Characterisation and Skin Distribution of Lecithin-Based Coenzyme Q10-Loaded Lipid Nanocapsules

The purpose of this study was to investigate the influence of the inner lipid ratio on the physicochemical properties and skin targeting of surfactant-free lecithin-based coenzyme Q10-loaded lipid nanocapsules (CoQ10-LNCs). The smaller particle size of CoQ10-LNCs was achieved by high pressure and a lower ratio of CoQ10/GTCC (Caprylic/capric triglyceride); however, the zeta potential of CoQ10-LNCs was above /− 60 mV/ with no distinct difference among them at different ratios of CoQ10/GTCC. Both the crystallisation point and the index decreased with the decreasing ratio of CoQ10/GTCC and smaller particle size; interestingly, the supercooled state of CoQ10-LNCs was observed at particle size below about 200 nm, as verified by differential scanning calorimetry (DSC) in one heating–cooling cycle. The lecithin monolayer sphere structure of CoQ10-LNCs was investigated by cryogenic transmission electron microscopy (Cryo-TEM). The skin penetration results revealed that the distribution of Nile red-loaded CoQ10-LNCs depended on the ratio of inner CoQ10/GTCC; moreover, epidermal targeting and superficial dermal targeting were achieved by the CoQ10-LNCs application. The highest fluorescence response was observed at a ratio of inner CoQ10/GTCC of 1:1. These observations suggest that lecithin-based LNCs could be used as a promising topical delivery vehicle for lipophilic compounds.     

负载辅酶Q10的两亲性聚肽-壳聚糖复合纳米粒的制备及性质研究

以两亲性聚肽-壳聚糖复合体系为壁材,采用自组装及离子凝聚法制备负载辅酶Q10的纳米粒子,探讨了制备纳米粒子的较佳条件,采用高分辨率透射电镜、动态激光散射仪、紫外光谱及Zeta电位分析仪对优化的聚肽-壳聚糖复合体系纳米粒子进行表征,结果表明:聚肽-壳聚糖复合体系纳米粒子是具有壳核结构的球型粒子;纳米粒子粒径大小在80 nm到300 nm之间,粒径分布较均匀,稳定性较好,粒子的载药率为3.46±0.3%,包封率为59.7±2.1%。24 h的累积释放为75%。     

辅酶Q10的合成

以还原型辅酶Q0(氢醌,Ⅳ)为母环,杂多酸为催化剂,与末端羟基取代的异戊二烯单元—(E)-1-苯磺酰基-2-甲基-4-羟基-2-丁烯(Ⅲ)进行偶联反应,溴苄酚羟基保护,制得6-(4-苯磺酰基-3-甲基-2-丁烯)-基-2,3-二甲氧基-5-甲基氢醌双苄醚(Ⅴ),收率75.0%,Ⅴ再与茄尼基溴(Ⅵ)偶联后制得接砜产物(Ⅶ),收率45.0%,Ⅶ经钠还原脱除苯磺酰基、硝酸铈铵氧化制得辅酶Q10(Ⅰ),收率30%。中间体与产物经测熔点、1HNMR和FTIR分析鉴定和确证结构。     

Effect of antioxidant-enriched foods on plasma Coenzyme Q10 and total antioxidant capacity

Twenty healthy subjects integrated their diet with two types of products sequentially: In the first phase (first 14 days), the volunteers were given the following food items not supplemented with vitamin E (vit. E) or Coenzyme Q₁₀ (CoQ₁₀): breakfast, 250 mL skimmed milk; mid-morning snack, 330 mL fruit juice; mid-afternoon snack, low-fat yogurt 125 g; after dinner, low-fat dessert 110 g. In the second phase, from day 14 to day 35, the same items were added with vit. E and CoQ₁₀ (a total daily supplementation of 19.4 mg CoQ₁₀ and 13.7 mg vit.E). After taking the supplemented products for 2 weeks, plasma CoQ₁₀ reached 1.30 µg/mL, nearly twice the initial values. These levels further increased after 3 weeks of supplementation. Vit. E levels significantly (p <0.001) increased only after 3 weeks of supplementation, reaching 7.9 ± 2.8 µg/mL, 39% more than the initial value. The total plasma fatty acid pattern did not change substantially throughout the study. The increase of the total antioxidant capacity of plasma was significant (p <0.001) for days 28 and 35, with values close to 0.24 ± 0.09 mM Trolox (a nearly 60% increase).     

辅酶Q10的合成

以茄尼醇和2，3－二甲氧基－5－甲基－1，4－苯醌为原料合成全反式辅酶Q10。首先以茄尼醇合成全反式侧链四十碳十烯－1－醇，再和2，3－二甲氧基－5－甲基－1，4－苯醌的还原产物在无水氯化锌存在下缩合。缩合反应收率为20％。     

Coenzyme Q10: Novel Formulations and Medical Trends

The aim of this review is to shed light over the most recent advances in Coenzyme Q₁₀ (CoQ₁₀) applications as well as to provide detailed information about the functions of this versatile molecule, which have proven to be of great interest in the medical field. Traditionally, CoQ₁₀ clinical use was based on its antioxidant properties; however, a wide range of highly interesting alternative functions have recently been discovered. In this line, CoQ₁₀ has shown pain-alleviating properties in fibromyalgia patients, a membrane-stabilizing function, immune system enhancing ability, or a fundamental role for insulin sensitivity, apart from potentially beneficial properties for familial hypercholesterolemia patients. In brief, it shows a remarkable amount of functions in addition to those yet to be discovered. Despite its multiple therapeutic applications, CoQ₁₀ is not commonly prescribed as a drug because of its low oral bioavailability, which compromises its efficacy. Hence, several formulations have been developed to face such inconvenience. These were initially designed as lipid nanoparticles for CoQ₁₀ encapsulation and distribution through biological membranes and eventually evolved towards chemical modifications of the molecule to decrease its hydrophobicity. Some of the most promising formulations will also be discussed in this review.     

Water-Soluble Coenzyme Q10 Inhibits Nuclear Translocation of Apoptosis Inducing Factor and Cell Death Caused by Mitochondrial Complex I Inhibition

The objectives of the study were to explore the mechanism of rotenone-induced cell damage and to examine the protective effects of water-soluble Coenzyme Q10 (CoQ10) on the toxic effects of rotenone. Murine hippocampal HT22 cells were cultured with mitochondrial complex I inhibitor rotenone. Water-soluble CoQ10 was added to the culture media 3 h prior to the rotenone incubation. Cell viability was determined by alamar blue, reactive oxygen species (ROS) production by dihydroethidine (DHE) and mitochondrial membrane potential by tetramethyl rhodamine methyl ester (TMRM). Cytochrome c, caspase-9 and apoptosis-inducing factor (AIF) were measured using Western blotting after 24 h rotenone incubation. Rotenone caused more than 50% of cell death, increased ROS production, AIF nuclear translocation and reduction in mitochondrial membrane potential, but failed to cause mitochondrial cytochrome c release and caspase-9 activation. Pretreatment with water-soluble CoQ10 enhanced cell viability, decreased ROS production, maintained mitochondrial membrane potential and prevented AIF nuclear translocation. The results suggest that rotenone activates a mitochondria-initiated, caspase-independent cell death pathway. Water-soluble CoQ10 reduces ROS accumulation, prevents the fall of mitochondrial membrane potential, and inhibits AIF translocation and subsequent cell death.     

Determination of Coenzyme Q<Subscript>9</Subscript> and Q<Subscript>10</Subscript> in Developing Palm Fruits

A series of biochemical changes took place as oil palm fruits developed into maturity. Among these are the active synthesis of glycerides as well as unsaponifiable compounds such as carotenes, vitamin E, sterols and squalene. Coenzyme Q₉ and Q₁₀ were found to be present at different stages of the oil palm fruits development. The presence of coenzyme Q₉ was detected as early as 4 weeks after anthesis (WAA) and its concentration diminished as the oil palm fruits ripen. Coenzyme Q₁₀ on the other hand, can only be detected from 12WAA onwards and its concentration remained at an elevated level throughout the remaining development period of the oil palm fruits. Their occurrence pattern suggested that there is a strong relationship between the concentration of coenzyme Q₉ and coenzyme Q₁₀ with the age of the oil palm fruits.     

Comparison of low‐temperature processes for oil and coenzyme Q10 extraction from mackerel and herring

Among the fat fish species available from Eastern Quebec (Canada), whole Atlantic mackerel (Scomber scombrus) and herring (Clupea harengus) represent abundant fishery resources which are currently under‐utilized. They have relatively high contents of oil and coenzyme Q10 (CoQ10) in their tissues, which could be valuable for nutraceutical applications. Therefore, two low‐temperature extraction processes were compared for the recovery of oil and CoQ10 from these resources, such as enzymatic hydrolysis using Protamex? and supercritical CO₂ (SCO₂) using fish lyophilizates. The results revealed that highest yields of oil and CoQ10 were obtained using the enzymatic hydrolysis process with mackerel. Whatever the process used, CoQ10 concentrations were higher in herring oil, due mainly to a more selective extraction of CoQ10 over that of the oil. The highest CoQ10 recovery rates (extraction efficiencies) were obtained using the enzymatic hydrolysis process with both types of fish, but also the SCO₂ process with herring under some conditions. For mackerel, the lower CoQ10 recovery rates obtained from the SCO₂ process were explained by its more important matrix effect. An economic assessment of both processes revealed that the enzymatic hydrolysis extraction process would be the most promising for up‐scaling the recovery of oil and CoQ10 from these resources.     

微生物发酵法生产辅酶Q10的研究进展

辅酶Q10由于具有多种生理功能而得到广泛应用,介绍了近年来微生物发酵法生产辅酶Q10在高产菌株的筛选、传统方法诱变育种、基因工程定向育种、发酵工艺优化、提取纯化工艺优化和定量分析等方面的研究进展.     

生物发酵产物中辅酶Q_(10)的快速分离柱高效液相色谱法测定

研究了用快速分离柱高效液相色谱法测定生物发酵产物中辅酶Q10的方法。发酵法得到的样品经匀浆后用四氢呋喃提取,提取液以ZORBAX Stab le Bound(4.6×50 mm,1.8μm)C18快速分离柱为固定相,m(甲醇)∶m(四氢呋喃)=4∶1为流动相分离,流速为2.0 mL/m in;在该色谱条件下,辅酶Q10在2.0 m in内可达到基线分离;用紫外二极管矩阵检测器检测。方法标准回收率为96%~102%,相对标准偏差为0.78%~1.22%。方法用于几种发酵样品中辅酶Q10的测定,结果满意。     

Chromosomal Engineering of Escherichia coli for Efficient Production of Coenzyme Q10

The plasmid-expression system is routinely plagued by potential plasmid instability. Chromosomal in-tegration is one powerful approach to overcome the problem. Herein we report a plasmid-free hyper-producer E. coli strain for coenzyme Q10 production. A series of integration expression vectors, pxKC3T5b and pxKT5b, were constructed for chemically inducible chromosomal evolution （multiple copy integration） and replicon-free and markerless chromosomal integration （single copy integration）, respectively. A coenzyme Q10 hyper-producer Es-cherichia coli TBW20134 was constructed by applying chemically inducible chromosomal evolution, replicon-free and markerless chromosomal integration as well as deletion of menaquinone biosynthetic pathway. The engineered E. coli TBW20134 produced 10.7 mg per gram of dry cell mass （DCM） of coenzyme Q10 when supplemented with 0.075 g·L^-1 of 4-hydroxy benzoic acid;this yield is unprecedented in E. coli and close to that of the commercial producer Agrobacterium tumefaciens. With this strain, the coenzyme Q10 production capacity was very stable after 30 sequential transfers and no antibiotics were required during the fermentation process. The strategy presented may be useful as a general approach for construction of stable production strains synthesizing natural products where various copy numbers for different genes are concerned.     

辅酶Q10的合成与应用

以茄尼醇、异戊二烯和辅酶Q0作为主要原料,通过溴化、加成和缩合3步反应合成了目标产物辅酶Q10.其中溴化和加成反应一次收率分别为95%和90%,缩合反应重复收率在70%左右.生产规模为5kg/次.并介绍了辅酶Q10的应用和市场需求.     

多策略代谢工程大肠杆菌高效生产辅酶Q10

Escherichia coli BW25113 was metabolically engineered for CoQ10 production by replacing ispB with ddsA from Gluconobacter suboxydans.Effects of precursor balance and reduced nicotinamide-adenine dinucleotide phosphate (NADPH) availability on CoQ10 production in E.coli were investigated.The knockout of pykFA along with pck overexpression could maintain a balance between glyceraldehyde 3-phosphate and pyruvate,increasing CoQ10 production.Replacement of native NAD-dependent gapA with NADP-dependent gapC from Clostridium acetobutylicum,together with the overexpression of gapC,could increase NADPH availability and then enhanced CoQ10 production.Three effects,overexpressions of various genes in CoQ biosynthesis and central metabolism,different vectors and culture conditions on CoQ10 production in E.coli,were all investigated.The investigation of different vectors indicated that low copy number vector may be more beneficial for CoQ10 production in E.coli.The recombinant E.coli (△ispB::ddsA,△pykFA and △gapA::gapC),harboring the two plasmids encoding pck,dxs,idi and ubiCA genes under the control of PT5 on pQE30,ispA,ddsA from Gluconobacter suboxydans and gapC from Clostridium acetobutylicum under the control of PBAD on pBAD33,could produce CoQ10 up to 3.24 mg·g-1 dry cell mass simply by changing medium from M9YG to SOB with phosphate salt and initial culture pH from 7.0 to 5.5.The yield is unprecedented and 1.33 times of the highest production so far in E.coli.     

辅酶Q10对H_2O_2损伤PC12细胞保护作用的实验研究

研究通过H2O2诱导的PC12细胞建立阿尔茨海默症氧化损伤细胞模型,以细胞活性、丙二醛含量、谷胱甘肽含量及谷胱甘肽转移酶活性水平为指标,检测Co Q10对PC12细胞氧化应激损伤的保护作用。结果表明Co Q10能够提高H2O2所致PC12细胞的细胞活力的降低,降低了由H2O2诱导的脂质过氧化程度,并且可以抑制H2O2诱导的PC12细胞GSH含量及GST活性的降低。Co Q10对H2O2损伤的PC12细胞具有保护作用,可能是通过降低脂质过氧化程度且提高谷胱甘肽含量及谷胱甘肽转移酶活性实现的。     

辅酶Q_(10)化学合成方法与工艺研究进展评述

辅酶Q10是一种具有重要生理和药理作用的生命活性物质,用途十分广泛。特别是近年来随着对其生理和药理作用认识的不断深入,辅酶Q10已经成为了一种重要的药品和保健品。笔者根据半个世纪以来有关辅酶Q10化学合成的各种文献报道,对辅酶Q10现有的各种化学合成路线及其工艺优劣进行了介绍和比较,重点介绍了从烟草中提取天然茄尼醇为原料的半化学合成辅酶Q10方法,并阐明了化学合成辅酶Q10的关键因素和发展方向。     

固体脂质对辅酶Q10脂质纳米囊的稳定性及透皮性能的影响

为了提高辅酶Q10（CoQ10）的透皮吸收效果，以辛酸癸酸甘油三酯（GTCC）为液体脂质，鲸蜡醇棕榈酸酯（CP）为固体脂质，制备得到了不同浓度固体脂质的CoQ10-脂质纳米囊，并对其稳定性及透皮性能进行了研究。结果表明，固体脂质的添加未对CoQ10-脂质纳米囊的粒径及储藏稳定性产生影响，粒径均为52nm左右，储藏180d后其粒径及外观均无明显变化。FT-IR结果表明，CoQ10与脂质纳米囊之间未发生化学反应且能被很好的包埋在脂质纳米囊中。通过闭合效应、体外透皮实验及激光共聚焦的观察发现，当CP质量浓度增加至70%和95%（以固液总脂质为基准，下同）时，CoQ10-脂质纳米囊能显著减少皮肤水分的蒸发，并更好的促进CoQ10的透皮吸收，使表皮层和真皮层的荧光强度明显增强。     

The Roles of Coenzyme Q in Disease: Direct and Indirect Involvement in Cellular Functions

Coenzyme Q (CoQ) is a key component of the respiratory chain of all eukaryotic cells. Its function is closely related to mitochondrial respiration, where it acts as an electron transporter. However, the cellular functions of coenzyme Q are multiple: it is present in all cell membranes, limiting the toxic effect of free radicals, it is a component of LDL, it is involved in the aging process, and its deficiency is linked to several diseases. Recently, it has been proposed that coenzyme Q contributes to suppressing ferroptosis, a type of iron-dependent programmed cell death characterized by lipid peroxidation. In this review, we report the latest hypotheses and theories analyzing the multiple functions of coenzyme Q. The complete knowledge of the various cellular CoQ functions is essential to provide a rational basis for its possible therapeutic use, not only in diseases characterized by primary CoQ deficiency, but also in large number of diseases in which its secondary deficiency has been found.     

高产辅酶Q_(10)光合细菌的分离与鉴定

由于辅酶Q10有特殊的生理功能,在治疗癌症等各种疾病方面有很好的疗效,是人体必需物质,无毒副作用。近年来在医药、食品添加剂和保健品方面得到了广泛应用,但目前辅酶Q10的合成效率还不是很高,且成本非常高,因此,这需要我们投入大量的研究工作,以满足市场上日益增长的需求。本课题以提高光合细菌(PSB)菌体辅酶Q10的产量为目的,从样品中分离到一株高产辅酶Q10的光合细菌,经初步鉴定为沼泽红假单胞菌。     

Preparation and Characterization of Catalase-Loaded Solid Lipid Nanoparticles Protecting Enzyme against Proteolysis

Catalase-loaded solid lipid nanoparticles (SLNs) were prepared by the double emulsion method (w/o/w) and solvent evaporation techniques, using acetone/methylene chloride (1:1) as an organic solvent, lecithin and triglyceride as oil phase and Poloxmer 188 as a surfactant. The optimized SLN was prepared by lecithin: triglyceride ratio (5%), 20-second + 30-second sonication, and 2% Poloxmer 188. The mean particle size of SLN was 296.0 ± 7.0 nm, polydispersity index range and zeta potential were 0.322-0.354 and -36.4 ± 0.6, respectively, and the encapsulation efficiency reached its maximum of 77.9 ± 1.56. Catalase distributed between the solid lipid and inner aqueous phase and gradually released from Poloxmer coated SLNs up to 20% within 20 h. Catalase-loaded SLN remained at 30% of H(2)O(2)-degrading activity after being incubated with Proteinase K for 24 h, while free catalase lost activity within 1 h.