{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,1]],"date-time":"2026-05-01T00:54:47Z","timestamp":1777596887897,"version":"3.51.4"},"reference-count":46,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2019,8,26]],"date-time":"2019-08-26T00:00:00Z","timestamp":1566777600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Chang Gung Memorial Hospital Research Foundation, Taiwan","award":["CMRPG2F0141-2, CMRPG3A0661, CMRPG2B0281, and CMRPG2D0091-3"],"award-info":[{"award-number":["CMRPG2F0141-2, CMRPG3A0661, CMRPG2B0281, and CMRPG2D0091-3"]}]},{"name":"Ministry of Science and Technology, Taipei, Taiwan","award":["MOST 105-2314-B-182A-133"],"award-info":[{"award-number":["MOST 105-2314-B-182A-133"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The purpose of this study was to investigate whether Ger-Gen-Chyn-Lian-Tang (GGCLT) suppresses oxidative stress, inflammation, and angiogenesis during experimental liver fibrosis through the hypoxia-inducible factor-1\u03b1 (HIF-1\u03b1)-mediated pathway. Male C57BL\/6 mice were randomly assigned to a sham-control or bile duct ligation (BDL) group with or without treatment with GGCLT at 30, 100, and 300 mg\/kg. Plasma alanine aminotransferase (ALT) levels were analyzed using a diagnostic kit. Liver histopathology and hepatic status parameters were measured. Compared to control mice, the BDL mice exhibited an enlargement in liver HIF-1\u03b1 levels, which was suppressed by 100 and 300 mg\/kg GGCLT treatments (control: BDL: BDL + GGCLT-100: BDL + GGCLT-300 = 0.95 \u00b1 0.07: 1.95 \u00b1 0.12: 1.43 \u00b1 0.05: 1.12 \u00b1 0.10 fold; p &lt; 0.05). GGCLT restrained the induction of hepatic hydroxyproline and malondialdehyde levels in the mice challenged with BDL, further increasing the hepatic glutathione levels. Furthermore, in response to increased hepatic inflammation and fibrogenesis, significant levels of ALT, nuclear factor kappa B, transforming growth factor-\u03b2, \u03b1-smooth muscle actin, matrix metalloproteinase-2 (MMP-2), MMP-9, and procollagen-III were found in BDL mice, which were attenuated with GGCLT. In addition, GGCLT reduced the induction of angiogenesis in the liver after BDL by inhibiting vascular endothelial growth factor (VEGF) and VEGF receptors 1 and 2. In conclusion, the anti-liver fibrosis effect of GGCLT, which suppresses hepatic oxidative stress and angiogenesis, may be dependent on an HIF-1\u03b1-mediated pathway.<\/jats:p>","DOI":"10.3390\/ijms20174181","type":"journal-article","created":{"date-parts":[[2019,8,26]],"date-time":"2019-08-26T10:54:53Z","timestamp":1566816893000},"page":"4181","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Positive Effects of Ger-Gen-Chyn-Lian-Tang on Cholestatic Liver Fibrosis in Bile Duct Ligation-Challenged Mice"],"prefix":"10.3390","volume":"20","author":[{"given":"Zi-Yu","family":"Chang","sequence":"first","affiliation":[{"name":"Department of Traditional Chinese Medicine, Chang Gung Memorial Hospital, Keelung 204, Taiwan"},{"name":"Institute of Traditional Medicine, School of Medicine, National Yang-Ming University, Taipei 112, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0871-7191","authenticated-orcid":false,"given":"Chin-Chang","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Traditional Chinese Medicine, Chang Gung Memorial Hospital, Keelung 204, Taiwan"},{"name":"Research Center for Chinese Herbal Medicine, Chang Gung University of Science and Technology, Taoyuan 333, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hsuan-Miao","family":"Liu","sequence":"additional","affiliation":[{"name":"Graduate Institute of Traditional Chinese Medicine, Chang Gung University, Taoyuan 333, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9656-0269","authenticated-orcid":false,"given":"Yuan-Chieh","family":"Yeh","sequence":"additional","affiliation":[{"name":"Department of Traditional Chinese Medicine, Chang Gung Memorial Hospital, Keelung 204, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6495-7221","authenticated-orcid":false,"given":"Tung-Yi","family":"Lin","sequence":"additional","affiliation":[{"name":"Department of Traditional Chinese Medicine, Chang Gung Memorial Hospital, Keelung 204, Taiwan"},{"name":"Graduate Institute of Traditional Chinese Medicine, Chang Gung University, Taoyuan 333, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4097-2085","authenticated-orcid":false,"given":"Tzung-Yan","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Traditional Chinese Medicine, Chang Gung Memorial Hospital, Keelung 204, Taiwan"},{"name":"Institute of Traditional Medicine, School of Medicine, National Yang-Ming University, Taipei 112, Taiwan"},{"name":"Graduate Institute of Traditional Chinese Medicine, Chang Gung University, Taoyuan 333, Taiwan"},{"name":"School of Traditional Chinese Medicine, Chang Gung University, Taoyuan 333, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5231-9932","authenticated-orcid":false,"given":"Tse-Hung","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Traditional Chinese Medicine, Chang Gung Memorial Hospital, Keelung 204, Taiwan"},{"name":"Research Center for Chinese Herbal Medicine, Chang Gung University of Science and Technology, Taoyuan 333, Taiwan"},{"name":"Graduate Institute of Traditional Chinese Medicine, Chang Gung University, Taoyuan 333, Taiwan"},{"name":"School of Traditional Chinese Medicine, Chang Gung University, Taoyuan 333, Taiwan"},{"name":"School of Nursing, National Taipei University of Nursing and Health Sciences, Taipei 112, Taiwan"},{"name":"Graduate Institute of Health Industry Technology, Chang Gung University of Science and Technology, Taoyuan 333, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2019,8,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"S38","DOI":"10.1016\/S0168-8278(02)00429-4","article-title":"Liver fibrosis\u2014from bench to bedside","volume":"38","author":"Friedman","year":"2003","journal-title":"J. Hepatol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"539","DOI":"10.1172\/JCI30542","article-title":"Models of liver fibrosis: Exploring the dynamic nature of inflammation and repair in a solid organ","volume":"117","author":"Iredale","year":"2007","journal-title":"J. Clin. Invest."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"703","DOI":"10.1007\/s00109-002-0380-0","article-title":"Hypoxia-induced angiogenesis in human hepatocellular carcinoma","volume":"80","author":"Kim","year":"2002","journal-title":"J. Mol. Med."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"258","DOI":"10.1055\/s-0030-1255355","article-title":"Hypoxia: A link between fibrogenesis, angiogenesis, and carcinogenesis in liver disease","volume":"30","author":"Rosmorduc","year":"2010","journal-title":"Semin. Liver Dis."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"795","DOI":"10.1111\/j.1600-0463.2007.apm_610.x","article-title":"Hypoxia-inducible factor-1alpha expression in experimental cirrhosis: Correlation with vascular endothelial growth factor expression and angiogenesis","volume":"115","author":"Bozova","year":"2007","journal-title":"Apmis"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2267","DOI":"10.1093\/carcin\/bgn216","article-title":"Redox mechanisms switch on hypoxia-dependent epithelial-mesenchymal transition in cancer cells","volume":"29","author":"Cannito","year":"2008","journal-title":"Carcinogenesis"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"976","DOI":"10.1016\/j.jhep.2010.07.004","article-title":"Intrahepatic angiogenesis and sinusoidal remodeling in chronic liver disease: New targets for the treatment of portal hypertension?","volume":"53","author":"Thabut","year":"2010","journal-title":"J. Hepatol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1041","DOI":"10.1002\/hep.27318","article-title":"Portal myofibroblasts promote vascular remodeling underlying cirrhosis formation through the release of microparticles","volume":"61","author":"Lemoinne","year":"2015","journal-title":"Hepatology"},{"key":"ref_9","first-page":"3810","article-title":"Hypoxia promotes fibrogenesis in vivo via HIF-1 stimulation of epithelial-to-mesenchymal transition","volume":"117","author":"Higgins","year":"2007","journal-title":"J. Clin. Invest."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1016\/j.febslet.2006.12.010","article-title":"Hypoxia induces the activation of human hepatic stellate cells LX-2 through TGF-beta signaling pathway","volume":"581","author":"Shi","year":"2007","journal-title":"FEBS Lett."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"683","DOI":"10.1016\/j.cellsig.2010.12.002","article-title":"Expression and functional analysis of endoglin in isolated liver cells and its involvement in fibrogenic Smad signalling","volume":"23","author":"Meurer","year":"2011","journal-title":"Cell Signal."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"251","DOI":"10.1111\/j.1365-2613.2008.00597.x","article-title":"Comparison of three research models of portal hypertension in mice: macroscopic, histological and portal pressure evaluation","volume":"89","author":"Geerts","year":"2008","journal-title":"Int. J. Exp. Pathol."},{"key":"ref_13","first-page":"305","article-title":"Prolonged bile duct obstruction: A new experimental model for cirrhosis in the rat","volume":"65","author":"Kountouras","year":"1984","journal-title":"Br. J. Exp. Pathol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"G323","DOI":"10.1152\/ajpgi.00394.2009","article-title":"Macrophage-mediated phagocytosis of apoptotic cholangiocytes contributes to reversal of experimental biliary fibrosis","volume":"298","author":"Popov","year":"2010","journal-title":"Am. J. Physiol. Gastrointest. Liver Physiol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"316","DOI":"10.1006\/jsre.1994.1151","article-title":"The pathophysiology of biliary obstruction and its effect on phagocytic and immune function","volume":"57","author":"Grogan","year":"1994","journal-title":"J. Surg. Res."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"236","DOI":"10.1016\/j.surg.2005.04.001","article-title":"Altered Kupffer cell function in biliary obstruction","volume":"138","author":"Minter","year":"2005","journal-title":"Surgery"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1157","DOI":"10.1016\/S0016-5085(00)70369-6","article-title":"Bile duct ligation in rats induces biliary expression of cytokine-induced neutrophil chemoattractant","volume":"118","author":"Saito","year":"2000","journal-title":"Gastroenterology"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1590\/S0041-87812003000100006","article-title":"Cholestasis in a murine experimental model: lesions include hepatocyte ischemic necrosis","volume":"58","author":"Prado","year":"2003","journal-title":"Rev. Hosp. Cl\u00edn."},{"key":"ref_19","first-page":"e52438","article-title":"Bile duct ligation in mice: induction of inflammatory liver injury and fibrosis by obstructive cholestasis","volume":"96","author":"Tag","year":"2015","journal-title":"J. Vis. Exp."},{"key":"ref_20","first-page":"265","article-title":"Proliferation and phenotypic modulation of portal fibroblasts in the early stages of cholestatic fibrosis in the rat","volume":"74","author":"Tuchweber","year":"1996","journal-title":"Lab. Invest."},{"key":"ref_21","first-page":"765","article-title":"Extracellular matrix deposition, lysyl oxidase expression, and myofibroblastic differentiation during the initial stages of cholestatic fibrosis in the rat","volume":"76","author":"Darby","year":"1997","journal-title":"Lab. Invest."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1080\/08941939.2016.1183734","article-title":"HIF-1 alpha as a Key Factor in Bile Duct Ligation-Induced Liver Fibrosis in Rats","volume":"30","author":"Moczydlowska","year":"2016","journal-title":"J. Invest. Surg."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"G582","DOI":"10.1152\/ajpgi.90368.2008","article-title":"Reduced liver fibrosis in hypoxia-inducible factor-1alpha-deficient mice","volume":"296","author":"Moon","year":"2009","journal-title":"Am. J. Physiol. Gastrointest. Liver Physiol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1010","DOI":"10.1053\/jhep.2002.32524","article-title":"Hypoxia-induced VEGF and collagen I expressions are associated with angiogenesis and fibrogenesis in experimental cirrhosis","volume":"35","author":"Corpechot","year":"2002","journal-title":"Hepatology"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1124\/jpet.111.189340","article-title":"Hypoxia-inducible factor activation in myeloid cells contributes to the development of liver fibrosis in cholestatic mice","volume":"341","author":"Copple","year":"2012","journal-title":"J. Pharmacol. Exp. Ther."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1038\/sj.onc.1201912","article-title":"Coordinated induction of VEGF receptors in mesenchymal cell types during rat hepatic wound healing","volume":"17","author":"Matli","year":"1998","journal-title":"Oncogene"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1347","DOI":"10.1136\/gut.52.9.1347","article-title":"Vascular endothelial growth factor and receptor interaction is a prerequisite for murine hepatic fibrogenesis","volume":"52","author":"Yoshiji","year":"2003","journal-title":"Gut"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1942","DOI":"10.2353\/ajpath.2007.060887","article-title":"Proangiogenic cytokines as hypoxia-dependent factors stimulating migration of human hepatic stellate cells","volume":"170","author":"Novo","year":"2007","journal-title":"Am. J. Pathol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1038\/embor.2009.276","article-title":"Integrin-TGF-beta crosstalk in fibrosis, cancer and wound healing","volume":"11","author":"Margadant","year":"2010","journal-title":"EMBO Rep."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1016\/j.jep.2012.04.034","article-title":"Anti-atherosclerotic action of Ger-Gen-Chyn-Lian-Tang and AMPK-dependent lipid lowering effect in hepatocytes","volume":"142","author":"Ho","year":"2012","journal-title":"J. Ethnopharmacol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"360","DOI":"10.1016\/j.jcma.2014.04.009","article-title":"Hepatoprotective effects of Ger-Gen-Chyn-Lian-Tang in thioacetamide-induced fibrosis in mice","volume":"77","author":"Chang","year":"2014","journal-title":"J. Chin. Med. Assoc."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1016\/S0003-2670(01)00900-X","article-title":"Microwave-assisted extraction of effective constituents from a Chinese herbal medicine Radix puerariae","volume":"436","author":"Guo","year":"2001","journal-title":"Analytica. Chimica. Acta."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"471","DOI":"10.1016\/S0039-9140(00)00514-2","article-title":"Determination of baicalein, baicalin and quercetin in Scutellariae Radix and its preparations by capillary electrophoresis with electrochemical detection","volume":"53","author":"Chen","year":"2000","journal-title":"Talanta"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1616","DOI":"10.1080\/14786411003757937","article-title":"Quality control for Coptidis rhizoma through the determination of five alkaloids by HPLC\u2013ELSD coupled with chemometrics","volume":"24","author":"Kong","year":"2010","journal-title":"Nat. Prod. Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1016\/j.jchromb.2004.10.026","article-title":"Simultaneous determination of glycyrrhizin, a marker component in radix Glycyrrhizae, and its major metabolite glycyrrhetic acid in human plasma by LC\u2013MS\/MS","volume":"814","author":"Lin","year":"2005","journal-title":"J. Chromatogr. B"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1129","DOI":"10.1007\/s11033-008-9288-2","article-title":"Protective effect of puerarin on diabetic retinopathy in rats","volume":"36","author":"Teng","year":"2009","journal-title":"Mol. Biol. Rep."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"8153","DOI":"10.3390\/ijms15058153","article-title":"Baicalin inhibits hypoxia-induced pulmonary artery smooth muscle cell proliferation via the AKT\/HIF-1\u03b1\/p27-associated pathway","volume":"15","author":"Zhang","year":"2014","journal-title":"Int. J. Mol. Sci."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"57","DOI":"10.3109\/01480545.2011.589437","article-title":"Antiangiogenic activity of berberine is mediated through the downregulation of hypoxia-inducible factor-1, VEGF, and proinflammatory mediators","volume":"35","author":"Hamsa","year":"2012","journal-title":"Drug Chem. Toxicol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"591","DOI":"10.1111\/j.1872-034X.2011.00953.x","article-title":"Hepatitis C virus core protein upregulates the expression of vascular endothelial growth factor via the nuclear factor-kappaB\/hypoxia-inducible factor-1alpha axis under hypoxic conditions","volume":"42","author":"Abe","year":"2012","journal-title":"Hepatol. Res."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1011","DOI":"10.1200\/JCO.2005.06.081","article-title":"Role of the vascular endothelial growth factor pathway in tumor growth and angiogenesis","volume":"23","author":"Hicklin","year":"2005","journal-title":"J. Clin. Oncol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1729","DOI":"10.1053\/j.gastro.2008.07.065","article-title":"Hepatic stellate cells secrete angiopoietin 1 that induces angiogenesis in liver fibrosis","volume":"135","author":"Taura","year":"2008","journal-title":"Gastroenterology"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1339","DOI":"10.1002\/hep.20965","article-title":"Upregulation of proinflammatory and proangiogenic cytokines by leptin in human hepatic stellate cells","volume":"42","author":"Aleffi","year":"2005","journal-title":"Hepatology"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1046\/j.0001-6772.2003.01229.x","article-title":"Evidence for intestinal oxidative stress in obstructive jaundice-induced gut barrier dysfunction in rats","volume":"180","author":"Assimakopoulos","year":"2004","journal-title":"Acta. Physiol. Scand."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1097\/01.sla.0000149306.35717.8b","article-title":"Bombesin and neurotensin reduce endotoxemia, intestinal oxidative stress, and apoptosis in experimental obstructive jaundice","volume":"241","author":"Assimakopoulos","year":"2005","journal-title":"Ann. Surg."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"477","DOI":"10.1111\/j.1478-3231.2006.01245.x","article-title":"Relationship between oxidative stress levels and activation state on a hepatic stellate cell line","volume":"26","author":"Guimaraes","year":"2006","journal-title":"Liver Int."},{"key":"ref_46","first-page":"49","article-title":"Antipuretic, antimicrobic action of gegen qinlian tang","volume":"12","author":"Tong","year":"1987","journal-title":"Zhong Yao Tong Bao"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/20\/17\/4181\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T13:14:08Z","timestamp":1760188448000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/20\/17\/4181"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,8,26]]},"references-count":46,"journal-issue":{"issue":"17","published-online":{"date-parts":[[2019,9]]}},"alternative-id":["ijms20174181"],"URL":"https:\/\/doi.org\/10.3390\/ijms20174181","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,8,26]]}}}