BEANS (PHASEOLUS VULGARIS L) EXTRACT AS ANTI DYSLIPIDEMIA: Decrease of total cholesterol, malondialdehyde, low density lipoprotein, and increase of high
density lipoprotein on rat wistar Sri Wahjuni
Chemistry Departement, Faculty of Math and Science Udayana University
ABSTRACT
Excessive fat consumption leads to increase of blood cholesterol level more than normal condition which is called as Dyslipidemia. Dyslipidemia is an abnormal lipoprotein metabolism, usually associated with lipoprotein over production or deficiency. Dyslipidemia is also often described as an event of hyperlipidemia and also as a risk factor for cardiovascular disease.
This is an experimental study for accessing the ability of beans extract as an anti dislipidemia. The anti dislipidemia were marked by decrease of total cholesterol, malondialdehid, Low Densyty Lipoprotein levels, and increase HDL of Wistar rat induced dislipisemia with high cholesterol diet for 16 week. This study applied randomized posttest only control group design. The samples were 24 Wistar Rat, randomized into 6 treatment: negative control group diet standard (Treatment group 1), positive control group with diet high cholesterol (Treatment cholesterol group 2), diet high cholesterol with beans ( Phaseolus vulgaris) 50 kg/bw (Treatment group 3),diet high cholesterol with beans (Phaseolus vulgaris L) Steenis) 100 kg/bw (Tretment group 4), diet high cholesterol with beans (Phaseolus vulgaris L) 150 kg/bw (Treatment group 5). diet drug simvastatin 0,18 mg/day /200 gram BB (Treatment group 6), After 16 weeks treatment, blood of rats were driven for total cholesterol and MDA assays. LDL and HDL . All of data analyzed by Anova to obtain the treatment different toward control by statistically with significance at α=0.05.
The result shows that extracts of beans (Phaseolus vulgaris L) in a dose of 150 mg/kg bw decrease total cholesterol of 23.88 %, MDA of 70.60 % LDL 38.09 % also increase the HDL of 59,63%, Extract beans (Phaseolus vulgaris L) has an ability to prevent cardiovascular disease. This is caused by the present of phytosterol in the beans extract which has been phyto-chemistry tested and analyzed by GC-MS such as stigmasterol substance.
Keywords: beans ( Phaseolus vulgaris L), dyslipidemia, Phytosterol, total cholesterol, LDL,HDL,MDA
INTRODUCTION
Nowdays many Indonesian people have the wrong diet, tend to like fast food that contains nutrients that are not balanced. In general, this fast food containing saturated fat and high salt with low fiber content. Excessive consumption of saturated fat, low carbohydrate, and less fiber from the dailydietisafactorcausingincreasedbloodcholesterol.High blood cholesterol (dyslipidemia) is not only experienced by obese people, but it can also occur in people who are thin and can afflict people who are young. Various walks of life, have to try to live a healthy lifestyle in order to keep cholesterol in the blood remained normal. In the body there is a normal cholesterol level is 160-200 mg (LIPI, 2009). Dilipidemia is a situation where an increase in blood cholesterol levels that exceed normal circumstances, accompanied by increased levels of total cholesterol, triglycerides, and LDL
cholesterol. Dilipidemia occur when total cholesterol concentration reaches ≥ 240 mg / dl, LDL ≥ 160 mg / dl, and triglycerides ≥ 150 mg / dl (Montgomery, 1993)
there is a normal cholesterol level is 160-200 mg (LIPI, 2009). Dilipidemia is a situation where an increase in blood cholesterol levels that exceed normal circumstances, accompanied by increased levels of total-cholesterol, triglycerides,and LDLcholesterol.Fat derived from food will undergo pencernaaan processes in the intestine into free fatty acids, triglycerides, phospholipids and cholesterol, then absorbed into the form of chylomicrons. Breakdown of chylomicrons circulate to the rest of the liver and separated into cholesterol. Most cholesterol is discharged into the bile as bile acids and in part again together with triglycerides to be allied with a particular protein (apoprotein) and form very low density lipoprotein (VLDL). VLDL very low density lipoproteins are further broken down by the enzyme lipoprotein LDL can not be the last 2-6 hours and immediately be converted to LDL. This process is called mechanism of dyslipidemia (Soeharto, 2004). . Demage Lipids in LDL-cholesterol in the blood due to dyslipidemia produce various
products of decomposition are relatively stable, especially aldehyde reactive α, β-unsaturated, such as malondialdehyde (MDA), 4-hydroxy-2-nonenal (HNE), heksanal, and 2-propenal (acrolein) (Uchida, 2003 Carani.et.al, 2004). Malodialdehid a (marker) one of the products of decomposition of per oksidadasi acids plural unsaturated fats (polyunsaturated fatty acids; PUFA).
Several studies medicinal plants tadisional that have been carried out include research conducted by Ratni (2013), gedi (Abelmoschus manihot) shows the effect of hypolipidemic that can lower
cholesterol levels in male rats Wistar with the content of flavonoids and steroid in gedi
(Abelmoschus manihot) , Utariningsih (2007) determines the hair decoction of maize (Zea mays) is effective in lowering cholesterol levels of mice, rats decreased levels of cholesterol using -sitosterol compounds contained in the hair of corn (Zea mays).
According to experts the researchers herbal, black cumin (Nigella sativa) play a role in reducing the excess cholesterol in the blood by active substance thymoquinone, omega 9, omega 3, omega 6, saponins and phytosterols (Edi Junaedi, 2011).Subekti (2006) using leaf katuk (Sauropus adrogynus L. Merr) to produce low cholesterol Japanese quail. Leaves katuk (Sauropus adrogynus L. Merr) phytosterol-containing compounds that can lower cholesterol in Japanese quail products. Wahyu (2013), using n-hexane extract of pomegranate fruit flesh is white (Punica granatum) which can lower blood cholesterol levels in rats (Rattus norvegicus L.) with a content of phytosterols in the flesh white pomegranate (Punica granatum). Based on research conducted by Jannah, et. al. (2013), the fruit extract of beans (Phaseolus vulgaris L.) contain phytosterol compounds. Phytosterols are natural plant sterols that contains a minimum of 70% -sitosterol. According Bonsdoff-Nikander (2005), a compound phytosterols can lower cholesterol through competition between cholesterol and phytosterols in a mixture of micelles, as well as by binding cholesterol in the digestive tract. There has been no research on the effectiveness of the test pieces of bean (Phaseolus vulgaris L.) in lowering cholesterol levels, so as to investigate the ability of ethanol extract of the fruit of beans (Phaseolus vulgaris L.) as an anti-hypercholesterolemia through reductions in total cholesterol, Low Density Lipoprotein (LDL) as well as an increase High Density Lipoprotein (HDL) in Wistar rats dyslipidemia.
RESULT AND DISCUSSION
Figure 1 Chromatograms of ethanol extractnof beans (Phaseolus vulgaris L.)
Table1. Substance detected in ethanol extract of beans
Peak Conpound tR m/z %area
1 Propane, 1-(1-ethoxydthoxy)-(CAS) 1-Ethoxy 3,07 45,00 6,63%
2 Ethanol, 2,2-diethoxy- 3,25 47,00 37,24%
3 Ethane, 1,1-diethoxy-(CAS) 1,1-Diethoxyethane 3,40 45,00 22,92%
4 Butane, 1,1-diethoxy-3-methyl- 3,57 103,00 2,02%
5 1-ethoxy-1-pentoxy-ethane 3,83 73,00 2,68%
6 1,1,3-Triethoxybutane 5,67 73,00 1,21%
7 2-Pentanone, 5,5-diethoxy- 6,49 103,00 1,43%
8 Ethanol, 2,2-diethoxy-(CAS) Glycolaldehyde 6,93 103,00 2,45% 9 1(2H)-Pyridinecarboxaldehyde, 3,4-dihydro-5-(2-piperidinyl) 10,83 110,00 0,78%
10 Propanoic acid, 2-methyl-,
1-(1,1-dimethylethy)-2-methyl-1,3-propanediyl ester (CAS) 12,38 71,00 1,96%
11 1-(+)-Ascorbic acid 2,6-dihexadecanoate 16,41 73,00 5,16% 12 Hexadecanoic acid, ethyl ester (CAS) Ethyl palmitate 16,75 88,00 2,03% 13 9,12-Octadecadienoic acid (Z,Z)- 18,05 67,00 4,88% 14 2H-Pyran, 2-(2-heptadecynyloxy)tetrahydro-(CAS) 18,11 55,00 3,99%
15 Octadecanoic acid 18,32 73,00 1,78%
16 9,12,15-Octadecatrienoic acid, ethyl ester, (Z,Z,Z)- 18,38 79,00 0,56% 17 9-Octadecenamide, (Z)-(CAS) OLEOAMIDE 20,06 59,00 0,61%
18 Hexadecanoic acid, 2-hydroxy-1-(hydrocymethyl)ethyl ester
(CAS) 21,29 98,00 0,67%
19 Stigmasterol 28,89 55,00 0,96%
In the ethanol extract of the fruit of beans (Phaseolus vulgaris L.) contain phytosterol
Gambar 2. Spektra Massa Stigmasterol
Table 2Pattern beheading
No m/z The possibility that the missing fragment
1. 412 M+ -
2. 394 M+-18 -H2O 3. 351 (M+-18)-43 -C3H7
Figure 3. Fragmentation stigmasterol
DYSLIPIDEMIA
Decrease of total Cholesterol level
Measurement of total cholesterol was conducted to determine the amount of cholesterol in the body. Results of the determination of total cholesterol shown in Table 3.
Table 3 Table average level of total cholesterol
Group Average level of total-cholest (mg/dL) Persentation decrease
Figure 4. Average level of total cholesterol
Description:
K1 = negative control group; K2 = positive control group P1 = group of ethanol extract of beans fruit 50 mg / kg bw
P2 = group of ethanol extract of fruit bean dose of 100 mg / kg bw P3 = group of ethanol extract of fruit bean dose 150 mg / kg bw KS = group of medication simvastatin dose of 0.18 mg / kg bw
Results of this study showed total cholesterol levels in the blood of Wistar rats
hypercholesterolemia decreased affected by the ethanol extract of the fruit of beans (Phaseolus vulgaris L.) with dose variation is a dose of 50 mg / kg BW, 100 mg / kg bw and 150 mg / Kg BB.
Tukey test results / HSD showed that the difference in total cholesterol levels were significantly occurred in the control group and all treatment groups with the value of the most significant differences in the treatment group of fruit ethanol extract of beans (Phaseolus vulgaris L.) with a dose of 150 mg / kg bw. This shows that the ethanol extract of the fruit of beans (Phaseolus vulgaris L.) with dose variation has an influence in the determination of total cholesterol Wistar
This is due to the content of phytosterols in fruit ethanol extract of beans (Phaseolus vulgaris L.) were detected as having significant influence stigmasterol.Antidislipidemia effect of
phytosterols depends on the amount of plant sterols and stanols that are used by beberapan doses (Jones, 2000). This research has the best results for lowering total cholesterol levels at doses of ethanol extract of fruit bean (Phaseolus vulgaris L.) 150 mg / kg bw. Drug simvastatin is used to lower the total cholesterol level has a value almost equal to the reduction in the ethanol extract of the fruit of beans (Phaseolus vulgaris L.) dose of 50 mg / Kg. Tangible results are also seen in the levels of total cholesterol medication simvastatin group approached the values of total cholesterol ethanol extract of the fruit group snaps dose 150 mg / kg bw.
Decrease Low Density Lipoprotein (LDL)
Measurement of Low Density Lipoprotein cholesterol (LDL) is performed to determine the risk of disease atherosclerosis. Atherosclerosis is a disease of blood vessel constriction by too much cholesterol amounts in the blood that form deposits on the walls of blood vessels (Almatsier, 2004). Cholesterol forming plaque in the blood vessel wall is cholesterol Low Density Lipoprotein (LDL). This is due to atherogenic LDL cholesterol has properties that memudahkanya attached to the inner wall of blood vessels (Heslet, 1996). LDL cholesterol is one of the causes of hypercholesterolemia. Eating a high fat can cause the concentration of cholesterol in the body increases and decreases the synthesis and activity of LDL receptors, so that the levels of LDL cholesterol in the body is
increased (Murray, 2006). LDL cholesterol determination results are shown in Table 4
Table 4 Table Average of LDL-cholesterol
Tukey test results / HSD in table4 above showed significant differences in levels of LDL occurs in controk group and all treatment groups with significant value palig difference in the treatment group of ethanol extract of beans (Phaseolus vulgaris L.) with a dose of 150 mg / kg bw. This shows that the ethanol extract of the fruit of beans (Phaseolus vulgaris L.) with dose variation has an influence in determining the levels of LDL Hypercholesterolemia Wistar rats. Decreased levels of LDL Wistar rats caused by the ethanol extract of the fruit of beans (Phaseolus vulgaris L.) contains phytosterols such as stigmasterol compounds that can lower blood cholesterol levels by inhibiting HMG CoA reductase will decrease the synthesis of cholesterol LDL (Price, 2006). In addition phytosterol is a sterol group also has antioxidant properties (Wang, 2002), which acts as a reducing LDL in the body (Radhika, 2011). Antioxidants can increase acid secretion empendu (Lamson, 2000).
In the heart, phytosterols and cholesterol that is secreted into bile by the transporter ABCG5 and ABCG8. The ability of phytosterols faster experience into the digestive secretion of the cholesterol, so more phytosterol enters the blood circulation and prevents the buildup of LDL cholesterol back to the blood circulation (Bonsdoff-Nikander, 2005). Phytosterols can lower LDL cholesterol levels the most effective at a dose of 150 mg / kg bw. Decrease in LDL cholesterol levels are influenced by a decrease in total cholesterol levels, as seen in total cholesterol levels in this study experienced a decrease as well as LDL cholesterol levels
Increase Kolesterol High Density Lipoprotein (HDL)
Measurement of High Density Lipoprotein cholesterol (HDL) was conducted to determine the transport of excess cholesterol in extrahepatic tissue and cell cleanser (scavenger cells) that will be brought back to the heart. In the extrahepatic tissues and cell cleanser (scavenger cells), HDL cholesterol will interact with the enzyme lecithin cholesterol acyl transferase release remnant and VLDL cholesterol to the liver and then excreted into the bile (Bahri, 2004).
HDL cholesterol levels in the blood are high very beneficial in reducing the risk of atherosclerosis, because the function of HDL cholesterol transports cholesterol from extrahepatic tissues to the liver so menncegah occurrence of calcification (Hartayo 2008). HDL cholesterol assay results are shown in Table 5.
Figure 4. Average level of HDL-cholesterol
HDL cholesterol levels in Table 5 shows that the Wistar rat blood hypercholesterolemia increased influenced oleh.pemberian fruit ethanol extract of beans (Phaseolus vulgaris L.) with dose variation is a dose of 50 mg / kg BW, 100 mg / kg bw and 150 mg / kg. Analysis of the results followed by one-way ANOVA test and Post Hoc Study by Tukey's test / HSD to mengaetahui group that has the same effect or different from the others. ANOVA analysis results which show the value of P <0.05 so H1 H¬0 accepted and rejected, which means there is a significant difference in HDL levels kelommpok control hypercholesterolemia with treatment group after administration of ethanol extract of the fruit of beans (Phaseolus vulgaris L.) with dose 50 mg / kg BW, 100 mg / kg BW, 150 mg / kg body weight and drug delivery simvastatin at a dose of 0.18 mg / kg bw. Tukey test results / HSD showed that significant differences in levels of HDL occurs in the control group and all treatment groups with the value of the most significant differences in the treatment group of fruit ethanol extract of beans (Phaseolus vulgaris L.) with a dose of 150 mg / kg bw. This shows that the ethanol extract of the fruit of beans (Phaseolus vulgaris L), with dose variation has an influence in the determination of HDL levels Wistar rat Hypercholesterolemia. In the group of dyslipidemia Wistar rats were given ethanol extract fruit perlakuaan beans (Phaseolus vulgaris L.) with a dose of 150 mg / Kg and perlakuaan medication simvastatin at a dose of 0.18 mg / kg bw overdosed, because it had higher levels of HDL cholesterol on average exceeded HDL cholesterol level of negative control group. This is due to the rich content of phytosterols in fruit ethanol extract of beans (Phaseolus vulgaris L.) that increase HDL cholesterol levels. HDL cholesterol levels increased significantly had the best results at a dose of 150mg/kgbw.
Decrease Malondialdehid level (MDA)
Tabel 6. Average of Malondialdehid
Group Average of malondialdehid (μM) Persentation decrease test results followed by one way ANOVA and Post Hoc Study by Tukey's test / HSD to
mengaetahui group that has the same effect or different from the others. ANOVA analysis results which show the value of P <0.05 so H1 H¬0 accepted and rejected, which means there is a
significant difference in HDL levels kelommpok control hypercholesterolemia with treatment group after administration of ethanol extract of the fruit of beans (Phaseolus vulgaris L.) with dose 50 mg / kg BW, 100 mg / kg BW, 150 mg / kg body weight and drug delivery simvastatin at a dose of 0.18 mg / kg Bw. Decrease in MDA levels in the blood may prevent decrease in membrane fluidity and cell damage. Based on the research that has been conducted in animals suggest that eating
unsaturated fatty acids cause a buildup chain unsaturated fatty acids in the body. Increased toxic products from the synthesis of unsaturated fatty acids (lipid peroxidation) generating
malondialdehyde.
CONCLUSSION AND FUTURE WORK Conclussions
1. Consumtion of the ethanol extract of the fruit of beans (Phaseolus vulgaris L.) can reduce total cholesterol in dyslipidemia Wistar rats at a dose of 150 mg / kg BW of 23.88%.
2. Consumtion of the ethanol extract of the fruit of beans (Phaseolus vulgaris L.) can reduce levels of Low Density Lipoprotein (LDL) on dyslipidemia Wistar rats at a dose of 150 mg / kg BW 38.09%.
4. extract the ethanol extract of the fruit of beans (Phaseolus vulgaris L.) can decrease
malondialdehyde in dyslipidemia Wistar rats at a dose of 150 mg / BW amounted to 70.60%. 5. The ethanol extract pieces of bean (Phaseolus vulgaris L.) contain phytosterol compounds based
analysis Gas Chromatography - Mass Spectrometry (GC-MS)
Future Work
The results obtained have suggested a few things, namely:
1. Need to do more research regarding the proper dosage to consume phytosterols which can lower blood cholesterol levels.
2. Further studies are needed to isolate and identify the compounds of phytosterols in fruit ethanol extract of beans (Phaseolus vulgaris L.) which can lower blood cholesterol
REFERENCES
Almatsier S. 2009. Prinsip dasar ilmu gizi. Jakarta: gramedia cetakan IX.
Bahri, A. 2004. Dislipidemia Sebagai Faktor Resiko Penyakit Jantung Koroner. e-USU Repositor.
Fakultas Kedokteran Universitas Sumatera Utara.
http://www.library.usu.ac.id/download/fk/gizibahri3. pdf. Diakses tanggal 17 mei 2015.
Bonsdorff-Nikander, A.V. 2005. Studies on Cholesterol-Lowering Microcrystalline Phytosterol Suspension in Oil. Helsinki : Faculty of Pharmacy of the University of Helsinki.
Carini.,M,.Aldini,Fcino,R.M.,2004. Mass Spectrometry for Detection of 4-hydroxy-trans-2-nonenal (HNE)
Adducts with peptides and Proteins [ Review].Mass.Spectrum Rev,23 :281-305.
Hartoyo, A., Dahrulsyah, N., Sripalupi dan Nugroho P. 2008. Pengaruh Fraksi Karbohidrat Kacang Komak (Lablab Purpureus (L) Sweet). Jurnal teknologi dan industri pangan, 19: 25-31.
Heslet, L. 1996. Kolesterol. Terjemahan Anton Adiwijoto. Jakarta : PT. Kesaint Blanc Indah.
Jannah, H., Sudarma, I.M., Andayani, Y. 2013. Analisis senyawa fitosterol dalam ekstrak buah buncis (Phaseolus vulgaris L.). Chem, Prog, Vol. 6, No.2.
Jones P.J., Raeini-Sarjaz M., Ntanios F.Y., Vanstone C.A., Feng J.Y., Parsons W.E. 2000. Modulation of plasma lipid levels and cholesterol kinetics by phytosterol versus phytostanol esters. J. Lipid Res., 2000, 41, 697–705.
Lamson., Davis., dan Matthew B. 2000. Antioxidants and cancer III. Quercetin, Alternative Medicine, Review Journal 5(3).
Montgomery, R. 1993. Biokimia Suatu Pendekatan Berorientasi Kasus. Yogyakarta: Gadjah Mada University Press.
Murray, R.K. 2006. Biokimia harper. Jakarta : buku kedokteran EGC.
Radhika, S., Smila K.H. dan Muthezhilan R. 2011. Antidiabetic and Hypolipidemic Activity of Punica granatum Linn on Alloxan Induced Rats. World Journal of Medical Sciences 6 (4): 178-182, 2011.
Ranti, G.C., Fatimawali, Wehantouw, F. 2013. Uji efektivitas ekstrak flavonoid dan steroid dari gedi (Abelmoschus manihot) sebagai anti obesitas dan hipolipedemik pada tikus putih jantan galur wistar. Jurnal Ilmiah Farmasi – UNSRAT, Vol.2 No. 02: 2302-249.
Soeharto, I. 2004. Jantung Koroner dan Serangan Jantung. Jakarta: Gramedia Pustaka Utama.
Subekti, S., Piliang, W.G., Manalu, W., Murdiati, T.B. 2006. Penggunaan Tepung Daun Katuk dan Ekstrak daun Katuk (Sauropus androgynus L. Merr) sebagai Substitusi Ransum yang Dapat Menghasilkan produk Puyuh Jepang Rendah Kolesterol. JITV Vol. 11 No.4 th 2006.
Uchida.,K.2003. 4-Hydroxy-2-nonenal a Product and Mediator of Oxidative Stress [ Review]. ProgLipid Res 42 : 318 – 43.
Utariningsih, D., Novita, W.R., Sari, R.P., Wati, E.M., Arifin, A.S., 2007. Dekok rambut jagung (Zea mays) efektif dalam menurunkan kadar kolesterol tikus putih (Rattus norvegicus). Malang : Universitas Muhammadiyah.
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