Effect of ich tri vuong capsule on plasma lipid level and degree of atherosclerosis in cholesterol - fed rabbits

Nội dung text: Effect of ich tri vuong capsule on plasma lipid level and degree of atherosclerosis in cholesterol - fed rabbits

1. JOURNAL OF MEDICAL RESEARCH EFFECT OF ICH TRI VUONG CAPSULE ON PLASMA LIPID LEVEL AND DEGREE OF ATHEROSCLEROSIS IN CHOLESTEROL-FED RABBITS Le Thi Minh Phuong1, Nguyen Tran Thi Giang Huong1, Do Thi Phuong1, Nguyen Thi Tuyet Nga2 1Faculty of Traditional Medicine, Hanoi Medical University, 2Military Institute of Traditional Medicine Ich Tri Vuong capsules (ITV) (components Ginkgo biloba, Radix Salviae miltiorrhi zae, Radix Astragali membranacei and Radix Angelicae sinensis) is used to treat diseases caused by atherosclerosis. This study was designed to assess the effects of ITV on plasma levels of total cholesterol, triglyceride, Low-density lipoprotein (LDL), High-density lipoprotein (HDL) and degrees of aorta atherosclerosis in rabbits with induced atherosclerosis by high cholesterol feeds. 50 New Zealand White rabbits were divided into 5 groups, 10 per group. Experiment conditions were carried out for 8 weeks. Normal group: fed with water 2 ml/kg/day. Control group: fed cholesterol 0.5 g/kg/day + water 2 ml/kg. Atorvastatin group: fed cholesterol 0.5 g/kg/day + Atorvastatin 5 mg/kg/day. ITV dose 1 group (60 mg/kg/day): fed cholesterol 0.5 g/kg/day+ ITV 60 mg/kg/day. ITV dose 2 group (180 mg/kg/day): fed cholesterol 0.5 g/kg/ngày + ITV 180 mg/kg/day. The results suggest that Ich Tri Vuong 180 mg/kg/day may decrease plasma levels of Total cholesterol and LDL and reduce atherosclerotic lesions of rabbits with induced atherosclerosis by high cholesterol feeds when compared to control group. These effects were equivalent to Atorvastatin 5 mg/kg/day and better than Ich Tri Vuong 60mg/kg/day. There were no significant differences in HDL and Triglyceride of ITV groups when compared to control group. Key works: Ich Tri Vuong, Atherosclerosis, plasma lipid I. INTRODUCTION Atherosclerosis is a disease of large and cells. Adherent monocytes migrate into medium arteries. Lesions begin as fatty the subendothelial space and differentiate streaks, caused by massive accumulation into macrophages. Uptake of oxLDL by of LDL oxidative product [1; 2] with high macrophages via scavenger receptors blood cholesterol LDL is increased in leads to foam cell formation. Interactions the plasma as well, and LDL is subject to between macrophage foam cells establish oxidative modifications in the subendothelial a chronic inflammatory process. Smooth space. Monocytes attache to endothelial muscle cells migrate to the side of inflammation. Necrosis of macrophage Corresponding author: Le Thi Minh Phuong, Faculty and smooth muscle cell-derived foam of Traditional Medicine, Hanoi Medical University cells leads to the formation of a necrotic Email: lethiminhphuong@hmu.edu.vn core and accumulation of extracellular Received: 05 June 2017 cholesterol, creating a plaque. Macrophage Accepted: 16 November 2017 secretion of matrix metalloproteinases and JMR 111 E2 (2) - 2018 27
2. JOURNAL OF MEDICAL RESEARCH neovascularization contribute to weakening Tri Vuong capsule has no acute and sub of the fibrous plaque. The ruptured plaque acute toxicities [9]. To hlear more about Ich is exposed to plasma components, Tri Vuong capsule is mechanism of action, causing the recruitment of platelets, and this study was designed to assess the effect the formation of a thrombus on the medial of Ich Tri Vuong capsule on plasma level of portion of the arterial wall. The thrombus total cholesterol, triglyceride, LDL, HDL and proliferates and secretes extracellular the degree of aorta atherosclerosis in an matrix proteins that form a fibrous plaque induced atherosclerosis by high cholesterol [1]. Atherosclerosis progresses slowly over feeds rabit model. years, and can cause stroke, myocardial II. SUBJECTS AND METHODS infarction, and peripheral artery disease. These are the mojor causes of disability 1. Animal and mortality. Therefore, treatment of lipid New Zealand and White rabbits (both metabolism disorders, especilly high special male and female gender, body weight 1.8 LDL, plays an important role in preventing – 2.5 kg) were supplied by Son Tay Goat atherosclerosis requires [2]. Recently, and Rabbit Research Center. The rabbits research show that many herbal medicines were fed with standard rabbit food in the that are used for treating diseases caused laboratory 7 days prior to the experiment. by atherosclerosis have an effect on lipid 2. Material metabolism and preventing atherosclerosis Ich Tri Vuong capsule [3; 4]. Ich Tri Vuong capsule (ITV) components Ich Tri Vuong capsule component include 40 mg Folium Ginkgo biloba extract, include: Folium Ginkgo biloba, Radix Salviae 150 mg extract of Radix Salviae miltiorrhi miltiorrhi zae, Radix Astragali membranacei, zae, and Radix Astragali membranacei, and Radix Angelicae sinensis. This capscule Radix Angelicae sinensis. ITV capsules is used in the treatment of chronic vertigo, were supplied by the National Institute of headache, and insomnia caused by cerebral Drug Quality Control and stored at room atherosclerosis. Ich tri vuong components temperature. Dose 1 in rabbit was 60 mg/ were used in the “Angelica decoction kg/day, dose 2 in rabbit was 180 mg/kg/day. for enriching plasma” (Radix Astragali Atorvastatin membranacei, Radix Angelicae sinensis), Atorvastatin 20 mg tablet, product a traditional medicine formula which of Standa Vietnam, stored at room effects plasma lipid and the expression temperature. Dose in rabbits was 5mg/kg/ of genes related to foam cell formation in day. the early stages of atherosclerosis [5]. Cholesterol oil Other components of Ich Tri Vuong, Folium Pure cholesterol (product of Merk- Ginkgo biloba and Radix Salviae miltiorrhi Germany) was dissolved in peanut oil zae, also effect plasma lipids in in-vitro and (product of Tuong An company). in-vivo [6 - 8]. Research has shown that Ich 28 JMR 111 E2 (2) - 2018
3. JOURNAL OF MEDICAL RESEARCH 3. Methods cholesterol (TC), high-density lipoprotein Rabbits were devided to 5 groups of 10 cholesterol (HDL-C) and low-density animals each. Experiment conditions were lipoprotein cholesterol (LDL-C) were carried out for 8 weeks. determined by commercial assay kits Normal group: fed water 2 ml/kg/day. (Hospitex Diagnostics Italy and DIALAB Control group: fed cholesterol oil 0.5 g/ GmbH Austria.) after 4 weeks and 8 weeks. kg/day, 1ml/kg, 2 hours later fed water 2 ml/ Quantification of gross atherosclerotic kg/day. lesions Atorvastatin group: fed cholesterol oil At the end of the 8 week - experiment, 0.5 g/kg/day, 1ml/kg, 2 hours later fed 30% of the rabbit aorta was collected Atorvastatin 5mg/kg/day. for analysis of the aortic lesions. The ITV dose 1 (60 mg/kg/day): fed progression of atherosclerosis was graded cholesterol oil 0.5 g/kg/day, 1ml/kg, 2 according to American Heart Association hours later fed Ich Tri Vuong 60 mg/kg/day guidelines dividing atherosclerotic lesions (equivalent to clinical dose). into I-VI morphological types [10]. ITV dose 2 (180 mg/kg/day): fed 4. Statistical analysis cholesterol oil 0.5 g/kg/day, 1ml/kg, 2 hours 2 way ANOVA test and Mann–Whitney U later fed Ich Tri Vuong 180 mg/kg/day (three test was used to analyze data, and p values time higher than clinical dose). less than 0.05 were considered statistically Determination of plasma lipid levels significant. The plasma triglycerides (TG), total III. RESULTS 8 weeks of the high cholesterol diet induced high blood lipids and atherosclerosis in rabbits of group 2, 3, 4, and 5 with significantly the increases TC, TG, LDL - C, and HDL - C plasma (p < 0.05), and lesions of aorta atherosclerosis seen on histological examination. Meanwhile, in group 1, there was no significant difference in lipids plasma levels with comparable to the begin (p > 0.05), and no lesion of atherosclerosis was seen in the aorta on histological examination. Plasma Lipids level To assess the effect of Ich Tri Vuong on plasma lipid level, we measured and compared plasma level of TC, TG, LDL - C, HDL - C at the beginning of the experiment (W0), after 4 weeks (W4) and after 8 weeks (W8). JMR 111 E2 (2) - 2018 29
4. JOURNAL OF MEDICAL RESEARCH Figure 1. Plasma level of Total Cholesterol (TC) In Figure 1, after 4 weeks and 8 weeks, plasma TC level of the Atorvastatin group (10.74 ± 8.18 mmol/l and 10.71 ± 6.17 mmol/l) was significantly lower than the control group (21.13 ± 9.98 mmol/l and 22.35 ± 10,31 mmol/l) and ITV dose 1 group (23.97 ± 14.28 mmol/l and 17.31 ± 9.56 mmol/l) (p < 0.05). ITV dose 2 group (13.91 ± 9.36 mmol/l and 12.76 ± 7,13 mmol/l) was significantly lower than Control group and had no significant difference to Atorvastatin (p > 0.05). The amount TC increased over 8 weeks in the Atorvastatin group (7.74 ± 5.84 mmol/l) and the ITV dose 2 (10.72 ± 6.77 mmol/l) group was significantly lower than the TC increase in the control group (19.57 ± 9.34 mmol/l) (p < 0.05). There was no significantly difference in the amount TC increased over 8 weeks in the ITV dose 2 group when compared to the Atorvastatin group (p > 0.05). Figure 2. Plasma level of LDL In Figure 2, after 4 weeks and 8 weeks, plasma LDL - C level of the Atorvastatin group 30 JMR 111 E2 (2) - 2018
5. JOURNAL OF MEDICAL RESEARCH (9.35 ± 7.31 mmol/l and 8.98 ± 5.35 mmol/l) were significantly lower than the control group (18.55 ± 9.18 mmol/l and 19.51 ± 9.66 mmol/l) and ITV dose 1 group (21.30 ± 3.12 mmol/l and 14.96 ± 8.58 mmol/l) (p < 0.05). ITV dose 2 group (11.96 ± 8.18 mmol/l and10.59 ± 6.23 mmol/l) was significantly lower than control group (p < 0.05) and had no significant difference to the Atorvastatin group (p > 0.05). After 8 weeks, increasing levels of plasma LDL level of Atorvastatin group (6.64 ± 5.13 mmol/l) and ITV dose 2 (9.11 ± 6.01 mmol/l) group were significantly lower than the control group (17.31 ± 8.82 mmol/l) (p < 0.05). The ITVdose 1 group had no significant difference in amount of LDL - C increase over 8 weekswhen compared to the control group (p > 0.05). There was no significant difference in ithe amount LDL - C increased over 8 weeks in the ITV dose 2 group when compared to the Atorvastatin group (p > 0.05). In Figure 3, after 4 weeks and 8 weeks, plasma HDL - C level of the Atorvastatin group (1.39 ± 0.89) and 1.73 ± 0.83 mmol/l) were significantly lower than the control group (2.59 ± 0.91 mmol/l and 2.84 ± 0.69 mmol/l) (p < 0.05). ITV dose 1 group (2.68 ± 1.26 mmol/l and 2.35 ± 1.01 mmol/l) and ITV dose 2 group (1.94 ± 1.20 mmol/l and 2.17 ± 0.94 mmol/l) had no significant difference to the Control group (p > 0.05). After 8 weeks, the amount that HDL - C increase over 8 weeks in the Atorvastatin group (1.10 ± 0.74 mmol/l) was significantly lower when the control group (2.26 ± 0.67 mmol/l) (p > 0.05) however the ratio of HDL/ LDL in the Atorvastain group (0.19) was higher than the Control group (0.15). There was no significant difference the amount HDL - C increase over 8 weeks in the ITV dose 2 group when compared to the Atorvastatin group (p > 0.05). Figure 3. Plasma level of HDL In figure 4, after 4 weeks and 8 weeks, plasma triglyceride level of 5 group hadno significantly difference (p > 0.05). JMR 111 E2 (2) - 2018 31
6. JOURNAL OF MEDICAL RESEARCH Figure 4. Plasma level of Triglyceride In figure 4, after 4 weeks and 8 weeks, plasma triglyceride level of all 4 groups had no significant difference when compared to the control (p > 0.05). Lesions of aortic atherosclerosis In Figure 5, after 8 weeks, all samples of Normal group and 2/3 sample of Atorvastin group and ITV dose 2 group had no aortic atherosclerotic gross lesions, 3/3 samples of Control group and 2/3 sample of ITV dose 1 had a gross lesion of aortic atherosclerosis. A Normal Control Atorvastatin ITV dose 1 ITV dose 2 32 JMR 111 E2 (2) - 2018
7. JOURNAL OF MEDICAL RESEARCH 100% 80% 60% Type III B 40% Type II 20% Normal 0% Normal group Control group Atorvastatin ITV dose 1 ITV dose 2 group group group Figure 5. Lesion of aortic atherosclerosis Figure 5: Lesion of aortic atherosclerosis (A) Aortic arch and histological examination of groups (B) Degree lesions in aortic arch sections histological examination In Figure 5, after 8 weeks, all samples of Normal group and 2/3 sample of Atorvastin group and ITV dose 2 group had no aortic atherosclerotic gross lesions, all samples of Control group and 2/3 sample of ITV dose 1 had a gross lesion of aortic atherosclerosis. Histological examination of aortic arch sections were examined to quantify the degree of atherosclerotic lesions. All samples of the Control group and 2/3 samples of ITV dose 1 group had type III lesions. There were fewer intimal lession in the Atorvastatin group (with 1/3 sample had type II of lesion, 2/3 sample was normal) and ITV dose 2 group (with 1/3 sample had type II of lesion, 2/3 sample was normal), when compared to the control group. IV. DISCUSSION Ich Tri Vuong capsule components difference in HDL when compared with the include Folium Ginkgo biloba, Radix Salviae control group. The increase over 8 week miltiorrhi zae, Radix Astragali membranacei, of plasma total cholesterol and LDL of the and Radix Angelicae sinensis. Capsules Atorvastatin group (7.74 ± 5.84 mmol/l have been used in the treatment chronic and 6.64 ± 5.13 mmol/l) and the ITV dose vertigo, headache and insomnia caused by 2 (10.72 ± 6.77 mmol/l and 9.11 ± 6.01 cerebral atherosclerosis. mmol/l) group were significantly lower than The results show that ITV dose 180 the control group (19.57 ± 9.34 mmol/l mg/kg/day and Atorvastatin 5 mg/kg/day and 17.31 ± 8.82 mmol/l) (p < 0.05). ITV significantly lower plasma level of total dose 60mg/kg/day had plasma levels of cholesterol and LDL when compared with Total cholesterol significantly lower than the control group. ITV dose 180 mg/kg/ the Control group, but had no significant day and 60 mg/kg/day had no significant difference in LDL compared with the Control JMR 111 E2 (2) - 2018 33
8. JOURNAL OF MEDICAL RESEARCH group. There was no significant difference of V. CONCLUSION the increase over 8 weeks of plasma lipids These results suggest that in an induced of the ITV dose 2 group when compared - atherosienrosis rabbit model Ich Tri Vuong with the Atorvastatin group (p > 0.05). 180 mg/kg/day may decrease plasma levels These effects of Ich Tri Vuong may be of Total cholesterol and LDL, and reduce due to effects of its components. Previous atherosclerosis lesion when compared to studies have shown that Ich Tri Vuong control group. The effect of ITV 180 mg/kg/ components had an effect on plasma day was equivalent to Atorvastatin 5 mg/kg/ lipids, expression of genes related to day and better than Ich Tri Vuong 60 mg/kg/ foam cell formation in the early stage of day. There was no significant difference in atherosclerosis in a diabetic rat model [5]. HDL and Triglyceride of ITV groups when Folium Ginkgo biloba and Radix Salviae compared to the control group. miltiorrhi of Ich Tri Vuong also had effects on plasma lipid in in-vitro and in-vivo model Acknowledgments [6 - 8]. We would like to thank the staff that Hypercholesterolemia is a major risk participated. This work was supported by a factor for atherosclerosis [1; 2]. In this research grant of Hanoi Medical University. research ITV dose 2 group and ITV dose 1 group decreased atherosclerosis lesion REFERENCE in histological examination of the aortic 1. James S. Forrester (2008), The arch, when compared with the Control Pathogenesis of Atherosclerosis and Plaque group. The decrease in atherosclerosis Instability, Atherosclerosis and Oxidant lesions effected by Ich Tri Vuong 180 mg/ Stress: A New Perspective, Springer, 1 – 7. kg/day was equivalent to that of Atorvastatin 2. Judith A. Berliner, Mohamad 5 mg/kg/day. Atherosclerotic lesions Navab, Alan M. Fogelman et al. (1995), begin as massive accumulations of LDL Atherosclerosis: basic mechanisms. oxidative productions. So the lowering Oxidation, inflammation, and genetics, Total cholesterol and LDL effects of Ich Circulation, 91(9), 2488 - 2496. Tri Vuong could mediate the reduction in 3. Hao Xu, Dazhuo Shi, Keji Chen atherosclerosis. Although Atorvastatin 5 (2012), Atherosclerosis: An Integrative mg/kg/day decreased LDL more than Ich East-WestMedicine Perspective, Evidence- Tri Vuong 180 mg/kg/day, the eddect on Based Complementary and Alternative atherosclerosis lesions of Atorvastatin 5 Medicine, Vol 2012, Article ID 148413, 4 mg/kg/day was equivalent to the effect of pages, doi:10.1155/2012/148413. Ich Tri Vuong 180 mg/kg/day. Therefore 4. Qing Liu, Jianping Li, Adam Ich Tri Vuong effect on atherosclerosis Hartstone-Rose et al. (2015), Chinese pathology may be mediated by more than Herbal Compounds for the Prevention and one pathway. Treatment of Atherosclerosis: Experimental 34 JMR 111 E2 (2) - 2018
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