Calcium‑sensing receptor promotes high glucose‑induced myocardial fibrosis via upregulation of the TGF‑β1/Smads pathway in cardiac fibroblasts

Yuan,Hui; Fan,Yuqi; Wang,Yuehong; Gao,Tielei; Shao,Yiying; Zhao,Bingbing; Li,Hongzhu; Xu,Changqing; Wei,Can

doi:10.3892/mmr.2019.10330

Calcium‑sensing receptor promotes high glucose‑induced myocardial fibrosis via upregulation of the TGF‑β1/Smads pathway in cardiac fibroblasts

Authors:
- Hui Yuan
- Yuqi Fan
- Yuehong Wang
- Tielei Gao
- Yiying Shao
- Bingbing Zhao
- Hongzhu Li
- Changqing Xu
- Can Wei
View Affiliations

Affiliations: Department of Pathophysiology, Harbin Medical University, Harbin, Heilongjiang 150086, P.R. China
Published online on: June 4, 2019 https://doi.org/10.3892/mmr.2019.10330
Pages: 1093-1102
Copyright: © Yuan et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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

Diabetic cardiomyopathy (DCM) is a major complication of diabetes and myocardial fibrosis is its major pathological feature. Calcium‑sensing receptor (CaSR) is a G protein‑coupled receptor and participates in the regulation of calcium homeostasis; it is implicated in a range of diseases, including myocardial ischemia/reperfusion injury, myocardial infarction and pulmonary hypertension. However, whether CaSR is associated with myocardial fibrosis in DCM has remained elusive. In the present study, type 1 diabetic (T1D) rats and primary neonatal rat cardiac fibroblasts (CFs) were used to observe changes in CaSR to assess its potential as an indicator of myocardial fibrosis. The in vivo experiments revealed that in the T1D and CaSR agonist (R568) groups, evident collagen (Col)‑I and ‑III deposition was present after 12 weeks. Furthermore, the in vitro experiment indicated that the levels of transforming growth factor (TGF)‑β1, phosphorylated (p‑) protein kinase C, p‑p38, p‑Smad2, TβRI, TβRII, along with the intracellular Ca2+ levels and the content of TGF‑β1 in the culture medium were significantly increased in a high‑glucose (HG) group and an R568‑treated group. Treatment with the CaSR inhibitor Calhex231 significantly inhibited the abovementioned changes. Collectively, the results indicated that the increase of CaSR expression in CFs may induce intracellular Ca2+ increases and the activation of TGF‑β1/Smads, and enhance the proliferation of CFs, along with the excessive deposition of Col, resulting in myocardial fibrosis. The present results indicate an important novel mechanism for HG‑induced myocardial fibrosis and suggest that CaSR may be a promising potential therapeutic target for DCM.

View Figures

View References

1	Echouffo-Tcheugui JB and Dagogo-Jack S: Preventing diabetes mellitus in developing countries. Nat Rev Endocrinol. 8:557–562. 2012. View Article : Google Scholar : PubMed/NCBI
2	Wang X, McLennan SV, Allen TJ, Tsoutsman T, Semsarian C and Twigg SM: Adverse effects of high glucose and free fatty acid on cardiomyocytes are mediated by connective tissue growth factor. Am J Physiol Cell Physiol. 297:C1490–C1500. 2009. View Article : Google Scholar : PubMed/NCBI
3	Westermeier F, Riquelme JA, Pavez M, Garrido V, Díaz A, Verdejo HE, Castro PF, García L and Lavandero S: New Molecular Insights of Insulin in Diabetic Cardiomyopathy. Front Physiol. 7:1252016. View Article : Google Scholar : PubMed/NCBI
4	Fowlkes V, Clark J, Fix C, Law BA, Morales MO, Qiao X, Ako-Asare K, Goldsmith JG, Carver W, Murray DB, et al: Type II diabetes promotes a myofibroblast phenotype in cardiac fibroblasts. Life Sci. 92:669–676. 2013. View Article : Google Scholar : PubMed/NCBI
5	Cavalera M, Wang J and Frangogiannis NG: Obesity, metabolic dysfunction, and cardiac fibrosis: Pathophysiological pathways, molecular mechanisms, and therapeutic opportunities. Transl Res. 164:323–335. 2014. View Article : Google Scholar : PubMed/NCBI
6	Hutchinson KR, Lord CK, West TA and Stewart JA Jr: Cardiac fibroblast-dependent extracellular matrix accumulation is associated with diastolic stiffness in type 2 diabetes. PLoS One. 8:e720802013. View Article : Google Scholar : PubMed/NCBI
7	Tharmalingam S and Hampson DR: The Calcium-Sensing Receptor and Integrins in Cellular Differentiation and Migration. Front Physiol. 7:1902016. View Article : Google Scholar : PubMed/NCBI
8	Hendy GN and Canaff L: Calcium-Sensing Receptor Gene: Regulation of Expression. Front Physiol. 7:3942016. View Article : Google Scholar : PubMed/NCBI
9	Peng X, Li HX, Shao HJ, Li GW, Sun J, Xi YH, Li HZ, Wang XY, Wang LN, Bai SZ, et al: Involvement of calcium-sensing receptors in hypoxia-induced vascular remodeling and pulmonary hypertension by promoting phenotypic modulation of small pulmonary arteries. Mol Cell Biochem. 396:87–98. 2014. View Article : Google Scholar : PubMed/NCBI
10	Xu C, Zhang W, Jiang C, Sun Y and Wang R: Involvement of calcium sensing receptor in myocardial ischemia/reperfusion injury and apoptosis. J Mol Cell Cardiol. 42:S80–S81. 2007. View Article : Google Scholar
11	Wang Y, Gao P, Wei C, Li H, Zhang L, Zhao Y, Wu B, Tian Y, Zhang W, Wu L, et al: Calcium sensing receptor protects high glucose-induced energy metabolism disorder via blocking gp78-ubiquitin proteasome pathway. Cell Death Dis. 8:e27992017. View Article : Google Scholar : PubMed/NCBI
12	Dong S, Li G, Zheng D, Wu J, Sun D, Yang F, Yu X, Li T, Sun A, Liu J, et al: A novel role for the calcium sensing receptor in rat diabetic encephalopathy. Cell Physiol Biochem. 35:38–50. 2015. View Article : Google Scholar : PubMed/NCBI
13	Liang CC, Park AY and Guan JL: In vitro scratch assay: A convenient and inexpensive method for analysis of cell migration in vitro. Nat Protoc. 2:329–333. 2007. View Article : Google Scholar : PubMed/NCBI
14	Adeghate E: Molecular and cellular basis of the aetiology and management of diabetic cardiomyopathy: A short review. Mol Cell Biochem. 261:187–191. 2004. View Article : Google Scholar : PubMed/NCBI
15	Lam S, Verhagen NAM, Strutz F, van der Pijl JW, Daha MR and van Kooten C: Glucose-induced fibronectin and collagen type III expression in renal fibroblasts can occur independent of TGF-β1. Kidney Int. 63:878–888. 2003. View Article : Google Scholar : PubMed/NCBI
16	Spector KS: Diabetic cardiomyopathy. Clin Cardiol. 21:885–887. 1998. View Article : Google Scholar : PubMed/NCBI
17	Kehlet SN, Willumsen N, Armbrecht G, Dietzel R, Brix S, Henriksen K and Karsdal MA: Age-related collagen turnover of the interstitial matrix and basement membrane: Implications of age- and sex-dependent remodeling of the extracellular matrix. PLoS One. 13:e01944582018. View Article : Google Scholar : PubMed/NCBI
18	Russo I and Frangogiannis NG: Diabetes-associated cardiac fibrosis: Cellular effectors, molecular mechanisms and therapeutic opportunities. J Mol Cell Cardiol. 90:84–93. 2016. View Article : Google Scholar : PubMed/NCBI
19	Loboda A, Sobczak M, Jozkowicz A and Dulak J: TGF-β1/Smads and miR-21 in Renal Fibrosis and Inflammation. Mediators Inflamm. 2016:83192832016. View Article : Google Scholar : PubMed/NCBI
20	Yao M, Wang X, Wang X, Zhang T, Chi Y and Gao F: The Notch pathway mediates the angiotensin II-induced synthesis of extracellular matrix components in podocytes. Int J Mol Med. 36:294–300. 2015. View Article : Google Scholar : PubMed/NCBI
21	Olson ER: Signaling mechanisms controlling the proliferation and differentiation of cardiac fibroblasts (unpublished PhD thesis). Kent State University, College of Biomedical Sciences. 2006.
22	Zhang X, Zhang T, Wu J, Yu X, Zheng D, Yang F, Li T, Wang L, Zhao Y, Dong S, et al: Calcium sensing receptor promotes cardiac fibroblast proliferation and extracellular matrix secretion. Cell Physiol Biochem. 33:557–568. 2014. View Article : Google Scholar : PubMed/NCBI
23	Park S, Ranjbarvaziri S, Lay FD, Zhao P, Miller MJ, Dhaliwal JS, Huertas-Vazquez A, Wu X, Qiao R, Soffer JM, et al: Genetic Regulation of Fibroblast Activation and Proliferation in Cardiac Fibrosis. Circulation. 138:1224–1235. 2018. View Article : Google Scholar : PubMed/NCBI
24	Du G, Fischer BE, Voss KO, Becker G, Taucher-Scholz G, Kraft G and Thiel G: The absence of an early calcium response to heavy-ion radiation in Mammalian cells. Radiat Res. 170:316–326. 2008. View Article : Google Scholar : PubMed/NCBI
25	Liu W, Wang X, Mei Z, Gong J, Huang L, Gao X, Zhao Y, Ma J and Qian L: BNIP3L promotes cardiac fibrosis in cardiac fibroblasts through [Ca2+]i-TGF-β-Smad2/3 pathway. Sci Rep. 7:19062017. View Article : Google Scholar : PubMed/NCBI
26	Zhang WH, Fu SB, Lu FH, Wu B, Gong DM, Pan ZW, Lv YJ, Zhao YJ, Li QF, Wang R, et al: Involvement of calcium-sensing receptor in ischemia/reperfusion-induced apoptosis in rat cardiomyocytes. Biochem Biophys Res Commun. 347:872–881. 2006. View Article : Google Scholar : PubMed/NCBI
27	Li GW, Miao HZ, Bo L, Wang GZ, Jin L, Lin Y, Deng ZH and Xiao W: Calcium-sensing receptor modulates pulmonary artery tension through G-protein-PLC-IP_3 pathways. Chin J Pathophysiol. 31:2015.(In Chinese).
28	Jabłońska-Trypuć A, Matejczyk M and Rosochacki S: Matrix metalloproteinases (MMPs), the main extracellular matrix (ECM) enzymes in collagen degradation, as a target for anticancer drugs. J Enzyme Inhib Med Chem. 31 (Suppl 1):177–183. 2016. View Article : Google Scholar : PubMed/NCBI
29	Nüchel J, Ghatak S, Zuk AV, Illerhaus A, Mörgelin M, Schönborn K, Blumbach K, Wickström SA, Krieg T, Sengle G, et al: TGFB1 is secreted through an unconventional pathway dependent on the autophagic machinery and cytoskeletal regulators. Autophagy. 14:465–486. 2018. View Article : Google Scholar : PubMed/NCBI
30	Wei Y, Meng T and Sun C: Protective effect of diltiazem on myocardial ischemic rats induced by isoproterenol. Mol Med Rep. 17:495–501. 2018.PubMed/NCBI
31	Ryu JM, Lee MY, Yun SP and Han HJ: High glucose regulates cyclin D1/E of human mesenchymal stem cells through TGF-beta1 expression via Ca2+/PKC/MAPKs and PI3K/Akt/mTOR signal pathways. J Cell Physiol. 224:59–70. 2010.PubMed/NCBI
32	Shi BH, Zong-Pei XU and Fan GW: The research progress on treatment of myocardial fibrosis by regulating TGF-β1/Smads pathway. Zhongguo Yaolixue Tongbao. (In Press).
33	Lan HY: Diverse roles of TGF-β/Smads in renal fibrosis and inflammation. Int J Biol Sci. 7:1056–1067. 2011. View Article : Google Scholar : PubMed/NCBI
34	Duran J, Troncoso MF, Lagos D, Ramos S, Marin G and Estrada M: GDF11 Modulates Ca2+-Dependent Smad2/3 Signaling to Prevent Cardiomyocyte Hypertrophy. Int J Mol Sci. 19:1333–1342. 2018. View Article : Google Scholar :