Effect of coenzyme Q10 supplementation on post-vitrification mouse embryo development

Objective:To investigate the effects of coenzyme Q10(CoQ10)supplementation on post-vitrification embryo development and gross morphology.Methods:Balb/c mouse embryos were cultured in potassium simplex optimised medium(KSOM)with varying CoQ10 concentrations[0(control),20,40,and 60μM].The most effective CoQ10 concentration(40μM)was selected for subsequent post-vitrification morphology study.Embryos were randomly divided into four groups:Group A(non-vitrified without CoQ10),Group B(non-vitrified with CoQ10),Group C(vitrified without CoQ10),and Group D(vitrified with CoQ10),followed by vitrification at the 8-cell stage.Survival rates and development until the blastocyst stage were evaluated through morphological examinations using ASEBIR's system,distinguishing normal and abnormal embryos.Results:Supplementation of 40μM CoQ10 significantly increased blastocyst formation(95%)compared to the control group(92%),20μM(62%),and 60μM(56%)(P<0.001).Following vitrification,Group D exhibited a significant increase in blastocyst formation(92%)compared to Group C(82%)(P<0.05).Morphological assessments indicated superior embryo quality in Group B over Group D during the cleavage stage,morula,and blastocyst(P<0.05).Conclusions:CoQ10 supplementation exhibits promising potential to enhance preimplantation embryo development,increase blastocyst formation rates,and improve embryo quality post-vitrification.This offers a promising approach to mitigate oxidative stress on embryos,potentially improving overall assisted reproductive technology outcomes.

COQ8A基因突变所致原发性辅酶Q10缺乏症1例

原发性辅酶Q10缺乏症属于常染色体隐性遗传病,由辅酶Q10生物合成过程中关键酶的编码基因发生突变导致。本文首次报道辅酶Q8A(coenzyme Q8A,COQ8A)基因一种新的杂合变异导致的原发性辅酶Q10缺乏症。患者为26岁男性,以缓慢起病的共济失调、运动不耐受、震颤以及癫痫发作为主要表现,诊断原发性辅酶Q10缺乏症,全外显子基因测序结果示COQ8A基因复合杂合突变[NM_020247.5:exon8:c.1009G>A(p.A337T)和NM_020247.5:exon8:c.1078delC(p.Q360Sfs*20)],予以补充辅酶Q10后症状有所改善。该病例提示当患者出现共济失调、运动障碍,尤其伴癫痫或认知功能障碍时,应排查可治性的原发性辅酶Q10缺乏症。

Multiple Strategies for Metabolic Engineering of Escherichia coli for Efficient Production of Coenzyme Q_（10）

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.

Advances in the Coenzyme Q10 Biosynthesis Pathway in Rhodobacter sphaeroides and the Enhancement of Coenzyme Q10 Production Based on Metabolic Engineering

Coenzyme Q10 is widely used in food,cosmetics and pharmaceuticals,possessing a broad market.Rhodobacter sphaeroides is enriched in natural coenzyme Q10 and is becoming an important microorganism for producing natural coenzyme Q10.The paper reviewed the biosynthesis pathways of coenzyme Q10 in R.sphaeroides and the advances in enhancement of coenzyme Q10 production in R.sphaeroides based on metabolic engineering.

Evidence on neuroprotective properties of coenzyme Q10 in the treatment of glaucoma

Glaucoma, the leading cause of visual impairment and irreversible blindness worldwide, is a multifactorial, progressive optic neuropathy characterized by loss of retinal ganglion cells, alterations of the optic nerve head, and specific visual field defects. Clinical evidence shows that intraocular pressure is the major risk factor of the treatable disease. However, in some patients, glaucoma develops and continues to progress despite normal intraocular pressure values, suggesting that other risk factors are involved in the disease. Consequently, neuroprotective treatments, focused on preventing retinal ganglion cells death by acting on different therapeutic strategies but not focused on intraocular pressure reduction, has therefore become of great interest. In this contest, coenzyme Q10, showing evidence in slowing or reversing pathological changes typical of the disease, has been proposed as a potential neuroprotective agent in glaucoma. In this review, we describe the possible mechanisms of action of coenzyme Q10 and the recent evidence in literature regarding the neuroprotective activity of the molecule.

Breeding of Coenzyme Q_(10) Produced Strain by Low-Energy Ion Implantation and Optimization of Coenzyme Q_(10) Fermentation

In order to increase the production efficiency of coenzyme Q10, the original strain Agrobacterium tumefaciens ATCC 4452 was mutated by means of Nitrogen ions implantation. A mutant strain, ATX 12, with high contents of coenzyme Q10 was selected. Subsequently, the conditions such as carbohydrate concentration, nitrogen source concentration, inoculum＇s size, seed age, aeration and temperature which might affect the production of CoQ10 were investigated in detail. Under optimal conditions, the maximum concentration of the intracellular CoQ10 reached 200.3 mg/L after 80 h fed-batch fermentation, about 245% increasing in CoQ10 production after ion implantation, compared to the original strain.

Effects of dietary coenzyme Q10 supplementation during gestation on the embryonic survival and reproductive performance of high‑parity sows

Background Fertility declines in high-parity sows.This study investigated whether parity-dependent declines in embryonic survival and reproductive performance could be restored by dietary coenzyme Q10(CoQ10)supplementation.Methods Two experiments were performed.In Exp.1,30 young sows that had completed their 2nd parity and 30 high-parity sows that had completed their 10^(th)parity,were fed either a control diet(CON)or a CON diet supple-mented with 1 g/kg CoQ10(+CoQ10)from mating until slaughter at day 28 of gestation.In Exp.2,a total of 314 post-weaning sows with two to nine parities were fed the CON or+CoQ10 diets from mating throughout gestation.Results In Exp.1,both young and high-parity sows had a similar number of corpora lutea,but high-parity sows had lower plasma CoQ10 concentrations,down-regulated genes involved with de novo CoQ10 synthesis in the endome-trium tissues,and greater levels of oxidative stress markers in plasma and endometrium tissues.High-parity sows had fewer total embryos and alive embryos,lower embryonic survival,and greater embryo mortality than young sows.Dietary CoQ10 supplementation increased the number of live embryos and the embryonic survival rate to levels simi-lar to those of young sows,as well as lowering the levels of oxidative stress markers.In Exp.2,sows showed a parity-dependent decline in plasma CoQ10 levels,and sows with more than four parities showed a progressive decline in the number of total births,live births,and piglets born effective.Dietary supplementation with CoQ10 increased the number of total births,live births,and born effective,and decreased the intra-litter covariation coefficients and the percentage of sows requiring farrowing assistance during parturition.Conclusions Dietary CoQ10 supplementation can improve the embryonic survival and reproductive performance of gestating sows with high parity,probably by improving the development of uterine function.

Validation and application of Vierordt’’s spectrophotometric method for simultaneous estimation of tamoxifen/coenzyme Q10 in their binary mixture and pharmaceutical dosage forms

For the sake of improving patient compliance and sustainability of chemotherapy healthcare system, both TC and CoQ10 were formulated as solid lipid nanoparticles (SLNs). The study was focused on establishing and validating a simple and reproducible spectrophotometric method for simultaneous determination of TC and CoQ10 in their binary mixture or pharmaceutical dosage forms. A new method based on simultaneous estimation of drug mixture without prior separation was developed. Validation parameters were checked with International Conference on Harmonization (ICH) guidelines. The accuracy and reproducibility of proposed method was statistically compared to HPLC. The TC and CoQ10 were quantified at absorptivity wavelengths of 236 nm and 275 nm, respectively. Calibration curves obeyed Beer’s law in range of 2–14 μg/ml with a correlation coefficient (R^2) of 0.999 in both methanol and simplified simulated intestinal fluid (SSIF). The %means recovery of TC and Co Q10 in pure state or binary mixture at various concentration levels were all around 100%.The low values of SD and %RSD (<2%) confirm high precision and accuracy of the proposed method. Formulated SLNs showed different %means recovery in range 81–92% for TC and 32–59% for CoQ10. The data obtained by applying simultaneous Vierordt’s equations showed no statistical significance in comparison to HPLC. Vierordt’s method was successfully applied as a simple, accurate, precise, and economical analysis method for estimating TC and CoQ10 concentrations in pure state, binary mixture and pharmaceutical dosage forms.

Plasma coenzyme Q10 levels in type 2 diabetic patients with retinopathy

AIMTo determine the relationship between proliferative diabetic retinopathy (PDRP) and plasma coenzyme Q10(CoQ10) concentration.

Chromosomal Engineering of Escherichia coli for Efficient Production of Coenzyme Q_(10)

The plasmid-expression system is routinely plagued by potential plasmid instability. Chromosomal integration 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 Escherichia 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-1of 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.

Coenzyme Q10 in neurodegenerative disorders: Potential benefit of Co Q10 supplementation for multiple system atrophy

Coenzyme Q10(Co Q10) is an essential cofactor in the mitochondrial respiratory pathway and also functions as a lipid-soluble antioxidant. Co Q10 deficiency has been implicated in many clinical disorders and aging. Primary Co Q10 deficiency is a group of recessively inherited diseases caused by mutations in any gene involved in the Co Q10 biosynthesis pathway. Although primary Co Q10 deficiency is rare, its diagnosis is important because it is potentially treatable with exogenous Co Q10. Multiple system atrophy(MSA) was recently shown to be linked to mutations in the COQ2 gene, one of the genes involved in the Co Q10 biosynthesis pathway. MSA is relatively common in adult-onset neurodegenerative diseases characterized by Parkinsonism, cerebellar ataxia and autonomic failures. Because COQ2 mutations are associated with an increased risk of MSA, oral Co Q10 supplementation may be beneficial for MSA, as for other primary Co Q10 deficiencies. Statins are 3-hydroxy-3-methylglutaryl coenzyme A inhibitors that inhibit the biosynthesis of cholesterol, as well as the synthesis of mevalonate, a critical intermediate in cholesterol synthesis. Statin therapy has been associ-ated with a variety of muscle complaints from myalgia to rhabdomyolysis. Statin treatment carries a potential risk of Co Q10 deficiency, although no definite evidence has implicated CQ10 deficiency as the cause of statinrelated myopathy.

Coenzyme Q10 as a therapeutic candidate for treating inherited photoreceptor degeneration

Inherited photoreceptor degeneration（IPD）：The human retina is a highly specialised tissue that enables the perception of light across a range of intensities and colours.It covers about65%of the inner surface of the eye and contains three layers of cells：the outer nuclear layer（ONL）containing the cell bodies and nuclei of the light-sensitive rod and cone photoreceptorswhose photopigment-containing outer segments form the photoreceptor layer; the inner nuclear layer （INL） containing bipolar, horizontal and amacrine cells; and the ganglion cell layer （GCL） from which the optic nerve arises. There are two layers of synaptic connections between these three layers： the photoreceptors synapse with second order neurons, mainly bi- polar cells, in the outer plexiform layer （OPL）, while in turn the bipolar cells form connections in the inner plexiform layer （IPL） with ganglion cells. The retinal pigment epithelium （RPE） lies directly behind the photoreceptor layer, is heavily pigmented to reduce scattering of light, and is essential for the nourishment, maintenance and metabolism of photoreceptors.

一株产CoQ_(10)的红假单胞菌的研究

从滇池分离到一株革兰氏阴性杆菌 ,于光照厌氧及黑暗好氧条件下生长良好 ,活细胞在 870、80 3 nm处有吸收峰 ,具平行片层状的光合内膜系统 ,可归属于红假单胞菌属 (Rhodopseudomonas) ;该菌体中提取分离得到的黄色结晶品 ,经 Craven试验、高效薄层层析、高效液相色谱、紫外光谱、红外光谱、核磁共振及质谱证实 ,为 Co Q1

CoQ10软胶囊在Beagle犬体内的相对生物利用度评价

目的:比较在 Beagle 犬体内3种辅酶 Q_(10)(CoQ_(10))制剂的相对生物利用度。方法:采用三制剂三周期(3×3拉丁方)自身对照交叉多剂量口服给药方式,周期间清洗1周。用高效液相色谱法测定6条健康 Beagle 犬口服 CoQ_(10)软胶囊(受试制剂A)、CoQ_(10)普通胶囊(参比制剂 B)和进口 CoQ_(10)软胶囊(参比制剂 C)后不同时间点血浆中 CoQ_(10)的浓度,扣除内源性 CoQ_(10)本底,绘制血药浓度-时间曲线,计算相对生物利用度。结果:受试犬口服 CoQ_(10)60mg 的受试制剂(A)和参比制剂(B,C)后,血浆中 CoQ_(10)AUC_(0-t)(μg·h·mL^(-1))分别为69.31±40.49,62.38±59.05,104.90±64.32。与参比制剂 B 与 C 相比,受试制剂中的相对生物利用度平均为(144.07±79.48)%和(78.60±54.28)%。结论:CoQ_(10)受试制剂 A 生物利用度高于参比制剂 B 而低于参比制剂 C。

Effect of levocarnitine+coenzyme Q10 adjuvant therapy on vasoactive molecules,endothelial injury and oxidative stress in patients with chronic heart failure

Objective: To study the effect of levocarnitine + coenzyme Q10 adjuvant therapy on vasoactive molecules, endothelial injury and oxidative stress in patients with chronic heart failure. Methods: A total of 90 patients with chronic heart failure who were treated in the hospital between December 2014 and December 2016 were collected and divided into control group and observation group by random number table method, 45 cases in each group. Control group received conventional therapy, and observation group received levocarnitine + coenzyme Q10 adjuvant therapy on the basis of conventional therapy. The differences in vasoactive molecule, endothelial injury and oxidative stress levels were compared between the two groups before and after treatment. Results: Before treatment, the differences in vasoactive molecule, endothelial injury and oxidative stress levels were not statistically significant between the two groups of patients. After treatment, serum vasoactive molecules ET-1, AngⅡ and TXB2 contents of observation group were lower than those of control group while NO content was higher than that of control group;endothelial function indexes FMD level was higher than that of control group;serum oxidative stress indexes SOD and T-AOC contents were higher than those of control group while MDA and ROS contents were lower than those of control group. Conclusion: Levocarnitine + coenzyme Q10 adjuvant therapy can optimize the vascular activity, and reduce the endothelial injury and systemic oxidative stress response in patients with chronic heart failure.

CoQ_（10）对醋氨酚肝毒性的保护作用探讨

＊Ｐ＜０．００１从表３可见，ＣｏＱ１０高低剂量组肝匀浆ＣｙｔＰ４５０含量与ｎＳ对照组比较没有明显差异（Ｐ＞０．０５），而苯巴比妥钠组小鼠肝匀浆ＣｙｔＰ４５０含量较ＮＳ对照组及高低剂量组显著升高，有非常显著性差异（Ｐ＜０．００１）。本巴比妥钠为肝药酶诱导剂，其阳性结果证明此实验方法可靠，也证明ＣｏＱ１０对肝细胞ＣｙｔＰ４５０含量没有影响。３讨论本实验结果表明：小鼠用ＣｏＱ１０作预处理，确能保护超量ＡＡＰ对肝脏的损伤作用；同时能使ＡＡＰ血清浓度明显升高，而对肝细胞ＣｙｔＰ４５０含量则无影响。据文献报道，超ｔＡＡＰ进入肝细胞微粒体后，通过肝药酶代谢产出大量毒性中间产物，因而引起肝损害甚至肝坏死［７］。ＣｏＱ１０能防止这一毒性效应，通过稳定肝细胞微粒体膜或抑制肝药酶活住，均可达此目的．上述实验结果．均提示ＣｏＱ１０的作用途径可能是稳定肝细胞微粒体膜，使ＡＡＰ进入微粒体数量减少，因而毒性中间代射产物生成减少，使肝细胞损伤减轻。

Effects of dietary CoQ₁₀and α-lipoic acid on CoQ₁₀levels in plasma and tissues of eggs laying hens

In this paper we described the effect of administrated CoQ10, and alfa-lipoic acid on the concentration of total CoQ10 inplasma end body tissues of eggs laying hens. Organisms raise a complex network of enzymes, metabolites and molecules with antioxidant activities in order to prevent oxidative damage of theirs bodies. Adequate blood concentrations of small weight molecules ingested with food and food additives are important for the proper functioning of the antioxidant defense. To test this hypothesis we prepared following experiment. Forty weeks old hens were selected from two genotypes;Ross 308 broiler mothers and Lohmann breed hens. Animals were fed for a period of 84 days. Concentrations of supplemented CoQ10 and ALAwere calculated from feed instruction tables so each hen received an average of approximately 5 mg of CoQ10 and 50 mg ofALAper kg of animal weight per day. During the experiment blood samples were taken and at the end of the experiment different body tissues (heart, liver, breast, legs) were collected and analyzed with originally developed HPLC-MS/MS method based selective ionization with LiCl on MRM scanning. We found a number of interesting and unexpected results. Supplemented CoQ10 increased concentrations of coenzyme CoQ10 inplasma and different hen’s tissues. Increased concentration of CoQ10 is the result of its transfer with chylomicrons from the digestive tract to various organs of the body and to the liver where exogenous and endogenous CoQ10 has been re-redistributed through lipoproteins. Supplemented ALA caused much greater concentration of CoQ10 indifferent tissues and plasma then CoQ10. Plausible explanation of our results is such that ALA may regenerates the antioxidants and accelerate the formation of endogenous CoQ10 which is distributed with lipoprotein carriers and increases overall concentration of CoQ10. Our experiments definitely show that Lipoic acid beside glutathione promotes also a synthesis of CoQ10 and increases the total concentration especially in liver and heart tissues.

Single intravenous injection of CoQ₁₀reduces infarct size in a rat model of ischemia and reperfusion injury

Maintenance of mitochondrial activity and antioxidant features of coenzyme Q10 (CoQ10) could be an effective background for treatment of acute myocardial ischemia. Dietary uptake of CoQ10 is limited to only a few percent. In urgent cases, parenteral administration of CoQ10 could provide fast increase of its plasma and myocardial levels. The aim was to evaluate whether a single intravenous (i.v.) injection of solubilized CoQ10 before ischemia/reperfusion (IR) could lead to replenishment of its myocardial levels and limits subsequent myocardial IR injury. Methods: 30 min prior to coronary artery occlusion rats received i.v. solubilized CoQ10 (30 mg/kg) or saline (1 ml/kg). After 30 min of ischemia and 120 min of reperfusion, infarct zone of left ventricle (LV) and quantity of CoQ10 in LV were determined. Cardiac rhythm was monitored through the whole experiment. Results: At the beginning of reperfusion, arrhythmias were recorded in 8 (from 9) in saline and 2 (from 9) in CoQ10-treated rats. Arrhythmias in CoQ10-treated rats arose later (40 ± 8 sec) and had less duration (26 ± 14 sec);14 ± 13 sec and 52 ± 17 sec in saline treated rats respectively. At the end of reperfusion CoQ10 treated rats revealed: 2 fold higher CoQ10 content in LV (p 10 were accompanied by less infarct size (r = ﹣0.77, p i.v. injection of CoQ10 effectively increased its myocardial levels and protected heart against IR injury by diminishing the size of the irreversibly damaged myocardium, decreasing frequency and duration of arrhythmias. The infarct zone inversely correlated with the quantity of CoQ10 in LV.

前体物质对辅酶Q_(10)生物合成的影响

首次研究了以辅酶Q10 (CoQ10 )主要侧链供给前体物质———茄尼醇和醌环供给前体———羟基苯甲酸和辅酶Q0 对粟酒裂殖酵母(Schizosaccharomycespromb 2 1794 - 2 3) 生产CoQ10 产量的影响,并对前体的转化工艺进行了初步研究。确定了适宜于粟酒裂殖酵母生长及高转化前体生成CoQ10 的条件为:酵母在2 8℃下,2 2 0r/min于发酵培养基中培养18h后,加入0 5 g/L茄尼醇继续发酵培养18h ,进行前体转化反应。结果表明,单独添加茄尼醇能达到最大产量33 1mg/L ,比对照样品增加了91% ,任2种前体物质共同添加都要比单独添加茄尼醇时产量低。茄尼醇和CoQ0 共同添加时,单位细胞胞外辅酶CoQ10 的产量达到最高的1 35mg/g ,比对照样品增加了117%。

利用广谱性和特异性组合诱变技术选育辅酶Q_(10)高产菌

为提高辅酶Q10产量,采用了对细胞进行全面的非特异性诱变和针对特定途径的特异性诱变相结合的育种策略.以放射型根瘤菌(Rhizobium radiobacter)WSH2601为出发菌株,选择UV射线和亚硝基胍作为诱变剂,在筛选获得放线菌素D抗性突变株的基础上,通过进一步的诱变处理,分别获得了放线菌素D和L-乙基硫氨酸双抗性突变株、放线菌素D和维生素K3双抗性突变株,以及抗放线菌素D和X-gal利用能力提高的突变株.与出发菌株相比,突变株辅酶Q10的产量提高幅度达25%～37%.其中一株放线菌素D和L-乙基硫氨酸双抗性突变株WSH-E25,胞内辅酶Q10含量和辅酶Q10总产量分别达到2.86 mg g DCW-1和40.0 mg L-1,均比出发菌株提高了37%,且遗传稳定性良好.