Tanshinone ⅡA inhibits cardiac fibrosis

doi:10.3969/j.issn.2095-4344.2013.20.015

Abstract

Abstract:

BACKGROUND: Transforming growth factor β-Smads is the critical path of cardiac fibrosis, and tanshinone ⅡA can limit cardiac fibrosis.
OBJECTIVE: To explore the effect of tanshinone ⅡA on the cardiac fibrosis induced by transforming growth factor-β1 and the possible mechanisms.
METHODS: Cardiac fibroblasts were isolated from cardiac tissues of neonatal Sprague-Dawley rats by the trypsin digestion and differential adhesion method. Passage 3 cells were treated with 5 μg/L transforming growth factor β1 alone or pretreated with tanshinone ⅡA at different concentrations (10-5 mol/L and 10-4 mol/L). Immunocytochemistry was used for cell identification, reverse transcription-PCR for detection of the mRNA expression of connective tissue growth factor and type Ⅰ collagen, western blotting for detection of the protein expression of Smad7 and phosphorylated Smad3, and immunohistochemistry and immunofluorescence for detection of the protein expression of phosphorylated Smad3, connective tissue growth factor and type Ⅰ collagen.
RESULTS AND CONCLUSION: Transforming growth factor-β1 induced the expression of connective tissue growth factor, type Ⅰ collagen, phosphorylated Smads and Smad7 in a time-dependant manner. The mRNA expression of connective tissue growth factor and type Ⅰ collagen was significantly increased 24 hours after transforming growth factor-β1 stimulation (P < 0.01 for all). The protein expression of phosphorylated Smad3 and Smad7 reached a peak 1 hour after transforming growth factor-β1 stimulation, much higher than the baseline level (P < 0.01 for all). Pretreatment with high concentration of tanshinone ⅡA resulted in a decrease in the expression of phosphorylated Smad3, connective tissue growth factor and type Ⅰ collagen (P< 0.01). The protein expression of Smad7 was substantially upregulated after pretreatment with two concentrations of tanshinone ⅡA as compared with that in the cells at 2 hours post transforming growth factor-β1 stimulation (P < 0.05, P < 0.01). Tanshinone ⅡA may exert an inhibitory effect on cardiac fibrosis by upregulating the expression of Smad7, suppressing the transforming growth factor-β1-induced phosphorylated Smad3 and partially blocking the transforming growth factor-β1-Smads signaling pathway.

Key words: tissue construction, tissue construction and traditional Chinese medicine, tanshinone ⅡA, cardiac fibroblasts, cardiac fibrosis, transforming growth factor beta 1, Smads3, Smads7, connective tissue growth factor, type Ⅰ collagen, signaling pathway, cell culture, provincial grants-supported paper

CLC Number:

R318

Zhou Dai-xing, Li Zhi-hui, Zhan Cheng-ye, Zhang Li-wei. Tanshinone ⅡA inhibits cardiac fibrosis[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(20): 3715-3722.

Figures/Tables 8

References

[1] Baudino T A,Carver W, Giles W, et al. Cardiac ?broblasts: friend or foe?. Am J Physiol Heart Circ Physiol.2006; 291(3): H1015-H1026.

[2] Bai D, Gao Q, Li C, et al.A conserved TGFβ1/HuR feedback circuit regulates the fibrogenic response in fibroblasts.Int J Biochem Cell Biol.2012;44(6):1031-1039.

[3] Drobic V, Cunnington RH, Bedosky KM, et al. Differential and combined effects of cardiotrophin-1 and transforming growth factor-β1 on cardiac myo?broblast proliferation and contraction. Am J Physiol Heart Circ Physiol.2007;293(2): H1053-H1064.

[4] Shyu KG, Wang BW, Wu GJ,et al.Mechanical stretch via transforming growth factor-β1 activates microRNA208a to regulate endoglin expression in cultured rat cardiac myoblasts. Chin Med J (Engl).2012;125(12):2205-2212.

[5] Frank V, Alian M. Transforming growth factor-β1and fibrosis. World J Gastroenterol.2007;13(22): 3056-3062.

[6] Leask A.Potential therapeutic targets for cardiac fibrosis: TGF-beta,angiotenangiotensin,endothelin,CCN2,and PDGF,partners in fibroblast activation.Circ Res.2010;106( 11): 1675-1680．

[7] Kania G, Blyszczuk P, STEIN S, et al. Heart-Infiltrating Prominin-1(+)/CD133(+) Progenitor Cells Represent the Cellular Source of Transforming Growth Factor beta-Mediated Cardiac Fibrosis in Experimental Autoimmune Myocarditis. Circ Res.2009;105(5): 462,U151.

[8] Glazer NL, Macy EM, Lumley T, et al.Transforming growth factor beta-1 and incidence of heart failure in older adults: The Cardiovascular Health Study[J].Circulation.2012;126(7):840-850.

[9] Zhang DM,Qin Y,Niu FL,et al.Liaoning Zhongyi Zazhi. 2008; 35(12):1934-1936.张冬梅,秦英,牛福玲,等,丹参酮ⅡA对AngⅡ诱导的心肌成纤维细胞增殖及Ⅰ型胶原合成的影响[J]. 辽宁中医杂志, 2008, 35(12):1934-1936.

[10] Sun XW,Cao L,Yu GH,et al.Disan Junyi Daxue Xuebao. 2007;29(7):585-587.孙兴旺,曹灵,于国华,等. 丹参酮Ⅱ-A磺酸钠对纤维化人肾间质成纤维细胞体外增殖及cyclin E蛋白表达的影响[J]. 第三军医大学学报, 2007, 29(7):585-587.

[11] Dobaczewski M,Bujak M,Li N,et al．Smad3 signaling critically regulates fibroblast phenotype and function in healing myocardial infarction.Circ Res.2010;107( 3):418-428．

[12] Moustaka SA, Souchelnytskyi S, Heldin CK. Smad regulation in transforming growth factor-βsignal transduction. Cell Science. 2001;114(6): 4359-4369.

[13] Cunnington RH, Nazari M, Dixon MC. C-Ski, Smurf2, and Arkadia as regulators of transforming growth factor-β signaling: new targets for managing myofibroblast function and cardiac fibrosis. Can J Physiol and Pharma.2009;87(10):764-772.

[14] He JH, Chen YL, Chen BL, et al. B-type natriuretic peptide attenuates cardiac hypertrophy via the transforming growth factor-beta1/smad7 pathway in vivo and in vitro. Clin and Exp Pharma and Physiol.2010;37(3):283-289.

[15] Yang F, Huang XR, Chuang AC, et al. Essential role for Smad3 in angiotensin II-induced tubular epithelial-mesenchymal transition. Pathol. 2010;221(4): 390-401.