大鼠活体注射台盼蓝后肝和肾的组织学与组织化学变化的观察

大鼠连续4天腹腔注射1%台盼蓝后,观察隔天、隔周、隔二周后肝和肾的组织结构及PAS、AlP、AcP、G-6-Pase、Mg~(2+)-ATPase和SDH的活性变化。结果发现:肝细胞和肾小管的上皮细胞中有台盼蓝颗粒;肝PAS反应阴性;隔天后肝AlP、AcP、G-6-Pase和SDH活性增强,Mg~(2+)-ATPase活性减弱;肾的上述组化反应活性都减弱;隔二周后肝和肾的上述组化反应接近对照。实验结果提示:活体注射台盼蓝对肝和肾未构成实质性损伤,隔天后的组化变化可能是一种生理适应性反应。     

PEROXIDASE ACTIVITY IN RAT LIVER MICROBODIES AFTER AMINO-TRIAZOLE INHIBITION

The in vivo effects of 3-amino-1,2,4-triazole (AT) on the fine structure of microbodies in hepatic cells of male rats has been studied by the peroxidase-staining technique. Within 1 hr of intraperitoneal injection AT abolishes microbody peroxidase-staining, and the return of staining coincides temporally with the known pattern of return of catalase activity following AT inhibition; this is further evidence that the peroxidase staining of microbodies is due to catalase activity. Peroxidase staining reappears in the microbody matrix without evidence of either massive degradation or rapid proliferation of the organelles. Furthermore, during the period of return of activity, ribosomal staining occurs adjacent to microbodies whose matrix shows little or no peroxidase staining. These observations are interpreted as evidence that (a) catalase is capable of entering preexisting microbodies without traversing the cisternae of the rough endoplasmic reticulum or the Golgi apparatus, and that (b) the ribosomal staining is probably not cytochemical diffusion artifact and may represent a localized site of synthesis or activation of catalase.     

急性给锂小鼠大脑皮层NOS活性与蛋白表达的时程变化

目的探讨急性给锂小鼠大脑皮层一氧化氮合酶(NOS)活性与蛋白表达的时程变化及其意义.方法选用昆明小鼠40只,分为对照组和腹腔注射1.5mmol/Kg氯化锂(LiCl)即刻、0.5h、1h、3h、6h、12h、24h组,每组5只.采用NADPH-d黄递酶组织化学和ABC免疫组化法,观察急性给锂后不同时程小鼠大脑皮层NOS和nNOS阳性神经元数目的变化.结果急性给锂即刻小鼠大脑皮层NOS和nNOS阳性神经元数目明显增加(P＜0.01),1h后达到高峰(P＜0.01),6h和12h恢复到正常水平(P＞0.05),24hNOS阳性神经元又明显增高(P＜0.01),nNOS阳性神经元处于正常水平(P＞0.05).结论本实验提示急性给锂对小鼠大脑皮层NOS和nNOS阳性神经元数目有一定影响,这种变化可能是锂影响脑发育及锂的神经毒性的机理之一.     

ALCOHOL DEHYDROGENASE ACTIVITY IN RAT BRAIN AND LIVER

Abstract— A significant level of alcohol dehydrogenase activity has been demonstrated in the soluble fraction of rat brain. The pH optimum, kinetic properties and response to inhibitors are similar to those of liver alcohol dehydrogenase. The nutritional state of the animal, such as that associated with feeding or fasting, appeared to have no effect on the levels of the alcohol dehydrogenase activities in either liver or brain. A cerebral mechanism for the metabolism of ethanol may be involved in local biochemical adjustments in tissues during exposure to alcohol and may play a significant role in the pathogenesis of the neural disorders which can accompany chronic alcohol ingestion or acute withdrawal.     

TYROSINE–ALPHA-KETOGLUTARATE AMINOTRANSFERASE ACTIVITY IN RAT LIVER AFTER SPINAL CORD SECTION

Spinal cord section brings about early and late changes in rat liver tyrosine-alpha-ketoglutarate aminotransferase activity. Early effects (4 h after surgery): spinal cord section at C₇ level causes an unreactiveness of rat liver tyrosine-alpha-ketoglutarate aminotransferase both to endogenously and exogenously elevated plasma glucocorticoid levels. Induction of tyrosine–alpha-ketoglutarate aminotransferase by hydrocortisone administration is almost completely inhibited. This unreactiveness of the rat liver enzyme to hydrocortisone is not due to delayed resorption of hydrocortisone by the peritoneum as tested with [³H]hydrocortisone, to changes in the secretion of hypophyseal hormones or to changes in the levels of glucose in blood or liver. L₃ level section of the spinal cord or sham operation results in a stress-like enzyme pattern (an increase at 4 h with return to normal level at 24 h). The stress elevation of tyrosine–alpha-ketoglutarate aminotransferase at 4 h after the operation is absent in C₇ level sectioned rats. This effect is not due to a decreased plasma corticosterone level since it is 4.1-fold higher in C₇ level sectioned rats and 2.7-fold higher in sham-operated controls (as measured 2.5 h after the operation). Late effects (24 h after the surgery): C₇ level section of the spinal cord brings about a nine-fold increase in a tyrosine–alpha-ketoglutarate aminotransferase activity in animals with intact adrenals and three-fold increase in adrenalectomized rats at 24 h after the operation. This increase is abolished almost completely by cycloheximide, irrespective of the time of administration. Experiments with actinomycin D, injected at different times after C₇ level section have shown that there exists a period of higher sensitivity of the amino-transferase toward the antibiotic (lasting about 3 h), followed by a period of lower sensitivity (lasting 16 h or longer). These results can be explained by assuming the existence of two tyrosine-alpha-ketoglutarate aminotransferase mRNAs with different lifetime. A direct participation of the CNS in the changes in enzymic activity observed after section of the spinal cord above the segments innervating the liver is suggested.     

济南假单胞制剂PJV对大鼠肝大颗粒淋巴细胞数量、酶活性及自然杀伤活性的影响

本文通过细胞分离、细胞化学染色、电镜观察及~3H-TdR释放试验体外检测细胞毒活性等方法,研究了在生物反应调节剂—济南假单胞制剂PJV作用下,大鼠肝大颗粒淋巴细胞(LGL)和枯否细胞(KC)的数量、形态及其免疫生物特性等变化。结果表明,PJV的应用可引起肝内LGL数量增长达4倍,其体外杀伤肿瘤细胞活性增长约2倍,细胞内ACP活性明显提高。肝KC数量及活性也有明显增加。本文还讨论了肝LGL与KC的相互关系。     

TYROSINE HYDROXYLASE ACTIVITY IN NORADRENERGIC NEURONS OF THE LOCUS COERULEUS AFTER RESERPINE ADMINISTRATION: SEQUENTIAL INCREASE IN CELL BODIES AND NERVE TERMINALS

The effect of a single systemic injection of reserpine on tyrosine hydroxylase activity in the locus coeruleus, cerebellum, hypothalamus, and hippocampus was examined. Increases in enzyme activity were seen in all four brain areas; the time-course of the changes, however, was different in each case. In the locus coeruleus the maximum change in enzyme activity was seen 3 days after drug administration; in the cerebellum, 7-11 days; in the hypothalamus, 8-11 days; and in the hippocampus, 21 days. Since tyrosine hydroxylase in the cerebellum and hippocampus is present in terminals of neurons whose cell bodies are located in the locus coeruleus, the delayed increase in enzyme activity in cerebellum and hippocampus probably depends upon the slow rate of transport of TH molecules in these neurons.     

EFFECTS OF STATIC AND ELF MAGNETIC FIELDS ON FREE-RADICAL PROCESSES IN RAT LIVER AND KIDNEY

The aim of the study was to evaluate the activities of the antioxidant enzymes superoxide dysmutase (SOD, EC. 1.15.1.1), catalase (CAT, EC. 1.11.1.6), and glutathione peroxidase (GSH-Px, EC. 1.11.1.9) and the levels of malonic dialdehyde (MDA), a phospholipid peroxidation product, in the liver and kidneys of male and female rats exposed to static (0.49-T) and extremely low-frequency (50-Hz, 0.018-T) magnetic fields. Extremely low frequency magnetic fields increased all enzyme activities and MDA levels in both the liver and kidneys. In contrast, static magnetic field did not produce changes. The results of the study suggest that ELF magnetic fields affect free-radical processes in the liver and kidney.     

BENZPYRENE HYDROXYLASE ACTIVITY IN ISOLATED PARENCHYMAL AND NONPARENCHYMAL CELLS OF RAT LIVER

Previous studies have implicated the reticuloendothelial cells of the liver in certain aspects of steroid metabolism. The similarity in the metabolism of steroids and polycyclic hydrocarbons suggested that the nonparenchymal cells possibly play a role in these areas. The present study presents evidence that at least one of the microsomal NADPH-requirig enzymes, benzpyrene hydroxylase, is present in nonparenchymal cells and, furthermore, is "inducible." In adult rats treated with 3-methylcholanthrene or β-naphthoflavone, the nonparenchymal cells exhibited increases in benzpyrene hydroxylase activity of 17-fold and five-fold, respectively. Treatment with phenobarbital resulted in only a slight increase in enzyme activity. Enzyme activity in parenchymal cells under similar conditions was increased sixfold and fivefold by 3-methylcholanthrene and β-naphthoflavone, respectively, but not by phenobarbital.     

CHARACTERIZATION AND LOCALIZATION OF ACID HYDROLASE ACTIVITY IN THE SYNAPTOSOMAL FRACTION FROM RAT CEREBRAL CORTEX

Synaptosomes were prepared from the cerebral cortex of adult rats by a rapid technique of centrifugation in a Ficoll-sucrose discontinuous gradient. The synaptosomal fraction contained 40 per cent of the total gradient activity of acid α-naphthyl phosphatase (EC 3.1.3.2). Quantitative electron microscopy of this fraction revealed rare, typical, extrasynaptosomal dense body lysosomes. pH-activity profiles of free and Triton X-100 (total) activities were prepared for α-naphthyl phosphatase, β-glucuronidase (EC 3.2.1.31), β-galactosidase (EC 3.2.1.23), arylsulfatase (EC 3.1.6.1) and N-acetylglucosaminidase (EC 3.2.1.30). The ratios of total to free activity varied in the order: arylsulfatase > β-galactosidase > β-glucuronidase > N-acetylglucosaminidase > acid phosphohydrolase. Incubation of synaptosomal fractions at pH 5 and 37°C produced significant activation of β-galactosidase and N-acetylglucosaminidase but no activation of cryptic lactate dehydrogenase (EC 1.1.1.27). Hyposmotic suspension and subfractionation of the synaptosomal fraction produced considerable solubilization of lactate dehydrogenase, arylsulfatase and β-galactosidase but only partial liberation of α-naphthyl phosphatase, the remainder being associated with synaptosomal membrane fragments. Incomplete equilibrium sedimentation of synaptosomes in a continuous sucrose gradient (0·55-1·5 M) provided a broad lactate dehydrogenase and Na + K ATPase (EC 3.6.1.4) peak (peak I) at low sucrose densities. β-Glucuronidase, β-glucosidase and α-naphthyl phosphatase were significantly present in peak I. Conversely, N-acetylglucosaminidase, arylsulphatase and β-galactosidase were predominantly located in denser particles sedimenting through 1·2 M sucrose (peak II). Electron microscopy confirmed the heterogeneity of this second peak and the presence of numerous extrasynapto-somal dense body lysosomes.     

SITES OF GLYCOGEN SYNTHESIS IN RAT LIVER CELLS AS SHOWN BY ELECTRON MICROSCOPE RADIOAUTOGRAPHY AFTER ADMINISTRATION OF GLUCOSE-H3

Glycogen synthesis was investigated by giving tritium (H³)-labeled glucose with carrier to fasted rats in vivo or incubating liver slices from fasted rats in vitro using a glucose-H³-containing medium. After 15 min or 1 hr, pieces of liver were fixed and radioautographed for light and electron microscopy. In vivo and in vitro, radioautographic reactions appeared over "glycogen areas" and over zones transitional between these areas and ergastoplasm. Treatment of sections by alpha amylase removed all but about 5% of the radioactivity, so that about 95% of it consisted of glycogen (synthesized during the 15 min or 1 hr elapsing after administration of glucose-H³). Within glycogen areas and transitional zones, most silver grains were over or very close to glycogen granules and smooth (or partly smooth) vesicles. Presumably, much of the label was added onto growing glycogen granules, in accord with the biochemical view that glycogen may serve as substrate for further glycogen synthesis. The few silver grains located far from glycogen granules—15% at the 15 min interval in vivo—approximated smooth (or partly smooth) vesicles of endoplasmic reticulum. This observation raised the possibility that smooth membranes play a role in glucose uptake at an early stage in de novo formation of glycogen granules.     

THE ALTERATION OF THE REGENERATIVE ACTIVITY AND THE CELL CYCLE OF PARTIALLY HEPATECTOMIZED RAT LIVER FOLLOWING ADMINISTRATION OF d-GALACTOSAMINE

A single injection of d-galactosamine given to rats at different times after partial hepatectomy (PH) changes the pattern of regenerative proliferation. When administered during the pre-replicative phase of regeneration, the onset of DNA synthesis and the increase in labelling index after injection of ³H-thymidine are delayed by about 12 hr. The injection of d-galactosamine at 24 hr after PH inhibits the drop in DNA synthesis occurring normally during the following 12 hr period. This was detected by a high labelling index and by an increased specific activity of DNA. The findings indicate a lengthening of the S phase, while G₂ and M remain normal. Two modes of action of d-galactosamine on the cell cycle are discussed.     

INDUCTION OF HEPATOCYTE GROWTH FACTOR IN THE LIVER, KIDNEY AND LUNG FOLLOWING TOTAL BODY IRRADIATION IN RAT

Hepatocyte factor (HGF) has been shown to have a pleiotropic function to act as a potent organotropic factor in the regeneration of injury in various organs, including the liver, kidney and lung. To examine the involvement of HGF in radiation injury, the authors analysed the changes in HGF mRNA and HGF protein levels in the rat organs (liver, lung, kidney) and plasma following 6 Gy of total body irradiation. Expression of HGF mRNA in the liver and kidney increased 6–48 h after total body irradiation and returned to previous values 1 week later. HGF protein levels in lung and liver showed 1.3-2 fold elevations 1–2 weeks after irradiation (P< 0.05). HGF levels in plasma stayed at undetectable levels up to 1 month after total body irradiation. The labelling index determined 2 weeks and 1 month after total body irradiation indicated no enhancement of regeneration. Thus, total body irradiation induced transient HGF elevation in these organs without enhancement of regeneration     

COMBINED CYTOCHEMICAL AND ELECTRON MICROSCOPIC DEMONSTRATION OF β-GLUCURONIDASE ACTIVITY IN RAT LIVER WITH THE USE OF A SIMULTANEOUS COUPLING AZO DYE TECHNIQUE

Rat liver was fixed in formal-cacodylate-sucrose and frozen sections were incubated in a simultaneous-coupling medium containing naphthol AS-BI glucuronide as substrate and hexazonium pararosanilin as the diazo reagent. By light microscopy, the sections demonstrated β-glucuronidase activity as red discrete granules in the pericanalicular cytoplasm and as a generalized cytoplasmic stain in the parenchymal cells. Brief treatment of sections in cold ethanol prior to incubation markedly enhanced the staining for the enzyme and made it possible to demonstrate sufficient amounts of the reaction product in sections embedded in epoxy resin following dehydration and propylene oxide treatment. Electron microscopy revealed that the reaction product was moderately electron opaque and deposited in greater amounts in the vacuolated dense bodies and occasionally in the dense bodies which did not show obvious vacuoles. In each dense body, the deposits occurred preferentially at the edge as well as in the area surrounding the vacuoles in the matrix. Control sections incubated in the presence of glucosaccharo-1:4-lactone were devoid of the reaction product. No deposits of the reaction product were found in the nucleus, mitochondria, or microbodies. The limitations of the present cytochemical technique for use in electron microscopy are briefly discussed.     

THE FINE STRUCTURAL LOCALIZATION OF ENDOGENOUS AND EXOGENOUS PEROXIDASE ACTIVITY IN KUPFFER CELLS OF RAT LIVER

Endogenous peroxidase activity has been demonstrated in sections of rat liver fixed briefly by glutaraldehyde perfusion and incubated in Graham and Karnovsky's medium for cytochemical demonstration of peroxidase activity (29). In 25–40% of sinusoidal cells, an electron-opaque reaction product is localized in segments of the endoplasmic reticulum, including the perinuclear cisternae, a few Golgi vesicles and saccules and in some large membrane-bounded granules. This staining is abolished after prolonged fixation or boiling of tissue sections in glutaraldehyde, and in the absence of H₂O₂ or DAB from the incubation medium. Furthermore, the reaction is inhibited completely by sodium azide and high concentrations of H₂O₂, and partially by KCN and aminotriazole. Among the different cells in hepatic sinusoids, the nonphagocytic "fat-storing" cells (39) are always peroxidase negative, whereas the lining cells in process of erythrophagocytosis are consistently peroxidase positive. The possible biological significance of endogenous peroxidase in Kupffer cells is discussed. In addition, the uptake of exogenous horseradish peroxidase by Kupffer cells has been investigated. The exogenous tracer protein, which in contrast to endogenous peroxidase of Kupffer cells is not inhibited by prolonged aldehyde fixation, is taken up by micropinocytosis and remains confined to the lysosomal system of Kupffer cells. The significance of these observations in respect to some recent studies suggesting localization of exogenous peroxidases in the endoplasmic reticulum of Kupffer cells and peritoneal macrophages (22, 23) is briefly discussed.     

ELECTRON MICROSCOPE STUDIES ON ENZYME ACTIVITY AND THE ISOLATION OF THIOHYDANTOIN-INDUCED MYELIN FIGURES IN RAT LIVER

Characteristic cytoplasmic inclusions (myelin figures), consisting of concentric multilaminar paired membranes surrounding one or more lipid bodies, were produced in rat liver parenchymal cells by incorporating high doses of an anticonvulsant agent (Bax 422Z) into the animals' diet. Enzymatic reaction product (presumably lead phosphate) was found around the central fat of these myelin figures in liver which had been fixed in glutaraldehyde, incubated in Wachstein and Meisel's medium containing adenosine triphosphate or inosine tri- or diphosphate, postosmicated, embedded in epoxy resin, and examined in the electron microscope. In an attempt to isolate myelin figures, fresh liver from medicated rats was homogenized and differentially centrifuged. Thin sections of osmium tetroxide-fixed, Epon-embedded pellets from each fraction were examined with the electron microscope. The concentric membranous whorls, which are probably derived from cisternae of the endoplasmic reticulum, broke up as the cells were disrupted and became inextricably mixed with the microsomal fraction. However, when liver previously fixed in formalin for 24 hours was homogenized, the myelin figures remained intact.     

水分胁迫对高粱等作物叶片中核糖核酸酶活力的影响

水分胁迫促使高粱,玉米,蚕豆叶片中核糖核酸酶(RNase)活力增加,其程度以下部老叶多于上部叶片。酶活力的增加不仅由于水分胁迫的直接影响,也与缺水促进叶子早衰有关。 亚胺环己酮(CHM)前处理对水分胁迫时高粱幼苗RNase活力的增加起强烈抑制作用;氯霉素(CP)前处理对脱水引起的RNase活力增加无明显抑制效果。 高粱叶细胞中的RNase活力约85％存在于细胞质的液相中,其余似以颗粒酶形式存在于各类细胞器内,不易因水分胁迫而被释出。因而水分胁迫下高粱RNase活力的增加主要决定于细胞质中的可溶性RNase。 根据酶的热稳定性和它对酸化-碱化处理的反应表明水分胁迫并未引起高梁RNase的性质发生变化。     

在生长、功能和萎退各期,大鼠乳腺磷酸酶的研究

前言虽然乳腺在生长、功能和萎退过程中形态上的变化是很大的(姚曾序、顾国彦,1957),可是和形态变化相伴随的化学和生物化学变化却研究得少。关于碱性磷酸酶(以下简称 AKP)方面,Kay(1925),Folley 和 Kay(1935)最早证明牛、羊和豚鼠乳腺含有相当量的这种酶。以后 Folley 和 Greenbaum(1947)进一步用生化方法研究了大鼠在怀     

A LIGHT AND ELECTRON MICROSCOPE STUDY OF THE MORPHOLOGICAL CHANGES INDUCED IN RAT LIVER CELLS BY THE AZO DYE 2-ME-DAB

The cytological changes induced in rat liver cells by the aminoazo dye 2-Me-DAB have been examined by light and electron microscopy. It is observed that this non-carcinogenic compound duplicates most of the morphological alterations produced by other hepatotoxins, some of which, such as the closely related aminoazo dye 3'-Me-DAB, are potent carcinogens. These non-specific effects involve both the granular and agranular forms of the endoplasmic reticulum as well as the glycogen content of hepatic cells. The arrays of cisternal profiles of the granular reticulum in normal hepatic cells become disorganized and the dispersed cisternae often appear fragmented and irregular. Large cytoplasmic inclusions, consisting of loosely organized tubules and vesicles, are also observed which result from a hypertrophy of the agranular reticulum. The glycogen in the cells progressively decreases in amount. The most specific effect of 2-Me-DAB is to induce an increase in the number of mitochondria per cell. Many of these organelles are characterized by the presence of a median double membrane continuous with the inner limiting membrane of the mitochondrial envelope. Evidence is presented in favor of the view that this partition is directly related to the phenomenon of mitochondrial division. It was noted also in the course of the experiment that an increasing number of cells appear which stain quite intensely with methylene blue and appear denser than normal under electron microscopy. The significance of these cells is not known.