Atorvastatin prevents Plasmodium falciparum cytoadherence and endothelial damage
- Zacharie Taoufiq1, 4Email author,
- Paco Pino1, 5,
- Nadine N'dilimabaka1,
- Issam Arrouss1,
- Serge Assi1, 6,
- Florent Soubrier2,
- Angelita Rebollo†1 and
- Dominique Mazier†1, 3Email author
© Taoufiq et al; licensee BioMed Central Ltd. 2011
Received: 30 November 2010
Accepted: 28 February 2011
Published: 28 February 2011
The adhesion of Plasmodium falciparum parasitized red blood cell (PRBC) to human endothelial cells (EC) induces inflammatory processes, coagulation cascades, oxidative stress and apoptosis. These pathological processes are suspected to be responsible for the blood-brain-barrier and other organs' endothelial dysfunctions observed in fatal cases of malaria. Atorvastatin, a drug that belongs to the lowering cholesterol molecule family of statins, has been shown to ameliorate endothelial functions and is widely used in patients with cardiovascular disorders.
The effect of this compound on PRBC induced endothelial impairments was assessed using endothelial co-culture models.
Atorvastatin pre-treatment of EC was found to reduce the expression of adhesion molecules and P. falciparum cytoadherence, to protect cells against PRBC-induced apoptosis and to enhance endothelial monolayer integrity during co-incubation with parasites.
These results might suggest a potential interest use of atorvastatin as a protective treatment to interfere with the pathophysiological cascades leading to severe malaria.
Malaria remains a major threat to public health with 40% of the world population currently at risk. Every year, an estimated 500 million cases of clinical malaria and at least one million deaths are reported [1, 2]. Fatal cases of malaria occur mainly in young children in Africa and consist of an acute neurological syndrome (cerebral malaria), either in isolation or concomitantly with multi-organ failure (pulmonary distress, acute renal failure) . Anti-malarial drugs, such as quinine and artemisinin derivatives are administered intravenously as an emergency treatment. However, although these drugs effectively and rapidly clear parasites from the blood, 15%-20% of patients still die, probably as the result of impaired host responses [4, 5]. Such a situation together with the difficulty in developing vaccines highlights the urgent need of novel strategies for complement therapeutics.
The severity of Plasmodium falciparum infection depends largely on the ability of parasitized red blood cells (PRBC) to adhere on endothelial cells (EC) and sequester in the capillary network of vital organs (e.g. brain, lungs, kidneys, liver.) Moreover, activation of endothelial cells resulting from the adhesion of infected erythrocytes leads to an overexpression of different mediators, such as adhesion molecules ("hyperadhesion" phenomenon), pro-inflammatory cytokines, coagulation factors and contributes by itself to the pathology . Among endothelial receptors, some are known to interact with PRBC, mainly via PfEMP1 (P. falciparum erythrocyte membrane protein 1), a highly polymorphic parasite ligand exported on the infected erythrocyte surface. A number of the cytoadherence receptors have been identified, such as ICAM-1, CD36, VCAM-1 or P-selectin . It was previously shown that P. falciparum adhesion to human endothelial cells can specifically trigger proinflammatory gene expression[8–10], oxidative stress  and caspases activation , further leading to perturbation of the endothelial barrier integrity [12, 13]. In addition PRBC adhesion to EC induces redox and rho-kinase dependent EC activation and apoptosis, which can be reversed respectively by the addition of anti-oxidants or fasudil rho-kinase inhibitor [12, 14].
Atorvastatin is an oral drug that lowers the level of cholesterol in the blood. All statins, including atorvastatin, prevent the production of cholesterol by blocking the enzyme HMGCoA reductase. However, there is increasing evidence that statins may also exert effects beyond cholesterol lowering. Many of these cholesterol-independent or "pleiotropic" vascular effects of statins appear to involve restoring or improving the endothelial function through increasing the bioavailability of nitric oxide, promoting re-endothelialization, reducing oxidative stress, and inhibiting inflammatory responses . Thus, the endothelium-dependent effects of statins are thought to contribute to many of the beneficial effects of statin therapy in cardiovascular disease . In addition to its lipids lowering effects, atorvastatin has been shown to promote nitric oxide (NO) production by decreasing caveolin-1 expression in EC, regardless of the level of extracellular LDL-cholesterol . Statins retard the initiation of atherosclerosis formation through the improvement of NO bioavailability by both up-regulation of endothelial-nitric oxide synthase (eNOS) mRNA and decrease of superoxide anion O2- production in EC . Statins modulate the adhesion cascade at multiple points by targeting both the endothelium and leukocytes and affect cell adhesion by inhibiting chemokine expression (MCP-1) in activated leukocytes and endothelial cells. There is also evidence that statins decrease ICAM-1 expression in stimulated EC and monocytes . The effects of atorvastatin on mature EC are correlated with the activation of the anti-apoptotic Akt pathway, as determined by the phosphorylation of Akt and eNOS . Therefore, activation of Akt represents a mechanism that can account for some of the beneficial side effects of statins.
Given the pleiotropic effects on endothelium of statins in general and atorvastatin in particular, atorvastatin was hypothesized to be useful as a protective drug against P. falciparum induced endothelial damages. Primary human lung endothelial cells (HLEC) were used here in co-culture with P. falciparum-infected erythrocytes Previous studies have already shown that PRBC adhesion triggers inflammation, oxidative stress and apoptosis within lung endothelial cells [9, 11, 12, 21, 22]. Moreover, acute respiratory distress as a complication of malaria infection is rare, but with a vey high rate of mortality [3, 23]. The data presented in this manuscript show that, concomitantly with increased expression of Akt within HLEC, atorvastatin reduces adhesion of P. falciparum-infected erythrocytes, and as a consequence, prevents the endothelial damages induced by PRBC.
Culture of human endothelial cell
Primary endothelial cells were isolated from human lung (HLEC) after enzymatic digestion and selected using a continuous gradient and immunomagnetic purification technique, as described elsewhere . Endothelial cells of ninth to twelfth passages derived from one batch were used for the experiments. Before use, cells were verified for their expression of ICAM-1, CD36, Von Willebrand factor, VCAM-1, CD31, E/P-selectin and CSA. HLEC were raised in M199 medium (Gibco), supplemented with 10 μg/ml of endothelial cell growth supplement (Upstate, NY) and 10% of fetal calf serum (Biowest), at 37°C with 5% CO2, using 8 chamber-Labtek (Nunc International, Napervil), 35 mm glass bottom dishes (MatTek corp.), 96-well plates (Costar), or Transwell insert supports (Corning LifeSciences).
Plasmodium falciparum culture
The P. falciparum 3D7 clone was used for these experiments. Infected erythrocytes were maintained in culture according to Trager and Jensen's technique in a suspension of erythrocytes in RPMI (Gibco) supplemented with 8.3 g/l of Hepes, 2.1 g/l of NaHCO, 0.1 mg.ml-1 of gentamycin, 2 g/l of dextrose, and 0.4% of Albumax II (Gibco, Invitrogen Corporation) . The 3D7 clone was characterized for adhesion phenotype as previously described  and adheres to ICAM-1 and CD36. For each experiment, parasite cultures were enriched in mature forms by Plasmagel floating . Briefly, erythrocytes were harvested from 5 to 10% parasitized cultures and centrifuged 5 minutes at 2000 rpm. Cells were resuspended in Plasmion® and incubated for 20 minutes at 37°C. The upper fraction containing mature trophozoites and schizonts was collected and washed three times in RPM, before adequate adjustment of the hematocrit and parasitemia.
Real time RT-PCR
HLECs were treated with atorvastatin (Pfizer) for 24 hours, and total RNA was prepared using RNeasy mini kit (Qiagen). RT and quantitative PCR were performed as previously described . Primers used were as follow: ICAM-1: forward 5'-GCAATGTGCAAGAAGATAGCCA-3' reverse 5'-GGGCAAGACCTCAGGTCATGT-3' VCAM-1: forward 5'-GAGTACGCAAACACTTTATGTCAATGT-3', reverse 5'-CTCGTCCTTTCGGGACCG-3', P-selectin forward 5'-AGACTCCCCACCAATGTGTGA-3', reverse 5'-CCACGAGTGTCAGAACAATCCA-3' CD36 forward 5'-TAATGGCACAGATGCAGCCT-3', reverse 5'-ACAGCATAGATGGACCTGCAA-3' HPRT1 forward 5'-AAAGGACCCCACGAAGTGTT-3' reverse 5'-TCAAGGGCATATCCTACAACAA-3'.
Plasmodium falciparum adhesion assay
HLEC were raised in Labtek until confluence. Suspensions of mature pRBC were deposited onto cells and incubated for one hour at 37°C with gentle shaking every 10 minutes. After the incubation, the unbound pRBC were removed and the preparation was fixed for 30 minutes at room temperature with 2% glutaraldehyde, before staining with Giemsa. The number of parasites adhering to 700 HLEC was counted by direct observation with light microscope.
Nucleosome release assay
The proportion of apoptotic cells was assessed by measuring the intracytoplasmic release of mono- and oligonucleosomes as, using Cell Death Dtection ELISA® (Roche) previously described . Briefly, HLEC were raised in 96-well plate (Costar) until confluence and exposed 24 hours to PRBC (hematocrit 5% parasitemia 50%) or RBC (hematocrit 5%). After 5 PBS washing steps, endothelial cells were lysed and cytoplasms were analysed for their nucleosomes content.
Endothelial barrier integrity assay
Confluent endothelial monolayer were obtained by seeding 30,000 HLEC on Transwell permeable support (polyester, 3 μm pores, 6.5 mm diameter) and raised in M199 medium supplemented as above during 36 hours. Endothelial monolayers were then exposed during 12 hours to PRBC suspensions at 1% parasitemia and 0.5% of hematocrit. Transwell compartments were then washed three times and cell monolayers on Transwell inserts were transferred to a new plate containing PBS. Evans Blue (0.5 mg/mL) (ICN Biomedicals) was then added in the 'upper compartments'. After 5 min of incubation at 37°C and 5% CO2, the 'under compartments' were collected for optic density analysis of diffused Evans Blue (630 nm) with a standard microplate reader (Bio-Tek™ EL311SX).
Cells (1 × 106) were lysed in Laemmli sample buffer and protein extracts were separated by SDS-PAGE, transferred to nitrocellulose, blocked (5% non-fat dry milk) in Tris-buffered saline (TBS: 20 mM Tris-HCl pH 7.5, 150 mM NaCl) plus 0.05% Tween-20 and incubated with the primary antibody over night at 4°C in TBS-0.5% non-fat dry milk. The membrane was washed and incubated with PO-conjugated secondary antibody for two hours at room temperature. Secondary antibodies on western blot membranes were revealed using the ECL system.
Immunofluorescence and confocal microscopy
HLEC were raised in 35 mm glass bottom dishes, before fixation with 1% paraformaldehyde for 5 min, permeabilized and then incubated with polyclonal anti-Akt antibody (Cell Signaling) for 2 h in PBS 3% BSA at room temperature. FITC-secondary antibody was added and incubated for 1 h at room temperature. After several washing steps, samples were incubated with methanol at -20°C for 10 min, mounted with Vectashield medium and analysed by confocal microscopy.
Differences between groups were analysed for statistical significance using the Games-Howell post-hoc (SPSS software). A p value at least less than 0.05 was considered significant.
Results and discussion
Atorvastatin prevents P. falciparum-induced endothelial adhesion molecules upregulation
Atorvastatin decreases P. falciparum cytoadherence and P. falciparum- induced endothelial apoptosis
Atorvastatin protects endothelial barrier integrity from P. falciparum- induced impairments
Atorvastatin increases Akt expression in endothelial cells exposed to P. falciparum
The data presented here demonstrate that atorvastatin can be used to reduce the cytoadherence of P. falciparum on endothelial cells, which is one of key events, along with the inflammatory burst, involved in the pathogenesis of human severe malaria cases. Atorvastatin can decrease the expression of adhesion molecules and also prevents the PRBC-induced overexpression of adhesion molecules. Moreover, atorvastatin shows an ability to strongly protect endothelial cell against P. falciparum-induced collateral damages, cell apoptosis and endothelial barrier permeabilization.
The observed cytoprotection effect on endothelial cells may mainly be due to the decrease of PRBC adhesion through the down-regulation of adhesion molecules, as PRBC contact was previously shown to specifically trigger pro-inflammatory and pro-apoptotic signaling cascades . Moreover, atorvastatin stimulates Akt expression within HLEC, which is the major actor of anti-apoptotic and endothelial cell survival pathways . Endothelial protection of atorvastatin appears then to be 'pleiotropic' via anti-adhesive, anti-inflammatory and anti-apoptotic synergistic effects. Indeed, PRBC cytoadherence is known to activate many deleterious cascades such as oxidative stress, via mitochondrial ROS (radical oxygen species) production, rho-kinase and NF-kappa B-dependent signaling to induce 'hyperadhesion' phenomenon, endothelial activation and apoptosis [8, 11, 12, 14, 28, 31]. When phosphorylated, Akt inactivates pro-apoptotic elements such as Bad, preventing mitochondrial ROS release, and caspase 9 . Statins can also increase the bioavailability and the physiological production of endothelial nitric oxide (NO) via Akt phosphorylation and downstream effector endothelial NO-synthase (eNOS), and via the stabilization of eNOS mRNA [30, 33]. NO has many crucial functions in vascular endothelium particularly in maintaining permanently anti-inflammatory and anti-adhesive endothelium homeostasis. NO-donors have currently been under investigation in cerebral malaria adjunctive emergency treatments, the efficiency of which remains controversial . Indeed, the deleterious scavenging of NO by ROS has not been considered with antioxidant co-treatment yet. As regard their molecular 'pleiotropic' effects, particularly with the antioxidant and the NO bioavailability promotion, statins may then be highly relevant to 'shield' the endothelium against both PRBC induced impairments and uncontrolled host responses in severe malaria cases.
There is increasing interest of using statins as complementary therapeutic to anti-plasmodial molecules. However, these studies with statins were performed on infected erythrocytes only and have shown interestingly to have a direct inhibitory effect on P. falciparum growth in vitro, using high doses ranging from 120 to 240 microM , from 5 to 40 microM  or from 2.5 to 10.8 microM . Atorvastatin was shown to be 10 times more active against P. falciparum compared to mevastatin, simvastatin, lovastatin, fluvastatin or pravastatin , suggesting that atorvastatin is the best candidate among statins for therapy prospects in malaria treatment. Simvastatin was also used in mouse model of cerebral malaria (C57BL/6 mice infected with Plasmodium berghei), but failed to decrease significantly parasitemia or to prevent death [38, 39]. Atorvastatin may have a beneficial effect on mice survival, only when administered in combination with artesunate . Indeed, atorvastatin was recently shown to reduce the IC50 of the anti-plasmodial activity of quinine, mefloquine and dihydroartemisinin [41–43]. However, the pathogenesis of cerebral malaria in murine model relies solely on the inflammatory response unlike the pathogenesis of human cerebral malaria. Indeed, the cytoadherence phenomenon does not exist in P. berghei mice infections [44, 45]. Atorvastatin was used in pre-treatment to specifically evaluate the potential effects on endothelial cells. Endothelial damage is almost universal in severe malaria pathogenesis and statins have proved their efficiency in clinical use for 20 years against cardiovascular diseases . Interestingly, significant endothelial protection effects were obtained with doses as low as 100 nanoM, 500 nanoM or 1 microM.
Although the 'pleiotropic' effects of statins are well documented, the precise molecular mechanism by which the PI3K/Akt/eNOS pathway is activated remains unknown [29, 46]. Since the cholesterol lowering property of statins by itself ameliorates the endothelial function, statins beneficial effects were for long attributed to the inhibition of HMG-CoA reductase and the cholesterol synthesis pathway. However, the endothelial function improvement was shown to occur earlier than cholesterol lowering and independently from the cholesterol rate in blood serum [47, 48]. Statins are inhibiting the synthesis of mevalonate, the second biochemical reaction in the cholesterol synthesis pathway. Mevalonate is also an important precursor of isoprenoid intermediates such as farnesyl pyrophosphate (FPP) or geranylgeranyl pyrophosphate (GGPP). Isoprenoids serve as cell surface inner membrane-anchoring molecules to functionalize Rho-GTPases, such as Rho (GGPP) and Ras (FPP) [49, 50], key signaling molecules, after geranylation and farnesylation post-translational modifications. Rho-GTPases act as molecular switches (active and inactive when bound to GTP and GDP, respectively), and are the first intermediates of the intracellular signaling engagement of these receptors with the downstream effector Rho-kinase. We have previously demonstrated that P. falciparum-induced endothelial collateral damages are precisely dependent on Rho-kinase signaling . Rho-kinase pathway by many aspects is antagonist to PI3K/Akt/eNOS endothelial cell survival pathway . In fact, strong endothelial protective effects, obtained with atorvastatin against P. falciparum, may likely be related to both rho-kinase pathway inhibition and Akt cell survival pathway promotion. However, it was previously reported that the rho-kinase inhibition by fasudil could not decrease significantly the number of parasites and P. falciparum cytoadherence [12, 51].
In conclusion, data from this report demonstrate that HMG-CoA reductase inhibitor atorvastatin prevents P. faciparum cytoadherence and confers the ability to the endothelium to resist against consequent cellular damages. In conclusion, this study suggests that atorvastatin may be a good candidate for further studies or clinical trials on the use of statins in malaria treatment. Moreover, the safety of statins on malaria treatment needs to be addressed, since these molecules were not originally designed for these therapeutic approaches.
List of abbreviations
red blood cells
parasitized red blood cells
human lung endothelial cells.
Acknowledgements and funding
We are grateful to David Walliker for the gift of the 3D7 clone, We also thank Kabongo Muanza for HLEC isolation, Maurel Tefit for technical assistance, and Ioannis Vouldoukis for critical manuscript discussions. Zacharie Taoufiq was financially supported by the Ministère de l'Education Nationale, de la Recherche et de la Technologie.and by la Fondation pour la Recherche Médicale (FRM).
- Snow RW, Guerra CA, Noor AM, Myint HY, Hay SI: The global distribution of clinical episodes of Plasmodium falciparum malaria. Nature. 2005, 434: 214-217. 10.1038/nature03342.PubMed CentralView ArticlePubMedGoogle Scholar
- UNICEF/WHO: World Malaria Report 2009. 2009, [http://www.whoint/malaria/world_malaria_report_2009]Google Scholar
- Marsh K, Forster D, Waruiru C, Mwangi I, Winstanley M, Marsh V, Newton C, Winstanley P, Warn P, Peshu N, Pasvol G, Snow R: Indicators of life-threatening malaria in African children. N Engl J Med. 1995, 332: 1399-1404. 10.1056/NEJM199505253322102.View ArticlePubMedGoogle Scholar
- Dondorp A, Nosten F, Stepniewska K, Day N, White N: Artesunate versus quinine for treatment of severe falciparum malaria: a randomised trial. Lancet. 2005, 366: 717-725. 10.1016/S0140-6736(05)67176-0.View ArticlePubMedGoogle Scholar
- Idro R, Jenkins NE, Newton CR: Pathogenesis, clinical features, and neurological outcome of cerebral malaria. Lancet Neurol. 2005, 4: 827-840. 10.1016/S1474-4422(05)70247-7.View ArticlePubMedGoogle Scholar
- van der Heyde HC, Nolan J, Combes V, Gramaglia I, Grau GE: A unified hypothesis for the genesis of cerebral malaria: sequestration, inflammation and hemostasis leading to microcirculatory dysfunction. Trends Parasitol. 2006, 22: 503-508. 10.1016/j.pt.2006.09.002.View ArticlePubMedGoogle Scholar
- Ho M, White NJ: Molecular mechanisms of cytoadherence in malaria. Am J Physiol. 1999, 276: 1231-1242.Google Scholar
- Jenkins N, Wu Y, Chakravorty S, Kai O, Marsh K, Craig A: Plasmodium falciparum intercellular adhesion molecule-1-based cytoadherence-related signaling in human endothelial cells. J Infect Dis. 2007, 196: 321-327. 10.1086/518795.PubMed CentralView ArticlePubMedGoogle Scholar
- Pino P, Vouldoukis I, Kolb JP, Mahmoudi N, Desportes-Livage I, Bricaire F, Danis M, Dugas B, Mazier D: Plasmodium falciparum--infected erythrocyte adhesion induces caspase activation and apoptosis in human endothelial cells. J Infect Dis. 2003, 187: 1283-1290. 10.1086/373992.View ArticlePubMedGoogle Scholar
- Tripathi AK, Sha W, Shulaev V, Stins MF, Sullivan DJ: Plasmodium falciparum-infected erythrocytes induce NF-kappaB regulated inflammatory pathways in human cerebral endothelium. Blood. 2009, 114: 4243-4252. 10.1182/blood-2009-06-226415.PubMed CentralView ArticlePubMedGoogle Scholar
- Pino P, Vouldoukis I, Dugas N, Hassani-Loppion G, Dugas B, Mazier D: Redox-dependent apoptosis in human endothelial cells after adhesion of Plasmodium falciparum-infected erythrocytes. Ann N Y Acad Sci. 2003, 1010: 582-586. 10.1196/annals.1299.109.View ArticlePubMedGoogle Scholar
- Taoufiq Z, Gay F, Balvanyos J, Ciceron L, Tefit M, Lechat P, Mazier D: Rho kinase inhibition in severe malaria: thwarting parasite-induced collateral damage to endothelia. J Infect Dis. 2008, 197: 1062-1073. 10.1086/528988.View ArticlePubMedGoogle Scholar
- Tripathi AK, Sullivan DJ, Stins MF: Plasmodium falciparum-infected erythrocytes decrease the integrity of human blood-brain barrier endothelial cell monolayers. J Infect Dis. 2007, 195: 942-950. 10.1086/512083.View ArticlePubMedGoogle Scholar
- Taoufiq Z, Pino P, Dugas N, Conti M, Tefit M, Mazier D, Vouldoukis I: Transient supplementation of superoxide dismutase protects endothelial cells against Plasmodium falciparum-induced oxidative stress. Mol Biochem Parasitol. 2006, 150: 166-173. 10.1016/j.molbiopara.2006.07.008.View ArticlePubMedGoogle Scholar
- Laufs U: Beyond lipid-lowering: effects of statins on endothelial nitric oxide. Eur J Clin Pharmacol. 2003, 58: 719-731.PubMedGoogle Scholar
- Rosenson RS: Pluripotential mechanisms of cardioprotection with HMG-CoA reductase inhibitor therapy. Am J Cardiovasc Drugs. 2001, 1: 411-420. 10.2165/00129784-200101060-00001.View ArticlePubMedGoogle Scholar
- Brouet A, Sonveaux P, Dessy C, Moniotte S, Balligand JL, Feron O: Hsp90 and caveolin are key targets for the proangiogenic nitric oxide-mediated effects of statins. Circ Res. 2001, 89: 866-873. 10.1161/hh2201.100319.View ArticlePubMedGoogle Scholar
- Sumi D, Hayashi T, Thakur NK, Jayachandran M, Asai Y, Kano H, Matsui H, Iguchi A: A HMG-CoA reductase inhibitor possesses a potent anti-atherosclerotic effect other than serum lipid lowering effects--the relevance of endothelial nitric oxide synthase and superoxide anion scavenging action. Atherosclerosis. 2001, 155: 347-357. 10.1016/S0021-9150(00)00597-9.View ArticlePubMedGoogle Scholar
- Romano M, Diomede L, Sironi M, Massimiliano L, Sottocorno M, Polentarutti N, Guglielmotti A, Albani D, Bruno A, Fruscella P, Salmona M, Vecchi A, Pinza M, Mantovani M: Inhibition of monocyte chemotactic protein-1 synthesis by statins. Lab Invest. 2000, 80: 1095-1100. 10.1038/labinvest.3780115.View ArticlePubMedGoogle Scholar
- Urbich C, Dernbach E, Zeiher AM, Dimmeler S: Double-edged role of statins in angiogenesis signaling. Circ Res. 2002, 90: 737-744. 10.1161/01.RES.0000014081.30867.F8.View ArticlePubMedGoogle Scholar
- Siau A, Toure FS, Ouwe-Missi-Oukem-Boyer O, Ciceron L, Mahmoudi N, Vaquero C, Froissard P, Bisvigou U, Bisser S, Coppee JY, Bischoff E, David PH, Mazier D: Whole-transcriptome analysis of Plasmodium falciparum field isolates: identification of new pathogenicity factors. J Infect Dis. 2007, 196: 1603-1612. 10.1086/522012.View ArticlePubMedGoogle Scholar
- Zang-Edou ES, Bisvigou U, Taoufiq Z, Lekoulou F, Lekana-Douki JB, Traore Y, Mazier D, Toure-Ndouo FS: Inhibition of Plasmodium falciparum field isolates-mediated endothelial cell apoptosis by Fasudil: therapeutic implications for severe malaria. PLoS One. 2010, 5: e13221-10.1371/journal.pone.0013221.PubMed CentralView ArticlePubMedGoogle Scholar
- Mishra SK, Mohanty S, Mohanty A, Das BS: Management of severe and complicated malaria. J Postgrad Med. 2006, 52: 281-287.PubMedGoogle Scholar
- Muanza K, Gay F, Behr C, Scherf A: Primary culture of human lung microvessel endothelial cells: a useful in vitro model for studying Plasmodium falciparum-infected erythrocyte cytoadherence. Res Immunol. 1996, 147: 149-163. 10.1016/0923-2494(96)83167-1.View ArticlePubMedGoogle Scholar
- Trager W: Cultivation of malaria parasites. Methods Cell Biol. 1994, 45: 7-26. full_text.View ArticlePubMedGoogle Scholar
- Goodyer ID, Johnson J, Eisenthal R, Hayes DJ: Purification of mature-stage Plasmodium falciparum by gelatine flotation. Ann Trop Med Parasitol. 1994, 88: 209-211.PubMedGoogle Scholar
- Brun M, Bourdoulous S, Couraud PO, Elion J, Krishnamoorthy R, Lapoumeroulie C: Hydroxyurea downregulates endothelin-1 gene expression and upregulates ICAM-1 gene expression in cultured human endothelial cells. Pharmacogenomics J. 2003, 3: 215-226. 10.1038/sj.tpj.6500176.View ArticlePubMedGoogle Scholar
- Tripathi AK, Sullivan DJ, Stins MF: Plasmodium falciparum-infected erythrocytes increase intercellular adhesion molecule 1 expression on brain endothelium through NF-kappaB. Infect Immun. 2006, 74: 3262-3270. 10.1128/IAI.01625-05.PubMed CentralView ArticlePubMedGoogle Scholar
- Mason JC: Statins and their role in vascular protection. Clin Sci (Lond). 2003, 105: 251-266. 10.1042/CS20030148.View ArticleGoogle Scholar
- Wolfrum S, Jensen KS, Liao JK: Endothelium-dependent effects of statins. Arterioscler Thromb Vasc Biol. 2003, 23: 729-736. 10.1161/01.ATV.0000063385.12476.A7.View ArticlePubMedGoogle Scholar
- Schofield L, Novakovic S, Gerold P, Schwarz RT, McConville MJ, Tachado SD: Glycosylphosphatidylinositol toxin of Plasmodium up-regulates intercellular adhesion molecule-1, vascular cell adhesion molecule-1, and E-selectin expression in vascular endothelial cells and increases leukocyte and parasite cytoadherence via tyrosine kinase-dependent signal transduction. J Immunol. 1996, 156: 1886-1896.PubMedGoogle Scholar
- Datta SR, Dudek H, Tao X, Masters S, Fu H, Gotoh Y, Greenberg ME: Akt phosphorylation of BAD couples survival signals to the cell-intrinsic death machinery. Cell. 1997, 91: 231-241. 10.1016/S0092-8674(00)80405-5.View ArticlePubMedGoogle Scholar
- Endres M, Laufs U, Liao JK, Moskowitz MA: Targeting eNOS for stroke protection. Trends Neurosci. 2004, 27: 283-289. 10.1016/j.tins.2004.03.009.View ArticlePubMedGoogle Scholar
- Sobolewski P, Gramaglia I, Frangos J, Intaglietta M, van der Heyde HC: Nitric oxide bioavailability in malaria. Trends Parasitol. 2005, 21 (9): 415-422. 10.1016/j.pt.2005.07.002.View ArticlePubMedGoogle Scholar
- Couto AS, Kimura EA, Peres VJ, Uhrig ML, Katzin AM: Active isoprenoid pathway in the intra-erythrocytic stages of Plasmodium falciparum: presence of dolichols of 11 and 12 isoprene units. Biochem J. 1999, 341: 629-637. 10.1042/0264-6021:3410629.PubMed CentralPubMedGoogle Scholar
- Pradines B, Torrentino-Madamet M, Fontaine A, Henry M, Baret E, Mosnier J, Briolant S, Fusai T, Rogier C: Atorvastatin is 10-fold more active in vitro than other statins against Plasmodium falciparum. Antimicrob Agents Chemother. 2007, 51: 2654-2655. 10.1128/AAC.01330-06.PubMed CentralView ArticlePubMedGoogle Scholar
- Parquet V, Briolant S, Torrentino-Madamet M, Henry M, Almeras L, Amalvict R, Baret E, Fusai T, Rogier C, Pradines B: Atorvastatin is a promising partner for antimalarial drugs in treatment of Plasmodium falciparum malaria. Antimicrob Agents Chemother. 2009, 53: 2248-2252. 10.1128/AAC.01462-08.PubMed CentralView ArticlePubMedGoogle Scholar
- Helmers AJ, Gowda DC, Kain KC, Liles WC: Statins fail to improve outcome in experimental cerebral malaria and potentiate Toll-like receptor-mediated cytokine production by murine macrophages. Am J Trop Med Hyg. 2009, 81: 631-637. 10.4269/ajtmh.2009.09-0204.View ArticlePubMedGoogle Scholar
- Kobbe R, Schreiber N, May J, Jacobs T: Simvastatin treatment shows no effect on the incidence of cerebral malaria or parasitemia during experimental malaria. Antimicrob Agents Chemother. 2008, 52: 1583-1584. 10.1128/AAC.01428-07.PubMed CentralView ArticlePubMedGoogle Scholar
- Bienvenu AL, Picot S: Statins alone are ineffective in cerebral malaria but potentiate artesunate. Antimicrob Agents Chemother. 2008, 52: 4203-4204. 10.1128/AAC.00513-08.PubMed CentralView ArticlePubMedGoogle Scholar
- Parquet V, Henry M, Wurtz N, Dormoi J, Briolant S, Gil M, Baret E, Amalvict R, Rogier C, Pradines B: Atorvastatin as a potential anti-malarial drug: in vitro synergy in combinational therapy with quinine against Plasmodium falciparum. Malar J. 2010, 9: 139-10.1186/1475-2875-9-139.PubMed CentralView ArticlePubMedGoogle Scholar
- Savini H, Souraud JB, Briolant S, Baret E, Amalvict R, Rogier C, Pradines B: Atorvastatin as a potential antimalarial drug: in vitro synergy in combinational therapy with dihydroartemisinin. Antimicrob Agents Chemother. 2010, 54: 966-967. 10.1128/AAC.01006-09.PubMed CentralView ArticlePubMedGoogle Scholar
- Wurtz N, Briolant S, Gil M, Parquet V, Henry M, Baret E, Amalvict R, Almeras L, Rogier C, Pradines B: Synergy of mefloquine activity with atorvastatin, but not chloroquine and monodesethylamodiaquine, and association with the pfmdr1 gene. J Antimicrob Chemother. 2010, 65: 1387-1394. 10.1093/jac/dkq173.View ArticlePubMedGoogle Scholar
- Lackner P, Beer R, Helbok R, Broessner G, Engelhardt K, Brenneis C, Schmutzhard E, Pfaller K: Scanning electron microscopy of the neuropathology of murine cerebral malaria. Malar J. 2006, 5: 116-10.1186/1475-2875-5-116.PubMed CentralView ArticlePubMedGoogle Scholar
- White NJ, Turner GD, Medana IM, Dondorp AM, Day NP: The murine cerebral malaria phenomenon. Trends Parasitol. 2009, 26: 11-15. 10.1016/j.pt.2009.10.007.View ArticlePubMedGoogle Scholar
- Zhou Q, Liao JK: Statins and cardiovascular diseases: from cholesterol lowering to pleiotropy. Curr Pharm Des. 2009, 15: 467-478. 10.2174/138161209787315684.PubMed CentralView ArticlePubMedGoogle Scholar
- MRC/BHF: MRC/BHF Heart Protection Study of cholesterol lowering with simvastatin in 20,536 high-risk individuals: a randomised placebo-controlled trial. Lancet. 2002, 360: 7-22. 10.1016/S0140-6736(02)09327-3.View ArticleGoogle Scholar
- O'Driscoll G, Green D, Taylor RR: Simvastatin, an HMG-coenzyme A reductase inhibitor, improves endothelial function within 1 month. Circulation. 1997, 95: 1126-1131.View ArticlePubMedGoogle Scholar
- Goldstein JL, Brown MS: Regulation of the mevalonate pathway. Nature. 1990, 343: 425-430. 10.1038/343425a0.View ArticlePubMedGoogle Scholar
- Van Aelst L, D'Souza-Schorey C: Rho GTPases and signaling networks. Genes Dev. 1997, 11: 2295-2322. 10.1101/gad.11.18.2295.View ArticlePubMedGoogle Scholar
- Waknine-Grinberg JH, McQuillan JA, Hunt N, Ginsburg H, Golenser J: Modulation of cerebral malaria by fasudil and other immune-modifying compounds. Exp Parasitol. 2010, 125: 141-146. 10.1016/j.exppara.2010.01.005.View ArticlePubMedGoogle Scholar
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.