《生物化学》重要章节——《糖酵解》课堂教学体会与思考

《生物化学》是生物类专业的一门重要学科基础课程。《糖酵解》属于《糖代谢》一章的其中一节。本节内容的学习需应用前一章《生物氧化与氧化磷酸化》的已学知识,也是《糖代谢》后续内容《三羧酸循环》及《糖异生》等的学习基础,因此是本章教学的重要环节,教学效果的好坏将影响到《糖代谢》一章的整体教学质量。课程教学团队在多年课堂教学实践基础上,对《糖酵解》一节的课堂教学体会及思考进行了总结,供大家交流与参考。     

A dynamic model linking cell growth to intracellular metabolism and extracellular by-product accumulation

Mathematical modeling of animal cell growth and metabolism is essential for the understanding and improvement of the production of biopharmaceuticals. Models can explain the dynamic behavior of cell growth and product formation, support the identification of the most relevant parameters for process design, and significantly reduce the number of experiments to be performed for process optimization. Few dynamic models have been established that describe both extracellular and intracellular dynamics of growth and metabolism of animal cells. In this study, a model was developed, which comprises a set of 33 ordinary differential equations to describe batch cultivations of suspension AGE1.HN.AAT cells considered for the production of α1-antitrypsin. This model combines a segregated cell growth model with a structured model of intracellular metabolism. Overall, it considers the viable cell concentration, mean cell diameter, viable cell volume, concentration of extracellular substrates, and intracellular concentrations of key metabolites from the central carbon metabolism. Furthermore, the release of metabolic by-products such as lactate and ammonium was estimated directly from the intracellular reactions. Based on the same set of parameters, this model simulates well the dynamics of four independent batch cultivations. Analysis of the simulated intracellular rates revealed at least two distinct cellular physiological states. The first physiological state was characterized by a high glycolytic rate and high lactate production. Whereas the second state was characterized by efficient adenosine triphosphate production, a low glycolytic rate, and reactions of the TCA cycle running in the reverse direction from α-ketoglutarate to citrate. Finally, we show possible applications of the model for cell line engineering and media optimization with two case studies.     

Glucose,glycolysis, and neurodegenerative diseases

Prolonged survival of a typical postmitotic neuron hinges on a balance between multiple processes, among these are a sustenance of ATP production and protection against reactive oxygen species. In neuropathological conditions, mitochondrial defects often lead to both a drop in ATP levels, as well as increase reactive oxygen species production from inefficient electron transport processes and NADPH-oxidases activities. The former often resulted in the phenomenon of compensatory aerobic glycolysis. The latter stretches the capacity of the cell's redox buffering capacity, and may lead to damages of key enzymes involved in energy metabolism. Several recent reports have indicated that enhancing glucose availability and uptake, as well as increasing glycolytic flux via pharmacological or genetic manipulation of glycolytic enzymes, could be protective in animal models of several major neurodegenerative diseases, including Parkinson's disease, Huntington's disease, and Amyotrophic lateral sclerosis. Activation of canonical Wnt signaling, which improves disease symptoms in mouse models of Alzheimer's disease also appears to work via an elevation of glycolytic enzymes and enhance glucose metabolism. Here, I discuss these findings and the possible underlying mechanisms of how an increase in glucose uptake and glycolysis could be neuroprotective. Increased glycolytic production of ATP would help alleviate energy deficiency, and ATP's hydrotropic effect may enhance solubility and clearance of toxic aggregates prevalent in many neurodegenerative diseases. Furthermore, channeling of glucose into the Pentose Phosphate Pathway would increase the redox buffering capacity of the cell.     

Development and validation of glycolysis-related prognostic score for prediction of prognosis and chemosensitivity of pancreatic ductal adenocarcinoma

Pancreatic ductal adenocarcinoma (PDAC) is a lethal malignancy with aggressive biological behaviour. Its rapid proliferation and tumour growth require reprogramming of glucose metabolism or the Warburg effect. However, the association between glycolysis-related genes with clinical features and prognosis of PDAC is still unknown. Here, we used the meta-analysis to correlate the hazard ratios (HR) of 106 glycolysis genes from MSigDB by the cox proportional hazards regression analysis in 6 clinical data sets of PDAC patients to form a training cohort, and a single group of PDAC patients from the TCGA, ICGC, Arrayexpress and GEO databases to form the validation cohort. Then, a glycolysis-related prognosis (GRP) score based on 29 glycolysis prognostic genes was established in 757 PDAC patients from the training composite cohort and validated in 267 ICGC-CA validation cohort (all P < .05). In addition, including PADC, the prognostic value was also confirmed in other 7 out of 30 pan-cancer cohorts. The GRP score was significantly related to specific metabolism pathways, immune genes and immune cells in the patients with PADC (all P < .05). Finally, by combining with immune cells, the GRP score also well-predicted the chemosensitivity of patients with PADC in the TCGA cohort (AUC = 0.709). In conclusion, this study developed a GRP score for patients with PDAC in predicting prognosis and chemosensitivity for PDAC.     

双孢蘑菇对高温胁迫的响应及耐热机理

本研究以双孢蘑菇Agaricus bisporus工厂化菌株A15和筛选得到的耐高温菌株A15-TH为研究对象,比较了高温胁迫对两个菌株菌丝生长的影响,并从氧化损伤修复及基础碳代谢-糖酵解途径两个角度探索双孢蘑菇对高温胁迫的响应及耐热机理。高温胁迫下,对照菌株A15的菌丝生长速度降低,菌丝分叉增加;而耐高温菌株A15-TH菌丝生长速度高于A15,菌丝形态优于对照菌株,表现出对高温具有一定的耐受性。对两个菌株高温胁迫下氧化损伤及抗氧化酶系统进行研究发现,高温胁迫30-90min导致对照菌株A15的三磷酸腺苷（ATP）含量下降54.4%-59.6%,线粒体复合物I、II、III活性升高,超氧阴离子（O₂^-）含量增加了34.9%-71.3%;此外高温胁迫降低了超氧化物歧化酶（SOD）的活性,影响了O₂^-的清除效率。耐高温菌株在受到高温胁迫后的氧化损伤及氧化修复效果与对照菌株不同,一方面体现在正常状态下维持较低的细胞能量代谢和较高的ROS合成量;另一方面抗氧化系统中sod1、sod2、cat1与对照菌株相比有不同程度的上调,SOD和过氧化氢酶（CAT）活性增强,可以更有效地清除过量的活性氧,减轻高温对菌丝的氧化损伤。尤其在高温胁迫120min时,A15的线粒体功能及抗氧化系统受到严重损伤,线粒体复合物I、II、III活性和CAT活性大幅度下降,但是A15-TH线粒体复合体I、III活性分别增加至正常状态下的1.4倍和8.9倍,CAT活性比对照菌株高128%,维持了正常的线粒体功能及对活性氧的有效清除。进一步研究发现高温胁迫下,双孢蘑菇菌丝的己糖激酶和丙酮酸激酶活性增加,糖酵解途径加快;耐高温菌株A15-TH在正常状态下和高温胁迫下,己糖激酶和丙酮酸激酶的活性均高于对照菌株A15,具有更活跃的碳代谢。     

Microbiological characterization of a coke oven contaminated site and evaluation of its potential for bioremediation

The soil microbial population of a coke oven site was investigated in order to evaluate its potential for bioremediation. The study was carried out in soil samples with distinct polynuclear aromatic hydrocarbon (PAH) contamination levels, comparing the population profiles constituted by total heterotrophic and PAH-utilizing strains. Isolation of degrading strains was performed with phenanthrene or pyrene as sole carbon sources. The ability to degrade other PAHs, such as anthracene, fluorene and fluoranthene was also investigated. The results showed a reduction of 30% in species diversity and microbial density drops one order of magnitude in contaminated samples. Furthermore, the number of PAH-utilizing colonies was higher in the contaminated area and about 20% of the isolates were able to degrade phenanthrene and pyrene, while this value decreased to 0.15% in uncontaminated samples. Three PAH-degrader strains were identified as: CDC gr. IV C-2, Aeromonas sp. and Pseudomonas vesicularis. The ability of these strains to degrade other PAHs was also investigated.     

过量铜、锌对外生菌根菌牛乳牛肝菌生物量、呼吸和糖酵解酶活性的影响

在过量铜或锌胁迫下,独立培养的外生菌根菌牛乳牛肝菌（Suillus     

The diabetic vasculature: Physiological mechanisms of dysfunction and influence of aerobic exercise training in animal models

Diabetes mellitus (DM) is associated with a number of complications of which chronic vascular complications are undoubtedly the most complex and significant consequence. With a significant impact on health care, 50–80% of people with diabetes die of cardiovascular disease (including coronary artery disease, stroke, peripheral vascular disease and other vascular disease), making it the major cause of morbidity and mortality in diabetic patients. A healthy lifestyle is essential in the management of DM, especially the inclusion of aerobic exercise, which has been shown effective in reducing the deleterious effects in vasculature. Interest in exercise studies has increased significantly with promising results that demonstrate a future for investigation. Considering the importance of this emerging field, the aim of this mini-review is to summarize and integrate animal studies investigating physiological mechanisms of vascular dysfunction and remodeling in type 1 diabetes mellitus (T1DM) and type 2 diabetes mellitus (T2DM) and how these are influenced by chronic aerobic exercise training.