Cholesterol profile due to granting of the rice bran types in rats
with diet used cooking oil
Eny Hartadiyati Wasikin Ha* Retno Sri Iswarib
aBiology Education Department, University of PGRI Semarang,
Jl. Lingga No. 1, Dr. Cipto-Semarang 50125, Indonesia
bBiology Department, Semarang State University, Kampus Sekaran Gunung Pati,
Semarang 50229, Indonesia
Abstract
The used cooking oil that heated repeatedly can contains free radicals. Free radicals can damage cellular biomolecules so that cholesterol levels can increase in blood serum. Rice bran contains anthocyanin as an antioxidant and antioxidants act as free radical scavengers . The aim of this study is to determine the influence of rice bran types on cholesterol profile with used cooking oil. This study used Posttest Only Control Group Design, 16 spraque dawley rats were divided into control group and added with used cooking oil but not added with rice bran. Otherwise, three experimental groups are added with used cooking oil and three types of rice bran respectively. The results showed there was a significant effect (p <0.05) of LDL cholesterol levels, but there was not significant effect (p> 0.05) of total cholesterol and HDL cholesterol levels. As the result black rice bran was able to provide the ratio of LDL / HDL better.
Keywords cholesterol profile, rice bran, used cooking oil
1. Introduction
One reason why dangerous to consume cooking oil that has been used many times is the presence of trans fatty acids (Sartika, 2008) and free radicals (Leong et al., 2008; Adam et al., 2009). Free radicals can cause oxidative stress (Nazrun et al., 2007; Dhibi, 2011), which turn into damage to cellular biomolecules body (Halliwell et al., 2007). Oxidative stress that continuously can cause a disease those are chronic and degenerative (Dorffman et al., 2009). One of degenerative disease caused by used cooking oil is atherosclerosis. Atherosclerosis is characterized by high serum cholesterol levels (Murray, 2009).
Antioxidants are protective agents that inactivate free radicals that can significantly minimize oxidative stress and eventually delay or prevent oxidative damage to biomolecules body (Hartadiyati, 2006).
The existence of endogenous antioxidant enzymes can provide a certain degree of protection against free radicals, but exogenous sources of dietary antioxidants is still an important need in minimizing oxidative stress. Presence of antioxidants: vitamins, phytochemicals such as carotenoids, isoflavones, saponins and polyphenols in fruits and vegetables have shown the ability to provide a protective effect.
Agricultural s litter such as rice bran contains antioxidants known as Vitamin E (tocopherols and tocotrienols), c-oryzanol and phenolic compounds (proanthocyanidins and anthocyanins in particular, especially in the red and black rice bran), then there are four compounds, including antioxidants such as cyanidin-3- galactoside , cyanidin-3- glukoside, cyanidin-3-rutinoside, and peonidin-3-glukoside (Min et al., 2014; Zhang, 2015). It also said that more anthocyanin and proanthocyanidin found in rice bran colored (Min et al., 2011). Empirically antioxidant group sterols (Hartadiyati, 2006) and phenol (Min et al., 2011; Min et al., 2014) are important compounds in the mechanism of free radical elimination. The role of red rice bran, black and white as hypocholesterolemia agent to minimize the impact of diet used cooking oil is an interesting issue to be studied.
Based on the above then research was done to analyze profile of cholesterol (total cholesterol, HDL cholesterol and LDL cholesterol) in blood serum of rats with diet used cooking oil after granting the types of rice bran (black rice bran, red rice bran and white rice bran); and also determine the types of rice bran that cause good cholesterol profile.
2. Materials and methods
This study uses Posttest Only Control Group Design. Sixteenmale sprague-dawley rats aged3monthsold (150-200 g)were divided randomly into 4groupsas follows :
P0 : control group were fed standard + 1.5 ml used cooking oil.
P1 : the treatment group were fed standard + 1.5 ml used cooking oil + 540 mg/head/day white rice bran
P2 : the treatment group were fed standard + 1.5 ml used cooking oil + 540 mg/head/day brown rice bran
P3 : the treatment group were fed standard + 1.5 ml used cooking oil + 540 mg/head/day black rice bran
The used cooking oil is made by heating 27 times of frying tofu. The rats were kept in individual cages and were treated for 21 days and feeding it ad libitum. Furthermore, the data retrieval cholesterol profile, consisting of total cholesterol, LDL cholesterol and HDL cholesterol.
3. Results and discussion
3.1 LDL cholesterol level
LDL cholesterol levels after granting the types of rice bran in rats blood serum with diet used cooking oil can be seen in Figure 1.
ANOVA statistical analysis showed significant differences in average LDL cholesterol levels between groups, F = 5.683 ; Sig = 0.012. Furthermore, a further test using LSD are shown in Table 1. Table 1 shows that all experiment groups were different significantly from the control group, but among the treatment groups were not different significantly.
Table 1.LSD test LDL cholesterol levels after granting the types of rice bran in rats blood serum with diet used cooking oil.
The granting of this type of white, red and black rice bran shows there is difference significantly (p < 0.05) to blood serum cholesterol LDL levels with diet used cooking oil. With a LSD test shows all granting type of rice bran (white, red and black) cause LDL cholesterol levels different significantly (p <0.05) with the controls (without giving rice bran), the results of the study also confirmed that the levels of LDL all treatment groups was significantly lower (p <0.05) than control .This is because the role of antioxidants in rice bran either white, red or black through a protection mechanism of oxidation by free radicals found in used cooking oil. Antioxidants protect cellular biomolecules including plasma lipoprotein thus protecting the LDL and HDL cholesterol in the blood plasma from free radical oxidation.
On the other hand lipoprotein liver cell membrane is also protected so that causing the LDL receptors can work well and accelerate the pace of entry of LDL into liver cells and affect low levels of LDL cholesterol in the blood plasma, further LDL will undergo metabolism. Endocytosis initiated from the binding of Apo B-100 to LDL receptors on the surface of the peripheral cell membrane, thus cell membranes form cytoplasmic vesicles by undergo invaginasi. Vesicles then fuse with lysosomes. LDL contained therein are degraded by lysosomal enzymes. Solving LDL in lysosomes include translocation apoproteins and cholesterol esters. LDL receptor is not destroyed but returned to the cell surface. Inflows cholesterol inhibits the activity of HMG CoA reductase, and stimulates the activity of LCAT (Murray, 2009). This lowers the cholesterol biosynthesis.
Further described by Min et al. (2011) that antioxidant indicators such as total phenol, total flavonoids, antioxidant capacity and radical scavanging correlated with the intensity of bran colors, but all three treatment types of rice bran (white, red and black) in mice with used cooking oil diet showed no significant difference (p> 0.05). This is likely due to the amount (g/kg body weight of mice) and duration of provision have not been able to significantly differentiate levels of LDL as a result of the provision of rice bran white, red and black.
3.2 HDL cholesterol level
Figure 2.Levels of HDL cholesterol after granting the types of rice bran in rats blood serum with diet used cooking oil.
HDL plays a role in reverse cholesterol transport process from extrahepatic tissues to the liver, cholesterol is known to be found in extrahepatic tissues via endocytosis LDL through the LDL receptor or via metabolism of VLDL. Excess of received cholesterol by extra hepatic tissues can move through the cell membrane to the recipient (acceptor) which is in fluid periseluler. Cholesterol acceptor has the form of small particles that contain a small amount of lipid and contain a lot of apo AI and phospholipid called pre- HDL synthesized in the liver cells. Lecithin-cholesterol acyltransferase (LCAT) reacts with these particles form an ester cholesterol and increase HDL particle size, called HDL. Large HDL particles can capture particles such as apo E. Furthermore, this particle "cleaned up" by the two receptors (LDL receptors) on liver cells (Murray, 2009; Asztalos, 2011).
As described LDL levels: antioxidants found in white rice bran, red and black will protect plasma lipoproteins, including HDL and lipoprotein membrane so that the above explains why the results showed HDL levels in all treatment groups was higher than the control group although statistically did not differ significantly (p >0.05). This is possible because the amount (g/kg body weight of rats) and duration of provition have not been able to significantly differentiate levels of LDL.
3.3 Total cholesterol level
Total cholesterol levels after granting the types of rice bran on blood serum of rats with diet used cooking oil can be seen in Figure 3. Further statistical analysis ANOVA showed there was not significant difference in average cholesterol levels between groups, F = 0.836; p = 0.5.
LDL levels due to the provision of white, red and black rice bran showed significant differences with the controls, but did not differ significantly among treatment groups as well as the levels of HDL. This cause total cholesterol levels did not differ significantly (p > 0.05).
3.4 Ratio LDL/HDL
To ascertain potential of rice bran types clearly carried by descriptions ratio of LDL/HDL, can be seen in Figure 4. The results showed black rice bran has a better effect on the appeal of red and white rice bran, based on the ratio of LDL/HDL, but have not been able to cause HDL levels higher than LDL. It is known that the ratio between LDL cholesterol and HDL cholesterol is very important as risk parameter of heart disease (Murray, 2009).
Figure 4.The ratio of LDL/HDL after granting the types of rice bran in rats blood serum with diet used cooking oil.
When viewed from the levels of anthocyanins as antioxidants, black rice bran contains more antioxidants than white rice bran and red rice bran. Antioxidants are phytochemicals that can minimize the oxidation that cause damage to important cellular biomolecules body, thus protecting the body's cellular biomolecules. Anthocyanin pigment have been reported to be highly effective in reducting cholesterol levels in the human body (Lee et al., 2008). This is as shown in the results of research that steeping of black rice bran effective as hepatoprotector (Hou et al., 2013) is certainly no exception for LDL receptors, thus affecting the levels of LDL, HDL and total cholesterol blood serum.
4. Conclusion and remarks
Granting the types of rice bran give positive effect on serum cholesterol profile spraque dawley rats with used cooking oil diet, based on:
There is significant difference (p < 0.05) of LDL cholesterol levels as a result of the provision of the types of rice bran in rats with diet used cooking oil, but between treatment groups (giving white rice bran, red and black) did not differ significantly (p > 0.05).
There is no significant difference (p > 0.05) between total cholesterol and HDL cholesterol levels as a result of the provision of the types of rice bran in rats with diet used cooking oil.
The black rice bran gives better effect than red and white rice bran, based on the ratio of LDL / HDL, but have not been able to cause HDL levels higher than LDL.
Suggestions from this study are to do more research on:
Granting of bran more than 21 days to see the potential of black rice bran until able to show HDL cholesterol is higher than LDL cholesterol.
The used cooking oil for frying other foods.
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