International Journal of PharmTech Research
CODEN (USA): IJPRIF, ISSN: 0974-4304, ISSN(Online): 2455-9563
Vol.10, No.02, pp 69-73, 2017
10. Young BC, Levine RJ, Karumanchi SA. Pathogenesis of preeclampsia, Annual Review of Pathology, 2010;5:173-92.
11. Wang A, Rana S, Karumanchi SA. Preeclampsia : the role of angiogenic factors in its pathogenesis, Physiology (Bethesda), 2009;24:147-58.
12. Rajakumar A, Conrad KP. Expression, ontogeny, and regulation of hypoxia-inducible transcription factors in the human placenta, Biology of Reproduction, 2000;63(2):559-69.
13. Irani RA, Zhang Y, Blackwell SC, Zhou CC, Ramin SM, Kellems RE, et al. The detrimental role of angiotensin receptor agonistic autoantibodies in intrauterine growth restriction seen in preeclampsia, Journal of Experimental Medicine, 2009;206(12):2809-22.
14. FAO. Curcumin- chemical and technical assessment 61st JECFA 2004.
15. Chattopadhyay I, Biswas K, Bandyopadhyay U, Banerjee JK. Turmeric and curcumin: Biological actions and medicinal applications, Current Science, 2004;1:44-153.
16. Jurenka JS. Anti-inflammatory properties of curcumin, a major constituent of Curcuma longa: a review of preclinical and clinical research, Alternative Medicne Review, 2009;14(2):141-53.
17. Hansson SR, Chen Y, Brodszki J, Chen M, Hernandez-Andrade E, Inman JM, et al. Gene expression profiling of human placentas from preeclamptic and normotensive pregnancies, Molecular Human Reproduction, 2006;12(3):169-79.
18. Subroto D. Curcumin decrease placental NF-κB expression and malonedialdehide serum in preeclamptic Mus musculus model. Surabaya: Universitas Airlangga; 2011.
19. Munaut C, Lorquet S, Pequeux C, Blacher S, Berndt S, Frankenne F, et al. Hypoxia is responsible for soluble vascular endothelial growth factor receptor-1 (VEGFR-1) but not for soluble endoglin induction in villous trophoblast, Human Reproduction, 2008;23(6):1407-15.
20. Pramatirta, A.Y., Laksono, B., Prima N.F., Anita D.A., Sofie R.K., Debbie S.R., Ani M.M., Erlina W. Effects of Low Dose Aspirin on Caspase 3, TNF-α and Apoptotic Index Levels in Preclampsia Maternal Serum-Induced Placental Trophoblast Cell Line In Vitro, International Journal of PharmTech Research, 2016;9(10):1-6.
21. Gunardi, J.I., Mose, J., Mieke, H.S., Anita, D.A., Prima, N.F., and Triyuli. Effects of Papua Ant Nests (Myrmecodia pendens) on Level of sFlt-1, PIGF, MDA and NO in Preeclampsia-induced HUVEC Cell Line, International Journal of PharmTech Research, 2016;9(6);424-435.
22. Wan, X.H., Li, Y.W. and Luo, X.P., 2007. Curcumin attenuated the lipid peroxidation and apoptotic liver injury in copper-overloaded rats. Zhonghua er ke za zhi= Chinese Journal of Pediatrics, 45(8), pp.604-608.
23. J. S. Wright, Predicting the antioxidant activity of curcumin and curcuminoids, Journal of Molecular Structures (Theochem) 591, 207–217 (2002).
24. Rahardjo, B., Widjajanto, E., Sujuti, H. and Keman, K., 2014. Curcumin decreased level of proinflammatory cytokines in monocyte cultures exposed to preeclamptic plasma by affecting the transcription factors NF-κB and PPAR-γ, Biomarkers and Genomic Medicine, 6(3), pp.105-115.
25. Gong, P., Liu, M., Hong, G., Li, Y., Xue, P., Zheng, M., Wu, M., Shen, L., Yang, M., Diao, Z. and Hu, Y., 2016. Curcumin improves LPS-induced preeclampsia-like phenotype in rat by inhibiting the TLR4 signaling pathway, Placenta, 41, pp.45-52.
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Variables tested in this study were normally distributed both in normal and preeclampsia serum treated with curcumin in various concentrations incubated for 24 h and 48 h (data are not shown). Effects of curcumin in various concentration, incubation time, and serums on level of MDA are presented in Figure 2. As shown in Figure 2, level of MDA decreased as in accordance with longer incubation time and increased curcumin concentration. Curcumin decreased level of MDA (p<0,001). Level of MDA in preeclampsia-induced HUVEC decreased from 18,703 μM to 8,577 μM after treated with curcumin of 62,5 μg/ml incubated for 24 h, and higher after 48 h from 18,872 μM to 8,667 μM.
Discussion
Oxidative stress is caused by imbalance of pro-oxidant and antioxidant. In preeclamptic patients, pro-oxidant decrease enzymatically in cell, followed by increase in lipid peroxida due to free radicals malondialdehyde (MDA). Furthermore, increase lipid peroxida leads to disturbance in cell integrity, uncontrolled lipid peroxida both in cell and tissues that promotes lysis of cell membrane and edhotels, vascular reactivation, and increase vascular permeability due to activation of neutrophil estalase. Neutrophil estalase can be used as marker in endothelial dysfunction on molecular level. In preeclampsia, MDA is present in plasma, small blood vessels, and desidua basalis.12, 13
Elevated free radicals in preeclampsia is associated with reduced cellular antioxidants. In the present study, curcumin decreased MDA level in preeclampsia-induced cell. This result is supported by previous study that curcumin significantly attenuated MDA level and recovered the GSH and SOD levels.22 Curcumin is known to protect biomembranes against peroxidative damage. Peroxidation of lipids is known to be a free-radical-mediated chain reaction, leading to the damage of the cell membranes, and the inhibition of peroxidation by curcumin is mainly attributed to the scavenging of the reactive free radicals involved in the peroxidation. Most of the antioxidants have either a phenolic functional group or a -diketone group. Curcumin is an unique antioxidant, which contains a variety of functional groups, including the B-diketo group, carbon–carbon double bonds, and phenyl rings containing varying amounts of hydroxyl and methoxy substituents. 23
Curcumin has been studied recently in treatment of preeclampsia. Curcumin treatment in various doses could decrease significantly pro-inflammatory cytokines levels in preeclamptic plasma-induced. After curcumin treatment, there was decreased level of nuclear NF-kB p50 and increased level of PPAR- significantly.24 In other study, LPS-curcumin-treated group had decreased blood pressure and urinary protein level, which was comparable to control group, as well as improved trophoblast invasion and spiral artery remodeling induced by LPS. Increased TLR4, NF-kB and IL-6, MCP-1 protein expressions in LPS-treated group were significantly decreased after curcumin administration.25
References
1. Duley L. The global impact of pre-eclampsia and eclampsia, Seminars in Perinatology, 2009;33(3):130-7.
2. Sibai B, Dekker G, Kupferminc M. Pre-eclampsia, Lancet, 2005;365(9461):785-99.
3. Baumwell S, Karumanchi SA. Pre-eclampsia: clinical manifestations and molecular mechanisms,. Nephron Clinical Practice, 2007;106(2):c72-81.
4. Tenhola S, Rahiala E, Halonen P, Vanninen E, Voutilainen R. Maternal preeclampsia predicts elevated blood pressure in 12-year-old children: evaluation by ambulatory blood pressure monitoring, Pediatric Research, 2006; 59(2):320-4.
5. Myatt L. Role of placenta in preeclampsia, Endocrine, 2002;19(1):103-11.
6. Huppertz B. Placental origins of preeclampsia: challenging the current hypothesis, Hypertension, 2008;51(4):970-5.
7. Broughton Pipkin F, Roberts JM. Hypertension in pregnancy, Journal of Human Hypertension, 2000; 14(10-11):705-24.
8. Levine RJ, Lam C, Qian C, Yu KF, Maynard SE, Sachs BP, Soluble endoglin and other circulating antiangiogenic factors in preeclampsia, New England Journal of Medicine, 2006;355(10):992-1005.
9. Lu F, Longo M, Tamayo E, Maner W, Al-Hendy A, Anderson GD, et al. The effect of over-expression of sFlt-1 on blood pressure and the occurrence of other manifestations of preeclampsia in unrestrained conscious pregnant mice, American Journal of Obstetrics and Gynecology, 2007;196(4):396 e1-7;
Pathogenesis of preeclampsia remains unclear. Placenta is believed to play the key role.5 Placenta is a source and central of any mediators in preeclampsia pathogenesis.6,7 Morever, placenta is also responsible in preeclampsia development, in which there is disturbance in placentation, poor invasion, and abnormal angiogenesis which is the main pathological manifestation.7 These events are results from oxidative stress found in placenta in preeclampsia. Studies show that soluble vascular endothelial growth factor receptor-1 (VEGFR-1) or known as soluble fms-like tyrosine kinase-1 (sFlt-1) and soluble endoglin (sEng), are mediator mechanism between ischemic placenta and oxidative stress with failure in local angiogenesis8,9, that leads to systemic vascular dysfunction.10 Oxidative strees can also stimulate release of cytokines, antiangiogenics, microparticles and other essential molecules in preeclampsia.11 Antiangiogenic plays role in early development of placental vascular and during trophoblast invasion, and hypoxia is the major regulator on its expression. 10 Previous studies show that oxidative stress induced sflt-1 concentraion. 11Elevated sflt-1 caused reduction in placental growth factor (PIGF) and vascular endothelial growth (VEG) signaling.
Oxidative stress is caused by imbalance of pro-oxidant and antioxidant. In preeclamptic patients, pro-oxidant decrease enzymatically in cell, followed by increase in lipid peroxida due to free radicals malondialdehyde (MDA). Furthermore, increase lipid peroxida leads to disturbance in cell integrity, uncontrolled lipid peroxida both in cell and tissues that promotes lysis of cell membrane and edhotels, vascular reactivation, and increase vascular permeability due to activation of neutrophil estalase. Neutrophil estalase can be used as marker in endothelial dysfunction on molecular level. In preeclampsia, MDA is present in plasma, small blood vessels, and desidua basalis.12,13
Elevated free radicals in preeclampsia is associated with reduced cellular antioxidants. Curcumin is a compound found in Curcuma longa.14 Curcumin has been known to posses many biological activities, such as antiinflamation and antioxidants.15 Thus, curcumin is considered as a potent approach in treating oxidative stress and inflammation-related diseases.15, 16Hansson, Chen17 proposed that curcumin can be utilized as alternative therapy for preeclampsia due to its ability in inhibiting regulator protein, as well as NFκB. Recent study found that curcumin reduced MDA level, as well as NFκB expression in preeclamptic rats.18 In the present study, we observed the effects of curcumin on MDA level in preeclampia-induced HUVEC cell line.
Materials and Method
Samples used were women at 32-42 weeks of gestational age which were diagnosed preeclampsia and normal pregnancy at the same gestational age from Dr. Hasan Sadikin General Hospital, Indonesia. Research subjects has fulfilled inclusion and exclusion criteria.
Cell culture
HUVEC cell line was grown in tissue culture flask (25 cm2) containing RPMI 1640 supplemented with 10% (v/v) FBS qualified (fetal bovine serum), antibiotic-anti-micotic 1% Penicillin G-Streptomycin Solution Stabilised dan (1% Fungizone Amphotericin B) and 1% gentamisin, then incubated at 37oC atmosphere 5% (v/v) CO2. Medium was replaced 2-3 times a day. Cell were the checked every 7 days or untill 90%. confluent 131 Cells viability was measured with trypan blue on haemocytometer under light microscope with 400x magnification.19-21
Measurement of MDA level
Cells of 6x105 cell/ml induced with both normal and preeclampsia serum, were placed into 96-well plate, then incubated at 370C 5% CO2 (v/v). Furthermore, each wells were washed 3-4 times with PBS 370C. Various concentrations of curcumin (0; 0,977; 1,953; 3,906; 7,813; 15,625; 31,25; 62,5; 125; 250 μg/ml) were then distributed to each well, incubated for 24 and 48 hours 370C 5% CO2 (v/v). Each well was washed with PBS pH 7,4 once for 5 minute, and then centrifuged for 20 minute at 3.000 rpm. Supernatants were carried for measurement of MDA level with Thiobarbituric Acid-reactive Substances (TBARS). Cells were given solution containing 15% w/v trichloroacetic acid, 0,375 w/v thiobarbituric acid, 0.25% hydrichloric acid and 0.2% triton X. Samples were then suspended with heating at 1000C for 15 minutes, then centrifuged at 4500 rpm for 10 min. Absorbance was read at 532 nm wavelength.20,21
Data analysis
Data was analyzed with T-test if normally distributed, and Mann Whitney test if not normally distributed. Data was quantitavely analyzed with ANOVA DMRT (Duncans’s Multiple Range Test) to determine the significance among variables in each treatment SPSS 14.
Results and Discussion
MDA levels
Figure 1. Effects of curcumin in various concentrations and incubation time on level of MDA
Figure 2. Ratio of MDA levels in preeclampsia-induced HUVEC based on incubation times and curcumin concentration
As shown in Figure 1, MDA level and treatment of curcumin in control and cell, was the highest among treatments. Treatment of curcumin showed effect both in normal and preeclampsia serum after 24 h nd 48 h incubation.
Effect of Curcumin in Decreasing MDA Level in
Preeclampsia-Induced Human Umbilical Vein Endothelial Cell (HUVEC)
Cut Meurah Yeni1*, Prima Nanda Fauziah2, Ani Melani Maskoen3,
Rovina Ruslami4, Johannes Mose5
1Department of Obstetric and Gynecology, Faculty of Medicine, Universitas Syiah Kuala, Banda Aceh, Indonesia
2Department of Medical Laboratory Technology, School of Health Sciences Jenderal Achmad Yani, Cimahi, Indonesia
3Department of Oral Biology, Faculty of Dentistry, Universitas Padjadjaran, Bandung, Indonesia
4Department of Pharmacology and Therapy, Faculty of Medicine, Universitas Padjadjaran, Bandung, Indonesia
5Department of Obstetric and Gynecology, Faculty of Medicine, Universitas Padjadjaran, Bandung, Indonesia
Abstract : Preeclampsia is multisystem disease occurred in 3-8% pregnancy, indicated by hypertension and proteinuria after 20 weeks of gestational age. In preeclamptic patients, pro-oxidant decrease enzymatically in cell, followed by increase in lipid peroxida due to free radicals malondialdehyde (MDA). Elevated free radicals in preeclampsia is associated with reduced cellular antioxidants. Curcumin has been known to posses many biological activities, such as antiinflamation and antioxidants. We evaluated effects of curcumin on MDA level in preeclampia-induced HUVEC cell line. In the present study, we observed the effects of curcumin on MDA level in preeclampia-induced HUVEC cell line. MDA level was measured with Thiobarbituric Acid-reactive Substances (TBARS). The result of the present study showed curcumin decreased MDA level in preeclampsia-induced cell.
Key words : curcumin, MDA, preeclampsia.
Introduction
Preeclampsia is multisystem disease occurred in 3-8% pregnancy, indicated by hypertension and proteinuria after 20 weeks of gestational age.1, 2 Preeclampsia causes both martenal and perinatal morbidity and mortality worldwide.1 Clinicial manifestasion of preeclampsia consists of hyperkoagulopati, hemolysis elevated liver enzyme and low platelets (HELLP), periportal hemorrhage in liver, ischemic lesion, fibrin cumualtion, subcapsular hemorrhage and intrahepatic hematoma or even hepatic rupture, acute renal failure and segmental glomeruloskeloris segmental.1-3 Moreover, preeclampsia causes cramp (eclampsia), headache, blurry vision, scotoma or even blidness, cerebral hemorrhage, brain damage due to ischemia or microinfark and fibrinoid necrosis in brain.2,3
International Journal of PharmTech Research, Vol.10, No.2, pp 69-73 (2017)
http://dx.doi.org/10.20902/IJPTR.2017.10110