Preparation of Subcellular Fractions from Granulation Tissue by Density Gradient Centrifugation.

Lehtinen, Pirjo; Kulonen, E.; Rämsby, S.; Stjernström, Nils E.; Enzell, Curt R.; Reid, William W.; Yanaihara, Noboru; Yanaihara, Chizuko

doi:10.3891/acta.chem.scand.33b-0327

Cited by 10 publications

(4 citation statements)

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“…The chemical assays were performed by routine methods described previously (3,4,(10)(11)(12)(13)(14). Ribonuclease was estimated according to Liu et al (15).…”

Section: Methodsmentioning

confidence: 99%

Fibroblast RNA and Macrophage Proteins (Including the Fibrogenic Factor) in Experimental Silicosis

Kulonen¹,

Aalto²,

Aho³

et al. 1983

Environmental Health Perspectives

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A hypothesis is presented for the action of silica-treated macrophages on protein synthesis in fibroblasts and also a method for the isolation of silica-attached materials in lung tissue. The increased protein synthesis in the fibroblasts is due, at least partly, to an increase in mRNA. Silica prevents the suppressing "macrophage effect" of macrophage-originated ribonuclease on fibroblasts. However, under certain conditions, collagen synthesis is stimulated by silica-treated macrophage preparations to such an extent that the effect cannot be explained by the inhibition of macrophage ribonuclease alone. We therefore postulate the existence of a fibrogenic factor, which is released by the macrophages. This factor has been demonstrated and can be purified from lung homogenate of SiO2-treated rats.

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“…The chemical assays were performed by routine methods described previously (3,4,(10)(11)(12)(13)(14). Ribonuclease was estimated according to Liu et al (15).…”

Section: Methodsmentioning

confidence: 99%

Fibroblast RNA and Macrophage Proteins (Including the Fibrogenic Factor) in Experimental Silicosis

Kulonen¹,

Aalto²,

Aho³

et al. 1983

Environmental Health Perspectives

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“…It was found to act on fibroblast polysomes (53), the yield ofwhich was increased and whose RNA was preserved from degradation with corresponding augmentation oftheir stability (55). Synthesis ofcollagen having occurred in rough endoplasmic reticulum, whose RNA was elevated (56), the protein was transferred in secretory vesicles to the extracllular fibrillary state (57). Collagen formation was not linked to tanscription, and collagenase was not involved (54); instead, a huge increase in RNA (particularly nuclear) translational capacity was detected in rat lung affected by experimental silicosis and was directed mainly to collagen synthesis (58).…”

Section: Collagen Productionmentioning

confidence: 99%

Minerals, fibrosis, and the lung.

Heppleston

1991

Environ Health Perspect

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Determinants of pulmonary fibrosis induced by inhaled mineral dusts include quantity retained, particle size, and surface area, together with their physical form and the reactive surface groups presented to alveolar cells. The outstanding problem is to ascertain how these factors exert their deleterious effects. Both compact and fibrous minerals inflict membrane damage, for which chemical mechanisms still leave uncertainty. A major weakness of cytotoxicity studies, even when lipid peroxidation and reactive oxygen species are considered, lies in tacitly assuming that membrane damage suffices to account for fibrogenesis, whereas the parallel occurrence of such manifestations does not necessarily imply causation. The two-phase procedure established that particles, both compact and fibrous, induce release of a macrophage factor that provokes fibroblasts into collagen synthesis. The amino acid composition of the macrophage fibrogenic factor was characterized and its intracellular action explained. Fibrous particles introduce complexities respecting type, durability, and dimensions. Asbestotic fibrosis is believed to depend on long fibers, but scrutiny of the evidence from experimental and human sources reveals that a role for short fibers needs to be entertained. Using the two-phase system, short fibers proved fibrogenic. Other mechanisms, agonistic and antagonistic, may participate. Growth factors may affect the fibroblast population and collagen production, with cytokines such as interleukin-1 and tumor necrosis factor exerting control. Immune involvement is best regarded as an epiphenomenon. Downregulation of fibrogenesis may follow collagenase release from macrophages and fibroblasts, while augmented type II cell secretion of lipid can interfere with the macrophage-particle reaction.

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“…The nuclei were recovered by centrifugation at 500 x.g for 5 min and washed once with 2 ml of buffer A. The combined supernatants were fractionated in a discontinuous gradient (8.54, 20, 45 and 65% sucrose) at 100000 χ g for 4 h. The interphase of 45/65% sucrose contained the rough endoplasmic reticulum and the surface of the gradient the soluble fractionl 15,16].…”

Section: Sub Cellular Fractionation Of the Incubated Granulationtissumentioning

confidence: 99%

Penetration of Various Mononuclear Ribonucleases into Rat Experimental Granulation-Tissue Fibroblasts and Their Intracellular Effects

Lehtinen¹,

Aho²,

Kulonen³

1981

Hoppe-Seyler´s Zeitschrift für physiologische Chemie

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The penetration of five different nuclear ribonucleases were not found particularly mononuclear ribonucleases into the subcellular in any subcellular compartment, particles of rat experimental granulation-tissue Both biologically active macrophage enzymes fibroblasts was compared, along with the effects deg raded the nuclear RNA of fibroblasts and of the enzymes on the fibroblast RNA released it to the soluble fraction in contrast to fractions.^Q biologically inactive macrophage enzyme Ribonucleases from normal and silica-treated and ribonuclease from normal human monocytes. rat peritoneal macrophages have been shown Instead the ribonuclease from normal human before to regulate the nucleic acid and protein monocytes seemed to degrade RNA in the soluble metabolism of rat experimental granulation-tissue fraction. There were no marked differences in fibroblasts. These biologically active enzymes the subcellular effects of ribonucleases from were taken into the fibroblasts in a greater amount normal and silica-treated macrophages. However, than the corresponding human monocyte enthe treatment of human monocytes with silica zymes and the biologically inactive rat macrochanged their ribonuclease so that it split the phage ribonuclease. The biologically active macnuclear RNA of fibroblast and released it to the rophage enzymes were incorporated mainly into soluble fraction in the same way as the biologithe nuclear fraction. The other three monocally active macrophage enzymes did. Penetration verschiedener mononuclearer Ribonucleasen in Fibroblasten aus experimentell gewonnenem Granulationsgewebe der Ratte und intrazelluläre Wirkungen dieser EnzymeZusammenfassung: Die Aufnahme von fünf verRibonucleasen aus normalen und Si0 2 -behandelschiedenen mononuclearen Ribonucleasen in die ten Peritoneal-Makrophagen der Ratte reguliesubzellulären Partikeln von Fibroblasten aus ren bekanntlich den Nucleinsäure-und Proteinexperimentell gewonnenem Granulationsgewebe Stoffwechsel der genannten Fibroblasten. Diese der Ratte wurde verglichen, ebenso wie die Wirbiologisch aktiven Enzyme werden von den Fikungen der Enzyme auf die Fibroblasten-RNAbroblasten in höherem Maße aufgenommen als die Fraktionen.entsprechenden menschlichen Monocyten-En-• Abbreviations: mph-RNase = ribonuclease from rat peritoneal macrophages; mon-RNase = ribonuclease from adhered human blood monocytes; prefix N for normal and SiÜ2 for silica-treated cells. Hepes = 4-(2-hydroxyethyl)-l-pi-(perazineethanesulphonic acid; PPO = 2,5-diphenyloxazole; POPOP = l,4-bis(5-phenyl-2-oxazolyl)benzene.

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Preparation of Subcellular Fractions from Granulation Tissue by Density Gradient Centrifugation.

Cited by 10 publications

References 0 publications

Fibroblast RNA and Macrophage Proteins (Including the Fibrogenic Factor) in Experimental Silicosis

Fibroblast RNA and Macrophage Proteins (Including the Fibrogenic Factor) in Experimental Silicosis

Minerals, fibrosis, and the lung.

Penetration of Various Mononuclear Ribonucleases into Rat Experimental Granulation-Tissue Fibroblasts and Their Intracellular Effects

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