to ensure the high fidelity of the cytokines, antibodies and vaccines produced in large-scale cultures. The monotonic relationship between the fluxes for heat and metabolism means that the enthalpy balance method can be employed to test the validity of the growth reaction for cells in culture. This showed that the crucial ratio between the substrates, glucose and glutamine, in the culture medium was incorrect at 5.5:1 instead of about 3:1, depending on the phase of the culture. Together with other changes to the medium composition, an improved formulation was made that ensured faster cell growth and greater specific rate (flux) of IFN-γ constitutive secretion while decreasing glucose utilisation and, most importantly, halving the excretion of lactate, that is toxic to the cells and harmful to the fidelity of their secondary products. Indirect calorimetry (oxygen uptake rate, OUR) is often favoured over the direct technique, but the former only measures aerobic metabolism. The environmental conditions in cultures favours lactate production even under fully aerobic conditions. Developments in measuring OUR mean that the stationary liquid phase balance can be used successfully to make the calorimetric:respirometric (CR) ratio a valuable tool in optimising cell culture to grow cells that synthesise the maximum amounts of the high fidelity secondary products.

Besides the value of heat flux in improving the cultures of animal cells producing heterologous products, three different techniques are examined that should be valuable in the testing the many compounds that are produced on a speculative basis as potential drugs. They are: (i) a thin-film thermopile transducer as an immunosensor; (ii) infra-red imaging of cells cultured in multi-well microtitre plates and (iii) integrated circuit (IC) calorimetry for small samples and low detection limit. One or more of these methods could well find favour with industry in the near future.     

细胞不同代谢类型的量热学研究 I.静息细胞有氧非生长代谢特征

The metabolic heat output of resting cells (Saccharobacter fermentatus WVB8512) has been determined by means of microcalorimetric method. The metabolic thermokinetic equation is
dp/dt=k0,k0=0
The thermochemical equations are
lntm=10.1812-6.5492×10-9C
dC/dP0=KCn n=2
The order of metabolism (n) is 2. The experimental results indicate that the heat output produced by each cell and the metabolic time depend on cell concentration and are inhibited by cell density. The metabolism presents a special kind of special inhibition effect.