Anti-hepatic fibrosis effects of a novel turtle shell decoction by inhibiting hepatic stellate cell proliferation and blocking TGF-β1/Smad signaling pathway in rats

Bai,Ganping; Yan,Guohe; Wang,Guojian; Wan,Ping; Zhang,Ronghua

doi:10.3892/or.2016.5078

Anti-hepatic fibrosis effects of a novel turtle shell decoction by inhibiting hepatic stellate cell proliferation and blocking TGF-β1/Smad signaling pathway in rats

Authors:
- Ganping Bai
- Guohe Yan
- Guojian Wang
- Ping Wan
- Ronghua Zhang
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Affiliations: Department of Integrated Chinese and Western Medicine, Southwest Hospital, The Third Military Medical University, Chongqing 400038, P.R. China, Institute of Combined Injuries, The State Key Laboratory of Trauma, Burns and Combined Injury, College of Preventive Medicine, The Third Military Medical University, Chongqing 400038, P.R. China
Published online on: September 9, 2016 https://doi.org/10.3892/or.2016.5078
Pages: 2902-2910

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Abstract

Hepatic fibrosis (HF), a wound-healing response to a variety of chronic stimuli, is characterized by the excessive synthesis of extracellular matrix (ECM) proteins by hepatic stellate cells (HSC) and eventually the development of hepatic cirrhosis. Turtle shell pill (TSP) is a common traditional Chinese medicine used for preventing and treating HF and early hepatic cirrhosis, but its side-effects and the shortage of ingredients limit its clinical application. In addition, its mechanism of action is not clear. In the present study, we first improved the original formula of TSP to produce a novel turtle shell decoction (NTSD) with less toxicity and easier accessible materials. In a carbon tetrachloride (CCl4)-induced HF rat model, we observed that NTSD and TSP had similar effects on the improvement of liver functions in rats, including a decrease in serum alanine amino transferase (ALT) and aspartate amino transferase (AST) serum concentrations and increased albumin content in addition to a marked attenuation of CCl4-induced liver damage and fibrosis. NTSD containing rat serum inhibited rat liver stellate cell line HSC-T6 cell proliferation and induced cell apoptosis in vitro. Moreover, the NTSD treatment significantly decreased the transforming growth factor beta 1 (TGF-β1) and Smad3 gene expression and increased inhibitory Smad7 gene expression in liver tissues of HF rats, suggesting that NTSD inhibited the ECM expression of HSC by downregulating the TGF-β1/Smad signaling pathway. The results of our rat model study revealed that NTSD showed good in vitro and in vivo anti-HF effects via proliferation inhibition and the induction of apoptosis of HSCs and blocked the TGF-β1/Smad signaling pathway.

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1	Mortality GBD: Global, regional, and national age-sex specific all-cause and cause-specific mortality for 240 causes of death, 1990–2013: A systematic analysis for the Global Burden of Disease Study 2013. Lancet. 385:117–171. 2015. View Article : Google Scholar
2	Henderson NC and Iredale JP: Liver fibrosis: Cellular mechanisms of progression and resolution. Clin Sci (Lond). 112:265–280. 2007. View Article : Google Scholar
3	Parsons CJ, Takashima M and Rippe RA: Molecular mechanisms of hepatic fibrogenesis. J Gastroenterol Hepatol. 22(Suppl 1): S79–S84. 2007. View Article : Google Scholar : PubMed/NCBI
4	Tsukada S, Parsons CJ and Rippe RA: Mechanisms of liver fibrosis. Clin Chim Acta. 364:33–60. 2006. View Article : Google Scholar
5	Cui W, Jin HB and Li ZW: Mechanism of the transforming growth factor-beta induction of fibronectin expression in hepatic stem-like cells. Braz J Med Biol Res. 43:36–42. 2010. View Article : Google Scholar
6	Yoshida K, Murata M, Yamaguchi T and Matsuzaki K: TGF-β/Smad signaling during hepatic fibro-carcinogenesis (Review). Int J Oncol. 45:1363–1371. 2014.PubMed/NCBI
7	Kaimori A, Potter J, Kaimori JY, Wang C, Mezey E and Koteish A: Transforming growth factor-beta1 induces an epithelial-to-mesenchymal transition state in mouse hepatocytes in vitro. J Biol Chem. 282:22089–22101. 2007. View Article : Google Scholar : PubMed/NCBI
8	Tahashi Y, Matsuzaki K, Date M, Yoshida K, Furukawa F, Sugano Y, Matsushita M, Himeno Y, Inagaki Y and Inoue K: Differential regulation of TGF-beta signal in hepatic stellate cells between acute and chronic rat liver injury. Hepatology. 35:49–61. 2002. View Article : Google Scholar : PubMed/NCBI
9	Wells RG: Cellular sources of extracellular matrix in hepatic fibrosis. Clin Liver Dis. 12:759–768. viii2008. View Article : Google Scholar : PubMed/NCBI
10	Zhang Q and Jin S: The modern clinical application and experimental research progress of turtule shell pills. J Hebei Tradit Chin Med Pharmacol. 21:35–36. 2006.
11	Lin W, Wei N, Gao B, Jiang G and Chang Y: Systematic evaluation of therapeutic effect of compound turtule shell pills against hepatic fibrosis. Chin J Gastroenterol Hepatol. 16:69–72. 2007.
12	Gao J, Tao J and Zhao C: The experimental study of turtle shell pills in prevention and treatment of hepatic fibrosis. Chin Arch Tradit Chin Med. 11:2462–2471. 2008.
13	Xi Z, Luan X and Li K: The study of dormant insect on inhibiting immune hepatic fibrosis in rats. Tradit Chin Med Res. 17:38–40. 2001.
14	Xu L and Liu P: The observation of effects on anti-hepatic fibrosis of peach extract-the study of immunohistochemistry and collagen metabollism. Tradit Chin Med Res. 5:14–16. 1993.
15	Lu Y, Ren X and Chen Y: Dynamic observation of the effects of turtle shell pills on liver collgen and serum pre-collagen III during the hepatic fibrosis process in rats. Henan Tradit Chin Med. 21:192001.
16	Moldovan GL, Pfander B and Jentsch S: PCNA, the maestro of the replication fork. Cell. 129:665–679. 2007. View Article : Google Scholar : PubMed/NCBI
17	Gressner AM, Weiskirchen R, Breitkopf K and Dooley S: Roles of TGF-beta in hepatic fibrosis. Front Biosci. 7:d793–d807. 2002. View Article : Google Scholar : PubMed/NCBI
18	Dai H, Zhang WX, He XL, Zhao RX, Fang L and Li CC: Effect of TGF-B1/Smad3 aignal pathway on airway remodelling in asthma rats with the regulation of Radix Astragali. Chin Arch Tradit Chin Med. 28:2494–2498. 2010.
19	Huang J, Zhang C, Zhan F and Zhang J: Effects of astragalan on liver fibrosis rat in TGF-β1/Smads signal pathway. China J Tradit Chin Med Pharm. 30:2184–2186. 2015.
20	Jiang W, Zhou ZS, Hu HB and Liu BJ: The Effect of Fuling, Yiyiren and Dongguazi on serum TGF-β1 and TNF-α levels in rats with pulmonary fibrosis. Med J Qilu. 28:237–240. 2013.
21	Niu SL, Wu ZL and Yao JJ: Hepatoprotective effect of Artemisia capillaris thumb flavones extract on chronic liver in-jury induced by carbon tetrachloride in rats. Med J Chin People's Armed Police Forces. 26:162–166. 2015.
22	Shang LZ, Wang F, Wang Q, et al: Effects and mechanism of chaihu shugan powder on TGF-β1/Smad signaling pathways in hepatic fibrosis model rats. Chin J Exp Tradit Med Formulae. 21:125–128. 2015.
23	Gao JR, Yao HP, Liu YW, et al: Turtle carapace decoction and drug containing serum on hepatic stellate cells. Chin Arch Tradit Chin Med. 31:2524–2528. 2013.
24	Zechini B, Pasquazzi C and Aceti A: Correlation of serum aminotransferases with HCV RNA levels and histological findings in patients with chronic hepatitis C: The role of serum aspartate transaminase in the evaluation of disease progression. Eur J Gastroenterol Hepatol. 16:891–896. 2004. View Article : Google Scholar : PubMed/NCBI
25	Pradat P, Alberti A, Poynard T, Esteban JI, Weiland O, Marcellin P, Badalamenti S and Trépo C: Predictive value of ALT levels for histologic findings in chronic hepatitis C: A European collaborative study. Hepatology. 36:973–977. 2002. View Article : Google Scholar : PubMed/NCBI
26	Natsume M, Tsuji H, Harada A, Akiyama M, Yano T, Ishikura H, Nakanishi I, Matsushima K, Kaneko S and Mukaida N: Attenuated liver fibrosis and depressed serum albumin levels in carbon tetrachloride-treated IL-6-deficient mice. J Leukoc Biol. 66:601–608. 1999.PubMed/NCBI
27	Hassan EM, El-Kherbawy GM, Ali MAM and Dewidar OM: The potential effect of special formulas on cirrhotic rats. Food Nutr Sci. 4:594–603. 2013. View Article : Google Scholar
28	Soon RK Jr and Yee HF Jr: Stellate cell contraction: Role, regulation, and potential therapeutic target. Clin Liver Dis. 12:791–803. viii2008. View Article : Google Scholar : PubMed/NCBI
29	Marra F: Hepatic stellate cells and the regulation of liver inflammation. J Hepatol. 31:1120–1130. 1999. View Article : Google Scholar : PubMed/NCBI
30	Kanzler S, Baumann M, Schirmacher P, Dries V, Bayer E, Gerken G, Dienes HP and Lohse AW: Prediction of progressive liver fibrosis in hepatitis C infection by serum and tissue levels of transforming growth factor-beta. J Viral Hepat. 8:430–437. 2001. View Article : Google Scholar : PubMed/NCBI
31	Date M, Matsuzaki K, Matsushita M, Tahashi Y, Furukawa F and Inoue K: Modulation of transforming growth factor beta function in hepatocytes and hepatic stellate cells in rat liver injury. Gut. 46:719–724. 2000. View Article : Google Scholar : PubMed/NCBI
32	Flanders KC: Smad3 as a mediator of the fibrotic response. Int J Exp Pathol. 85:47–64. 2004. View Article : Google Scholar : PubMed/NCBI
33	Latella G, Vetuschi A, Sferra R, Catitti V, D'Angelo A, Zanninelli G, Flanders KC and Gaudio E: Targeted disruption of Smad3 confers resistance to the development of dimethylnitrosamine-induced hepatic fibrosis in mice. Liver Int. 29:997–1009. 2009. View Article : Google Scholar : PubMed/NCBI
34	Dooley S, Hamzavi J, Breitkopf K, Wiercinska E, Said HM, Lorenzen J, Ten Dijke P and Gressner AM: Smad7 prevents activation of hepatic stellate cells and liver fibrosis in rats. Gastroenterology. 125:178–191. 2003. View Article : Google Scholar : PubMed/NCBI
35	Fu MY, He YJ, Lv X, Liu ZH, Shen Y, Ye GR, Deng YM and Shu JC: Transforming growth factor-β1 reduces apoptosis via autophagy activation in hepatic stellate cells. Mol Med Rep. 10:1282–1288. 2014.PubMed/NCBI