scholarly journals Prostaglandin E1 Induces c-Fos and Myc Proteins and Protects Rat Hippocampal Cells against Hypoxic Injury

1994 ◽  
Vol 14 (1) ◽  
pp. 150-155 ◽  
Author(s):  
Hideo Otsuki ◽  
Kazuo Yamada ◽  
Takamichi Yuguchi ◽  
Mamoru Taneda ◽  
Toru Hayakawa

We investigated the effects of prostaglandin (PG) E1 on the hypoxic injury of fetal rat hippocampal cells. Primary hippocampal cell cultures (embryonic day 18) were established and maintained. After 72 h in culture, PGE1 was added to the serum-free medium at a final concentration of 10−5-10−9 M. Cultures were divided into two groups: The normoxia group was in culture for another 48 h, and the hypoxia group was exposed to 24 h of hypoxia followed by continuation of culture for another 24 h. As a quantitative measure of cell death, lactate dehydrogenase (LDH) activity was estimated in the culture medium. The LDH activity, released by the hypoxic insult, was significantly smaller with PGE1 treatment at 10−6, 10−7, and 10−8 M (p < 0.01) and 10−9 M (p < 0.05) compared with the control. No differences in the LDH activities were observed in the normoxia group. Glial culture was not affected by the hypoxia. Western blot analysis showed an increased induction of 62-kDa c-Fos and 58, 60, and 66 kDa Myc proteins in rat hippocampal cells with 10−7 M PGE1 treatment. We conclude that PGE1 at concentrations of 10−6-10−9 M protects rat hippocampal neurons against hypoxic insult.

1991 ◽  
Vol 27 (8) ◽  
pp. 625-632 ◽  
Author(s):  
D. Jassal ◽  
R. N. N. Han ◽  
I. Caniggia ◽  
M. Post ◽  
A. K. Tanswell

1988 ◽  
Vol 90 (4) ◽  
pp. 683-689 ◽  
Author(s):  
A. Kimura ◽  
T. Kawaguchi ◽  
T. Ono ◽  
A. Sakuma ◽  
Y. Yokoya ◽  
...  

Two variants (74AD and 74FL) established from rat ascites hepatoma AH7974 were examined for the production of glycosaminoglycans in culture. There was no difference between the adhesive (74AD) and the floating (74FL) variants in quantity of glycosaminoglycans produced by their cultivation in minimum essential medium supplemented with 10% foetal calf serum. However, they were distinctly different in the distribution patterns of heparan sulphate. In 74FL, about 70% of total heparan sulphate was found in the culture medium in soluble form, whereas in 74AD, only 7% was found in the medium and the rest was in the cell-substratum complex. In a serum-free medium, 74AD cells grew without adhering to the substratum. After cultivation, more than 90% of total heparan sulphate was found in the cell-associated fractions and the rest in the substratum fractions. No heparan sulphate was detected in the culture medium. On the other hand, 74FL cells released heparan sulphate to the serum-free medium as much as to the serum-containing medium. The increase in amount of heparan sulphate in the culture medium of 74FL cells was supposed to be caused by failure of the cells to deposit heparan sulphate at the cell surface and not caused by increased production. Cell-substratum adhesion mechanisms involving cell surface heparan sulphate (heparan sulphate proteoglycan) and some serum intermediate(s) are discussed for 74AD cells.


1971 ◽  
Vol 26 (10) ◽  
pp. 1045-1048 ◽  
Author(s):  
Dieter F. Hülser ◽  
Werner Frank

Normal embryonic rat cells incubated in serum-free medium accumulate in G1-phase of the cell cycle. On addition of a growth-stimulating protein isolated from fetal calf serum they are triggered to proceed through the cycle, and they resume DNA-synthesis 15 to 20 hours later. In this paper it is demonstrated that the surface membrane potential difference (PD) decreases immediately after changing serum-free medium against culture medium containing either calf serum or the isolated serum protein; the original PD is restored 2 to 3 hours later. Serumprotein without growthstimulating activity does not affect the PD.A permanent rat cell line which grows independently of serum also has been tested. The PD of these cells is not significantly influenced by calf serum.


1984 ◽  
Vol 103 (2) ◽  
pp. 195-203 ◽  
Author(s):  
D. J. Hill ◽  
R. D. G. Milner

ABSTRACT The actions of partially purified porcine platelet-derived growth factor (PDGF) and highly purified multiplication-stimulating activity (MSA) II and MSA III-2, which are somatomedins, were investigated on the incorporation of [3H]thymidine and [35S]sulphate by fetal rat costal cartilage in vitro. This was compared with their effects in the presence of 1% fetal calf serum (FCS) on the uptake of thymidine by growth-arrested fetal rat fibroblasts. Platelet-derived growth factor at concentrations of 0·21–21 μg/l enhanced the incorporation of both isotopes by fetal cartilage in the presence of 1% FCS, but had an inconsistent action on thymidine uptake and no significant action on sulphate uptake in serum-free medium. Platelet-derived growth factor promoted thymidine uptake by growth-arrested, isolated fetal rat fibroblasts. Multiplication-stimulating activity II (10–100 μg/l) stimulated the uptake of thymidine and sulphate by fetal cartilage in medium containing 1% FCS but had no consistent action in serum-free medium, although MSA II and PDGF had a synergistic effect on thymidine uptake in the absence of serum. Multiplication-stimulating activity III-2 had no consistent action on thymidine or sulphate incorporation by fetal cartilage in either serum-free or serum-supplemented medium. However, the same preparation of MSA III-2 stimulated the uptake of [3H]thymidine into fetal rat fibroblasts with a half-maximal response at a concentration of 5–10 μg/l. The results identify PDGF as a possible mitogenic agent for fetal rat connective tissues in vitro and show a differential sensitivity of fetal cartilage to MSA peptides. J. Endocr. (1984) 103, 195–203


1972 ◽  
Vol 27 (12) ◽  
pp. 1500-1507 ◽  
Author(s):  
Rafael Manso-Martinez ◽  
Werner Frank

3H-thymidine incorporation by secondary cultures of embryonic rat cells and by cultures of a permanent rat cell line is strongly inhibited a few minutes after addition of hydroxyurea (HU) to the culture medium (2 × 10-3 ᴍ and 5 × 10-4 ᴍ, resp.); normal embryonic rat cells are considerably less sensitive to the drug than are permanent cells. About 76 percent of the population of a secondary culture can be synchronized at the beginning of the S-period, if the cells at first are accumulated in G1 phase by incubation in serum-free medium and subsequently are stimulated by the addition of ten percent calf serum in the presence of 2 × 10-3 ᴍ HU.The size of DNA replicated in the presence of HU has been analysed by centrifugation in alkaline sucrose gradients. The results indicate that under these conditions DNA subunits are synthesized with sedimentation coefficients between 21S and 30S. These intermediates cannot be joined together, probably because a ligase is inhibited by HU.Secondary culture incubated in medium-containing 1 × 10-4 ᴍ HU incorporate twice as much 3H-thymidine as do control cells in HU-free medium. This is due to an increased number of cells in the S phase, as was shown by autoradiography. The kinetics of thymidine incorporation indicate that the duration of G1 phase is shortened by small doses of HU


2019 ◽  
Vol 2019 ◽  
pp. 1-9 ◽  
Author(s):  
Miaomiao Chen ◽  
Xiaoshuang Zuo ◽  
Qinqin Xu ◽  
Rong Wang ◽  
Suhua Fan ◽  
...  

Nanodiamonds (NDs) have been recognized as emerging carbon-based delivery vehicles due to their biocompatibility. NDs were reported to be nontoxic and suited for biomedical applications in the complete cell culture medium. However, in this study, the cytotoxicity of NDs in serum-free medium was studied and indicated that serum proteins in cell culture medium played significant effect on the toxicity of NDs. Therefore, the interaction mechanism between NDs and a serum protein (human serum albumin, HSA) was first investigated by fluorescence quenching technique and circular dichroism (CD) spectrometry. The results suggest that HSA strongly bonds on the NDs surface to form “protein corona,” which not only prevents the aggregation of NDs and improves its stability but also inhibits the cytotoxicity of NDs. In serum-free medium, NDs exhibited obvious toxicity toward the human lung epithelial cell line (BEAS-2B) and showed concentration-dependent cytotoxicity. In the presence of bovine serum albumin (BSA), which shares structural homology and similar properties with HSA, toxicity of NDs was apparently inhibited. Therefore, the interaction between serum protein and NDs should be considered in the understanding of the biological effects of NDs exposure in biomedical applications.


Parasitology ◽  
1987 ◽  
Vol 95 (1) ◽  
pp. 165-171 ◽  
Author(s):  
D. C. Jenkins ◽  
P. Topley ◽  
E. B. Rapson

SUMMARYThe cidal properties of some phenolic, halogenated diphenyl, salicylanilide, benzimidazole and diaminophenoxyalkane anthelmintics, against 6-week-old worms of Fasciola hepatica were assessed in vitro. In a conventional fluke culture medium containing RPMI 1640, supplemented with serum with or without rabbit erythrocytes or pink-ghosts, only the halogenated diphenyl and salicylanilide compounds showed activity at concentrations equal to or less than 100 μm. However, when basal, serum and cell-free RPMI 1640 was used, all compounds other than diamphenethide were highly active, their minimum lethal concentrations being some 25–125 times lower under these conditions. The inclusion of rabbit liver microsomes in the basal culture medium resulted in diamphenethide exhibiting cidal activity equivalent to that seen when its free-amine active metabolite was assayed. The possibility that the activity of many of these compounds was masked in vitro because of their serum binding properties is discussed. Recommendations are made that in vitro screens for new fasciolicides should be carried out in serum-free medium and that additional replicates containing mammalian liver microsomes and liver cytosolic extracts be included as means for the metabolic activation of certain otherwise undetectable prodrugs.


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