[an error occurred while processing this directive]

Journal of Chinese Integrative Medicine ›› 2012, Vol. 10 ›› Issue (2): 193-199.doi: 10.3736/jcim20120210

• Original Experimental Research • Previous Articles     Next Articles

Effects of the mixture of Swertia pseudochinensis Hara and Silybum marianum Gaertn extracts on CCl4-induced liver injury in rats with non-alcoholic fatty liver disease

Mao Zhi-min1,Song Hai-yan1,Yang Li-li1,Liu Tao1,Li Dong-fei1,Zheng Pei-yong1,Liu Ping2,Ji Guang1,2()   

  1. 1. Institute of Digestive Disease, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China
    2. E-Institute of Shanghai Municipal Education Commission, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
  • Received:2011-09-01 Accepted:2011-10-26 Online:2012-02-20 Published:2018-10-09

RichHTML

PDF (PC)

275

Like

OBJECTIVE: To study the mechanism of liver injury induced by carbon tetrachloride (CCl4) in rats with non-alcoholic fatty liver disease (NAFLD), and the therapeutic effects of the extract mixture of Dangyao (Swertia pseudochinensis Hara) and Shuifeiji (Silybum marianum Gaertn) on NAFLD rats with liver injury.
METHODS: Male Wistar rats were randomized into normal control group, CCl4 group, high-fat diet group, high-fat diet plus CCl4 injection group (model group), diammonium glycyrrhizinate group and extract mixture group. Except the normal control and CCl4 groups, rats were fed with high-fat diet (88% normal chow, 10% lard and 2% cholesterol) to induce NAFLD. Diammonium glycyrrhizinate and extracts were given by gavage. After eight weeks, a nonlethal dose of CCl4 was injected intraperitoneally to all rats except the normal and high-fat diet groups. And 48 h later, all rats were sacrificed, and serum and liver tissues were collected for further study. Paraffin-processed liver tissue was stained with hematoxylin-eosin (HE) to observe the pathological changes. Serum alamine aminotrasferas (ALT) and aspartate aminotransferase (AST) were measured. The levels of triacylglycerol (TAG), malondialdehyde (MDA) and glutathione (GSH) in liver tissues were also examined. Expression of uncoupling protein 2 (UCP2) was determined by reverse transcription-polymerase chain reaction and Western blotting.
RESULTS: Liver sections stained with HE showed that the histopathological changes in the normal control group and the CCl4 group were mild; massive hepatosteatosis diffusing in lobules was shown in the high-fat diet groups; steatosis, hepatocellular ballooning degeneration and inflammatory infiltration were severe around the central vein in sections of the model group. Compared with the model group, hepatosteatosis and ballooning were significantly attenuated in the treatment groups. Levels of serum ALT and AST, contents of TAG and MDA and the UCP2 expression in liver tissues of the model group increased obviously, while the level of liver GSH decreased. Compared with rats in the model group, the above biomarkers in the treatment groups were improved significantly.
CONCLUSION: The mixture of Dangyao and Shuifeiji extracts can decrease the susceptibility and degree of liver injury induced by hepatotoxin in rats with NAFLD. Regulation of the balance of pro- and anti-oxidative stress factors is involved in the mechanism.

Key words: fatty live, non-alcoholic, liver injury, carbon tetrachloride, oxidative stress, plant extracts, rats

Figure 1

Liver tissues of rats in different groups by hematoxylin and eosin staining （Light microscopy, ×200） A: Normal group; B: CCl4 group; C: High-fat diet group; D: High-fat diet plus CCl4 group; E: Dangyao and Shuifeiji group; F: Diammonium glycyrrhizinate group."

Table 1

Serum ALT and AST levels and TAG, MDA and GSH contents of liver tissues of rats in different groups(x±s)"

Group n ALT (U/L) AST (U/L) TAG (mg/g liver) MDA (μmol/g protein) GSH (mg/g protein)
Nomal control 10 32.92±4.17* 41.79±7.16* 0.77±0.26* 5.30±1.17* 106.57±10.62*
CCl4 10 39.84±4.43*△ 50.17±1.61△ 0.93±0.12*△ 5.55±0.89*△ 104.15±11.32*△
High-fat diet 10 57.33±10.01△ 65.18±19.43△ 1.73±0.16△ 8.05±1.53△ 71.59±13.54△
High-fat diet plus CCl4 10 82.90±15.80* 84.14±7.90* 2.11±0.12* 10.11±1.13* 28.10±9.07*
Dangyao and Shuifeiji 10 43.28±3.63*△ 38.42±13.35*△ 1.20±0.12*△ 6.50±0.67△ 51.48±17.21*△
Diammonium glycyrrhizinate 10 49.64±12.84△ 31.97±7.27*△ 1.18±0.13*△ 7.58±0.64△ 51.96±15.77*△

Figure 2

Expressions of UCP2 mRNA (a) and protein (b) in liver tissues of rats in different groups A: Normal group; B: CCl4 group; C: High-fat diet group; D: High-fat diet plus CCl4 group; E: Dangyao and Shuifeiji group; F: Diammonium glycyrrhizinate group. UCP2: uncoupling protein 2; GAPDH: glyceraldehyde-3-phosphate dehydrogenase."

Table 2

UCP2 expression of rat liver tissues in different groups(x±s)"

Group n UCP2 mRNA expression
(Ratio to β-actin)		 UCP2 protein expression
(Ratio to GAPDH)
Nomal control 3 2.27±0.05*△ 1.73±0.01*△
CCl4 3 2.36±0.09*△ 1.92±0.02*△
High-fat diet 3 3.71±0.02△ 2.73±0.02△
High-fat diet plus CCl4 3 5.22±0.02* 3.06±0.04*
Dangyao and Shuifeiji 3 2.41±0.06*△ 1.88±0.01*△
Diammonium glycyrrhizinate 3 2.40±0.05*△ 1.86±0.03*△
[1] The Chinese National Workshop on Fatty Liver and Alcoholic Liver Disease for the Chinese Liver Disease Association. Guidelines for management of nonalcoholic fatty liver disease: an updated and revised edition. Zhonghua Gan Zang Bing Za Zhi. 2010; 18(3): 163-166. Chinese.
doi: 10.3760/cma.j.issn.1007-3418.2010.03.003
中华医学会肝脏病学分会脂肪肝和酒精性肝病学组. 非酒精性脂肪性肝病诊疗指南( 2010年修订版).中华肝脏病杂志. 2010; 18(3):163-166.
doi: 10.3760/cma.j.issn.1007-3418.2010.03.003
[2] Xie P, Zhou XX, Zhang Q . Pathogenesis and treatment of non-alcoholic fatty liver disease. J Chin Inter Med. 2010 ; 8(3) : 201-209. Chinese with abstract in English.
谢萍, 周新喜, 张琴 . 非酒精性脂肪肝的发病机制和治疗.中西医结合学报. 2010; 8(3):201-209.
[3] Malik A, Cheah PL, Hilmi IN, Chan SP, Goh KL . Non-alcoholic fatty liver disease in Malaysia: a demographic, anthropometric, metabolic and histological study. J Dig Dis. 2007; 8(1) : 58-64.
doi: 10.1111/cdd.2007.8.issue-1
[4] Donthamsetty S, Bhave VS, Mitra MS, Latendresse JR, Mehendale HM . Nonalcoholic fatty liver sensitizes rats to carbon tetrachloride hepatotoxicity. Hepatology. 2007; 45(2) : 391-403.
doi: 10.1002/hep.21530 pmid: 17256749
[5] Fan JG, Ding XD, Wang GL, Xu ZJ, Tian LY, Zheng XY . Expression of uncoupling protein 2 and its relationship to the content of adenosine triphosphate in the nonalcoholic fatty livers of rats fed a high-fat diet. Zhonghua Gan Zang Bing Za Zhi. 2005; 13(5): 374-377. Chinese with abstract in English.
范建髙, 丁晓东, 王国良, 徐正婕, 田丽艳, 郑晓英 . 非酒精性脂肪性肝病大鼠肝脏解偶联蛋白2表达及其与能量贮备的关系.中华肝脏病杂志. 2005; 13(5):374-377.
[6] Jiang XL, Zhang J, Liu SL, Wang GH . Oxidative stress and enzymatic antioxidant status in patients with hepatosteatosis. Sichuan Yi Xue. 2009 ; 30(12) : 1958-1960. Chinese with abstract in English.
蒋兴亮, 张均, 刘素兰, 王光华 . 非酒精性脂肪肝患者氧化应激和抗氧化酶状态.四川医学. 2009; 30(12):1958-1960.
[7] Singh R, Wang Y, Schattenberg JM, Xiang Y, Czaja MJ . Chronic oxidative stress sensitizes hepatocytes to death from 4-hydroxynonenal by JNK/c-Jun overactivation. Am J Physiol Gastrointest Liver Physiol. 2009 ; 297(5) : G907-G917.
doi: 10.1152/ajpgi.00151.2009
[8] Adams LA, Lindor KD . Nonalcoholic fatty liver disease. Ann Epidemiol. 2007; 17(11):863-869.
doi: 10.1016/j.annepidem.2007.05.013
[9] Krauss S, Zhang CY, Lowell BB . The mitochondrial uncoupling-protein homologues. Nat Rev Mol Cell Biol. 2005; 6(3) : 248-261.
doi: 10.1038/nrm1572 pmid: 15738989
[10] Wang W, Zhou JY . Uncoupling protein 2 and alcoholic liver disease. Guo Ji Nei Ke Xue Za Zhi. 2008; 35(5): 295-298. Chinese with abstract in English.
王玮, 周俊英 . 解偶联蛋白2与酒精性肝病.国际内科学杂志. 2008; 35(5):295-298.
[11] Ricquier D, Bouillaud F . Mitochondrial uncoupling proteins: from mitochondria to the regulation of energy balance. J Physiol. 2000; 529(Pt 1):3-10.
doi: 10.1111/j.1469-7793.2000.00003.x pmid: 11080246
[12] Fleury C, Neverova M, Collins S, Raimbault S, Champigny O, Levi-Meyrueis C, Bouillaud F, Seldin MF, Surwit RS, Ricquier D, Warden CH . Uncoupling protein-2 : a novel gene linked to obesity and hyperinsu- linemia. Nat Genet. 1997; 15(3):269-272.
doi: 10.1038/ng0397-269 pmid: 9054939
[13] Chan CB, Saleh MC, Koshkin V, Wheeler MB . Uncoupling protein 2 and islet function. Diabetes. 2004; 53(Suppll):S136-S142.
doi: 10.2337/diabetes.53.2007.S136 pmid: 14749279
[14] Diehl AM, Hoek JB . Mitochondrial uncoupling: role of uncoupling protein anion carriers and relationship to thermogenesis and weight control athe benefits of losing control” . JBioenergBiomembr. 1999; 31(5):493-506.
[15] Rashid A, Wu TC, Huang CC, Chen CH, Lin HZ, Yang SQ, Lee FY, Diehl AM . Mitochondrial proteins that regulate apoptosis and necrosis are induced in mouse fatty liver. Hepatology. 1999; 29(4):1131-1138.
doi: 10.1002/(ISSN)1527-3350
[16] Chavin KD, Yang SQ, Lin HZ, Chatham J, Chacko VP, Hoek JB, Walajtys-Rode E, Rashid A, Chen CH, Huang CC . Obesity induces expression of uncoupling protein-2 in hepatocytes and promotes liver ATP depletion. J Biol Chem. 1999; 274(9):5692-5700.
doi: 10.1074/jbc.274.9.5692
[17] Zheng PY, Ma ZS, Liu T, Xing LJ, Ji G . Effect of Jiangzhi Granulas on serum leptin and liver leptin receptor mRNA of nonalcoholic fatty liver disease rat. Zhongguo Zhong Yi Yao Xin Xi Za Zhi. 2008; 15(6): 36-38. Chinese with abstract in Enlgish.
郑培永, 马赞颂, 柳涛, 邢练军, 季光 . 降脂颗粒对非酒精性脂肪肝模型大鼠血清痩素及肝脏痩素受体 mRNA的影响.中国中医药信息杂志. 2008; 15(6):36-38.
[18] MaZS, Liu T, Zheng PY, Xing LJ, Ji G . Effect of Chinese medicine Jiangzhi Granula on hepatic lipid of nonalcoholic fatty liver disease rats. Zhonghua Zhong Yi Yao Xue Kan. 2007; 25(5): 942-944. Chinese with abstract in English.
doi: 10.3969/j.issn.1673-7717.2007.05.032
马赞颂, 柳涛, 郑培永, 邢练军, 季光 . 降脂颗粒对非酒精性脂肪肝大鼠肝脏脂质的影响.中华中医药学刊. 2007; 25(5) : 942-944.
doi: 10.3969/j.issn.1673-7717.2007.05.032
[19] Zheng PY, Ma ZS, Hua YQ, Liu T, Xing LJ, Ji G . Effects of puerarin on expressions of leptin receptor mRNA, phosphorylated Janus kinase 2/phosphorylated signal transducers and activators of transcription 3 proteins in the liver of rats with non-alcoholic fatty liver. J Chin Inter Med. 2008; 6(9): 921-927. Chinese with abstract in English.
郑培永, 马赞颂, 花永强, 柳涛, 邢练军, 季光 . 葛根素对非酒精性脂肪性肝病大鼠肝脏痩素受体mRNA和磷酸化Janus激酶2/磷酸化信号转导与转录激活因子 3的影响.中西医结合学报. 2008; 6(9):921-927.
[20] Zhou DC, Piao HS, Qin WJ, Ye ZG . Research progress of Swertia pseudochinensis Hara. Shizhen Guo Yi Guo Yao. 2011; 22(2): 462-465. Chinese.
周大成, 朴惠善, 秦文杰, 叶祖光 . 当药的研究进展. 时珍国医国药. 2011,22(2):462-465.
[21] Zhou DC, Piao HS, Zhang SY, Zhang GP . Protective effects of Swertia pseudochinensis Hara extract on experimental liver injury. Zhongguo Shi Yan Fang Ji Xue Za Zhi. 2010; 16(17): 125-128. Chinese with abstract in English.
doi: 10.3969/j.issn.1005-9903.2010.17.038
周大成, 朴惠善, 张思玉, 张广平 . 当药提取物对鼠试验性肝损伤的保护作用. 中国实验方剂学杂志. 2010; 16(17):125-128.
doi: 10.3969/j.issn.1005-9903.2010.17.038
[22] Yu LC, Gu CH . Research progress of pharmacological effect of Silymarin. Zhongguo Yi Yuan Yao Xue Za Zhi. 2001; 21(8): 493-494. Chinese.
于乐成, 顾长海 . 水飞萷素药理学效应研究进展. 中国医院药学杂志. 2001; 21(8):493-494
[1] Na Wu , Lu Song , Xin-xin Yang, Jiang-lei Wei , Zhen-guo Liu. Effects of Chinese herbal medicine Tianqi Pingchan Granule on G protein-coupled receptor kinase 6 involved in the prevention of levodopa-induced dyskinesia in rats with Parkinson disease. Journal of Chinese Integrative Medicine, 2012, 10(9): 1018-1024.
[2] Lawrence A. Olatunji, Taofeek O. Usman, Joseph O. Adebayo, Victoria A. Olatunji . Effects of aqueous extract of Hibiscus sabdariffa on renal Na+-K+-ATPase and Ca2+-Mg2+-ATPase activities in Wistar rats. Journal of Chinese Integrative Medicine, 2012, 10(9): 1049-1055.
[3] Dhirender Kaushik, Ajay Kumar, Pawan Kaushik, Avatar C. Rana. Anticonvulsant activity of alcoholic extract of bark of Pinus roxburghii Sarg. Journal of Chinese Integrative Medicine, 2012, 10(9): 1056-1060.
[4] Chan Chen, Chang-xun Chen, Xi-min Wu, Rui Wang, Yi-ming Li. Effects of extracts of Radix Scrophulariae on blood pressure in spontaneously hypertensive rats and the underlying mechanisms. Journal of Chinese Integrative Medicine, 2012, 10(9): 1009-1017.
[5] Jian-sheng Li, Ya Li, Su-yun Li, Yuan-yuan Wang, Li Deng, Yan-ge Tian, Su-li Jiang , Ying Wang. Long-term effects of Tiaobu Feishen therapies on systemic and local inflammation responses in rats with stable chronic obstructive pulmonary disease. Journal of Chinese Integrative Medicine, 2012, 10(9): 1039-1048.
[6] Karuna A. Talpate, Uma A. Bhosale, Mandar R. Zambare. Clitorea ternatea, a herb from Indian folklore, improves streptozotocin-induced diabetes and diabetes-induced cognitive decline in rats. Journal of Chinese Integrative Medicine, 2012, 10(8): 939-947.
[7] Samuel Joshua Pragasam, Vachana Murunikara, Evan Prince Sabina, Mahaboobkhan Rasool. Antiperoxidative potential of p-coumaric acid, a common dietary phenol, in adjuvant-induced arthritis in rats. Journal of Chinese Integrative Medicine, 2012, 10(8): 932-938.
[8] Zi-hui Li, Jian Wang, Rong-lin Cai, You-wen Wang, Jian-peng Hu. Establishment and evaluation of a rat model of ulcerative colitis with syndrome of dampness stagnancy due to spleen deficiency. Journal of Chinese Integrative Medicine, 2012, 10(8): 918-924.
[9] Meng Li, Ling Hu, Rong-lin Cai, Zi-jian Wu, Ke-ming Wang. Effects of electro-acupuncture at PC6 and BL15 on nerve electrical activity in spinal dorsal root and norepinephrine and dopamine contents in paraventricular nucleus of hypothalamus in rats with acute myocardial ischemia. Journal of Chinese Integrative Medicine, 2012, 10(8): 874-879.
[10] Xin-min Li, Hui Chen, Yan-yan Ren, Yan-li Lu. Effects of Chinese herbal medicine Xifeng Capsule on multidrug resistance-associated protein 1 expression in hippocampus and cortex of rats with lithium-pilocarpine-induced epilepsy. Journal of Chinese Integrative Medicine, 2012, 10(8): 911-917.
[11] Hong Wang , Jing-tian Ou , Xin Zhao , Ying Guoz , Hao-ping Mao. Vasodilator effect of oroxylin A on thoracic aorta isolated from rats. Journal of Chinese Integrative Medicine, 2012, 10(8): 880-885.
[12] C. Mouli Kalla, Vijaya Tartte, Dattatreya Rao Sirpurkar. Effectiveness of flavonoid-rich leaf extract of Acalypha indica in reversing experimental myocardial ischemia: biochemical and histopathological evidence. Journal of Chinese Integrative Medicine, 2012, 10(7): 784-792.
[13] Feng Xin-ying, Zhu Min, Zhang Qi-qi, Chen Yi-ping, Li Wen-wei. Selective protection of nigral dopaminergic neurons by echinacoside in a rat model of Parkinson disease induced by rotenone. Journal of Chinese Integrative Medicine, 2012, 10(7): 777-783.
[14] Chuan Chen, Hui-ying Chi, Zhi-hua Yu, Jiu-lin Chen . Effects of Chinese herbal medicine Shoushen Granule on telomere length and telomerase activity of peripheral white blood cells and vascular cells in rats with atherosclerosis. Journal of Chinese Integrative Medicine, 2012, 10(6): 667-673.
[15] Hui-qing Zhang , Zi-fei Yin , Jia-yu Sheng , Zhu-qian Jiang , Bao-yu Wu , Yong-hua Su. A comparison study of pharmacokinetics between bufalin-loaded bovine serum albumin nanoparticles and bufalin in rats. Journal of Chinese Integrative Medicine, 2012, 10(6): 674-680.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
[1] Dong Yang, Yong-ping Du, Qing Shen, Wei Chen, Yan Yu, Guang-lei Chen. Expression of alpha-smooth muscle actin in renal tubulointerstitium in patients with kidney collateral stasis. Journal of Chinese Integrative Medicine, 2008, 6(1): 41-44
[2] Hai-feng Wei, Bai-liu Ya, Ling Zhao, Cui-fei Ye, Li Zhang, Lin Li. Evaluation of tongue manifestation of blood stasis syndrome and its relationship with blood rheological disorder in a rat model of transient brain ischemia. Journal of Chinese Integrative Medicine, 2008, 6(1): 73-76
[3] Xi Lin, Jian-ping Liu. Herbal medicines for viral myocarditis. Journal of Chinese Integrative Medicine, 2008, 6(1): 76
[4] Xi Lin, Jian-ping Liu. Tai chi for treating rheumatoid arthritis. Journal of Chinese Integrative Medicine, 2008, 6(1): 82
[5] Liang-ping Hu, Hui Gao. Discrimination of errors in statistical analysis of medical papers published in the first issue of 2006 in Journal of Chinese Integrative Medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 98-106
[6] Yan-bo Zhu , Qi Wang, Cheng-yu Wu, Guo-ming Pang, Jian-xiong Zhao, Shi-lin Shen, Zhong-yuan Xia , Xue Yan . Logistic regression analysis on relationships between traditional Chinese medicine constitutional types and overweight or obesity. Journal of Chinese Integrative Medicine, 2010, 8(11): 1023-1035
[7] Wei Xu, Meng Shi, Jian-gang Liu, Cheng-long Wang . Collagen protein expressions in ischemic myocardium of rats with acute myocardial infarction and effects of qi-tonifying, yin-tonifying and blood-activating herbs and detoxifying and blood-activating herbs. Journal of Chinese Integrative Medicine, 2010, 8(11): 1041-1047
[8] Tao Wang , Feng Qin. Effects of Chinese herbal medicine Xiaoyao Powder on monoamine neurotransmitters in hippocampus of rats with postpartum depression. Journal of Chinese Integrative Medicine, 2010, 8(11): 1075-1079
[9] Ying Xu , Chang-chun Zeng , Xiu-yu Cai , Rong-ping Guo , Guang Nie , Ying Jin. Chromaticity and optical spectrum colorimetry of the tongue color in different syndromes of primary hepatic carcinoma. Journal of Chinese Integrative Medicine, 2012, 10(11): 1263-1271
[10] Xiang-ying Mao , Qin Bian , Zi-yin Shen. Analysis of the osteogenetic effects exerted on mesenchymal stem cell strain C3H10T1/2 by icariin via MAPK signaling pathway in vitro. Journal of Chinese Integrative Medicine, 2012, 10(11): 1272-1278
[an error occurred while processing this directive]