Search JIM Advanced Search

Journal of Chinese Integrative Medicine ›› 2012, Vol. 10 ›› Issue (10): 1155-.doi: 10.3736/jcim20121013

Previous Articles     Next Articles

Danshensu inhibits acetaldehyde-induced proliferation and activation of hepatic stellate cell-T6

Li Zhang1, Tao Wu2, Jun-ming Chen3, Li-li Yang1, Hai-yan Song1, Guang Ji1()   

  1. 1. Institute of Digestive Diseases, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China
    2. Center of Chinese Medicine Therapy and Systems Biology, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
    3. Department of Traditional Chinese Medicine, Shanghai Sixth People’s Hospital, Shanghai Jiaotong University, Shanghai 200233, China;
  • Received:2012-04-09 Accepted:2012-05-22 Online:2012-10-20 Published:2018-10-15
  • Contact: Guang Ji E-mail:jiliver@vip.sina.com

Objective: To evaluate the effects of danshensu, the main component of the extract of Chinese medicineSalvia miltiorrhiza, on the proliferation and activation of hepatic stellate cells (HSCs). .

Methods: The activation of HSC-T6 was induced by exposure to acetaldehyde. In the meantime, different doses of danshensu were added to the culture medium. After 24 h of treatment with danshensu in acetaldehyde, the viability of HSC-T6 cells was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, the cell cycle was determined through flow cytometry, and the gene transcription levels of plasminogen activator inhibitor-1 (PAI-1), transforming growth factor-β1 (TGF-β1), urokinase-type plasminogen activator (uPA) and matrix metalloproteinase-2 (MMP-2) were analyzed by real-time quantitative polymerase chain reaction.RESULTS: The proliferation of HSCs induced by 200 μmol/L acetaldehyde could be significantly inhibited by danshensu, and the percentage of HSCs in S phase was significantly increased as compared with the control cells (P<0.05), which were respectively evidenced by MTT assay and flow cytometry. Danshensu down-regulated the mRNA expression of TGF-β1 and PAI-1 and up-regulated the uPA transcription level (P<0.01), while the transcription level of MMP-2 was not significantly affected in HSC-T6.

Conclusion: Danshensu can inhibit the proliferation and activation of HSC-T6, as well as regulate some cytokines involved in extracellular matrix accumulation, which offers a potential therapeutic alternative for liver fibrosis.

Key words: danshensu, hepatocytes, cell proliferation, extracellular matrix, in vitro

"

Primer Primer sequence
PAI-1 Forward 5′-GTGGTTCGGCACAATCCAACAGAG-3′
Reverse 5′-GCAATGGAGGACGATACAAGAGGT-3′
uPA Forward 5′-AAGGTCTGGGATACTAGAGCC-3′
Reverse 5′-CAGGATATCCTCAGTCTCCACC-3′
TGF-β1 Forward 5′-GCCCTGGACACCAACTATTGC-3′
Reverse 5′-GCTGCACTTGCAGGAGCGCAC-3′
MMP-2 Forward 5′-ACTCCCACTTTGATGACGAC-3′
Reverse 5′-GAAGAGGAAAGGGAACTTGC-3′
GAPDH Forward 5′-GGTCGGAGTCAACGGATTTG-3′
Reverse 5′-ATGAGCCCCCAGCCTTCTCCA-3′

Figure 1

Viability of HSC-T6 cells affected by different doses of acetaldehyde and danshensuA: line graph showing the percentage of viable cells subjected to acetaldehyde (0, 100, 200, and 300 μmol/L); B: line graph showing the percentage of viable cells exposed to danshensu (0, 150, 250, 615 and 1 000 μmol/L) respectively. Values were measured in three independent experiments done in triplicate and mean ± standard error was plotted. HSC: hepatic stellate cell. "

Figure 2

Effects of Danshensu on the cell cycle of HSC-T6 HSC-T6 cultured with DMEM as control, and cells cultured with 200 μmol/L acetaldehyde as model. Cells were exposed to 200 μmol/L acetaldehyde with danshensu in different concentrations (100, 125 and 150 μmol/L) for 24 h. Cells were suspended, fixed and stained with PI, then were analyzed by flow cytometry. HSC: hepatic stellate cell; DMEM: Dulbecco’s Modified Eagle’s Medium; D: danshensu; PI: propidium iodide."

Figure 3

Effects of danshensu on PAI-1, TGF-β1, uPA and MMP-2 expressionGene transcription levels were measured by real-time quantitative polymerase chain reaction, with GAPDH expression level as the native control. Experiment for each sample was carried out in triplicate and data were expressed as mean±standard error. a: PAI-1 mRNA expression; b: TGF-β1 mRNA expression; c: cytokine uPA gene expression; d: cytokine MMP-2 gene expression. **P<0.01, vs control group; △△P<0.01, vs model group. Ace: acetaldehyde; D: danshensu; PAI-1: plasminogen activator inhibitor-1; uPA: urokinase-type plasminogen activator; MMP-2: matrix metalloproteinase-2; TGF-β1: transforming growth factor-β1; GAPDH: glyceraldehyde-3-phosphate dehydrogenase."

Figure 4

The chemical structure of danshensu"

[1] Chinese Association of Integrative Medicine. Guideline for the diagnosis and treatment of liver fibrosis with integrative medicine[J]. J Chin Integr Med, 2006,4(6):551-555
doi: 10.7661/CJIM.2006.11.1052
中国中西医结合学会肝病专业委员会. 肝纤维化中西医结合诊疗指南[J]. 中西医结合学报, 2006,4(6):551-555
doi: 10.7661/CJIM.2006.11.1052
[2] Borg BB, Seetharam A, Subramanian V, Basha HI, Lisker-Melman M, Korenblat K, Anderson CD, Shenoy S, Chapman WC, Crippin JS, Mohanakumar T . Immune response to extracellular matrix collagen in chronic hepatitis C-induced liver fibrosis[J]. Liver Transpl, 2011,17(7):814-823
doi: 10.1002/lt.22303 pmid: 3700355
[3] Marquez-Aguirre A, Sandoval-Rodriguez A, Gonzalez-Cuevas J, Bueno-Topete M, Navarro-Partida J, Arellano-Olivera I, Lucano-Landeros S, Armendariz-Borunda J . Adenoviral delivery of dominant-negative transforming growth factor beta type Ⅱ receptor up-regulates transcriptional repressor SKI-like oncogene, decreases matrix metalloproteinase 2 in hepatic stellate cell and prevents liver fibrosis in rats[J]. J Gene Med, 2009,11(3):207-219
doi: 10.1002/jgm.v11:3
[4] Peng WH, Tien YC, Huang CY, Huang TH, Liao JC, Kuo CL, Lin YC . Fraxinus rhynchophylla ethanol extract attenuates carbon tetrachloride-induced liver fibrosis in rats via down-regulating the expressions of uPA, MMP-2, MMP-9 and TIMP-1[J]. J Ethnopharmacol, 2010,127(3):606-613
doi: 10.1016/j.jep.2009.12.016
[5] Du L, Miao X, Jiang Y, Jia H, Tian Q, Shen J, Liu Y . An effective strategy for the synthesis of biocompatible gold nanoparticles using danshensu antioxidant: prevention of cytotoxicity via attenuation of free radical formation. Nanotoxicology. 2012 Feb 7. [Epub ahead of print]
[6] Zheng YY, Dai LL, Wang WB, Jia LP, Zhou X . Effect and mechanism of Tanshensu on fibrotic rats[J]. Zhonghua Gan Zang Bing Za Zhi, 2003,11(5):288-290
doi: 10.3760/j.issn:1007-3418.2003.05.009
郑元义, 戴立里, 王文兵, 贾丽萍, 周贤 . 丹参素治疗肝纤维化及其作用机制研究[J]. 中华肝脏病杂志, 2003,11(5):288-290
doi: 10.3760/j.issn:1007-3418.2003.05.009
[7] Sánchez-Muñoz D, Castellano-Megías VM, Romero-Gómez M . Expression of bcl-2 in ductular proliferation is related to periportal hepatic stellate cell activation and fibrosis progression in patients with autoimmune cholestasis[J]. Dig Liver Dis, 2007,39(3):262-266
doi: 10.1016/j.dld.2006.12.004
[8] González-Cuevas J, Navarro-Partida J, Marquez-Aguirre AL, Bueno-Topete MR, Beas-Zarate C, Armendáriz-Borunda J . Ethylenediaminetetraacetic acid induces antioxidant and anti-inflammatory activities in experimental liver fibrosis[J]. Redox Rep, 2011,16(2):62-70
doi: 10.1179/174329211X13002357050851
[9] Di Sario A, Candelaresi C, Omenetti A, Benedetti A . Vitamin E in chronic liver diseases and liver fibrosis[J]. Vitam Horm, 2007, 76551-573
doi: 10.1016/S0083-6729(07)76021-1 pmid: 17628189
[10] Liu J, Tan H, Sun Y, Zhou S, Cao J, Wang F . The preventive effects of heparin-superoxide dismutase on carbon tetrachloride-induced acute liver failure and hepatic fibrosis in mice[J]. Mol Cell Biochem, 2009,327(1-2):219-228
doi: 10.1007/s11010-009-0060-2
[11] Ren R, Wang T, Jiang N, Liu T, Du Y, Li C, Zhang L, Fu F . Protective effects of Danshensu on liver injury induced by omethoate in rats[J]. Toxicol Mech Methods, 2010,20(8):510-514
doi: 10.3109/15376516.2010.514963 pmid: 20843270
[12] Tian T, Xu LM . Effects of Salviae miltiorrhizae and salvianolic acid B on microcirculation of liver in mice with portal hypertension[J]. J Chin Integr Med, 2009,7(2):151-156
田甜, 徐列明 . 丹参及其组分丹参酚酸B盐对门静脉高压小鼠肝脏微循环的影响[J]. 中西医结合学报, 2009,7(2):151-156
[13] Zhang LJ, Chen L, Lu Y, Wu JM, Xu B, Sun ZG, Zheng SZ, Wang AY . Danshensu has anti-tumor activity in B16F10 melanoma by inhibiting angiogenesis and tumor cell invasion[J]. Eur J Pharmacol, 2010,643(2-3):195-201
doi: 10.1016/j.ejphar.2010.06.045
[14] Tang Y, Wang M, Chen C, Le X, Sun S, Yin Y . Cardiovascular protection with danshensu in spontaneously hypertensive rats[J]. Biol Pharm Bull, 2011,34(10):1596-601
doi: 10.1248/bpb.34.1596 pmid: 21963501
[15] Wang T, Fu F, Han B, Zhang L, Zhang X . Danshensu ameliorates the cognitive decline in streptozotocin-induced diabetic mice by attenuating advanced glycation end product-mediated neuroinflammation[J]. J Neuroimmunol, 2012,245(1-2):79-86
doi: 10.1016/j.jneuroim.2012.02.008
[16] Li G, Gao Y, Li S, Li C, Zhu X, Li M, Liu Z . Study on toxicity of danshensu in beagle dogs after 3-month continuous intravenous infusion[J]. Toxicol Mech Methods, 2009,19(6-7):441-446
doi: 10.1080/15376510903150623 pmid: 19778246
[17] Potter JJ, Mezey E . Acetaldehyde increases endogenous adiponectin and fibrogenesis in hepatic stellate cells but exogenous adiponectin inhibits fibrogenesis[J]. Alcohol Clin Exp Res, 2007,31(12):2092-2100
doi: 10.1111/acer.2007.31.issue-12
[18] Ha MH, Wei L, Rao HY, Liu F, Wang XY, Feng B, Fei R, Chen HS, Cong X . Effect of interferon-γ on hepatic stellate cells stimulated by acetaldehyde[J]. Hepatogastroenterology, 2008,55(84):1059-1065
doi: 10.1002/1529-0131(199801)41:13.0.CO;2-4 pmid: 18705328
[19] Wu ZH, Dai LL, Yu BB, Li D . Effect of danshensu on JNK and NF-κB signal transduction of rat hepatic stellate cells induced by interleukin-1 β[J]. Nan Fang Yi Ke Da Xue Xue Bao, 2009,29(5):914-917
doi: 10.3321/j.issn:1673-4254.2009.05.021
吴宗辉, 戴立里, 余斌斌, 李冬 . 丹参素对IL-1β刺激的大鼠肝星状细胞JNK、NF-κB信号通道的影响[J]. 南方医科大学学报, 2009,29(5):914-917
doi: 10.3321/j.issn:1673-4254.2009.05.021
[20] Cheng K, Yang N, Mahato RI . TGF-β1 gene silencing for treating liver fibrosis[J]. Mol Pharm, 2009,6(3):772-779
doi: 10.1021/mp9000469 pmid: 1607348
[21] Wang H, Zhang Y, Heuckeroth RO . PAI-1 deficiency reduces liver fibrosis after bile duct ligation in mice through activation of tPA[J]. FEBS Lett, 2007,581(16):3098-3104
doi: 10.1016/j.febslet.2007.05.049
[22] Salas AL, Montezuma TD, Fariña GG, Reyes-Esparza J, Rodríguez-Fragoso L . Genistein modifies liver fibrosis and improves liver function by inducing uPA expression and proteolytic activity in CCl4-treated rats[J]. Pharmacology, 2008,81(1):41-49
doi: 10.1159/000107968
[23] Galli A, Svegliati-Baroni G, Ceni E, Milani S, Ridolfi F, Salzano R, Tarocchi M, Grappone C, Pellegrini G, Benedetti A, Surrenti C, Casini A . Oxidative stress stimulates proliferation and invasiveness of hepatic stellate cells via a MMP2-mediated mechanism[J]. Hepatology, 2005,41(5):1074-1084
doi: 10.1002/(ISSN)1527-3350
[1] Sherman Gu, Arthur Yin Fan. Controversial conclusions from two randomized controlled trials for acupuncture's effects on polycystic ovary syndrome or in vitro fertilization support. Journal of Integrative Medicine, 2020, 18(2): 89-91.
[2] Sana Zaki, Nasreen Jahan, Mohd Kalim, Ghausia Islam. In vitro antilithiatic activity of the hydro-alcoholic extract of Cinnamomum zeylanicum Blume bark on calcium oxalate crystallization. Journal of Integrative Medicine, 2019, 17(4): 273-281.
[3] Syed Nasir Abbas Bukhari, Fahad Hussain, Hnin Ei Thu, Zahid Hussain. Synergistic effects of combined therapy of curcumin and Fructus Ligustri Lucidi for treatment of osteoporosis: cellular and molecular evidence of enhanced bone formation. Journal of Integrative Medicine, 2019, 17(1): 38-45.
[4] Sahar Dehdari, Homa Hajimehdipoor, Somayeh Esmaeili, Rasool Choopani, Seyed Alireza Mortazavi. Traditional and modern aspects of hemorrhoid treatment in Iran: A review. Journal of Integrative Medicine, 2018, 16(2): 90-98.
[5] Yu-fan Shen, Xue Zhang, Ying Wang, Fan-fan Cao, Georges Uzan, Bin Peng, Deng-hai Zhang. Celastrol targets IRAKs to block Toll-like receptor 4-mediated nuclear factor-κB activation. Journal of Integrative Medicine, 2016, 14(3): 203-208.
[6] Qing-qing Zang, Lu Zhang, Ning Gao, Cheng Huang. Ophiopogonin D inhibits cell proliferation, causes cell cycle arrest at G2/M, and induces apoptosis in human breast carcinoma MCF-7 cells. Journal of Integrative Medicine, 2016, 14(1): 51-59.
[7] Mukherjee Avinaba, Boujedaini Naoual, Rahman Khuda-Bukhsh Anisur. Homeopathic Thuja 30C ameliorates benzo(a)pyrene-induced DNA damage, stress and viability of perfused lung cells of mice in vitro. Journal of Chinese Integrative Medicine, 2013, 11(6): 397-404.
[8] Dao-fang Ding, Xiao-feng Li, Hao Xu, Zhen Wang, Qian-qian Liang, Chen-guang Li, Yong-jun Wang. Mechanism of resveratrol on the promotion of induced pluripotent stem cells. Journal of Chinese Integrative Medicine, 2013, 11(6): 389-396.
[9] Yong Xiao, Zeng-chun Ma, Yu-guang Wang, Hong-ling Tan, Xiang-ling Tang, Qian-de Liang, Cheng-rong Xiao, Yue Gao. Cardioprotection of Shenfu preparata on cardiac myocytes through cytochrome P450 2J3. Journal of Chinese Integrative Medicine, 2013, 11(5): 327-336.
[10] Bolleddula Jayaprakasam, Nan Yang, Ming-Chun Wen, Rong Wang, Joseph Goldfarb, Hugh Sampson, Xiu-Min Li. Constituents of the anti-asthma herbal formula ASHMITM synergistically inhibit IL-4 and IL-5 secretion by murine Th2 memory cells, and eotaxin by human lung fibroblasts in vitro. Journal of Chinese Integrative Medicine, 2013, 11(3): 195-205.
[11] Hong Wang , Jing-tian Ou , Xin Zhao , Ying Guoz , Hao-ping Mao. Vasodilator effect of oroxylin A on thoracic aorta isolated from rats. Journal of Chinese Integrative Medicine, 2012, 10(8): 880-885.
[12] Yathiraj Ashwini, Mohamed Rafiq, Gollapalle L. Viswanatha, Subbanna Rajesh, Selvam Senthilraja, Mohammed Azeemuddin, Suryakanth D. Anturlikar, Paramesh Rangesh, Pralhad S. Patki . Polyherbal formulation Bresol® protects the mast cells against compound 48/80-induced disruption and histamine release: A non-immunological mechanism of mast cell stabilization. Journal of Chinese Integrative Medicine, 2012, 10(6): 690-694.
[13] Dao-fang Ding , Song-pu Wei, Xiao-feng Li , Xiao-gang Zhang , Hong-sheng Zhan , Tie-li Duan , Yue-long Cao. Inhibition effect of osthole on proliferation of rat chondrocytes. Journal of Chinese Integrative Medicine, 2012, 10(12): 1413-1418.
[14] Mukherjee Avinaba, Sikdar Sourav, Bishayee Kausik, Paul Avijit, Ghosh Samrat, Boujedaini Naoual, Rahman Khuda-Bukhsh Anisur. Ethanolic extract of Thuja occidentalis blocks proliferation of A549 cells and induces apoptosis in vitro. Journal of Chinese Integrative Medicine, 2012, 10(12): 1451-1459.
[15] Arul Raj Chinthamony, Sophia Dominic, Ragavendran Paramasivam, Starlin Thangarajan, Ahalliya Rathi Muthian, Kanniappan Gopalakrishnan Velliyur. Leaf extract of Alpinia purpurata (Vieill.) K. Schum screened for its phytochemical constituents and antibacterial and anticancer activities. Journal of Chinese Integrative Medicine, 2012, 10(12): 1460-1464.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
[1] Dong Yang, Yong-ping Du, Qing Shen, Wei Chen, Yan Yu, Guang-lei Chen. Expression of alpha-smooth muscle actin in renal tubulointerstitium in patients with kidney collateral stasis. Journal of Chinese Integrative Medicine, 2008, 6(1): 41-44
[2] Hai-feng Wei, Bai-liu Ya, Ling Zhao, Cui-fei Ye, Li Zhang, Lin Li. Evaluation of tongue manifestation of blood stasis syndrome and its relationship with blood rheological disorder in a rat model of transient brain ischemia. Journal of Chinese Integrative Medicine, 2008, 6(1): 73-76
[3] Xi Lin, Jian-ping Liu. Herbal medicines for viral myocarditis. Journal of Chinese Integrative Medicine, 2008, 6(1): 76
[4] Xi Lin, Jian-ping Liu. Tai chi for treating rheumatoid arthritis. Journal of Chinese Integrative Medicine, 2008, 6(1): 82
[5] Liang-ping Hu, Hui Gao. Discrimination of errors in statistical analysis of medical papers published in the first issue of 2006 in Journal of Chinese Integrative Medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 98-106
[6] Yan-bo Zhu , Qi Wang, Cheng-yu Wu, Guo-ming Pang, Jian-xiong Zhao, Shi-lin Shen, Zhong-yuan Xia , Xue Yan . Logistic regression analysis on relationships between traditional Chinese medicine constitutional types and overweight or obesity. Journal of Chinese Integrative Medicine, 2010, 8(11): 1023-1035
[7] Wei Xu, Meng Shi, Jian-gang Liu, Cheng-long Wang . Collagen protein expressions in ischemic myocardium of rats with acute myocardial infarction and effects of qi-tonifying, yin-tonifying and blood-activating herbs and detoxifying and blood-activating herbs. Journal of Chinese Integrative Medicine, 2010, 8(11): 1041-1047
[8] Tao Wang , Feng Qin. Effects of Chinese herbal medicine Xiaoyao Powder on monoamine neurotransmitters in hippocampus of rats with postpartum depression. Journal of Chinese Integrative Medicine, 2010, 8(11): 1075-1079
[9] Ying Xu , Chang-chun Zeng , Xiu-yu Cai , Rong-ping Guo , Guang Nie , Ying Jin. Chromaticity and optical spectrum colorimetry of the tongue color in different syndromes of primary hepatic carcinoma. Journal of Chinese Integrative Medicine, 2012, 10(11): 1263-1271
[10] Xiang-ying Mao , Qin Bian , Zi-yin Shen. Analysis of the osteogenetic effects exerted on mesenchymal stem cell strain C3H10T1/2 by icariin via MAPK signaling pathway in vitro. Journal of Chinese Integrative Medicine, 2012, 10(11): 1272-1278