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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

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*△
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