- Email: [email protected]
The antioxidant capacities and total phenol contents of some medicinal plants in Iran
S330
Abstracts
Keywords: Antioxidant activity, Pistacia atlantica, lipid peroxidation, superoxide dismutase, catalase doi:10.1016/j.clinbiochem.2011.08.812
Poster – [A-10-225-5] Acute and chronic toxicity of the jaft aqueous extracts a part of oak fruit in Iran Ali Mirzaei, Nooshin Mirzaei, Mahsa Mirzaei Medicinal Plant Research Center, Yasouj University of Medical Sciences, Yasouj, Iran E-mail addresses: [email protected] (A. Mirzaei), [email protected] (N. Mirzaei), [email protected] (M. Mirzaei) Introduction: Oak (Quercus species) Fruit has two covering external and internal layers that locally internal layer of fruit known as Jaft in some part of Iran. Materials and Methods: A study was conducted to investigate the safety of an aqueous extract of Jaft by acute toxicity methods. Acute toxicity (LD50) was estimated intraperitonal route (i.p.) and oral route (p.o.) in two separate groups. For the sub-chronic study, Twenty (20) adults Wistar rats (250–300 g) were randomly divided into four groups (I–IV) of 5 animals for each group. Animals were received Jaft- extract daily by gavages doses of 100, 500 and 1000 mg/kg for 28 days. At the end of experiment blood was collected for hematology and biochemical tests. Results: The result of acute toxicity values demonstrate that LD50 of the Jaft extract in the (i.p.) and (p.o.) procedures were > 3000 and > 5000 mg/kg respectively. All hematological parameters, in the experiment groups except (White Blood Cell) did not significantly differ compared to the control group. There were no statically significant observed in serum cholesterol, triglyceride, Blood Urea Nitrogen concentrations and alkaline phosphatase (ALP), glutamate pyruvate transaminase (GPT) and glutamate oxaloacetate transaminase (GOT) activities in all treatment groups compare to control group. No gross changes were found in the organs of rats following extract administration. Conclusion: These results suggest that the aqueous Jaft extract could be used with safety especially by oral route. Keywords: Acute toxicity, Chronic toxicity, Aqueous extract of jaft, Biochemical parameters, Hematological parameters doi:10.1016/j.clinbiochem.2011.08.813
Poster – [A-10-225-6] Antioxidant activities and hepatooprotectivity of Jaft a part of oak fruit Mehdi Akbartabar Tori, Ali Mirzaei, Mahsa Mirzaei, Nooshin Mirzaei Medicinal Plant Research Center, Yasouj University of Medical Sciences, Yasouj, Iran E-mail addresses: [email protected] (M.A. Tori), [email protected] (A. Mirzaei), [email protected] (M. Mirzaei), [email protected] (N. Mirzaei) Introduction: Aqueous extract of internal layer of oak fruit (Jaft) was evaluated for in vitro antioxidant activities and hepatoprotective in carbon tetrachloride induced liver damage in rats. Material and Methods: For in vitro determination of antioxidant activity used FRAP, DPPH, ABTS and PMB and for analysis of antioxidant components applied total phenol and flavonoids tests. For study of hepatoprotectivity thirty six male wistar rats were
chosen. Group (I, II and III) served as negative controls given olive oil, distilled water (D.W.) and jaft extract (500 mg/kg) respectively. Group(IV) as toxic group received CCl4 1 ml/kg, (i.p.) as single dose at every 72 h. The Protective groups (V) and (VI) one hour after the injection of carbon tetrachloride at every 72 h.received aqueous jaft extract (250 and 500 mg/kg) body weight orally respectively. Results: Jaft extract displayed significant antioxidant activity in different solvent systems as manifest by different (DPPH, ABTS, PMB and FRAP) assay systems. This study showed that in vitro efficiency was supported by a significant dose dependent hepatoprotection (at 250 mg/kg and 500 mg/kg dose) by decreasing the activity of serum enzymes and bilirubin (P < .01). CCl4 induced a significant rise in aspartate amino transferase (AST), alanine amino transferase (ALT), alkaline phosphatase (ALP), total bilirubin and with a reduction of total protein (P < 0.01). Treatment of rats with different doses of Jaft extract (250 and 500 mg/kg) significantly (P < 0.001) altered serum marker enzymes to near normal against CCl4 treated rats. Conclusion: in this study jaft extract is not hepatotoxic itself and demonstrated hepatoprotective effects. Keywords: Antioxidant activity, Jaft: hepatoprotective, Enzyme markers, Oak fruit doi:10.1016/j.clinbiochem.2011.08.814
Poster – [A-10-225-7] The antioxidant capacities and total phenol contents of some medicinal plants in Iran Mirzaei Ali, Ghasemi, Mirzaei Mahsa, Tavakol Kokhodan Laleh E-mail addresses: [email protected] (M. Ali), [email protected] (Ghasemi), [email protected] (M. Mahsa), [email protected] (T.K. Laleh) Introduction: Free radicals are highly reactive molecules that may cause great damage to cell membranes and DNA and result in inducing oxidation DNA mutations leading to cancer, degenerative, and other diseases. Derived plant antioxidants may prevent damages to free radicals. Material and methods: The stems and flower samples of plants were air-dried, finely ground and were extracted by ethanol:water (70:30) for 48 h. Extracts were filtered and dried under vacuum. The antioxidant activity of five ethanolic extracts of medicinal plants (Descurainia sophia, Plantago major, Trachyspermum copticum L., Coriandrum sativum and Trigonella foenum-graecum) from Iran were analyzed by five methods (1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, 2,2,azinobis-(3-ethylbenzthiazoline-6-sulphonic acid (ABTS) radical cation, Ferric-reducing antioxidant power assay (FRAP), phosphomolybdenum (PMB) and reducing power (RP). In addition, for determination of antioxidant components, total phenolic content was also analyzed. Results: The total phenolic content of plants varied from 74 to 154.3 mg Gallic acid/g extract as measured by Folin–Ciocalteau method. Values of DPPH ranged from 15.5 to 19.6 μmol trolex/g of extract. FRAP activity ranged from 124.2 to 753 μmol of Fe(II)/g of extract. Antioxidant activity of the Plantago major was always higher compared to the other plants' extracts with total phenol content and antioxidant capacity in DPPH, ABTS, and FRAP, (154.33 mg GAE/g, 1856 μmol trolox, 750 μmol trolox and 1169 μmol of Fe(II)/g, respectively). The range of total antioxidant activity by phosphomolybdenum method was 513.3 to 870 μmol trolox/g. The reducing ability of the tested extracts was between 0.31 and 1.26. Plantago major has also the highest activity in both tests. Conclusion: This study clearly demonstrated that Plantago major crude extract exhibits significant antioxidant activity.
Abstracts
Keywords: Antioxidant activity, Medicinal herbs, Plantago major, Total phenol doi:10.1016/j.clinbiochem.2011.08.815
Poster – [A-10-225-8] In vitro antioxidant activity by different methods in some vegetables in Yasouj, Iran Ebrahim Adibfar, Ali Mirzaei, Reza Haji Hossani, Sadeghi Haibatoallah Medicinal Plant Reserch Center, Yasouj Medical Sciences University, Yasouj, Iran E-mail addresses: [email protected] (E. Adibfar), [email protected] (A. Mirzaei) Introduction and Objective: Antioxidants are compounds that protect the body against damage caused by oxidative stress. There are different types of radical inhibitors in the living body that most are providing from the diet such as fruits, vegetables and beverages. In this study, was compared antioxidant activity, total phenols and flavonoids in some vegetable consumption in Yasouj. Materials and Methods: In this experimental study, the samples (Parsley, Coriander, Spinach, Tomato, and eggplant) were collected from the village of Imam Zadeh Jafar and Yasouj. Methanol extracts were prepared and then total phenol and flavonoids were assessed by methods Folin-Ciocalteu and Zyshen respectively. Antioxidant activity were estimated by using Diphenylpicrylhydrazyl (DPPH), Trolox equivalent antioxidant capacity (TEAC), ferric-reducing antioxidant power(FRAP), phosphomolybdat, nitric oxide. Data obtained were analyzed by software spss (version 17) and oneway ANOVA. Results: parsley and eggplant had the highest and lowest amount of total phenol respectively, while spinach and eggplant had the most and the least amount of flavonoids respectively. Antioxidant activity of coriander and tomato were maximum and minimum by method diphenylpicrylhydrazyl respectively. In both system of FRAP and TEAC methods Spinach and tomato had highest and lowest antioxidant activity respectively. In the nitric oxide assay coriander was highest and spinach was lowest while spinach was highest antioxidant activity by method Phosphomolybdat. Conclusion: This study showed that spinach and coriander had the highest antioxidant activity.
Keywords: Vegetables, Antioxidant, Total phenols, DPPH, FRAP
doi:10.1016/j.clinbiochem.2011.08.816
Poster – [A-10-233-1] Effect of the spinach (Spinacia oleracea) on blood glucose and insulin levels in diabetic rats Amir Reza Karami Bonari Faculty of Veterinary Medicine Islamic Azad University, Shabestar Branch, Shabestar, Iran E-mail address: [email protected] Introduction: Diabetes mellitus is a common endocrine disorder characterized by hyperglycemia. Plant medicine is used worldwide in the traditional treatment of some diseases. Natural medicines are much safer than synthetic drugs. spinach, originally found in Eastern Europe and Asia, can now be found in every corner of the globe. It is a fleshy-leafed annual that forms a heavy rosette of broad crinkly, tender leaves, where the high content of vitamins and minerals are
S331
found. Spinach contains of chlorophyll and oxalic acid. Studies have shown that Spinach extract has antioxidant effect and it could reduce the level of blood cholesterol. We assessed the antidiabetic efficacy of Spinach. Methods: Forty wistar rats, 10 non-diabetic and 30 induced diabetic rats by alloxan (150 mg/kg b.w , i.p), were used. Diabetic rats were divided equally into 3 groups: group 1: treated with saline (2 ml/kg b.w,p.o, diabetic control), group 2: treated with spinach (400 mg/kg b.w,p.o.), group 3: treated with chlorpropamide (250 mg/kg b.w,p.o.). All drugs were given for 30 days. Results and discussion: Spinach and chlorpropamide decreased significantly the blood glucose and increased significantly insulin concentrations (P < 0.05), in comparison with the diabetic control rats. The antidiabetic activity of sphinage at a dose of 400 mg/kg body weight was comparable to standard drug, chlorpropamide. Based on these results, the principal action of sphinage is on B cells, stimulating insulin secretion and thus reducing plasma glucose. Keywords: Spinage, Chlorpropamide, Insulin, Rats doi:10.1016/j.clinbiochem.2011.08.817
Poster – [A-10-240-3] Study of the effects of walnut leaf on the levels of a number of Blood Biochemical Factors in normal male rats fed with high cholesterol diet Eghbali Hadis, Hosseini-zijoud Seyyed Mostafa, Nabati Saeedeh, Mahmoodi Mehdi Rafsanjan, Iran E-mail addresses: [email protected] (E. Hadis), [email protected] (H.-z.S. Mostafa), [email protected] (N. Saeedeh), [email protected] (M. Mehdi) Introduction: Hypercholesterolemia is a major risk factor for coronary artery disease and thus for myocardial infarction and stroke. High serum low density lipoprotein (LDL) levels particularly pose a significantly increased risk for heart disease. Traditionally herbal plants and their derivative are used to lessen complications of metabolic disorders, including hypercholesterolemia. In the present study we investigated the impact of walnut leaf (Juglans regia) on the levels of a number of Blood Biochemical Factors in normal male rats fed with high cholesterol diet. Methods: We prepared powder of walnut leaf and mixed it with usual rat food as plate. Rats were divided to 5 groups and treated for 45 days: the first group was treated with usual food. The second group was treated with 1% cholesterol. The third group was treated with 1% walnut leaf and 1% cholesterol. The forth group was treated with 2% walnut leaf and 1% cholesterol. Fifth group treated with 5% walnut leaf and 1% cholesterol. We collected blood samples before and after treatment period and evaluated biochemical factors. Results: Our finding showed that in second group urea, TG, cholesterol and LDL increased and in forth group creatinine, TG and LDL decreased and in fifth group FBS, urea, TG, cholesterol and LDL decreased and HDL increased significantly (P<0.05). We also did not observe any significant difference in the studied factors in other groups. Discussion: Based on the results of this study it can be concluded that walnut leaf is a hypolipidemic agent that can reduce lipid profile as has been mentioned in traditional medicine. Keywords: Walnut leaf, Hypercholesterolemia, Cholesterol, Rat doi:10.1016/j.clinbiochem.2011.08.818
Abstracts
Keywords: Antioxidant activity, Pistacia atlantica, lipid peroxidation, superoxide dismutase, catalase doi:10.1016/j.clinbiochem.2011.08.812
Poster – [A-10-225-5] Acute and chronic toxicity of the jaft aqueous extracts a part of oak fruit in Iran Ali Mirzaei, Nooshin Mirzaei, Mahsa Mirzaei Medicinal Plant Research Center, Yasouj University of Medical Sciences, Yasouj, Iran E-mail addresses: [email protected] (A. Mirzaei), [email protected] (N. Mirzaei), [email protected] (M. Mirzaei) Introduction: Oak (Quercus species) Fruit has two covering external and internal layers that locally internal layer of fruit known as Jaft in some part of Iran. Materials and Methods: A study was conducted to investigate the safety of an aqueous extract of Jaft by acute toxicity methods. Acute toxicity (LD50) was estimated intraperitonal route (i.p.) and oral route (p.o.) in two separate groups. For the sub-chronic study, Twenty (20) adults Wistar rats (250–300 g) were randomly divided into four groups (I–IV) of 5 animals for each group. Animals were received Jaft- extract daily by gavages doses of 100, 500 and 1000 mg/kg for 28 days. At the end of experiment blood was collected for hematology and biochemical tests. Results: The result of acute toxicity values demonstrate that LD50 of the Jaft extract in the (i.p.) and (p.o.) procedures were > 3000 and > 5000 mg/kg respectively. All hematological parameters, in the experiment groups except (White Blood Cell) did not significantly differ compared to the control group. There were no statically significant observed in serum cholesterol, triglyceride, Blood Urea Nitrogen concentrations and alkaline phosphatase (ALP), glutamate pyruvate transaminase (GPT) and glutamate oxaloacetate transaminase (GOT) activities in all treatment groups compare to control group. No gross changes were found in the organs of rats following extract administration. Conclusion: These results suggest that the aqueous Jaft extract could be used with safety especially by oral route. Keywords: Acute toxicity, Chronic toxicity, Aqueous extract of jaft, Biochemical parameters, Hematological parameters doi:10.1016/j.clinbiochem.2011.08.813
Poster – [A-10-225-6] Antioxidant activities and hepatooprotectivity of Jaft a part of oak fruit Mehdi Akbartabar Tori, Ali Mirzaei, Mahsa Mirzaei, Nooshin Mirzaei Medicinal Plant Research Center, Yasouj University of Medical Sciences, Yasouj, Iran E-mail addresses: [email protected] (M.A. Tori), [email protected] (A. Mirzaei), [email protected] (M. Mirzaei), [email protected] (N. Mirzaei) Introduction: Aqueous extract of internal layer of oak fruit (Jaft) was evaluated for in vitro antioxidant activities and hepatoprotective in carbon tetrachloride induced liver damage in rats. Material and Methods: For in vitro determination of antioxidant activity used FRAP, DPPH, ABTS and PMB and for analysis of antioxidant components applied total phenol and flavonoids tests. For study of hepatoprotectivity thirty six male wistar rats were
chosen. Group (I, II and III) served as negative controls given olive oil, distilled water (D.W.) and jaft extract (500 mg/kg) respectively. Group(IV) as toxic group received CCl4 1 ml/kg, (i.p.) as single dose at every 72 h. The Protective groups (V) and (VI) one hour after the injection of carbon tetrachloride at every 72 h.received aqueous jaft extract (250 and 500 mg/kg) body weight orally respectively. Results: Jaft extract displayed significant antioxidant activity in different solvent systems as manifest by different (DPPH, ABTS, PMB and FRAP) assay systems. This study showed that in vitro efficiency was supported by a significant dose dependent hepatoprotection (at 250 mg/kg and 500 mg/kg dose) by decreasing the activity of serum enzymes and bilirubin (P < .01). CCl4 induced a significant rise in aspartate amino transferase (AST), alanine amino transferase (ALT), alkaline phosphatase (ALP), total bilirubin and with a reduction of total protein (P < 0.01). Treatment of rats with different doses of Jaft extract (250 and 500 mg/kg) significantly (P < 0.001) altered serum marker enzymes to near normal against CCl4 treated rats. Conclusion: in this study jaft extract is not hepatotoxic itself and demonstrated hepatoprotective effects. Keywords: Antioxidant activity, Jaft: hepatoprotective, Enzyme markers, Oak fruit doi:10.1016/j.clinbiochem.2011.08.814
Poster – [A-10-225-7] The antioxidant capacities and total phenol contents of some medicinal plants in Iran Mirzaei Ali, Ghasemi, Mirzaei Mahsa, Tavakol Kokhodan Laleh E-mail addresses: [email protected] (M. Ali), [email protected] (Ghasemi), [email protected] (M. Mahsa), [email protected] (T.K. Laleh) Introduction: Free radicals are highly reactive molecules that may cause great damage to cell membranes and DNA and result in inducing oxidation DNA mutations leading to cancer, degenerative, and other diseases. Derived plant antioxidants may prevent damages to free radicals. Material and methods: The stems and flower samples of plants were air-dried, finely ground and were extracted by ethanol:water (70:30) for 48 h. Extracts were filtered and dried under vacuum. The antioxidant activity of five ethanolic extracts of medicinal plants (Descurainia sophia, Plantago major, Trachyspermum copticum L., Coriandrum sativum and Trigonella foenum-graecum) from Iran were analyzed by five methods (1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, 2,2,azinobis-(3-ethylbenzthiazoline-6-sulphonic acid (ABTS) radical cation, Ferric-reducing antioxidant power assay (FRAP), phosphomolybdenum (PMB) and reducing power (RP). In addition, for determination of antioxidant components, total phenolic content was also analyzed. Results: The total phenolic content of plants varied from 74 to 154.3 mg Gallic acid/g extract as measured by Folin–Ciocalteau method. Values of DPPH ranged from 15.5 to 19.6 μmol trolex/g of extract. FRAP activity ranged from 124.2 to 753 μmol of Fe(II)/g of extract. Antioxidant activity of the Plantago major was always higher compared to the other plants' extracts with total phenol content and antioxidant capacity in DPPH, ABTS, and FRAP, (154.33 mg GAE/g, 1856 μmol trolox, 750 μmol trolox and 1169 μmol of Fe(II)/g, respectively). The range of total antioxidant activity by phosphomolybdenum method was 513.3 to 870 μmol trolox/g. The reducing ability of the tested extracts was between 0.31 and 1.26. Plantago major has also the highest activity in both tests. Conclusion: This study clearly demonstrated that Plantago major crude extract exhibits significant antioxidant activity.
Abstracts
Keywords: Antioxidant activity, Medicinal herbs, Plantago major, Total phenol doi:10.1016/j.clinbiochem.2011.08.815
Poster – [A-10-225-8] In vitro antioxidant activity by different methods in some vegetables in Yasouj, Iran Ebrahim Adibfar, Ali Mirzaei, Reza Haji Hossani, Sadeghi Haibatoallah Medicinal Plant Reserch Center, Yasouj Medical Sciences University, Yasouj, Iran E-mail addresses: [email protected] (E. Adibfar), [email protected] (A. Mirzaei) Introduction and Objective: Antioxidants are compounds that protect the body against damage caused by oxidative stress. There are different types of radical inhibitors in the living body that most are providing from the diet such as fruits, vegetables and beverages. In this study, was compared antioxidant activity, total phenols and flavonoids in some vegetable consumption in Yasouj. Materials and Methods: In this experimental study, the samples (Parsley, Coriander, Spinach, Tomato, and eggplant) were collected from the village of Imam Zadeh Jafar and Yasouj. Methanol extracts were prepared and then total phenol and flavonoids were assessed by methods Folin-Ciocalteu and Zyshen respectively. Antioxidant activity were estimated by using Diphenylpicrylhydrazyl (DPPH), Trolox equivalent antioxidant capacity (TEAC), ferric-reducing antioxidant power(FRAP), phosphomolybdat, nitric oxide. Data obtained were analyzed by software spss (version 17) and oneway ANOVA. Results: parsley and eggplant had the highest and lowest amount of total phenol respectively, while spinach and eggplant had the most and the least amount of flavonoids respectively. Antioxidant activity of coriander and tomato were maximum and minimum by method diphenylpicrylhydrazyl respectively. In both system of FRAP and TEAC methods Spinach and tomato had highest and lowest antioxidant activity respectively. In the nitric oxide assay coriander was highest and spinach was lowest while spinach was highest antioxidant activity by method Phosphomolybdat. Conclusion: This study showed that spinach and coriander had the highest antioxidant activity.
Keywords: Vegetables, Antioxidant, Total phenols, DPPH, FRAP
doi:10.1016/j.clinbiochem.2011.08.816
Poster – [A-10-233-1] Effect of the spinach (Spinacia oleracea) on blood glucose and insulin levels in diabetic rats Amir Reza Karami Bonari Faculty of Veterinary Medicine Islamic Azad University, Shabestar Branch, Shabestar, Iran E-mail address: [email protected] Introduction: Diabetes mellitus is a common endocrine disorder characterized by hyperglycemia. Plant medicine is used worldwide in the traditional treatment of some diseases. Natural medicines are much safer than synthetic drugs. spinach, originally found in Eastern Europe and Asia, can now be found in every corner of the globe. It is a fleshy-leafed annual that forms a heavy rosette of broad crinkly, tender leaves, where the high content of vitamins and minerals are
S331
found. Spinach contains of chlorophyll and oxalic acid. Studies have shown that Spinach extract has antioxidant effect and it could reduce the level of blood cholesterol. We assessed the antidiabetic efficacy of Spinach. Methods: Forty wistar rats, 10 non-diabetic and 30 induced diabetic rats by alloxan (150 mg/kg b.w , i.p), were used. Diabetic rats were divided equally into 3 groups: group 1: treated with saline (2 ml/kg b.w,p.o, diabetic control), group 2: treated with spinach (400 mg/kg b.w,p.o.), group 3: treated with chlorpropamide (250 mg/kg b.w,p.o.). All drugs were given for 30 days. Results and discussion: Spinach and chlorpropamide decreased significantly the blood glucose and increased significantly insulin concentrations (P < 0.05), in comparison with the diabetic control rats. The antidiabetic activity of sphinage at a dose of 400 mg/kg body weight was comparable to standard drug, chlorpropamide. Based on these results, the principal action of sphinage is on B cells, stimulating insulin secretion and thus reducing plasma glucose. Keywords: Spinage, Chlorpropamide, Insulin, Rats doi:10.1016/j.clinbiochem.2011.08.817
Poster – [A-10-240-3] Study of the effects of walnut leaf on the levels of a number of Blood Biochemical Factors in normal male rats fed with high cholesterol diet Eghbali Hadis, Hosseini-zijoud Seyyed Mostafa, Nabati Saeedeh, Mahmoodi Mehdi Rafsanjan, Iran E-mail addresses: [email protected] (E. Hadis), [email protected] (H.-z.S. Mostafa), [email protected] (N. Saeedeh), [email protected] (M. Mehdi) Introduction: Hypercholesterolemia is a major risk factor for coronary artery disease and thus for myocardial infarction and stroke. High serum low density lipoprotein (LDL) levels particularly pose a significantly increased risk for heart disease. Traditionally herbal plants and their derivative are used to lessen complications of metabolic disorders, including hypercholesterolemia. In the present study we investigated the impact of walnut leaf (Juglans regia) on the levels of a number of Blood Biochemical Factors in normal male rats fed with high cholesterol diet. Methods: We prepared powder of walnut leaf and mixed it with usual rat food as plate. Rats were divided to 5 groups and treated for 45 days: the first group was treated with usual food. The second group was treated with 1% cholesterol. The third group was treated with 1% walnut leaf and 1% cholesterol. The forth group was treated with 2% walnut leaf and 1% cholesterol. Fifth group treated with 5% walnut leaf and 1% cholesterol. We collected blood samples before and after treatment period and evaluated biochemical factors. Results: Our finding showed that in second group urea, TG, cholesterol and LDL increased and in forth group creatinine, TG and LDL decreased and in fifth group FBS, urea, TG, cholesterol and LDL decreased and HDL increased significantly (P<0.05). We also did not observe any significant difference in the studied factors in other groups. Discussion: Based on the results of this study it can be concluded that walnut leaf is a hypolipidemic agent that can reduce lipid profile as has been mentioned in traditional medicine. Keywords: Walnut leaf, Hypercholesterolemia, Cholesterol, Rat doi:10.1016/j.clinbiochem.2011.08.818