辅酶Q_(10)的毒性试验及对溶血空斑形成细胞的刺激效应

辅酶Q_10（以下简称CoQ_(10)）是生物体内普遍存在的一类生物活性物质,具有重要的生理、生化作用。它是细胞呼吸链中主要的递氢体,在生物能量转化中占重要地位,它又能作为非特异性免疫增强剂,增强机体的防御能力,因此人们对于CoQ_(10)的临床研究越来越受到重视（中国科学院昆明动物研究所,1979）。 我们结合我国具体情况,独创地从提取细胞色素丙的猪心残渣中分离制备了CoQ_(10), （许以盛等,1985）,并与有关生产及临床单位协作,于1977年12月通过了鉴定,为我国的生化药物填补了一项空白。     

提取细胞色素C的研究

本文报道了以猪心为原料提取细胞色素Ｃ的小批量实验结果。通过十批次的实验,经含量、活性和收率等指标的测定,平均收率为１９８．５６ｍｇ／ｋｇ,活性在９６～１１０％之间。此收率稍高于国内报道的提取细胞色素Ｃ最高水平（１９８．２３ｍｇ／ｋｇ）,超过省内生产厂家的标准。提示用本文采用提取精制细胞色素Ｃ的方法是可行的。     

鸡心线粒体辅酶Q-细胞色素c还原酶的分离及性质研究

鸡心线粒体内膜辅酶Q-细胞色素c还原酶在抗霉素A稳定下经TritonX-100增溶、羟基磷灰石分离,凝胶过滤而纯化。所获得的辅酶Q-细胞色素c还原酶以二聚体形式与TritonX-100生成单分散相的蛋白——去垢剂复合物。经十二烷基磺酸钠-聚丙烯酰胺凝胶电泳鉴定,它由八个不同的亚基组成,其表观分子量各为:45,000;41,000;36,000;31,000;25,000;15,500;12,500;10,000。光谱分析测定,细胞色素b和细胞色素c_1的克分子比为2:1。采用同样方法也巳从猪心线粒体制备了辅酶Q-细胞色素c还原酶-Triton复合物。两者具有相似的吸收光谱。但猪心还原酶丢失了分子量25,000的亚基。     

环境影响因子对玉米芽中辅酶Q10含量的影响

探讨了玉米芽中影响辅酶Q₁₀含量的环境因子以及辅酶Q₁₀的提取新方法。结果表明:光和水环境因子影响较大,在受到水或光因子胁迫时,玉米芽中辅酶Q₁₀有较明显的增长趋势;玉米芽中辅酶Q₁₀的初步提取方法为采用70%乙醇同鲜活玉米芽混合进行鲜磨匀浆萃取,固液分离后残渣再用氯仿超声提取3次为最佳提取方法。     

动态吸附法生产牛心细胞色素丙粗品

国内以牛心为原料生产细胞色素而粗品的厂家占一定比例,而且普遍采用静态吸附法传统工艺[1],产率一般在120mp／kg左右（原料以全牛心重量计）。采用动态吸附法于实验室制备猪心细胞色素两可大幅度提高收率[2]。我们报道了牛心细胞色素丙粗品批量生产中采用动态吸附法,得到了5个连续批次的高收率,平均收率为160mg/kg。1材料与方法1．1材料1．1．1原料牛心,未去脂肪、结缔组织,购自本地向联厂冷库。1．1．2试剂硫酸朝阳轻化工研究所试剂厂;氨水朝阳轻化工研究所试剂厂;氯化钠沈阳市试剂三厂;三氯乙酸广州化学试剂厂;氢氧化钠锦州化…     

动物学研究第6卷总目录

第1 期从提取细胞色素丙后的猪心残渣中制备辅酶Q;。的研究 —”””””””’”””””··’·’…·’……··’…·’…··’·’………………·许以盛 胡 钧 张汉云(1)中国白纹伊蚊亚组伊蚊的研究 1.幼虫—…………………………………… .r击宝 /4(9)西沙群岛常见蚊蝇调查初报—………………………………………·黄为良 肖 森(19)华西雨蛙一新亚种描述………………………………………………利思敏 杨大同(23)团扇鳃科(Platyrhinidac)和团扇锻属(Platyrhina)分类特征问题的商榷……………… ’………··’…’……··…     

人参辅酶Q_(10)防晒霜对紫外线损伤小鼠的保护作用

目的研究人参辅酶Q_(10)防晒霜对紫外线损伤小鼠的保护作用。方法 36只小鼠随机分组,皮肤脱毛后分别涂抹相应霜剂,正常对照组不做任何处理,其他组进行紫外线照射,连续8周后,小鼠眼球取血处死进行指标检测。结果与正常组小鼠比较,模型组小鼠皮肤粗糙有皱纹、且角质层脱落,T-SOD与GSH-Px活力均明显下降,白细胞数与MDA含量明显增高(P0.05);与模型组比较,人参辅酶Q_(10)防晒霜组小鼠皮肤光滑、角质层没有脱落,T-SOD活力、红细胞与血红蛋白含量上升,白细胞数量、MDA含量与明显下降(P0.05),阳性对照组小鼠皮肤光滑、角质层没有脱落,GSH-Px活力上升,白细胞数量、MDA含量下降(P0.05)。结论人参辅酶Q_(10)防晒霜具有抗紫外线损伤的作用,其预防效果与甲氧基肉桂酸辛酯辅酶Q_(10)霜剂相当。     

酵母发酵生产辅酶Q10提取方法研究进展

辅酶Q10是存在于哺乳动物中,能与酶蛋白形成复合物以发挥酶学活性的有机小分子化合物,目前广泛应用于医药、日化、保健、食品等不同领域。辅酶Q10来源丰富,其中酵母是其工业生产的主要来源之一。广受关注的酵母发酵生产辅酶Q10的提取分离手段不断革新,产量不断增加,处理方式更加环保,应用日渐拓宽。本文就近年来国内外酵母发酵生产辅酶Q10的提取分离方法进行了综述,包括酵母菌种的类型与优化、辅酶Q10提取检测方法、工业生产放大工艺以及与产品质量相关的各个影响因素,并对酵母发酵生产辅酶Q10的前景进行了展望。     

紫细菌光合色素指纹图谱的建立与色素分析

【目的】探索快速高效的色素样品制备方法;为建立紫细菌全色素TLC和HPLC标准指纹图谱和数据库提供研究方法和思路;为色素代谢与调控等研究提供快速的色素分析方法。【方法】选择沼泽红假单胞菌(Rhodopesudomonas palustris CQV97)为材料,采用改良丙酮甲醇提取法、TLC重复展层法、图像灰度分析法、吸收光谱法、HPLC和MS法进行色素分析。【结果】甲醇或丙酮可选择性地提取样品的细菌叶绿素和类胡萝卜素,通过对丙酮甲醇法的改良,使色素提取总量提高了13.5%。建立了CQV97菌株全色素的TLC和HPLC指纹图谱,二者均含有11种色素组分。图像灰度分析法估算了TLC指纹图谱各色素组分的表观相对含量。以TLC图谱的各色素组分为外标物,明确了HPLC图谱中各色素组分的保留时间(Rt)与TLC图谱中各色素组分的迁移率(Rf)之间的对应关系。结合色素代谢途径、光谱分析和MS,定性分析了指纹图谱中11种色素组分。TLC或HPLC分析结果表明,理论样品与对照样品色素组分和含量一致,而实际样品与对照样品色素组分一致,但含量不同,重复测定3次,样品中色素相对含量的RSD均小于5%。【结论】改良的丙酮甲醇法可以快速高效地提取光合细菌的色素。采用TLC重复展层法和HPLC法建立的全色素指纹图谱色素组成一致,重复稳定性好,各具特色。TLC和HPLC两种指纹图谱分析方法均能进行光合色素的快速分析,适合于紫细菌色素合成途径中主要积累色素的组分和含量变化规律研究。     

三种天然果蔬色素稳定性研究

以新鲜菠菜、橘皮、番茄为原料,采用溶剂萃取法提取叶绿素、橘黄素、番茄红素,以吸光度为判断指标,研究了温度、光照、pH及金属离子对3种天然色素稳定性的影响。结果表明：橘皮色素稳定性较好,较耐热和耐光,pH稳定性较好;叶绿素耐热性和耐光性较差,酸性条件下较稳定;番茄红素稳定性最差,有一定的耐热性,不耐光,光照和紫外光下都易分解,不耐酸碱,酸性条件更不稳定。3种色素抗Mg2+、Na+、Ca2+等3种金属离子干扰能力较强,Cu2+、Fe3+对3种色素稳定性影响较大。     

Dietary coenzyme Q(10) supplement renders swine hearts resistant to ischemia-reperfusion injury

To examine whether nutritional supplementation of coenzyme Q(10) (CoQ(10)) can reduce myocardial ischemia-reperfusion injury, a group of swine was fed a regular diet supplemented with CoQ(10) (5 mg x kg(-1) x day(-1)) for 30 days. Another group of pigs that were fed a regular diet supplemented with placebo served as a control. After 30 days, isolated in situ pig hearts were prepared and hearts were perfused with a cardiopulmonary pump system. Each heart was subjected to 15 min of regional ischemia by snaring of the left anterior descending coronary artery, followed by 60 min of hypothermic cardioplegic global ischemia and 120 min of reperfusion. After the experiments were completed, myocardial infarct size was measured by triphenyltrazolium chloride staining methods. Postischemic left ventricular contractile function was better recovered in the CoQ(10) group than in the control group of pigs. CoQ(10)-fed pigs revealed less myocardial infarction and less creatine kinase release from the coronary effluent compared with control pigs. The experimental group also demonstrated a smaller amount of malonaldehyde in the coronary effluent and a higher content of the endogenous antioxidants ascorbate and thiol. Significant induction of the expression of ubiquitin mRNA was also found in the hearts of the CoQ(10)-fed group. The results of this study demonstrate that nutritional supplementation of CoQ(10) renders the hearts resistant to ischemia-reperfusion injury, probably by reducing the oxidative stress.     

氨基酰化酶中金属锌离子的功能作用

 氨基酰化酶是含锌金属酶。该酶每摩尔蛋白中含2摩尔Zn(Ⅱ)离子。金属鳌合剂与酶作用,通过竞争螯合Zn(Ⅱ)离子使酶活力下降。残余活力与残留金属含量呈正相关。竞争螯合的结果,生成不含金属的脱辅基酶蛋白,并导致酶活力的丧失。脱辅基酶由于加入Zn(Ⅱ)离子而恢复其活力。实验表明金属锌离子是氨基酰化酶催化活力所必需。与Zn(Ⅱ)离子相似,Co(Ⅱ)离子也可与脱辅基酶相结合并使之复活。 在190—240nm区域内对比了天然酶、脱辅基酶蛋白与Co(Ⅱ)置换氨基酰化酶的圆二色谱。远紫外圆二色谱表明,与天然酶相比,在脱辅基酶中由于金属离子的丧失导致主链构象发生变化,其中α螺旋增加约7％。因而锌离子(钴离子)对蛋白主链的反应最适构象有一定的稳定作用。脱辅基酶与Co(Ⅱ)离子结合,酶的主链构象恢复至与天然酶几近相同。可认为这是促使酶复活的内在因素。     

Palmitic and erucic acid metabolism in isolated perfused hearts from weanling pigs

Hearts from 4 week-old weanling pigs were capable of continuous work output when perfused with Krebs-Henseleit buffer containing 11 mM glucose. Perfused hearts metabolized either glucose or fatty acids, but optimum work output was achieved by a combination of glucose plus physiological concentrations (0.1 mM) of either palmitate or erucate. Higher concentrations of free fatty acids increased their rate of oxidation but also resulted in a large accumulation of neutral lipids in the myocardium, as well as a tendency to increased acetylation and acylation of coenzyme A and carnitine. When hearts were perfused with 1 mM fatty acids, the work output declined below control values. Erucic acid is known to be poorly oxidized by isolated rat heart mitochondria and, to a lesser degree, by perfused rat hearts. In addition, it has been reported that erucic acid acts as an uncoupler of oxidative phosphorylation. In isolated perfused pig hearts used in the present study, erucic acid oxidation rates were as high as palmitate oxidation rates. When energy coupling was measured by 31P-NMR, the steady-state levels of ATP and phosphocreatine during erucic acid perfusion did not change noticeably from those during glucose perfusion. It was concluded that the severe decrease in oxidation rates and ATP production resulting from the exposure of isolated pig and heart mitochondria to erucic acid are not replicated in the intact pig heart.     

THE PREDOMINANT ROLE OF THE SPLEEN IN LYMPHOCYTE RECIRCULATION

Lymphocyte recirculation through the isolated pig spleen was studied by means of a perfusion system which kept the organ alive for a prolonged period of time. By changing the perfusate to a leucocyte-enriched or cell-free perfusate and taking serial arterial and venous samples, the numbers of lymphocytes which homed to or were released from the spleen were measured. In all experiments more lymphocytes homed than were released per minute. There was no apparent difference when autologous or allogeneic cells were used. The number of lymphocytes released depended on the number of lymphocytes homed previously. During the phase of constant release up to 3-3 × 10⁶ lymphocytes were released per gram spleen per minute. From these values it can be extrapolated that up to 270 × 10⁹ lymphocytes recirculate through the isolated pig spleen per day. Based on kinetic data from other species it is estimated that in the entire pig a total number of 300–400 × 10⁹ lymphocytes recirculate per day. Thus, it can be concluded that the spleen is the most important organ for lymphocyte recirculation in the pig.     

Influence of gel and powdered formulations of coenzyme Q10 on metabolic parameters in rats

The healthful benefits of two commercially available formulations of coenzyme Q10 (Co Q10), one in gel and the other in a powdered form, on a variety of metabolic parameters in Sprague–Dawley rats (SD) were compared to control. The principal metabolic parameters examined were systolic blood pressure (SBP), DNA fragmentation, and free radical formation in hepatic and renal tissues. Compared to control, the powdered formulation significantly decreased SBP in the normotensive SD, whereas both commercial formulations lowered hepatic and renal DNA fragmentation and free radical formation. The gel-formulation lowered hepatic DNA fragmentation more than the powdered-formulation. In conclusion, both gel- and powdered-formulations of Co Q10 significantly influenced the metabolic parameters assessed in a favorable fashion, with the powdered-formulation more effective on SBP and the gel-formulation more effective on overcoming hepatic DNA fragmentation. From the data, we conclude that the choice of the formulation containing Co Q10 to be used should be based on the desired healthful benefits.     

Essentiality of coenzyme Q for the oxidation of -glycerophosphate by pig brain mitochondria

Serial extraction of lyophilized pig brain mitochondria with cold pentane resulted in complete loss of α-glycerophosphate oxidase activity. On titration with coenzyme Q₁₀ the activity was fully recovered. On comparing the decline of α-glycerophosphate, NADH, and succinoxidase activities during serial extraction with pentane, α-glycerophosphate oxidation was always the first to be lost. Extraction of coenzyme Q₁₀ from lyophilized brain mitochondria with pentane does not affect the activities of α-glycerophosphate or NADH dehydrogenase, but succinate dehydrogenase is partially inactivated. Reversible inactivation of the α-glycerophosphate oxidase system on depletion of the coenzyme Q content is taken as evidence that coenzyme Q is an obligatory component of this system. In accord with the conclusion that coenzyme Q is probably the physiological oxidant of α-glycerophosphate dehydrogenase, in antimycin-treated brain mitochondria α-glycerophosphate causes full activation of endogenous succinate dehydrogenase, in analogy to the previously observed activation by NAD-linked substrates in liver and heart mitochondria and by NADH in submitochondrial particles.     

Reinvestigation of coenzyme Q10 isolation from Sporidiobolus johnsonii

There is considerable current interest in coenzyme Q10 (CoQ10) from a medical perspective. CoQ10 has been shown to alleviate the side effects of statin drugs, for instance, and so there is a push to find naturally high producers of the compound. Sporidiobolus johnsonii (S. johnsonii) has been reported to produce CoQ10 in studies that used only standards on thin‐layer chromatography (TLC) and also suggested the production of coenzyme Q9 (CoQ9). This work set out to verify CoQ9/CoQ10 production in S. johnsonii and quantify as appropriate. We show that S. johnsonii produces CoQ10 but found no evidence for CoQ9 biosynthesis. The specific production of CoQ10 was noted at 10 mg/g dry cell weight (DCW) in media supplemented with 4‐hydroxybenzoic acid (HBA). This makes S. johnsonii a naturally high CoQ10 producer. New methods for extraction and purification of CoQ10 are also discussed, and identification of a closely eluting side product under normal phase isolation is reported.     

HYPERTROPHY OF THE HUMAN HEART AT THE LEVEL OF FINE STRUCTURE : An Analysis and Two Postulates

Muscle cells in the left ventricular walls of four markedly hypertrophied human hearts (above 600 gm) were compared with muscle cells in four non-hypertrophied hearts (up to 310 gm). Blocks of tissue obtained postmortem within 6 hours were processed for light and electron microscopy under conditions suitable for good preservation of myofibrils. A lattice parameter, q_h, was defined as the number of myosin filaments per square micron in either H zones or A bands. By the use of methods of electron microscopy, q_h was determined for perpendicular cross-sections of A bands in a large number of well preserved myofibrils of muscle cells in both groups of hearts. Statistical evaluation of the distributions of values of q_h revealed no significant difference between the two groups. Thus, the myofilament lattices in hypertrophied cells were geometrically within normal limits. Planimetric measurements of cross-sectional areas of muscle fibers were made, using photomicrographs obtained from one representative hypertrophied heart and from one control. The size-frequency distribution of the measurements showed a marked difference between the two hearts, and confirmed the presence of hypertrophy of muscle cells. Counts of the number of myofibrils per muscle cell were determined for samples from the same two hearts, evaluated statistically, and found to be significantly higher for the hypertrophied heart. It is proposed (a) that myofibrils in hypertrophied heart muscle cells have filament lattices with geometrical arrangement and macromolecular parameters that are the same as those found in myofibrils of normal heart muscle cells; and (b) that in hypertrophy the number of myofilaments increases through formation of new myofibrils, and possibly also by addition of filaments to preexisting myofibrils.     

THE DISTRIBUTION OF SOLUBLE AND MEMBRANE-BOUND FORMS OF GLUTAMINASE IN PIG BRAIN

Abstract— About 10% of the glutaminase activity associated with pig brain mitochondria was readily extractable by a variety of techniques but the remainder was very resistant to extraction. These two forms, which have been termed the soluble and membrane-bound forms respectively, have been shown to differ in their responses to activation by phosphate and phosphate-borate containing buffers. Submitochondrial fractionation studies indicated that the soluble form was located in the mitochondrial inner matrix whereas the membrane-bound form was associated with the inner membrane. The mitochondria associated with the synaptosomes were found to contain only the membrane-bound form of the enzyme whereas both forms were present in the free brain mitochondria.