In vitro reversal of deformity and inhibition of aggregation of sickle red blood cells by two Congolese herbal medicines

2012 ◽  
Vol 6 (31) ◽  
Author(s):  
Marie Miezi Nsimba,
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
Herbal Medicines ◽  
Sickle Red Blood Cells

Plasma Hemoglobin and Heme Trigger Weibel Palade Body Exocytosis and Vaso-Occlusion in Transgenic Sickle Mice

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 896-896 ◽  
Author(s):  
John D Belcher ◽  
Julia Nguyen ◽  
Chunsheng Chen ◽  
Ann Smith ◽  
Abdu I Alayash ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Red Blood Cells ◽  
Blood Cells ◽  
Research Funding ◽  
Umbilical Vein ◽  
Pulmonary Veins ◽  
Heme Oxygenase 1 ◽  
Sickle Red Blood Cells

Abstract Abstract 896 Sickle cell disease (SCD) is a devastating hemolytic disease characterized by recurring episodes of painful vaso-occlusive crises and endothelial dysfunction. We hypothesize that hemoglobin (Hb) or hemin (ferri-protoporphyrin IX), released from HbS derived from hemolyzed sickle red blood cells, is fundamental to vaso-occlusion and vasculopathy in SCD. Utilizing intravital microscopy and dorsal skin fold chambers implanted on mice we previously demonstrated transient stasis in subcutaneous venules in response to hypoxia in transgenic sickle, but not normal mice. In the current studies we used NY1DD sickle mice; Townes-AA, -AS, and -SS mice; and C57 normal mice to test whether Hb or heme induce vaso-occlusion in the absence of a hypoxic stimulus. Four groups of 3–6 mice were given a bolus infusion (0.012 ml/g i.v.) of the following: 1) saline; 2) stroma-free HbA (32 umols heme/kg); 3) hemin (32 umols/kg); or 4) water (to induce intravascular hemolysis). In NY1DD mice, Hb, hemin or water induced a range of 36–46% stasis at 1 and 4 hours post-infusion compared to only 4–7% stasis at 1 and 4 hours in saline controls (Fig 1A, p<0.001 for all groups vs saline). In contrast, Hb, hemin or saline induced only 0–6% stasis in C57 mice (Fig 1B, p<0.001 for Hb or hemin in C57 vs NY1DD). Similarly, Townes-AA, -AS and –SS mice had a range of 5–15%, 27–37% and 40–50% stasis, respectively, in response to HbA (p<0.05 for all Townes pairs). The response to Hb or hemin was dose dependent with as little as 0.32 umols heme/kg, equivalent to ∼5 uM heme, a level found in SCD patients. These data support that heme, derived from Hb, promotes vascular stasis in sickle mice. The plasma of SCD patients is deficient in the two primary Hb/heme scavenging proteins haptoglobin (Hp) and hemopexin (Hpx). Therefore we infused equimolar Hp-Hb or Hpx-hemin into NY1DD mice and inhibited Hb- and heme-induced stasis at 1 hour by 79 and 88%, respectively (p<0.001). To examine whether nitrogen derivatives contribute to the mechanism of Hb- or heme-induced stasis, we measured plasma nitrite and nitrate (NOx) in NY1DD sickle mice 4 hours after infusion of Hb or hemin. NOx levels decreased in mice infused with Hb, but not in mice infused with hemin suggesting that NO consumption does not play a role in heme-induced stasis. Previous studies have demonstrated a role for P-selectin in vaso-occlusion in sickle mice. We tested the ability of heme to trigger Weibel Palade body (WPB) exocytosis in cultured primary human umbilical vein endothelial cells (HUVEC). Cells were treated with 10 uM hemin for 2, 5, 10, 15, 30 or 60 minutes and then fixed and stained (without permeabilization) for surface expression of P-selectin and von Willebrand factor (vWF). Cells treated with 100 uM histamine and vehicle served as positive and negative controls, respectively. There was rapid expression of P-selectin as well as vWF strings on the surface of HUVEC within 5 minutes of hemin addition. This is the first report that heme is an agonist for WPB exocytosis. Recently, heme has been shown to be an extracellular inflammatory signaling molecule with strict binding specificity for toll-like receptor-4 (TLR4). A specific small molecule inhibitor of TLR4 (TAK-242) completely prevented heme-induced P-selectin expression in vitro. In vivo the pulmonary veins and arteries of sickle mice injected with hemin expressed surface P-selectin within 15 minutes. Supporting this novel mechanism, blocking antibodies to P-selectin or the drug TAK-242 inhibited heme-induced stasis and thus provide a potential therapy for vaso-occlusion. These data strongly support that heme, released from hemolyzed sickle red blood cells, is fundamental to vaso-occlusion and vasculopathy in SCD. We speculate that removal of Hb and heme with Hp and Hpx, or as we've previously shown, detoxifying heme with heme oxygenase-1, would decrease the oxidative stress, inflammation and vaso-occlusion in SCD that cause endothelial cell dysfunction. Novel therapies focusing on the consequences of endothelial cell/heme interactions such as TLR4 or P-selectin antagonists, in addition to Hp and Hpx modulators should be considered in SCD. Disclosures: Belcher: Sangart, Inc: Research Funding. Nguyen:Sangart Inc: Research Funding. Chen:Sangart, Inc: Research Funding. Vercellotti:Sangart, Inc: Research Funding.

Dipyridamole inhibits sickling-induced cation fluxes in sickle red blood cells

Blood ◽  
2001 ◽  
Vol 97 (12) ◽  
pp. 3976-3983 ◽  
Author(s):  
Clinton H. Joiner ◽  
Maorong Jiang ◽  
William J. Claussen ◽  
Nancy J. Roszell ◽  
Zahida Yasin ◽  
...  
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
K Channel ◽  
Ionophore A23187 ◽  
Anion Conductance ◽  
Rbc Membrane ◽  
Cellular Dehydration ◽  
Sickle Red Blood Cells

Sickling-induced cation fluxes contribute to cellular dehydration of sickle red blood cells (SS RBCs), which in turn potentiates sickling. This study examined the inhibition by dipyridamole of the sickling-induced fluxes of Na+, K+, and Ca++ in vitro. At 2% hematocrit, 10 μM dipyridamole inhibited 65% of the increase in net fluxes of Na+ and K+ produced by deoxygenation of SS RBCs. Sickle-induced Ca++ influx, assayed as 45Ca++uptake in quin-2–loaded SS RBCs, was also partially blocked by dipyridamole, with a dose response similar to that of Na+and K+ fluxes. In addition, dipyridamole inhibited the Ca++-activated K+ flux (via the Gardos pathway) in SS RBCs, measured as net K+ efflux in oxygenated cells exposed to ionophore A23187 in the presence of external Ca++, but this effect resulted from reduced anion conductance, rather than from a direct effect on the K+channel. The degree of inhibition of sickling-induced fluxes was dependent on hematocrit, and up to 30% of dipyridamole was bound to RBC membranes at 2% hematocrit. RBC membrane content of dipyridamole was measured fluorometrically and correlated with sickling-induced flux inhibition at various concentrations of drug. Membrane drug content in patients taking dipyridamole for other clinical indications was similar to that producing inhibition of sickling-induced fluxes in vitro. These data suggest that dipyridamole might inhibit sickling-induced fluxes of Na+, K+, and Ca++ in vivo and therefore have potential as a pharmacological agent to reduce SS RBC dehydration.

Sickle red blood cells are more susceptible to in vitro haemolysis when exposed to normal saline versus Plasma‐Lyte A

Vox Sanguinis ◽  
2019 ◽  
Vol 114 (4) ◽  
pp. 325-329
Author(s):  
Neil Blumberg ◽  
Kelly Henrichs ◽  
Jill Cholette ◽  
Anthony Pietropaoli ◽  
Sherry Spinelli ◽  
...  
Keyword(s):  
Red Blood Cells ◽  
Normal Saline ◽  
Blood Cells ◽  
Sickle Red Blood Cells

In-vitro anti-sickling and membrane stability potentials of Mishenland polyherbal extract on sickle red blood cells

2019 ◽  
Vol 44 (1) ◽  
pp. 65 ◽  
Author(s):  
MusiliuA Oyenike ◽  
HelenB Akpan ◽  
OlatoyeJ Otulana ◽  
AdebayoK Adefule ◽  
KamoruA Adedokun ◽  
...  
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
Membrane Stability ◽  
Sickle Red Blood Cells

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

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).

Effect of liposomal curcumin on red blood cells in vitro

2013 ◽  
Vol 1 (Suppl. 1) ◽  
pp. A4.1
Author(s):  
Angela Storka
Keyword(s):  
Red Blood Cells ◽  
Blood Cells

scholarly journals THE SYNTHESIS OF PROTOPORPHYRIN IN VITRO BY RED BLOOD CELLS OF THE DUCK

1950 ◽  
Vol 183 (2) ◽  
pp. 757-765 ◽  
Author(s):  
David Shemin ◽  
Irving M. London ◽  
D. Rittenberg
Keyword(s):  
Red Blood Cells ◽  
Blood Cells

Nucleated red blood cells participate in myocardial regeneration in the toad Bufo Gargarizan Gargarizan

2021 ◽  
pp. 153537022110132
Author(s):  
Shu-Qin Liu ◽  
Xiao-Ye Hou ◽  
Feng Zhao ◽  
Xiao-Ge Zhao
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
Cardiac Injury ◽  
Myocardial Regeneration ◽  
Matrix Metalloproteinase 2 ◽  
Enzyme Linked Immunosorbent Assay ◽  
Cardiac Tissues ◽  
Nucleated Red Blood Cells ◽  
Model Animal

Heart regeneration is negligible in humans and mammals but remarkable in some ectotherms. Humans and mammals lack nucleated red blood cells (NRBCs), while ectotherms have sufficient NRBCs. This study used Bufo gargarizan gargarizan, a Chinese toad subspecies, as a model animal to verify our hypothesis that NRBCs participate in myocardial regeneration. NRBC infiltration into myocardium was seen in the healthy toad hearts. Heart needle-injury was used as an enlarged model of physiological cardiomyocyte loss. It recovered quickly and scarlessly. NRBC infiltration increased during the recovery. Transwell assay was done to in vitro explore effects of myocardial injury on NRBCs. In the transwell system, NRBCs could infiltrate into cardiac pieces and could transdifferentiate toward cardiomyocytes. Heart apex cautery caused approximately 5% of the ventricle to be injured to varying degrees. In the mildly to moderately injured regions, NRBC infiltration increased and myocardial regeneration started soon after the inflammatory response; the severely damaged region underwent inflammation, scarring, and vascularity before NRBC infiltration and myocardial regeneration, and recovered scarlessly in four months. NRBCs were seen in the newly formed myocardium. Enzyme-linked immunosorbent assay and Western blotting showed that the levels of tumor necrosis factor-α, interleukin- 1β, 6, and11, cardiotrophin-1, vascular endothelial growth factor, erythropoietin, matrix metalloproteinase- 2 and 9 in the serum and/or cardiac tissues fluctuated in different patterns during the cardiac injury-regeneration. Cardiotrophin-1 could induce toad NRBC transdifferentiation toward cardiomyocytes in vitro. Taken together, the results suggest that the NRBC is a cell source for cardiomyocyte renewal/regeneration in the toad; cardiomyocyte loss triggers a series of biological processes, facilitating NRBC infiltration and transition to cardiomyocytes. This finding may guide a new direction for improving human myocardial regeneration.

scholarly journals Multimodal Diagnostics of Microrheologic Alterations in Blood of Coronary Heart Disease and Diabetic Patients

Diagnostics ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 76
Author(s):  
Anastasia Maslianitsyna ◽  
Petr Ermolinskiy ◽  
Andrei Lugovtsov ◽  
Alexandra Pigurenko ◽  
Maria Sasonko ◽  
...  
Keyword(s):  
Coronary Heart Disease ◽  
Heart Disease ◽  
Red Blood Cells ◽  
Blood Cells ◽  
Optical Methods ◽  
Diabetic Patients ◽  
Aggregation Index ◽  
Significant Difference

Coronary heart disease (CHD) has serious implications for human health and needs to be diagnosed as early as possible. In this article in vivo and in vitro optical methods are used to study blood properties related to the aggregation of red blood cells in patients with CHD and comorbidities such as type 2 diabetes mellitus (T2DM). The results show not only a significant difference of the aggregation in patients compared to healthy people, but also a correspondence between in vivo and in vitro parameters. Red blood cells aggregate in CHD patients faster and more numerously; in particular the aggregation index increases by 20 ± 7%. The presence of T2DM also significantly elevates aggregation in CHD patients. This work demonstrates multimodal diagnostics and monitoring of patients with socially significant pathologies.

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