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Published in: BMC Complementary Medicine and Therapies 1/2018

Open Access 01-12-2018 | Research article

Protective effects of Liuweiwuling tablets on carbon tetrachloride-induced hepatic fibrosis in rats

Authors: Huimin Liu, Zhenfang Zhang, Huangwanyin Hu, Congen Zhang, Ming Niu, Ruishen Li, Jiabo Wang, Zhaofang Bai, Xiaohe Xiao

Published in: BMC Complementary Medicine and Therapies | Issue 1/2018

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Abstract

Background

Liuweiwuling tablets (LWWL) are an herbal product that exerts remarkable effects on liver protection and aminotransferase levels, and they have been approved by the Chinese State Food and Drug Administration (CFDA). Clinical studies have found that LWWL can inhibit collagen production and reduce the levels of liver fibrosis markers in the serum. Thus, LWWL is expected to have beneficial effects in the treatment of liver fibrosis. The purpose of this study was to evaluate the pharmacological effects of LWWL.

Methods

Hepatic fibrosis was induced in rats via carbon tetrachloride (CCl4) treatment. The rats were treated twice weekly for 8 weeks with either 2 mL·kg− 1 body weight of a 50% solution of CCl4 in olive oil or olive oil alone by oral gavage. A subset of rats received daily intraperitoneal injections of either colchicine (0.2 mg/kg per day), LWWL (0.4, 1.6, or 6.4 g/kg per day), or vehicle (N = 12 for all groups) during weeks 9–12. The rats were sacrificed after 12 weeks. Pathological changes in hepatic tissue were examined using hematoxylin and eosin (H&E) and Sirius Red staining. Immunohistochemistry was performed to observe α-smooth muscle actin (α-SMA) and collagen type I (collagen I) protein expression. Western blotting was also used to detect α-SMA protein expression. Real-time quantitative reverse-transcription polymerase chain reaction (RT-qPCR) was used to detect transforming growth factor-1 (TGF-β1), platelet-derived growth factor (PDGF), tissue inhibitor of metalloproteinase-1 (TIMP1), and tissue inhibitor of metalloproteinase-2 (TIMP2) mRNA expression.

Results

LWWL significantly reversed histological fibrosis and liver injury, reduced the hydroxyproline content in liver tissue, and decreased α-SMA and collagen I expression. LWWL also suppressed hepatic stellate cell (HSC) activation by reducing the expression of the profibrogenic factors TGF-β1 and PDGF. The expression levels of TIMP1 and TIMP2, which regulate extracellular matrix (ECM) degradation, were decreased after CCl4 injury in LWWL-treated rats.

Conclusions

These data suggest that LWWL may serve as a promising therapeutic agent to reduce fibrogenesis.
Literature
1.
go back to reference Wree A, Eguchi A, Mcgeough MD, Pena CA, Johnson CD, Canbay A, et al. NLRP3 inflammasome activation results in hepatocyte pyroptosis, liver inflammation, and fibrosis in mice. Hepatology. 2014;59:898.CrossRefPubMed Wree A, Eguchi A, Mcgeough MD, Pena CA, Johnson CD, Canbay A, et al. NLRP3 inflammasome activation results in hepatocyte pyroptosis, liver inflammation, and fibrosis in mice. Hepatology. 2014;59:898.CrossRefPubMed
3.
go back to reference Yang C, Zeisberg M, Mosterman B, Sudhakar A, Yerramalla U, Holthaus K, et al. Liver fibrosis: insights into migration of hepatic stellate cells in response to extracellular matrix and growth factors. Gastroenterology. 2003;124:147–59.CrossRefPubMed Yang C, Zeisberg M, Mosterman B, Sudhakar A, Yerramalla U, Holthaus K, et al. Liver fibrosis: insights into migration of hepatic stellate cells in response to extracellular matrix and growth factors. Gastroenterology. 2003;124:147–59.CrossRefPubMed
4.
go back to reference Hemmann S, Graf J, Roderfeld M, Roeb E. Expression of MMPs and TIMPs in liver fibrosis-a systematic review with special emphasis on anti-fibrotic strategies. J Hepatol. 2007;46:955–75.CrossRefPubMed Hemmann S, Graf J, Roderfeld M, Roeb E. Expression of MMPs and TIMPs in liver fibrosis-a systematic review with special emphasis on anti-fibrotic strategies. J Hepatol. 2007;46:955–75.CrossRefPubMed
5.
go back to reference Schuppan D, Ruehl M, Somasundaram R, Hahn EG. Matrix as a modulator of hepatic Fibrogenesis. Semin Liver Dis. 2001;21:351–72.CrossRefPubMed Schuppan D, Ruehl M, Somasundaram R, Hahn EG. Matrix as a modulator of hepatic Fibrogenesis. Semin Liver Dis. 2001;21:351–72.CrossRefPubMed
6.
go back to reference Heldin CH, Miyazono K, Ten DP. TGF-beta signalling from cell membrane to nucleus through SMAD proteins. Nature. 1997;390:465–71.CrossRefPubMed Heldin CH, Miyazono K, Ten DP. TGF-beta signalling from cell membrane to nucleus through SMAD proteins. Nature. 1997;390:465–71.CrossRefPubMed
7.
go back to reference Liu B, Xuan Z, Zhang FC, Zong JB, Zhang W, Zhao Y. Aberrant TGF-β1 signaling contributes to the development of primary biliary cirrhosis in murine model. World J Gastroenterol. 2013;19:5828–36.CrossRefPubMedPubMedCentral Liu B, Xuan Z, Zhang FC, Zong JB, Zhang W, Zhao Y. Aberrant TGF-β1 signaling contributes to the development of primary biliary cirrhosis in murine model. World J Gastroenterol. 2013;19:5828–36.CrossRefPubMedPubMedCentral
8.
go back to reference Han YP, Zhou L, Wang J, Xiong S, Garner WL, French SW, et al. Essential role of matrix metalloproteinases in interleukin-1-induced myofibroblastic activation of hepatic stellate cell in collagen. J Biol Chem. 2004;279:4820–8.CrossRefPubMed Han YP, Zhou L, Wang J, Xiong S, Garner WL, French SW, et al. Essential role of matrix metalloproteinases in interleukin-1-induced myofibroblastic activation of hepatic stellate cell in collagen. J Biol Chem. 2004;279:4820–8.CrossRefPubMed
9.
10.
go back to reference Adachi T, Togashi H, Suzuki A, Kasai S, Ito J, Sugahara K, et al. NAD (P) H oxidase plays a crucial role in PDGF-induced proliferation of hepatic stellate cells. Hepatology. 2005;41:1272–81.CrossRefPubMed Adachi T, Togashi H, Suzuki A, Kasai S, Ito J, Sugahara K, et al. NAD (P) H oxidase plays a crucial role in PDGF-induced proliferation of hepatic stellate cells. Hepatology. 2005;41:1272–81.CrossRefPubMed
11.
go back to reference Zhang DY, Friedman SL. Fibrosis-dependent mechanisms of Hepatocarcinogenesis. Hepatology. 2012;56:769–75.CrossRefPubMed Zhang DY, Friedman SL. Fibrosis-dependent mechanisms of Hepatocarcinogenesis. Hepatology. 2012;56:769–75.CrossRefPubMed
12.
go back to reference Wu JL. Therapeutic effect of Liuweiwuling tablets on hepatic fibrosis. Chinese J Liver Dis. 2011;3:8–10. Wu JL. Therapeutic effect of Liuweiwuling tablets on hepatic fibrosis. Chinese J Liver Dis. 2011;3:8–10.
13.
go back to reference Zhao SF, Kan QC. Protective effects of Liuweiwulingpian on hepatic fibrosis induced by carbon tetrachloride in rats. Chin Pharmacol Bull. 2011;27:872–5. Zhao SF, Kan QC. Protective effects of Liuweiwulingpian on hepatic fibrosis induced by carbon tetrachloride in rats. Chin Pharmacol Bull. 2011;27:872–5.
14.
go back to reference Poo JL, Feldmann G, Moreau A, Gaudin C, Lebrec D. Early colchicine administration reduces hepatic fibrosis and portal hypertension in rats with bile duct ligation. J Hepatol. 1993;19:90–4.CrossRefPubMed Poo JL, Feldmann G, Moreau A, Gaudin C, Lebrec D. Early colchicine administration reduces hepatic fibrosis and portal hypertension in rats with bile duct ligation. J Hepatol. 1993;19:90–4.CrossRefPubMed
15.
go back to reference Nikolaidis N, Kountouras J, Giouleme O, Tzarou V, Chatzizisi O, et al. Colchicine treatment of liver fibrosis. Hepato-Gastroenterology. 2006;53:281–5.PubMed Nikolaidis N, Kountouras J, Giouleme O, Tzarou V, Chatzizisi O, et al. Colchicine treatment of liver fibrosis. Hepato-Gastroenterology. 2006;53:281–5.PubMed
16.
go back to reference Gressner AM, Weiskirchen R. Modern pathogenetic concepts of liver fibrosis suggest stellate cells and TGF-beta as major players and therapeutic targets. J Cell Mol Med. 2006;10:76–99.CrossRefPubMed Gressner AM, Weiskirchen R. Modern pathogenetic concepts of liver fibrosis suggest stellate cells and TGF-beta as major players and therapeutic targets. J Cell Mol Med. 2006;10:76–99.CrossRefPubMed
17.
go back to reference Stefanovic B, Stefanovic L, Schnabl B. TRAM2 protein interacts with endoplasmic reticulum Ca2+ pump Serca2b and is necessary for collagen type I synthesis. Mol Cell Biol. 2004;24:1758–68.CrossRefPubMedPubMedCentral Stefanovic B, Stefanovic L, Schnabl B. TRAM2 protein interacts with endoplasmic reticulum Ca2+ pump Serca2b and is necessary for collagen type I synthesis. Mol Cell Biol. 2004;24:1758–68.CrossRefPubMedPubMedCentral
19.
go back to reference Tahashi Y, Matsuzaki K, Date M, Yoshida K, Furukawa F, Sugano Y, et al. Differential regulation of TGF-beta signal in hepatic stellate cells between acute and chronic rat liver injury. Hepatology. 2002;35:49–61.CrossRefPubMed Tahashi Y, Matsuzaki K, Date M, Yoshida K, Furukawa F, Sugano Y, et al. Differential regulation of TGF-beta signal in hepatic stellate cells between acute and chronic rat liver injury. Hepatology. 2002;35:49–61.CrossRefPubMed
20.
go back to reference Fallowfield JA. Therapeutic targets in liver fibrosis. Am J Physiol Gastrointest Liver Physiol. 2011;300:G709–15.CrossRefPubMed Fallowfield JA. Therapeutic targets in liver fibrosis. Am J Physiol Gastrointest Liver Physiol. 2011;300:G709–15.CrossRefPubMed
21.
go back to reference Benyon RC, Iredale JP, Goddard S, Winwood PJ, Arthur MJ. Expression of tissue inhibitor of metalloproteinases 1 and 2 is increased in fibrotic human liver. Gastroenterology. 1996;110:821–31.CrossRefPubMed Benyon RC, Iredale JP, Goddard S, Winwood PJ, Arthur MJ. Expression of tissue inhibitor of metalloproteinases 1 and 2 is increased in fibrotic human liver. Gastroenterology. 1996;110:821–31.CrossRefPubMed
22.
go back to reference Iredale JP, Benyon RC, Arthur MJ, Ferris WF, Alcolado R, Winwood PJ, et al. Tissue inhibitor of metalloproteinase-1 messenger RNA expression is enhanced relative to interstitial collagenase messenger RNA in experimental liver injury and fibrosis. Hepatology. 1996;24:176–84.CrossRefPubMed Iredale JP, Benyon RC, Arthur MJ, Ferris WF, Alcolado R, Winwood PJ, et al. Tissue inhibitor of metalloproteinase-1 messenger RNA expression is enhanced relative to interstitial collagenase messenger RNA in experimental liver injury and fibrosis. Hepatology. 1996;24:176–84.CrossRefPubMed
23.
go back to reference Knittel T, Mehde M, Kobold D, Saile B, Dinter C, Ramadori G. Expression patterns of matrix metalloproteinases and their inhibitors in parenchymal and non-parenchymal cells of rat liver: regulation by TNF-alpha and TGF-beta1. J Hepatol. 1999;30:48–60.CrossRefPubMed Knittel T, Mehde M, Kobold D, Saile B, Dinter C, Ramadori G. Expression patterns of matrix metalloproteinases and their inhibitors in parenchymal and non-parenchymal cells of rat liver: regulation by TNF-alpha and TGF-beta1. J Hepatol. 1999;30:48–60.CrossRefPubMed
24.
go back to reference Wang Z, Juttermann R, Soloway PD. TIMP-2 is required for efficient activation of proMMP-2 in vivo. J Biol Chem. 2000;275:26411–5.CrossRefPubMed Wang Z, Juttermann R, Soloway PD. TIMP-2 is required for efficient activation of proMMP-2 in vivo. J Biol Chem. 2000;275:26411–5.CrossRefPubMed
25.
go back to reference Qian Q, Liu X, He W, An Y, Chen Q, Wu J, et al. TG accumulation inhibitory effects of Jinqi formula by AMPK signaling pathway. J Ethnopharmacol. 2012;143:41–8.CrossRefPubMed Qian Q, Liu X, He W, An Y, Chen Q, Wu J, et al. TG accumulation inhibitory effects of Jinqi formula by AMPK signaling pathway. J Ethnopharmacol. 2012;143:41–8.CrossRefPubMed
26.
go back to reference Du CY, Choi RC, Zheng KY, Dong TT, Lau DT, Tsim KW. Yu ping Feng san, an ancient Chinese herbal decoction containing Astragali Radix, Atractylodis Macrocephalae Rhizoma and Saposhnikoviae Radix, regulates the release of cytokines in murine macrophages. PLoS One. 2013;8:e78622.CrossRefPubMedPubMedCentral Du CY, Choi RC, Zheng KY, Dong TT, Lau DT, Tsim KW. Yu ping Feng san, an ancient Chinese herbal decoction containing Astragali Radix, Atractylodis Macrocephalae Rhizoma and Saposhnikoviae Radix, regulates the release of cytokines in murine macrophages. PLoS One. 2013;8:e78622.CrossRefPubMedPubMedCentral
27.
go back to reference Guo SG, Zhang W, Jiang T, Dai M, Zhang LF, Meng YC, et al. Influence of serum collected from rat perfused with compound Biejiaruangan drug on hepatic stellate cells. World J Gastroenterol. 2004;10:1487–94.CrossRefPubMedPubMedCentral Guo SG, Zhang W, Jiang T, Dai M, Zhang LF, Meng YC, et al. Influence of serum collected from rat perfused with compound Biejiaruangan drug on hepatic stellate cells. World J Gastroenterol. 2004;10:1487–94.CrossRefPubMedPubMedCentral
28.
go back to reference Wang QL, Yuan JL, Tao YY, Yue Z, Ping L, Liu CH. Fuzheng Huayu recipe and vitamin E reverse renal interstitial fibrosis through counteracting TGF-β1-induced epithelial-to-mesenchymal transition. J Ethnopharmacol. 2010;127:631–40.CrossRefPubMed Wang QL, Yuan JL, Tao YY, Yue Z, Ping L, Liu CH. Fuzheng Huayu recipe and vitamin E reverse renal interstitial fibrosis through counteracting TGF-β1-induced epithelial-to-mesenchymal transition. J Ethnopharmacol. 2010;127:631–40.CrossRefPubMed
29.
go back to reference Yang MD, Chiang YM, Higashiyama R, Asahina K, Mann DA, Mann J, et al. Rosmarinic acid and baicalin epigenetically de-repress Pparγ in hepatic stellate cells for their anti-fibrotic effect. Hepatology. 2012;55:1271–81.CrossRefPubMed Yang MD, Chiang YM, Higashiyama R, Asahina K, Mann DA, Mann J, et al. Rosmarinic acid and baicalin epigenetically de-repress Pparγ in hepatic stellate cells for their anti-fibrotic effect. Hepatology. 2012;55:1271–81.CrossRefPubMed
30.
go back to reference Zhang L, Schuppan D. Traditional Chinese medicine (TCM) for fibrotic liver disease: hope and hype. J Hepatol. 2014;61:166–8.CrossRefPubMed Zhang L, Schuppan D. Traditional Chinese medicine (TCM) for fibrotic liver disease: hope and hype. J Hepatol. 2014;61:166–8.CrossRefPubMed
31.
go back to reference Tan Y, Lv ZP, Bai XC, Liu XY, Zhang XF. Traditional Chinese medicine Bao Gan Ning increase phosphorylation of CREB in liver fibrosis in vivo and in vitro. J Ethnopharmacol. 2006;105(1–2):69–75.CrossRefPubMed Tan Y, Lv ZP, Bai XC, Liu XY, Zhang XF. Traditional Chinese medicine Bao Gan Ning increase phosphorylation of CREB in liver fibrosis in vivo and in vitro. J Ethnopharmacol. 2006;105(1–2):69–75.CrossRefPubMed
32.
go back to reference Hsieh CC, Fang HL, Lina WC. Inhibitory effect of Solanum nigrum on thioac etamide-induced liver fibrosis in mice. J Ethnopharmacol. 2008;119(1):117–21.CrossRefPubMed Hsieh CC, Fang HL, Lina WC. Inhibitory effect of Solanum nigrum on thioac etamide-induced liver fibrosis in mice. J Ethnopharmacol. 2008;119(1):117–21.CrossRefPubMed
34.
go back to reference Yang C, et al. Liver fibrosis: insights into migration of hepatic stellate cells in response to extracellular matrix and growth factors. Gastroenterology. 2003;124:147–59.CrossRefPubMed Yang C, et al. Liver fibrosis: insights into migration of hepatic stellate cells in response to extracellular matrix and growth factors. Gastroenterology. 2003;124:147–59.CrossRefPubMed
35.
go back to reference Ogawa K, Chen F, Kuang C, Chen Y. Suppression of matrix metalloproteinase-9 transcription by transforming growth factor-b is mediated by a nuclear factor-kB site. Biochem J. 2004;381:413–22.CrossRefPubMedPubMedCentral Ogawa K, Chen F, Kuang C, Chen Y. Suppression of matrix metalloproteinase-9 transcription by transforming growth factor-b is mediated by a nuclear factor-kB site. Biochem J. 2004;381:413–22.CrossRefPubMedPubMedCentral
36.
go back to reference Bissell DM, Wang S-S, Jarnagin WR, Roll FJ. Cell-specific expression of transforming growth factor-b in rat liver. Evidence for autocrine regulation of hepatocyte proliferation. J Clin Invest. 1995;96:447–55.CrossRefPubMedPubMedCentral Bissell DM, Wang S-S, Jarnagin WR, Roll FJ. Cell-specific expression of transforming growth factor-b in rat liver. Evidence for autocrine regulation of hepatocyte proliferation. J Clin Invest. 1995;96:447–55.CrossRefPubMedPubMedCentral
37.
38.
go back to reference Yan C, Boyd DD. Regulation of matrix metalloproteinase gene expression. J Cell Physiol. 2007;211:299–356.CrossRef Yan C, Boyd DD. Regulation of matrix metalloproteinase gene expression. J Cell Physiol. 2007;211:299–356.CrossRef
39.
go back to reference Krstic J, Santibanez JF. Transforming growth factor-beta and matrix metalloproteinases: functional interactions in tumor stroma-infiltrating myeloid cells. ScientificWorldJournal. 2014;2014:521754.CrossRefPubMedPubMedCentral Krstic J, Santibanez JF. Transforming growth factor-beta and matrix metalloproteinases: functional interactions in tumor stroma-infiltrating myeloid cells. ScientificWorldJournal. 2014;2014:521754.CrossRefPubMedPubMedCentral
40.
go back to reference Tang H, Gao L, Mao J, He H, Liu J, Cai X, Lin H, Wu T. Salidroside protects against bleomycin-induced pulmonary fibrosis: activation of Nrf2-antioxidant signaling, and inhibition of NF-κB and TGF-β1/Smad-2/−3 pathways. Cell Stress Chaperones. 2016;21:239–49.CrossRefPubMed Tang H, Gao L, Mao J, He H, Liu J, Cai X, Lin H, Wu T. Salidroside protects against bleomycin-induced pulmonary fibrosis: activation of Nrf2-antioxidant signaling, and inhibition of NF-κB and TGF-β1/Smad-2/−3 pathways. Cell Stress Chaperones. 2016;21:239–49.CrossRefPubMed
Metadata
Title
Protective effects of Liuweiwuling tablets on carbon tetrachloride-induced hepatic fibrosis in rats
Authors
Huimin Liu
Zhenfang Zhang
Huangwanyin Hu
Congen Zhang
Ming Niu
Ruishen Li
Jiabo Wang
Zhaofang Bai
Xiaohe Xiao
Publication date
01-12-2018
Publisher
BioMed Central
Published in
BMC Complementary Medicine and Therapies / Issue 1/2018
Electronic ISSN: 2662-7671
DOI
https://doi.org/10.1186/s12906-018-2276-8

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