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Journal of Chinese Integrative Medicine ›› 2013, Vol. 11 ›› Issue (4): 262-268.doi: 10.3736/jintegrmed2013034

• Research Article • Previous Articles     Next Articles

Bioactivity of five components of Chinese herbal formula Jiangzhi granules against hepatocellular steatosis

Hai-yan Songa, Li Zhanga, Jie-lu Pana, Li-li Yanga, Guang Jia,b()   

  1. a. Institute of Digestive Disease, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China
    b. E-institute of Shanghai Municipal Education Commission, Shanghai University of Traditional ChineseMedicine, Shanghai 200032, China
  • Received:2013-03-24 Accepted:2013-05-15 Online:2013-07-10 Published:2013-07-15

Objective

This study aims to evaluate the bioactivity of five components of the traditional Chinese medicine complex prescription Jiangzhi granules against hepatocellular steatosis.


Methods

The five major components, including protopanaxadiol, tanshinone IIA, emodin, chlorogenic acid, and nuciferine, were extracted from Jiangzhi granules. Their cytotoxicity was assessed to determine the safe dose of each component for HepG2 cells. HepG2 cellular steatosis was induced using 1 mmol/L of free fatty acids (FFAs) for 24 h, and then treated with each component at high, intermediate, and low doses (500, 50, and 5 μmol/L), respectively for another 24 h. The effects on HepG2 steatosis were observed directly under optical phase microscopy, or through oil red O staining and Nile red assays. In addition, the levels of reactive oxygen species (ROS) in the steatotic HepG2 cells with and without high-dose protopanaxadiol treatment were measured using fluorescent dye 2′,7′-dichlorodihydrofluorescein diacetate staining.


Results

No obvious cytotoxicity was observed in the HepG2 cells incubated with each of the five components at up to 500 μmol/L. At 24 h after incubation with FFAs, the HepG2 cells swelled and many lipid droplets accumulated. The lipid content was attenuated after 24 h of incubation with protopanaxadiol, tanshinone IIA, and emodin at 500 or 50 μmol/L (P < 0.05), especially with 500 μmol/L protopanaxadiol (P < 0.01). In addition, the ROS level was elevated in steatotic cells, but decreased after intervention with 500 μmol/L protopanaxadiol (P < 0.05).


Conclusion

Protopanaxadiol, tanshinone IIA, and emodin alleviate hepatocellular steatosis in a dose-dependent manner, and oxidative stress regulation may partially contribute to the effects of protopanaxadiol.

Key words: Hepatocellular steatosis, Jiangzhi granules, Plant extracts, Oxidative stress

Figure 1

Effects of the high- and intermediate-dose protopanaxadiol, tanshinone IIA and emodin on HepG2 cellular steatosis (Oil red O staining under a light microscope, ×200) A: Control; B: Model; C: High-dose protopanaxadiol; D: Intermediate-dose protopanaxadiol; E: High-dose tanshinone IIA; F: Intermediate-dose tanshinone IIA; G: High-dose emodin; H: Intermediate-dose emodin."

Figure 2

Effects of the five components on HepG2 cellular steatosisThe steatotic HepG2 cells was incubated with high, intermediate or low dose of the five components, and the intracellular fat accumulation was determined by Nile red fluorescence intensity, which was shown here as the fold of control, n=3; **P<0.01, vs control group; △P<0.05, △△P<0.01, vs model group."

Figure 3

Effects of high-dose protopanaxadiol on HepG2 cellular reactive oxygen species generation (fluorescence micro-scopy with excitation at 488 nm, ×200)A: Control; B: Model; C: High-dose protopanaxadiol."

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