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1.
Metabolic function of hepatic hexokinase in perch, Perca fluviatilis   总被引:1,自引:0,他引:1  
Feeding previously fasted perch with a carbohydrate‐rich pelleted diet resulted in a rapid transient insulin response followed by a transient increase in the activity of the hepatic low‐affinity hexokinase (glucokinase, GK). The activity of the high‐affinity hexokinase (hexokinase, HK) increased more slowly and steadily to about seven times the fasting value on the 10th day after start of feeding, in parallel with an increase in the hepatic glycogen content. An insulin response was also observed when fasted perch were fed a diet consisting of boiled fillet of cod (a high‐protein, very low‐fat and practically carbohydrate‐free diet). In this case no GK activity was observed, whereas the glycogen content increased steadily from the start of feeding to about the same level as when fish were given the carbohydrate‐rich diet. There was a delayed increase in the activity of HK, starting on about the sixth day (after the glycogen stores were filled), concomitant with increased activities of lipogenic enzymes (glucose‐6‐phosphate dehydrogenase, ATP citrate lyase and malic enzyme). The results are in line with the hypothesis that the mitochondria‐bound hepatic HK has a particular anabolic function in glycogen and fat synthesis.  相似文献   
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A marine furanoterpenoid derivative, 10-acetylirciformonin B (10AB), was found to inhibit the proliferation of leukemia, hepatoma, and colon cancer cell lines, with selective and significant potency against leukemia cells. It induced DNA damage and apoptosis in leukemia HL 60 cells. To fully understand the mechanism behind the 10AB apoptotic induction against HL 60 cells, we extended our previous findings and further explored the precise molecular targets of 10AB. We found that the use of 10AB increased apoptosis by 8.9%–87.6% and caused disruption of mitochondrial membrane potential (MMP) by 15.2%–95.2% in a dose-dependent manner, as demonstrated by annexin-V/PI and JC-1 staining assays, respectively. Moreover, our findings indicated that the pretreatment of HL 60 cells with N-acetyl-l-cysteine (NAC), a reactive oxygen species (ROS) scavenger, diminished MMP disruption and apoptosis induced by 10AB, suggesting that ROS overproduction plays a crucial rule in the cytotoxic activity of 10AB. The results of a cell-free system assay indicated that 10AB could act as a topoisomerase catalytic inhibitor through the inhibition of topoisomerase IIα. On the protein level, the expression of the anti-apoptotic proteins Bcl-xL and Bcl-2, caspase inhibitors XIAP and survivin, as well as hexokinase II were inhibited by the use of 10AB. On the other hand, the expression of the pro-apoptotic protein Bax was increased after 10AB treatment. Taken together, our results suggest that 10AB-induced apoptosis is mediated through the overproduction of ROS and the disruption of mitochondrial metabolism.  相似文献   
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  目的  研究桂花Osmanthus fragrans己糖激酶家族成员序列特征及表达变化规律。  方法  选取3个不同花色桂花品种‘堰虹桂’O. fragrans ‘Yanhong Gui’、‘玉玲珑’O. fragrans ‘Yulinglong’、‘金球桂’O. fragrans ‘Jinqiu Gui’,从转录组数据中筛选得到HXK同源基因,进行序列分析和系统进化树分析,并利用实时荧光定量PCR检测不同品种桂花不同组织及不同发育阶段的OfHXKs基因表达情况。  结果  筛选得到OfHXK1~OfHXK4基因,分析表明不同品种桂花OfHXK1、OfHXK3和OfHXK4基因核苷酸序列相似性均高于99%。OfHXKs基因编码461~510个氨基酸残基,均包含2个保守的磷酸化作用位点和1个糖结合位点。OfHXK1和OfHXK2具有N端膜锚定结构,与拟南芥Arabidopsis thaliana AtHXK1与AtHXK2聚为一支;无跨膜区域的OfHXK3与AtHXK3亲缘关系较近,推测二者具有催化作用但无糖信号传导功能;OfHXK4蛋白腺苷结合位点处多出11个氨基酸残基,与AtHKL1和AtHKL2亲缘关系较近。4个桂花HXK基因成员在桂花1年生茎、2年生茎、嫩叶、成熟叶和花序中均有表达。随着花开放的进程,整体上OfHXK1、OfHXK3和OfHXK4基因的表达量呈现先上升后下降的趋势,而OfHXK2基因在3个品种花序发育过程中表达模式不同。  结论  根据序列分析与进化树分析,推测桂花OfHXK1~OfHXK4均具有催化己糖磷酸化的功能,OfHXK1和OfHXK2具有糖信号感知和转导的功能。OfHXK1、OfHXK3和OfHXK4基因随花序发育呈现有规律升降的表达模式,可能与其参与糖类物质代谢有关。图6表4参30  相似文献   
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An 8-week growth trial was carried out in a semi-recirculation system to investigate the effect of high dietary starch levels on the growth performance, blood chemistry, starch utilization and body composition of gibel carp ( Carassius auratus var. gibelio). Five isonitrogenous and isocarloric experimental diets were formulated to contain different starch levels (24%, 28%, 32%, 36% and 40% respectively). Triplicate groups of fish (24 fish per tank with an average body weight, of 8.5 g) were assigned to each diet. The results showed that dietary carbohydrate levels significantly affected the growth performance, hepatopancreatic lipid content, pyruvate kinase (PK) activity and whole-body lipid content. Growth performance, body crude lipid and plasma glucose concentrations showed a decreasing trend with an increase in dietary starch from 24% to 40%. Pyruvate kinase activities and hepatopancreatic lipid content showed an increasing trend with the dietary starch increasing from 24% to 32%, and then a decreasing trend with the dietary starch increasing from 32% to 40%. No significant difference in the hepatopancreatic hexokinase (HK) activity, plasma triglyceride contents, body crude protein, ash and calcium (Ca) and phosphorus (P) contents was observed between different treatments. In conclusion, higher dietary starch levels (32–40%) significantly ( P <0.05) decreased the growth of gibel carp in the present study.  相似文献   
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Abstract

The synthetic activity of protein and changes in the activity profiles of enzymes during cold acclimation were studied in leaves, stems, and crowns of a hardy winter wheat cultivar ( Triticum aestivum L. cv. Horoshirikomugi) and a less hardy winter wheat cultivar (Triticum aestivum L. cv. Norin 61). The results showed that one of the genetic differences between the two cuitivars is found in the activities of protein synthesis under low temperatures. The profiles of the changes in enzymatic activities showed that the adjustment of enzymatic activities was a tissue-specific event to accommodate environmental changes. The synthesis of the enzymes involved in a peroxide scavenging system such as hexokinase, glucose-6-phosphate dehydrogenase, dehydroascorbate reductase, ascorbate peroxidase, and so on started at the acclimation stage, while the phosphoglucose isomerase, which appears to be a maintenance-type enzyme in the wintering wheat, showed a small increase in the activity at later stages of acclimation period in their stems of both cuitivars. When the field was covered with snow, the catalase activity in the leaves started to decrease abruptly, while catalase activities in the stems and crowns did not exhibit appreciable changes during the life under snow. The life-cycle dependent changes in enzymatic activities appear to occur in late March or early April under snow.  相似文献   
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山药多糖对2型糖尿病大鼠降糖机理的研究   总被引:2,自引:0,他引:2  
为探讨山药多糖对2型糖尿病的降糖机理,采用高热量饮食结合腹腔注射小剂量链脲佐菌素(STZ)的方法制备试验性2型糖尿病大鼠模型,以不同剂量山药多糖[(100、70和40 mg/(kg.d)]和二甲双胍100 mg/(kg.d)灌胃治疗4周后,检测血清胰岛素及胰高血糖素、己糖激酶(HK)、琥珀酸脱氢酶(SDH)和苹果酸脱氢酶(MDH)的活性。结果表明,与糖尿病模型组比较,山药多糖具有明显的降血糖作用;治疗组HK、SDH、MDH活性显著提高,提示山药多糖对2型糖尿病的治疗机制之一可能是山药多糖提高了糖代谢关键的酶活性。  相似文献   
9.
郭春倩  田洁 《浙江农业学报》2022,34(9):1925-1934
为明确AsHXK2基因在根际促生菌缓解干旱胁迫下的作用,采用TA克隆方法克隆得到乐都紫皮大蒜AsHXK2基因序列,并对其基本特性进行生物信息学分析。通过实时荧光定量PCR(qRT-PCR)分析了AsHXK2基因在大蒜不同组织和干旱胁迫、根际促生菌等条件下的表达情况。结果表明,AsHXK2开放阅读框全长1 500 bp,可编码499个氨基酸,分子量约为57.77 ku,理论等电点为6.37;亚细胞定位预测结果显示,AsHXK2蛋白主要定位于叶绿体,含有一个特异位点,属于己糖激酶2家族。在进化关系上,大蒜AsHXK2与天门冬科的芦笋HXK2亲缘关系最为接近。qRT-PCR分析表明,AsHXK2基因表达具有明显的组织特异性,其中鳞芽中的表达量最高。此外,不同组织中AsHXK2对根际促生菌作用下的干旱胁迫表现出不同的响应模式。干旱胁迫能够显著上调大蒜根、叶和鳞芽的AsHXK2表达水平;而在干旱胁迫施加根际促生菌的处理下,仅在叶中AsHXK2表达量显著高于干旱胁迫,表明叶片中AsHXK2对根际促生菌缓解干旱胁迫的响应较为明显。本研究为进一步探究根际促生菌缓解大蒜干旱胁迫的作用机制奠定了基础。  相似文献   
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为获得微生物工程菌株来源的己糖激酶(Hexokinase,HK)并提高HK的发酵水平,对已构建产HK的大肠工程菌株BW25113-pYBE/schk进行发酵工艺优化,为HK法血糖检测试剂盒的配制提供技术支撑。本研究采用正交试验对菌株发酵工艺进行优化,并对超声破壁获取粗酶及粗酶的纯化工艺进行研究。结果表明:工程菌株BW25113-pYBE/schk的最佳发酵培养条件为:装液量80mL/500mL,发酵液初始pH 6.5,发酵温度37℃,接种量2%,诱导剂添加时间为接种后1h,转速180r/min。超声破壁最佳工艺为:功率200 W,工作1.5s间歇1s,破壁5min。在最佳发酵和破壁工艺条件下,HK的发酵活力稳定在157U/mL,经过Ni-Agarose His标签蛋白试剂纯化获得60kD的HK。本研究建立了高产己糖激酶大肠杆菌工程菌株的发酵工艺,有利于降低己糖激酶法血糖试剂盒的成本,为糖尿病患者的准确诊断及普及提供便利条件。  相似文献   
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