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. 2015 Feb 1;8(2):1292-301.
eCollection 2015.

Effects of 18α-glycyrrhizin on TGF-β1/Smad signaling pathway in rats with carbon tetrachloride-induced liver fibrosis

Ying Qu  1 , Lei Zong  1 , Mingyi Xu  1 , Yuwei Dong  1 , Lungen Lu  1
Affiliations

Effects of 18α-glycyrrhizin on TGF-β1/Smad signaling pathway in rats with carbon tetrachloride-induced liver fibrosis

Ying Qu et al. Int J Clin Exp Pathol. .

Abstract

Background: Glycyrrhizin has various pharmacological effects including hepato-protection. This study aimed to investigate the potential mechanism underlying the protective effects of 18α-glycyrrhizin (18α-GL) in rats with carbon tetrachloride (CCl4) induced liver fibrosis.

Methods: Male Sprague-Dawley (SD) rats were randomly divided into control group, fibrosis group, 25 mg/kg 18α-GL group and 12.5 mg/kg 18α-GL group. Rats in experimental groups were subcutaneously injected with 40% CCl4 twice weekly for 8 weeks. Immunohistochemical examination was carried out to detect the protein expressions of collagen I, collagen III, TGF-β1, p-Smad2, p-Smad3, Smad 7 and SP-1, in the liver, and the mRNA and protein expressions of these genes were determined in the liver by real time PCR and Western blot assay, respectively.

Results: 18α-GL ameliorated histological changes and significantly suppressed collagen deposition. 18α-GL significantly decreased the mRNA expressions of TGF-β1, Smad2, Smad3 and SP-1 in the liver. Immunohistochemical staining revealed that TGF-β1, p-Smad2, p-Smad3 and SP-1 expressions reduced following 18α-GL therapy. Western blot assay showed p-Smad2, p-Smad3, smad2 and smad3 expressions decreased after 18α-GL treatment. The mRNA and protein expression of Smad7 remained unchanged.

Conclusion: 18α-GL is able to attenuate CCl4 induced liver fibrosis in rat.

Keywords: 18α-glycyrrhizin; Smad; TGF-β1; collagen; liver fibrosis.

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Figures

Figure 1
Figure 1
Histological examination of the liver in different groups. H&E staining: (A-D) Masson staining: (E-H) Representative photographs of control group (A, E), liver fibrosis (B, F), 12.5 mg/kg 18α-GL group (C, G) and 25 mg/kg 18α-GL group (D, H) (100×).
Figure 2
Figure 2
Effects of 18α-GL on hypdroxyproline in THE rat liver. Data are expressed as means ± standard deviation *P<0.05 vs fibrosis group.
Figure 3
Figure 3
Effects of 18α-GL on the collagen I and III expression in Rat Liver (immunohistochemistry). Collagen I (A-D) and collagen III (E-H) expression of control group (A, E), liver fibrosis group (B, F), 12.5 mg/kg 18α-GL group (C, G) and 25 mg/kg 18α-GL group (D, H). Positive cells had brown granules (100×). (I, J) Ratio of collagen I and III expression. Data are expressed as means ± standard deviation *P<0.05 vs liver fibrosis group.
Figure 4
Figure 4
Effects of 18αGL on the TGF/Smad mRNA expression. qPCR was performed to detect the mRNA expression of TGF-β1, Smad2/3/7 and SP-1. Data are expressed as means ± standard deviation *P<0.05 vs liver fibrosis group.
Figure 5
Figure 5
Immunohistochemical staining of TGF-β/Smad in the rat liver after 18α-GL treatment. Immunohistochemistry was performed to detect the protein expression of TGF-β1 (A-D), p-smad2 (E-H), p-Smad3 (I-L), Smad7 (M-P) and SP-1 (Q-T) in control group, liver fibrosis group, and 12.5 mg/kg 18α-GL group and 25 mg/kg 18α-GL group. Positive cells had brown granules (100×).
Figure 6
Figure 6
Western blot assay of Smad2/3 and p-smad2/3 expression in the rat liver after 18α-GL treatment.
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