Increased activities of glycogenolytic enzymes in liver after splanchnic-nerve stimulation

Electrical stimulation of the splanchnic nerve of rabbits caused a marked increase, within 30 seconds after the onset of stimulation, in liver-glycogen phosphorylase and glucose-6-phosphatase activities. The increased activity of liver phosphorylase after splanchnic-nerve stimulation was likewise observed in adrenalectomized and pancreatectomized rabbits. Glycogen content of the liver decreased only slightly after 5-minute stimulation.     

Insulin stimulation of nucleoside triphosphatase activity in isolated nuclear envelopes

The activity of nucleoside triphosphatase, an enzyme that regulates nuclear messenger RNA transport, was measured in highly purified nuclear envelopes isolated from rat liver. Addition of picomolar concentrations of insulin to freshly prepared nuclear envelopes directly increased the enzyme activity. The major effect of insulin on this enzyme was to increase the maximum velocity of its activity; no significant effects were seen on the affinity constant. These studies raise the possibility, therefore, that the nuclear envelope is a site where insulin regulates nuclear functions.     

FGF-21促进HepG2细胞摄取葡萄糖研究

肝脏是代谢的中枢性器官,在糖脂代谢中扮演重要角色。FGF-21是近年来发现的一种治疗糖尿病新型药物,研究其对肝脏糖代谢影响及机制将为FGF-21成药性提供理论依据。以Hep G2细胞为肝细胞模型,探究FGF-21对Hep G2细胞葡萄糖吸收影响及作用机制。FGF-21处理Hep G2细胞,采用葡萄糖氧化酶-过氧化物酶(GOD-POD)法检测细胞对葡萄糖摄取情况,并检查胰岛素与FGF-21协同作用,蒽酮法检测细胞内糖原含量,半定量和实时荧光定量PCR检测FGF-21对葡萄糖转运蛋白(GLUTs)m RNA表达影响。结果表明,FGF-21可促进Hep G2细胞摄取葡萄糖,与胰岛素具有一定协同作用,增加糖原合成。半定量PCR结果显示在FGF-21作用下,仅GLUT1m RNA表达有所增加。实时荧光定量PCR检测FGF-21作用时间对GLUT1 m RNA表达量影响,发现FGF-21作用6 h时GLUT1 m RNA表达量倍数增加最大。说明FGF-21可通过增加GLUT1 m RNA表达促进Hep G2细胞消耗葡萄糖,参与糖原合成。     

蜂胶中4种黄酮对HepG2细胞胰岛素抵抗的改善作用

为探究蜂胶中4种黄酮成分(高良姜素、短叶松素、松属素、柯因)对胰岛素抵抗的改善作用。通过高浓度胰岛素诱导的方式建立胰岛素抵抗HepG2细胞模型;设立正常组、模型组、高良姜素处理组、短叶松素处理组、松属素处理组、柯因处理组,测定各试验组对HepG2细胞增殖、葡萄糖消耗量、糖原含量、已糖激酶(hexokinase,HK)和丙酮酸激酶(pyruvate kinase,PK)的影响。结果显示:4种黄酮成分在有效浓度范围内对胰岛素抵抗HepG2细胞增殖均无显著影响(P0.05);柯因、短叶松素作用效果不明显(P0.05);高良姜素和松属素均能不同程度地提高IR-HepG2细胞葡萄糖消耗量(glucose consumption,GC)、肝糖原含量、HK和PK活力(P0.05)。上述结果表明蜂胶中高良姜素和松属素能较好地调节IR-HepG2糖代谢,改善胰岛素抵抗,为蜂胶产品的深度开发提供参考。     

泰克多的柞蚕饲料添加效应

泰克多填食对柞蚕的发育经过无影响,其生物效应在于能够提高蛹体重和茧层率。这可能是由于此剂通过某种途径和机制增强了柞蚕肠壁对食物营养成分的吸收活性,并对体内一些重要酶系特别是蛋白质合成酶系的活动有进一步的促进。     

鸡树条荚蒾果多酚改善胰岛素抵抗HepG2细胞的糖代谢效应

目的探讨鸡树条荚蒾果多酚对HepG2细胞胰岛素抵抗（IR）模型的改善作用,以评价其降血糖活性。方法本研究采用高浓度胰岛素诱导,建立体外IR模型,并进行模型稳定性（细胞活性法）及可靠性（Z因子法）评估。试验设空白组、IR模型组、阳性对照（二甲双胍）组和荚蒾果多酚组,MTT法检测细胞活性;葡萄糖氧化酶法检测培养液葡萄糖含量,计算葡萄糖消 耗量;蒽酮法测定糖原含量;比色法检测细胞己糖激酶（HK）、丙酮酸激酶（PK）活性,酶联免疫吸附试验（ELISA）检测细胞磷酸烯醇式丙酮酸激酶（PEPCK）、葡萄糖六磷酸酶（G6PC）活性。结果10? 6 mol/L的胰岛素诱导处理HepG2细胞24 h是产生胰岛素抵抗模型的最适条件,且IR模型在12 ~ 36 h内有较高的稳定性和可靠性。0.10 ~ 1.00 mg/mL 荚蒾果多酚组的葡萄糖消耗量显著高于模型组（P < 0.05）,24 h、0.50 mg/mL组的糖消耗量最高,为（3.49 ± 0.11）mmol/L,消耗率可达88.81%（P < 0.01）。与模型组对比,荚蒾果多酚可提高糖原的含量33.65%（P < 0.01）,HK、PK活性可分别提高43.36%（P < 0.05）、48.41%（P < 0.01）,对G6PC、PEPCK活性抑制率为分别为22.86%（P < 0.01）、17.33%（P < 0.05）。结论鸡树条荚蒾果多酚可提高IR- HepG2细胞的HK、PK活性,加快糖酵解,增加糖原含量;抑制G6PC、PEPCK活性,从而减少细胞内源性葡萄糖的产生。所以鸡树条荚蒾果多酚对胰岛素抵抗细胞的治疗具有一定效果。      

Insulin mediators from rat skeletal muscle have differential effects on insulin-sensitive pathways of intact adipocytes

The effects of partially purified insulin-generated mediators from rat skeletal muscle were compared to those of insulin on intact adipocytes. Insulin and insulin mediator stimulated both pyruvate dehydrogenase and glycogen synthase activity of intact adipocytes. In contrast, insulin stimulated glucose oxidation and 3-O-methylglucose transport, whereas insulin-generated mediators had no effect. Insulin-generated mediators cannot account for all the pleiotropic effects of insulin, especially membrane-controlled processes.     

Pineal N-acetyltransferase activity: effect of sympathetic stimulation

Stimulation of preganglionic sympathetic fibers to the superior cervical ganglia elevates the activity of pineal N-acetyltransferase. After the stimulation-induced rise in enzyme activity, a return toward baseline levels occurs whether or not nerve stimulation is continued. The ability of pineal N-acetyltransferase activity to fall in the presence of stimulation may account for the persistence of its rhythm in blinded animals.     

褐飞虱GSK-3调控糖原与海藻糖代谢的潜在功能

【目的】昆虫胰岛素信号途径能够介导糖原合成酶激酶3（glycogen synthase kinase 3,简称GSK-3或GSK3）调控体内糖原及海藻糖等糖代谢过程,从而控制昆虫的各项生命活动。论文旨在探究糖原合成酶激酶在褐飞虱（Nilaparvata lugens）体内对糖原与海藻糖代谢的调控作用。【方法】首先,基于GSK-3的cDNA编码序列,利用ExPASy工具翻译GSK-3氨基酸序列,预测蛋白分子量大小及等电点（pI）;然后利用SignaIP4.1Server对其信号肽进行分析。其次,以笔者实验室饲养的褐飞虱为研究对象,从4龄开始,每12 h取材,取至成虫48 h。利用Trizol法提取褐飞虱总RNA,根据反转录试剂盒合成第一链DNA,以18S作为内参基因,通过实时荧光定量PCR（qRT-PCR）检测褐飞虱GSK-3在不同龄期mRNA水平上的相对表达量。然后利用RNAi技术,向褐飞虱体内显微注射双链RNA(dsRNA)抑制GSK-3,以注射dsGFP的褐飞虱作为对照组。注射后48 h利用qRT-PCR技术检测GSK-3的表达情况,确定抑制效果。另外,取注射后48 h虫体,分别测定褐飞虱体内海藻糖、葡萄糖、糖原含量及海藻糖酶（trehalase,TRE）活性变化。最后采用qRT-PCR检测胰岛素信号通路胰岛素受体基因(insulin receptor,InR)、类胰岛素多肽基因(insulin-like peptides,Ilps)及海藻糖代谢途径TRE、海藻糖合成酶基因(trehalose-6-phophate synthase,TPS)、糖原磷酸化酶基因(glycogen phosphorylase,GP)、糖原合成酶基因(glycogen synthase,GS)中相关基因的表达,分析GSK-3在胰岛素信号通路及海藻糖代谢途径中的调控作用。【结果】褐飞虱GSK-3开放阅读框为1 914 bp,编码637个氨基酸;预测蛋白分子量为69.25 kD,等电点为9.15,为偏碱性蛋白,无信号肽结构,序列高度保守。发育表达模式结果显示GSK-3在不同发育阶段表达不一致,5龄若虫蜕皮前后低表达。GSK-3的dsRNA注射后48 h,与对照组dsGFP相比,GSK-3表达极显著下降,表明RNA干扰效果明显。糖原含量和两类海藻糖酶活性显著下降,而海藻糖含量显著上升,推测糖原和葡萄糖转化为海藻糖,作为其生理活动的能量来源。qRT-PCR检测发现,当GSK-3表达抑制后48 h,TRE1-2的表达量显著下降,而TRE1-1和TRE2的表达量极显著下降。另外,2个TPS基因、GS以及GP的表达量均极显著下降;胰岛素信号通路的2个InR基因和4个Ilps基因的表达同样被抑制,间接表明InR能够调控GSK-3的表达。【结论】褐飞虱GSK-3低表达后能够通过调控胰岛素信号通路及海藻糖代谢途径相关基因表达来调控糖原及海藻糖代谢。相关研究结果有助于更加全面地探索褐飞虱等昆虫糖原合成酶激酶调控海藻糖及糖类物质平衡的潜在分子机理。     

The Effect of Different Nitrogen Form on Key Enzyme Activity of Sugarbeet（Vulgaris L.） Carbon and Nitrogen Metabolism

This article analyses the effect of the proportion of the different nitrogen forms on key enzyme activity of carbon and nitrogen metabolism under the condition of nutritional water while Tian Yan-7 was used as experimental material.The result showed that nitrate reductase(NR) activity in the leaves gradually enhanced with the increase of NO3^-,No matter in root or leaves,glutamina synthetase(GS) activity first enhanced with increasing NH4^ when NH4^ was lower than that of NO3^-,and GS activity was the highest when NH4^ and NO3^- was equal,then GS activity declined with NH4^ increasing further,In the anaphase of growth,synthetic activity in root of sucrose synthetase(SS) in the mixed NH4^ and NO3^- was obviously highr than or NO3^- alone,Both of the root and sugar yields were the highest when the proportion of NH4^ and NO3^- was 1:1.     

Serum dopamine- -hydroxylase: decrease after chemical sympathectomy

Dopamine-beta- hydroxylase is an enzyme that is localized to catecholamine-containing vesicles in sympathetic nerves and the adrenal medulla, and is also found in the serum. Treatment of rats with 6-hydroxydopamine, a drug which destroys sympathetic nerve terminals, leads to a decrease in serum dopamine-beta-hydroxylase activity. The decrease is not due to an effect on the adrenal medulla or to an increase in circulating inhibitor or inhibitors of enzyme. These data represent evidence that at least a portion of the circulating dopamine-beta-hydroxylase activity arises from sympathetic nerve terminals.     

不同耐氮肥烤烟品种质外体NH4+浓度差异和有关生理指标分析

 【目的】研究不同耐氮肥烤烟品种叶片质外体NH4+浓度差异与耐氮肥性关系,为筛选对氮素适应范围广的烤烟品种提供理论依据。【方法】以耐氮肥的烟草品种中烟90、中耐氮肥的K326和对氮肥敏感的NC89为试验材料,用盆栽法研究它们之间的质外体NH4+浓度差异。【结果】生育后期3个烤烟品种叶片质外体的NH4+浓度均呈现逐步升高的趋势,中烟90的质外体NH4+浓度最大,NC89最小.谷氨酰胺合成酶 (GS)活性和总氮含量下降明显,质外体pH值急剧减小,谷氨酸脱氢酶(NADH-GDH)活性缓慢上升,质外体pH始终低于叶片pH;质外体NH4+浓度与GS活性和总氮都呈显著负相关,与GDH活性呈显著正相关。【结论】 烤烟品种耐肥性与其生育后期质外体NH4+浓度,GS活性和GDH活性有关,耐肥性强的烤烟品种其质外体NH4+浓度高,GS活性低,GDH活性高,NH4+挥发多。     

FGF19 as a postprandial, insulin-independent activator of hepatic protein and glycogen synthesis

Fibroblast growth factor (FGF) 19 is an enterokine synthesized and released when bile acids are taken up into the ileum. We show that FGF19 stimulates hepatic protein and glycogen synthesis but does not induce lipogenesis. The effects of FGF19 are independent of the activity of either insulin or the protein kinase Akt and, instead, are mediated through a mitogen-activated protein kinase signaling pathway that activates components of the protein translation machinery and stimulates glycogen synthase activity. Mice lacking FGF15 (the mouse FGF19 ortholog) fail to properly maintain blood concentrations of glucose and normal postprandial amounts of liver glycogen. FGF19 treatment restored the loss of glycogen in diabetic animals lacking insulin. Thus, FGF19 activates a physiologically important, insulin-independent endocrine pathway that regulates hepatic protein and glycogen metabolism.     

影响两系杂交稻结实期茎鞘贮藏碳水化合物转运的生理因素研究

选用三种类型两系杂交稻组合,研究灌浆结实期茎、鞘贮藏碳水化合物转运活性及其影响因素。结果表明：茎、鞘高含量可溶性糖是转运活跃的特征;淀粉酶活性高有利淀粉降解转运;分枝酶（ Ｑ 酶） 可能影响淀粉的降解和转运效率;蔗糖合成酶（ Ｓ Ｓ） 、蔗糖磷酸合成酶（ Ｓ Ｐ Ｓ） 对蔗糖含量和转运有影响。     

Effect of adenosine 3',5'-monophosphate on neuronal pacemaker activity: a voltage clamp analysis

Bursting pacemaker activity in nerve cells can be modified for long periods by synaptic input of short duration. There is evidence that cyclic nucleotides may play a role in these modifications. The predominant effect of elevated levels of adenosine 3',5'-monophosphate in Aplysia neurons was an increased slope conductance to hyperpolarizing pulses, evident in voltage clamp records. A similar increase in slope conductance was seen as one component of maximum strength synaptic stimulation, which is consistent with the idea that cyclic nucleotides are important in the expression of synaptic alteration of bursting pacemaker activity.     

不同施氮水平对小麦旗叶氮素代谢相关酶活性的影响

选用新冬18号小麦品种,进行3个施氮水平的土壤施肥处理,再选取不同水平处理下的小麦旗叶作为供试材料,分别测定小麦旗叶中的谷氨酰胺合成酶、硝酸还原酶、内肽酶,氨肽酶、羧肽酶这5种氮素代谢关键酶的酶活力。研究结果显示,在小麦开花之后14 d,施氮水平为240 kg/hm2可有效增加小麦旗叶中可溶性蛋白质含量和主要氮代谢相关酶的活性。在此水平下,旗叶中可溶性蛋白质含量与谷氨酰胺合成酶及硝酸还原酶活性变化趋势相同,相比施氮水平为350,180 kg/hm2更能减缓小麦开花之后14 d的蛋白质水解,可进一步促进蛋白质的合成。与此同时,在开花之后第21天,高水平施氮量处理下的小麦旗叶中可溶性蛋白质含量则与内肽酶、氨肽酶等水解酶活性变化趋势相反。说明在240 kg/hm2施氮水平下,开花后期其施氮量能有效促进氮素分解代谢相关酶活性,旗叶可溶性蛋白含量降低,进而为小麦植株中氮素的再分配和利用起到了协助调节作用,提高后期小麦籽粒蛋白质的品质。     

"Transdifferentiation" of C6 glial cells in culture

The activities of cyclic nucleotide phosphohydrolase, an enzyme marker for oligodendrocytes, and glutamine synthetase, an enzyme marker for astrocytes, were studied at early (21 to 26) and late (82 to 88) cell passages. The activity of cyclic nucleotide phosphohydrolase was markedly high and that of glutamine synthetase was low in the early passages, but this relation was reversed in the late passages. These findings suggest a "transdifferentiation" of C6 glial cells with passage in culture.     

Narcotic drugs: effects on the serotonin biosynthetic systems of the brain

The effects of short- and long-term administration of morphine on the activity of two measurable forms of rat brain tryptophan hydroxylase were studied. Morphine administration produced an immediate decrease and a longterm increase in the nerve ending (particulate) enzyme activity but did not change the cell body (soluble) enzyme activity. Cocaine administration demnonstrated a short-term decrcease in measurable nerve eniding enzyme activity that was due to the inhibition of the high affinity uptake (the Michaelis constant, K(m) is 10-(5) molar) of trytophan, the serotonin precursor. Cocaine did not aflect the low affinity uptake K(m) = 10-(5) molar) of tryptophan. Both the uptake of the precursor and the enizymiie activity appeared to be drug-sensitive regullatory processes in the biosynthlesis of serotonin.     

胆碱对鹅体内脂质代谢及肝脏FAS基因mRNA表达的影响

【目的】探讨胆碱对青农灰鹅体内脂质转运、代谢的作用及脂肪酸合成酶FAS（fatty acid synthase）基因表达的影响,确定鹅日粮中胆碱的最适添加量。【方法】将180只1日龄青农灰鹅随机分为6个处理组,每处理组3个重复,每个重复10只。试验在玉米-豆粕型饲粮的基础上各组的胆碱添加量分别为0、600、1 200、1 800、2 400和3 000 mg•kg-1,试验期15周。屠宰后测定肝脏和血清总胆固醇、甘油三酯,血清低密度脂蛋白胆固醇、高密度脂蛋白胆固醇、葡萄糖、胰岛素、胰高血糖素水平,肝脏脂蛋白酯酶、肝脂酶及血清胆碱酯酶活性和肝脏脂肪酸合成酶（FAS）基因表达量。【结果】①4周龄时,饲粮中添加胆碱显著降低肝脏和血清总胆固醇、甘油三酯、血清低密度脂蛋白胆固醇、胰岛素水平和血清胆碱酯酶活性（P＜0.05或P＜0.01）;显著提高肝脏脂蛋白酯酶、肝脂酶活性和血清胰高血糖素水平（P＜0.05）;但对血清高密度脂蛋白胆固醇、葡萄糖含量影响不显著（P＞0.05）。15周龄时,饲粮中添加胆碱显著降低肝脏和血清总胆固醇、甘油三酯、低密度脂蛋白胆固醇含量与胰岛素水平及血清胆碱酯酶活性（P＜0.05或P＜0.01）;显著提高肝脏脂蛋白酯酶、肝脂酶活性和血清高密度脂蛋白胆固醇、胰高血糖素水平（P＜0.05）;但对血清葡萄糖含量影响不显著（P＞0.05）。②饲粮中添加胆碱显著提高肝脏FAS基因的表达量（P＜0.01）,且肝脏脂肪酸合成酶基因表达量随胆碱添加水平呈先下降后上升趋势。【结论】综合考虑胆碱对鹅肝脏脂质代谢的影响,1—4和5—15周龄鹅饲粮中胆碱适宜添加量分别为1 200—1 800和1 200 mg•kg-1。     

Carbohydrate supply as a regulator of rat liver phosphoenolpyruvate carboxykinase activity

Administration of glucose, fructose, and glycerol to fasted rats produced a significant depression of liver phosphoenolpyruvate carboxykinase activity within 4 to 8 hours; galactose and ribose were much less effective. All the compounds yielded appreciable quantities of liver glycogen. The depression of phosphoenolpyruvate carboxykinase activity by glucose and glycerol was diminished by the concomitant administration of 2-deoxyglucose. The latter depressed glycogen formation from administered carbohydrate in muscle but not in liver. In rats made diabetic by alloxan, depression of elevated phosphoenolpyruvate carboxykinase activity by insulin was dependent upon a dietary source of carbohydrate. These results were interpreted to indicate that depression of certain gluconeogenic enzymes after carbohydrate ingestion is initiated by the metabolism of carbohydrate in some extrahepatic site.