Influence of proinflamatory macrophages (M-1) on proliferative activity and PPAR-γ expression in neoplastic rat hepatocytes in vitro

We sought to analyse the proliferative activity and PPARγ expression in neoplastic and non-neoplastic rat hepatocytes, exposed to immunologically trained macrophages M1 (Mf-M1). Ten-week-old female Wistar rats were divided into two groups: I - control (n=5) and II neoplstic (n=5). To induce HCC in the neoplastic group, genotoxic diethylnitrosamine (DEN) was administered after a partial hepatectomy (PH). Hepatocytes were isolated by liver perfusion method and the mononuclear blood cells were isolated using Lymphoprep density-gradient centrifugation. After differentiation, blood cells were treated with barley-derived β-glucan (BBG) (10 μg/ml). Adhered heaptocytes and Mf were cultured in 3D Quasi-Vivo System during 24 h. Then, hepatic proliferation and PPAR γ expression were analysed after 72 h and the 1st ,the 2nd and the 3rd week of incubation. The proliferation index of control hepatocytes ranged between 0.41±0.04 – 0.43±0.03 after 72 h and after the 3rd week of incubation respectively. Exposure of these cells to Mf resulted in marked increase of IP after the 1st, the 2nd and the 3rd week of incubation. When hepatocytes were influenced by proinflammatory Mf- M1, their proliferation was maintained at control stage. DEN-obtained hepatocytes, not influenced by macrophages, exhibit enhanced proliferation. When these cells where co-cultured with Mf-M1, marked (P≤0.05) inhibition of cell proliferation was observed. In such condition, a high negative relationship (r= -0.93) between proliferative activity of the hepatocytes and the PPARγ concentration was observed. We conclude that immunologically trained macrophages M1, are capable to activation of PPARγ in rat neoplastic hepatocytes derived from experimentally induced HCC. In turn intensified expression of PPARγ may inhibited proliferation of neoplastic cell in vitro.

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Proliferative and oxidative response of hepatocytes (Hep) and hepatic stellate cells (HSC) isolated from rats exposed to ketogenic diet

Polish Journal of Veterinary Sciences ◽

10.2478/pjvs-2014-0102 ◽

2014 ◽

Vol 17 (4) ◽

pp. 703-711 ◽

Cited By ~ 1

Author(s):

M. Wójcik ◽

J. Wessely-Szponder ◽

U. Kosior-Korzecka

Keyword(s):

Ketogenic Diet ◽

Hepatic Stellate Cells ◽

Proliferative Activity ◽

Gradient Centrifugation ◽

Stellate Cells ◽

No Production ◽

Proliferative Effect ◽

The Media ◽

Plasma Ffa

Abstract Ketogenic diet (KD) is considered in the context of its anti-epileptic effects, but its influence on liver dysfunction has not been elucidated yet. The study was aimed to investigate the activity of hepatocytes (Hep) and hepatic stellate cells (HSC) isolated from rats fed with KD, in respect of NO and superoxide generation by these cells as well as their proliferative activity in vitro. We also sought to characterize the plasma FFA profiles in control and ketogenic rats. Hep and HSC were isolated by the collagenase perfusion method and separated by the Percoll gradient centrifugation. After the 4th, 8th and 12th day of incubation, the media were collected for further analysis. NO generation increased within the time of incubation both in Hep and HSC isolated from KD-rats. In HSC group NO production raised significantly from 2.65 ± 0.07 μM/106 cells on 4th day of incubation to 5.49 ± 1.2 μM/106 cells on 12th day of incubation. In respect to O2-. generation experimental Hep and HSC provide considerably higher quantities of this free radical. until 12th day of incubation (2.5 ± 0.07 and 3.2 ± 0.3 nM/106 cells, respectively). Although KD exerts anti-proliferative effect on hepatocytes, in respect to HSC it intensifies their proliferative activity. Furthermore, as we estimated on the basis of NO and O2-. generation both Hep and HSC exposed to KD are the source of free radicals

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DETERMINATION OF BIOCOMPATIBILITY AND CYTOTOXICITY OF POROUS TITANIUM-BASED MATERIALS IN EXPERIMENT

Science and Innovations in Medicine ◽

10.35693/2500-1388-2017-0-3-18-22 ◽

2017 ◽

Vol 2 (3) ◽

pp. 18-22

Author(s):

AV V Kolsanov ◽

AN N Nikolaenko ◽

VV V Ivanov ◽

SA A Prikhodko ◽

PV V Platonov

Keyword(s):

Electron Microscopy ◽

Scanning Electron Microscopy ◽

Proliferative Activity ◽

Dermal Fibroblasts ◽

Titanium Silicide ◽

Proliferation Index ◽

Contact Method ◽

Test Materials ◽

Scanning Electron

Aim - to evaluate the proliferative activity of dermal fibroblast cultures in the presence of composite materials based on titanium silicides in vitro. Materials and methods. To assess the proliferative activity of dermal fibroblasts in vitro, the following materials were used: titanium silicide, titanium carbosilicide oxidized in vacuum and without vacuum, titanium VT-00 (comparison group). Testing of proliferative activity was carried out by the direct contact method. The proliferation index, the doubling time and the number of culture doubling during the cultivation period were calculated. Attachment of dermal fibroblasts to the surface of the test materials and their presence on it during cultivation was assessed by scanning electron microscopy. Results. The study of the morphofunctional characteristics of dermal fibroblasts cultured in the presence of the test samples of material showed that during the entire experiment no major changes occurred in any of the series, the cells retained the monolayer growth characteristic of fibroblasts, preferably spindleshaped with 2-4 shoots. Moreover, all cultures of dermal fibroblasts underwent the same number of doublings during the experiment and reached saturation density 7 days after sowing, which indicates good proliferative activity of cells in the presence of test materials. The results of scanning electron microscopy demonstrate the high affinity of human dermal fibroblasts for both titanium silicide and titanium carbosilicides. Conclusion. Absence of morphofunctional changes in dermal fibroblasts and active proliferation testify to the absence of cytotoxicity of the investigated alloys, and the ability of cells to adhere to the surface of materials indicates their good biocompatibility.

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L-Ascorbate protects rat hepatocytes against sodium arsenite—induced cytotoxicity and oxidative damage

Human & Experimental Toxicology ◽

10.1177/0960327109357215 ◽

2009 ◽

Vol 29 (2) ◽

pp. 103-111 ◽

Cited By ~ 23

Author(s):

Asit Kumar Bera ◽

Tanmoy Rana ◽

Subhashree Das ◽

Subhasish Bandyopadhyay ◽

Debasis Bhattacharya ◽

...

Keyword(s):

Oxidative Stress ◽

Cell Proliferation ◽

Sodium Arsenite ◽

In Vitro Study ◽

Rat Hepatocytes ◽

Proliferation Index ◽

No Production ◽

Cell Proliferation Index ◽

And Oxidative Stress

Sodium arsenite—exposed hepatocytes of rat showed higher production of nitric oxide (NO) and increased lipid peroxidation (LPO) level vis-a-vis activity of superoxide dismutase (SOD) and catalase (CAT) were significantly lowered. Subsequently, the cell proliferation index (CPI) and cell viability were also reduced. Treatment with L-ascorbate was found effective in normalizing the arsenic-induced alteration of SOD and CAT activity and LPO level in rat hepatocytes. These observations indicated that L-ascorbate also has potent cytoprotective role as it could reduce the NO production and normalize the cell proliferation and viability of hepatocytes. Therefore, the in vitro study suggested that ascorbic acid is helpful to ameliorate the arsenic-induced cytotoxicity and oxidative stress of rat hepatocytes.

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Detection of Viral Proteins after Infection of Cultured Hepatocytes with Rabbit Hemorrhagic Disease Virus

Journal of Virology ◽

10.1128/jvi.72.5.4492-4497.1998 ◽

1998 ◽

Vol 72 (5) ◽

pp. 4492-4497 ◽

Cited By ~ 38

Author(s):

Matthias König ◽

Heinz-Jürgen Thiel ◽

Gregor Meyers

Keyword(s):

Disease Virus ◽

Genome Organization ◽

Gradient Centrifugation ◽

Liver Perfusion ◽

Viral Proteins ◽

Rabbit Hemorrhagic Disease Virus ◽

Hemorrhagic Disease ◽

Rabbit Hemorrhagic Disease ◽

Infected Cells

ABSTRACT The calicivirus rabbit hemorrhagic disease virus (RHDV), which replicates predominantly in the livers of infected rabbits, cannot be propagated in tissue culture. To enable the performance of in vitro studies, rabbit hepatocytes were isolated by liver perfusion and gradient centrifugation. After inoculation with purified RHDV, more than 50% of the cells proved to be infected. Protein analyses led to the detection of 13 RHDV-specific polypeptides within the infected cells. These proteins were assigned to defined regions of the viral genome, resulting in a refined model of RHDV genome organization.

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Study of the IL-4 and IL-4δ2 interaction with the IL-4R receptors of the I and II types in silico and the effect on the functional activity of mononuclear cells in vitro

Bulletin of Siberian Medicine ◽

10.20538/1682-0363-2010-3-51-56 ◽

2010 ◽

Vol 9 (3) ◽

pp. 51-56

Author(s):

K. V. Goremykin ◽

A. N. Silkov ◽

B. V. Shilov ◽

V. Yu. Serebrov ◽

A. E. Sazonov ◽

...

Keyword(s):

Blood Cells ◽

Proliferative Activity ◽

Mononuclear Cells ◽

Splice Form ◽

Antagonistic Effects ◽

Computer Methods ◽

Full Form ◽

Alternatively Spliced ◽

Mononuclear Blood Cells

Alternatively spliced interleukins are very actively studied over the last years. Splice form of IL-4 — IL-4δ2 has antagonistic effects to its full form on proliferative activity of human mononuclear blood cells and their IL-6 production. Antagonistic effects between IL-4 and IL-4δ2 were confirmed and explained in this study due to combination of biochemical and computer methods.

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Immunoenzymatic demonstration of the simultaneous presence of transferrin, hemopexin and albumin in a same hepatocyte and their ultrastructural localization

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100081309 ◽

1978 ◽

Vol 36 (2) ◽

pp. 88-89

Author(s):

M. Kraemer ◽

J. Foucrier ◽

J. Vassy ◽

M.T. Chalumeau

Keyword(s):

Plasma Proteins ◽

Rat Hepatocytes ◽

Ultrastructural Localization ◽

Carrier Proteins ◽

Normal Cells ◽

Adult Rat ◽

Adult Rat Hepatocytes ◽

Monospecific Antisera ◽

Different Levels

Some authors using immunofluorescent techniques had already suggested that some hepatocytes are able to synthetize several plasma proteins. In vitro studies on normal cells or on cells issued of murine hepatomas raise the same conclusion. These works could be indications of an hepatocyte functionnal non-specialization, meanwhile the authors never give direct topographic proofs suitable with this hypothesis.The use of immunoenzymatic techniques after obtention of monospecific antisera had seemed to us useful to bring forward a better knowledge of this problem. We have studied three carrier proteins (transferrin = Tf, hemopexin = Hx, albumin = Alb) operating at different levels in iron metabolism by demonstrating and localizing the adult rat hepatocytes involved in their synthesis.Immunological, histological and ultrastructural methods have been described in a previous work.

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Ultrastructural observations on the in vitro cytoadherence of Plasmodium falciparum infected red blood cells

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100162132 ◽

1990 ◽

Vol 48 (3) ◽

pp. 914-915

Author(s):

D.J.P. Ferguson ◽

A.R. Berendt ◽

J. Tansey ◽

K. Marsh ◽

C.I. Newbold

Keyword(s):

Plasmodium Falciparum ◽

Red Blood Cells ◽

Blood Cells ◽

Umbilical Vein ◽

Cell Types ◽

Amelanotic Melanoma ◽

Cytoplasmic Process ◽

Umbilical Vein Endothelial Cells

In human malaria, the most serious clinical manifestation is cerebral malaria (CM) due to infection with Plasmodium falciparum. The pathology of CM is thought to relate to the fact that red blood cells containing mature forms of the parasite (PRBC) cytoadhere or sequester to post capillary venules of various tissues including the brain. This in vivo phenomenon has been studied in vitro by examining the cytoadherence of PRBCs to various cell types and purified proteins. To date, three Ijiost receptor molecules have been identified; CD36, ICAM-1 and thrombospondin. The specific changes in the PRBC membrane which mediate cytoadherence are less well understood, but they include the sub-membranous deposition of electron-dense material resulting in surface deformations called knobs. Knobs were thought to be essential for cytoadherence, lput recent work has shown that certain knob-negative (K-) lines can cytoadhere. In the present study, we have used electron microscopy to re-examine the interactions between K+ PRBCs and both C32 amelanotic melanoma cells and human umbilical vein endothelial cells (HUVEC).We confirm previous data demonstrating that C32 cells possess numerous microvilli which adhere to the PRBC, mainly via the knobs (Fig. 1). In contrast, the HUVEC were relatively smooth and the PRBCs appeared partially flattened onto the cell surface (Fig. 2). Furthermore, many of the PRBCs exhibited an invagination of the limiting membrane in the attachment zone, often containing a cytoplasmic process from the endothelial cell (Fig. 2).

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Mechanism of the Antiplatelet Action of Dipyridamole in Whole Blood: Modulation of Adenosine Concentration and Activity

Thrombosis and Haemostasis ◽

10.1055/s-0038-1661437 ◽

1986 ◽

Vol 55 (01) ◽

pp. 012-018 ◽

Cited By ~ 86

Author(s):

Paolo Gresele ◽

Jef Arnout ◽

Hans Deckmyn ◽

Jos Vermylen

Keyword(s):

Platelet Aggregation ◽

Adenosine Receptor ◽

Whole Blood ◽

Blood Cells ◽

Inhibitory Action ◽

Phosphodiesterase Inhibitor ◽

Inhibitory Effect ◽

Adenosine Concentration ◽

Pharmacological Studies

SummaryDipyridamole inhibits platelet aggregation in whole blood at lower concentrations than in plasma. The blood cells responsible for increased effectiveness in blood are the erythrocytes. Using the impedance aggregometer we have carried out a series of pharmacological studies in vitro to elucidate the mechanism of action of dipyridamole in whole blood. Adenosine deaminase, an enzyme breaking down adenosine, reverses the inhibitory action of dipyridamole. Two different adenosine receptor antagonists, 5’-deoxy-5’-methylthioadenosine and theophylline, also partially neutralize the activity of dipyridamole in blood. Enprofylline, a phosphodiesterase inhibitor with almost no adenosine receptor antagonistic properties, potentiates the inhibition of platelet aggregation by dipyridamole. An inhibitory effect similar to that of dipyridamole can be obtained combining a pure adenosine uptake inhibitor (RE 102 BS) with a pure phosphodiesterase inhibitor (MX-MB 82 or enprofylline). Mixing the blood during preincubation with dipyridamole increases the degree of inhibition. Lowering the haematocrit slightly reduces the effectiveness.Although we did not carry out direct measurements of adenosine levels, the results of our pharmacological studies clearly show that dipyridamole inhibits platelet aggregation in whole blood by blocking the reuptake of adenosine formed from precursors released by red blood cells following microtrauma. Its slight phosphodiesterase inhibitory action potentiates the effects of adenosine on platelets.

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