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Journal of Chinese Integrative Medicine ›› 2012, Vol. 10 ›› Issue (4): 454-461.doi: 10.3736/jcim20120415

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Salvianolic acid B inhibits ERK signal transduction pathway activated by transforming growth factor-β1 in rat hepatic stellate cells

Ya-fang Song1,2,3, Zhi-gang Lü1,2, Lie-ming Xu1,2,4,5()   

  1. 1. Department of Liver Cirrhosis, Shuguang Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
    2. Institute of Liver Diseases, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
    3. Institue of Stomach and Spleen Diseases, Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China
    4. Key Laboratory of Liver and Kidney Diseases, Shanghai 201203, China
    5. Key Laboratory of Clinical Traditional Chinese Medicine, Shanghai 201203, China
  • Received:2011-08-18 Accepted:2011-09-15 Online:2012-04-20 Published:2018-04-15

Objective: To investigate the effects of salvianolic acid B (SA-B) on extracellular signal-regulated kinase (ERK) signal transduction pathway activated by transforming growth factor-β1 (TGF-β1) in rat hepatic stellate cells (HSCs).
Methods: HSCs were isolated from male Sprague-Dawley rats by in situ perfusion and Nycodenz density-gradient centrifugation method. TGF-β1 and SA-B were directly added to the serum-free medium of HSCs. Total and phosphorylated ERK, MEK, Raf and α-smooth muscle actin (α-SMA) and typeⅠcollagen were assayed by Western blotting.
Results: Phosphorylation of MEK in HSCs with or without TGF-β1 was inhibited by SA-B; however, phosphorylation of Raf in HSCs with or without TGF-β1 was not inhibited by SA-B. Expression of α-SMA in HSCs with TGF-β1 was inhibited by SA-B. Combination of SA-B and the inhibitors of ERK (PD98059) can effectively inhibit the expression of α-SMA. SA-B also inhibited synthesization of typeⅠcollagen in HSCs with or without TGF-β1.
Conclusion: The action point of SA-B inhibiting ERK signaling induced by TGF-β1 in HSCs is the inhibition of the phosphorylation of MEK. SA-B reduces the increase of expression of α-SMA and protein synthesization of type Ⅰ collagen induced by TGF-β1 by means of inhibiting ERK signaling in activated HSCs of rats.

Key words: salvianolic acid B, hepatic stellate cells, transforming growth factor-β1, extracellular signal-regulated MAP kinases, rats

Figure 1

Expressions of ERK and p-ERK1/2, MEK and p-MEK, and Raf-1 and p-Raf-1 in HSCs The levels of ERK and p-ERK1/2, MEK and p-MEK and Raf-1 and p-Raf-1 were determined by Western blot using specific antibodies. Loading control β-actin was determined by Western blot analysis as indicated. Data are expressed as percentage of the control or β-actin; n=3; *P<0.05, **P<0.01, vs control group; △△P<0.01, vs TGF-β1 group. A: Expression of ERK and p-ERK1/2 in HSC. B: Expression of MEK and p-MEK in HSC. C: Expression of Raf-1 and p-Raf-1 in HSC. SA-B: salvianolic acid-B; HSC: hepatic stellate cell; TGF-β1: transforming growth factor-β1; ERK: extracellular signal-regulated kinase."

Figure 2

Effects of different concentrations of PD98059 on the phosphorylation of ERK in HSCs The levels of p-ERK were determined by Western blot using specific antibodies. Data are expressed as percentage of the control; n=3; **P<0.01, vs control group; △△P<0.01, vsTGF-β1 group. HSC: hepatic stellate cell; TGF-β1: transforming growth factor-β1; ERK: extracellular signal-regulated kinase."

Figure 3

Effects of SA-B and PD98059 on the expression of α-SMA in HSCs The levels of α-SMA protein were determined by Western blotting using specific antibodies. Data are expressed as percentage of the β-actin; n=3; *P<0.05, vs control group; △△P<0.01, vs TGF-β1 group. SA-B: salvianolic acid-B; HSC: hepatic stellate cell; TGF-β1: transforming growth factor-β1; α-SMA: α-smooth muscle actin."

Figure 4

Expression of type Ⅰ collagen in HSCs in each group The levels of type Ⅰ collagen were determined by Western blotting using specific antibodies. Data are expressed as percentage of the β-actin; n=3; **P<0.01, vs control group; △△P<0.01, vs TGF-β1 group. SA-B: salvianolic acid-B; HSC: hepatic stellate cell; TGF-β1: transforming growth factor-β1."

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