Betulinic acid prevents high glucose‑induced expression of extracellular matrix protein in cardiac fibroblasts by inhibiting the TGF‑β1/Smad signaling pathway

Jiang,Ling; Chen,Feng‑Xia; Zang,Shu‑Ting; Yang,Qiao‑Fang

doi:10.3892/mmr.2017.7323

Betulinic acid prevents high glucose‑induced expression of extracellular matrix protein in cardiac fibroblasts by inhibiting the TGF‑β1/Smad signaling pathway

Authors:
- Ling Jiang
- Feng‑Xia Chen
- Shu‑Ting Zang
- Qiao‑Fang Yang
View Affiliations

Affiliations: Hypertension Department, Henan Provincial People's Hospital, Zhengzhou, Henan 450003, P.R. China, Department of Oncology, Henan Provincial People's Hospital, Zhengzhou, Henan 450003, P.R. China, Emergency Intensive Care Unit, Henan Provincial People's Hospital, Zhengzhou, Henan 450003, P.R. China, Department of Cardiology, Henan Provincial People's Hospital, Zhengzhou, Henan 450003, P.R. China
Published online on: August 22, 2017 https://doi.org/10.3892/mmr.2017.7323
Pages: 6320-6325

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The proliferation and differentiation of cardiac fibroblasts (CFs) is central to cardiac fibrosis. Betulinic acid (BA) is an active compound isolated from the bark of the birch tree Betula spp. (Betulaceae) and has been shown to attenuate hepatic fibrosis. However, the effect of BA on the high glucose‑induced fibrosis response in CFs remains to be elucidated, therefore, the present study investigated the effect of BA on high glucose‑induced CFs and examined the possible mechanism underlying the effect of BA on CF transdifferentiation. CFs were pre‑incubated with various concentrations of BA for 24 h and then stimulated with high glucose (25 mM) for various times. Cell proliferation was evaluated using an MTT assay. The mRNA expression levels of α‑smooth muscle actin (SMA) and transforming growth factor (TGF)‑β1 were determined using reverse transcription‑quantitative polymerase chain reaction analysis. The protein expression levels of α‑SMA, collagen I, collagen III, fibronectin, TGF‑β1, small mothers against decapentaplegic (Smad)2/3, phosphorylated (p)‑Smad2 and p‑Smad3 and were detected using western blot analysis. The data revealed that BA attenuated the CF proliferation and myofibroblast differentiation induced by high glucose. In addition, BA inhibited the expression of extracellular matrix (ECM) in the CFs induced by high glucose. It was also found that BA inhibited the high glucose‑induced phosphorylation of Smad2/3 in the CFs. Taken together, BA suppressed the high glucose‑induced increase in the proliferation of CFs and expression of ECM via inhibition of the TGF‑β1/Smad signaling pathway. Thus, BA may offer therapeutic potential towards the treatment of cardiac fibrosis.

View Figures

View References

1	Kong P, Christia P and Frangogiannis NG: The pathogenesis of cardiac fibrosis. Cell Mol Life Sci. 71:549–574. 2014. View Article : Google Scholar : PubMed/NCBI
2	Krenning G, Zeisberg EM and Kalluri R: The origin of fibroblasts and mechanism of cardiac fibrosis. J Cell Physiol. 225:631–637. 2010. View Article : Google Scholar : PubMed/NCBI
3	Teunissen BE, Smeets PJ, Willemsen PH, De Windt LJ, Van der Vusse GJ and Van Bilsen M: Activation of PPARdelta inhibits cardiac fibroblast proliferation and the transdifferentiation into myofibroblasts. Cardiovasc Res. 75:519–529. 2007. View Article : Google Scholar : PubMed/NCBI
4	Fan D, Takawale A, Lee J and Kassiri Z: Cardiac fibroblasts, fibrosis and extracellular matrix remodeling in heart disease. Fibrogenesis Tissue Repair. 5:152012. View Article : Google Scholar : PubMed/NCBI
5	Tian K, Liu Z, Wang J, Xu S, You T and Liu P: Sirtuin-6 inhibits cardiac fibroblasts differentiation into myofibroblasts via inactivation of nuclear factor κB signaling. Transl Res. 165:374–386. 2015. View Article : Google Scholar : PubMed/NCBI
6	Bugyei-Twum A, Advani A, Advani SL, Zhang Y, Thai K, Kelly DJ and Connelly KA: High glucose induces Smad activation via the transcriptional coregulator p300 and contributes to cardiac fibrosis and hypertrophy. Cardiovasc Diabetol. 13:892014. View Article : Google Scholar : PubMed/NCBI
7	Tang M, Zhang W, Lin H, Jiang H, Dai H and Zhang Y: High glucose promotes the production of collagen types I and III by cardiac fibroblasts through a pathway dependent on extracellular-signal-regulated kinase 1/2. Mol Cell Biochem. 301:109–114. 2007. View Article : Google Scholar : PubMed/NCBI
8	Kasperczyk H, La Ferla-Brühl K, Westhoff MA, Behrend L, Zwacka RM, Debatin KM and Fulda S: Betulinic acid as new activator of NF-κB: Molecular mechanisms and implications for cancer therapy. Oncogene. 24:6945–6956. 2005. View Article : Google Scholar : PubMed/NCBI
9	Yogeeswari P and Sriram D: Betulinic acid and its derivatives: A review on their biological properties. Curr Med Chem. 12:657–666. 2005. View Article : Google Scholar : PubMed/NCBI
10	Pavlova N, Savinova O, Nikolaeva S, Boreko E and Flekhter O: Antiviral activity of betulin, betulinic and betulonic acids against some enveloped and non-enveloped viruses. Fitoterapia. 74:489–492. 2003. View Article : Google Scholar : PubMed/NCBI
11	Xia A, Xue Z, Li Y, Wang W, Xia J, Wei T, Cao J and Zhou W: Cardioprotective effect of betulinic acid on myocardial ischemia reperfusion injury in rats. Evid Based Complement Alternat Med. 2014:5737452014. View Article : Google Scholar : PubMed/NCBI
12	Wan Y, Wu YL, Lian LH, Xie WX, Li X, Ouyang BQ, Bai T, Li Q, Yang N and Nan JX: The anti-fibrotic effect of betulinic acid is mediated through the inhibition of NF-κB nuclear protein translocation. Chem-Biol Interact. 195:215–223. 2012. View Article : Google Scholar : PubMed/NCBI
13	Turner NA, Das A, Warburton P, O'Regan DJ, Ball SG and Porter KE: Interleukin-1alpha stimulates proinflammatory cytokine expression in human cardiac myofibroblasts. Am J Physiol Heart Circ Physiol. 297:H1117–H1127. 2009. View Article : Google Scholar : PubMed/NCBI
14	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
15	Chen HN, Wang DJ, Ren MY, Wang QL and Sui SJ: TWEAK/Fn14 promotes the proliferation and collagen synthesis of rat cardiac fibroblasts via the NF-кB pathway. Mol Biol Rep. 39:8231–8241. 2012. View Article : Google Scholar : PubMed/NCBI
16	Driesen RB, Nagaraju CK, Abi-Char J, Coenen T, Lijnen PJ, Fagard RH, Sipido KR and Petrov VV: Reversible and irreversible differentiation of cardiac fibroblasts. Cardiovasc Res. 101:411–422. 2013. View Article : Google Scholar : PubMed/NCBI
17	Wang P, Li H, Wang Y, Chen H and Zhang P: Effects of recombinant human relaxin upon proliferation of cardiac fibroblast and synthesis of collagen under high glucose condition. J Endocrinol Invest. 32:242–247. 2009. View Article : Google Scholar : PubMed/NCBI
18	Neumann S, Huse K, Semrau R, Diegeler A, Gebhardt R, Buniatian GH and Scholz GH: Aldosterone and D-glucose stimulate the proliferation of human cardiac myofibroblasts in vitro. Hypertension. 39:756–760. 2002. View Article : Google Scholar : PubMed/NCBI
19	Zhou H, Zhang KX, Li YJ, Guo BY and Wang M and Wang M: Fasudil hydrochloride hydrate, a Rho-kinase inhibitor, suppresses high glucose-induced proliferation and collagen synthesis in rat cardiac fibroblasts. Clin Exp Pharmacol Physiol. 38:387–394. 2011. View Article : Google Scholar : PubMed/NCBI
20	Porter KE and Turner NA: Cardiac fibroblasts: At the heart of myocardial remodeling. Pharmacol Ther. 123:255–278. 2009. View Article : Google Scholar : PubMed/NCBI
21	Fedak PW, Bai L, Turnbull J, Ngu J, Narine K and Duff HJ: Cell therapy limits myofibroblast differentiation and structural cardiac remodeling basic fibroblast growth factor-mediated paracrine mechanism. Circ Heart Fail. 5:349–356. 2012. View Article : Google Scholar : PubMed/NCBI
22	Wang X, Gao G, Liu J, Guo R, Lin Y, Chu Y, Han F, Zhang W and Bai Y: Identification of differentially expressed genes induced by angiotensin II in rat cardiac fibroblasts. Clin Exp Pharmacol Physiol. 33:41–46. 2006. View Article : Google Scholar : PubMed/NCBI
23	Lim H and Zhu Y: Role of transforming growth factor-beta in the progression of heart failure. Cell Mol Life Sci. 63:2584–2596. 2006. View Article : Google Scholar : PubMed/NCBI
24	Bujak M and Frangogiannis NG: The role of TGF-beta signaling in myocardial infarction and cardiac remodeling. Cardiovasc Res. 74:184–195. 2007. View Article : Google Scholar : PubMed/NCBI
25	Kuwahara F, Kai H, Tokuda K, Kai M, Takeshita A, Egashira K and Imaizumi T: Transforming growth factor-beta function blocking prevents myocardial fibrosis and diastolic dysfunction in pressure-overloaded rats. Circulation. 106:130–135. 2002. View Article : Google Scholar : PubMed/NCBI
26	Singh VP, Baker KM and Kumar R: Activation of the intracellular renin-angiotensin system in cardiac fibroblasts by high glucose: Role in extracellular matrix production. Am J Physiol Heart Circ Physiol. 294:H1675–H1684. 2008. View Article : Google Scholar : PubMed/NCBI
27	Liu J, Zhuo X, Liu W, Wan Z, Liang X, Gao S, Yuan Z and Wu Y: Resveratrol inhibits high glucose induced collagen upregulation in cardiac fibroblasts through regulating TGF-β1-Smad3 signaling pathway. Chem Biol Interact. 227:45–52. 2015. View Article : Google Scholar : PubMed/NCBI