TY - JOUR
T1 - Klotho Modulates Pro-Fibrotic Activities in Human Atrial Fibroblasts through Inhibition of Phospholipase C Signaling and Suppression of Store-Operated Calcium Entry
AU - Hung, Yuan
AU - Chung, Cheng Chih
AU - Chen, Yao Chang
AU - Kao, Yu Hsun
AU - Lin, Wei Shiang
AU - Chen, Shih Ann
AU - Chen, Yi Jen
N1 - Funding Information:
Funding: This work was supported by grants from the Ministry of Science and Technology (MOST 105-2314-B-281-004-MY2, MOST 105-2314-B-038-059-MY3, MOST 108-2314-B-016-049, MOST 108-2314-B-016-049, MOST 109-2314-B-016-001-MY2, MOST 109-2314-B-038-124-MY3, MOST 110-2314-B-038-107-MY3, and MOST 110-2314-B-016-037-MY3), Taipei Medical University-Wan Fang Hospital (105-swf-02, 105-wf-eva-08, 105-wf-eva-14, 106-eva-02, 106-eva-06, and 106-swf-01), Tri-Service General Hospital (TSGH-C108-008-S01, TSGH-C02-108019, TSGH-C02-109019, TSGH-C01-110013, TSGH-C05-111040-03, and TSGH-E-111197), and the National Defense Medical Center, Taiwan (MAB-107-044).
Publisher Copyright:
© 2022 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2022/7
Y1 - 2022/7
N2 - Background: Atrial fibroblasts activation causes atrial fibrosis, which is one major patho-physiological contributor to atrial fibrillation (AF) genesis. Klotho is a pleiotropic protein with remarkable cardiovascular effects, including anti-inflammatory, anti-oxidative, and anti-apoptotic effects. This study investigated whether Klotho can modulate the activity of human atrial fibroblasts and provides an anti-fibrotic effect. Methods: Cell migration assay and proliferation assay were used to investigate fibrogenesis activities in single human atrial fibroblasts with or without treatment of Klotho (10 and 100 pM, 48 h). Calcium fluorescence imaging, the whole-cell patch-clamp, and Western blotting were performed in human atrial fibroblasts treated with and without Klotho (100 pM, 48 h) to evaluate the store-operated calcium entry (SOCE), transient receptor potential (TRP) currents, and downstream signaling. Results: High dose of Klotho (100 pM, 48 h) significantly reduced the migration of human atrial fibroblasts without alternating their proliferation; in addition, treatment of Klotho (100 pM, 48 h) also decreased SOCE and TRP currents. In the presence of BI-749327 (a selective canonical TRP 6 channel inhibitor, 1 µM, 48 h), Klotho (100 pM, 48 h) could not inhibit fibroblast migration nor suppress the TRP currents. Klotho-treated fibroblasts (100 pM, 48 h) had lower phosphorylated phospholipase C (PLC) (p-PLCβ3 Ser537) expression than the control. The PLC inhibitor, U73122 (1 µM, 48 h), reduced the migration, decreased SOCE and TRP currents, and lowered p-PLCβ3 in atrial fibroblasts, similar to Klotho. In the presence of the U73122 (1 µM, 48 h), Klotho (100 pM, 48 h) could not further modulate the migration and collagen synthesis nor suppress the TRP currents in human atrial fibroblasts. Conclusions: Klotho inhibited pro-fibrotic activities and SOCE by inhibiting the PLC signaling and suppressing the TRP currents, which may provide a novel insight into atrial fibrosis and arrhythmogenesis.
AB - Background: Atrial fibroblasts activation causes atrial fibrosis, which is one major patho-physiological contributor to atrial fibrillation (AF) genesis. Klotho is a pleiotropic protein with remarkable cardiovascular effects, including anti-inflammatory, anti-oxidative, and anti-apoptotic effects. This study investigated whether Klotho can modulate the activity of human atrial fibroblasts and provides an anti-fibrotic effect. Methods: Cell migration assay and proliferation assay were used to investigate fibrogenesis activities in single human atrial fibroblasts with or without treatment of Klotho (10 and 100 pM, 48 h). Calcium fluorescence imaging, the whole-cell patch-clamp, and Western blotting were performed in human atrial fibroblasts treated with and without Klotho (100 pM, 48 h) to evaluate the store-operated calcium entry (SOCE), transient receptor potential (TRP) currents, and downstream signaling. Results: High dose of Klotho (100 pM, 48 h) significantly reduced the migration of human atrial fibroblasts without alternating their proliferation; in addition, treatment of Klotho (100 pM, 48 h) also decreased SOCE and TRP currents. In the presence of BI-749327 (a selective canonical TRP 6 channel inhibitor, 1 µM, 48 h), Klotho (100 pM, 48 h) could not inhibit fibroblast migration nor suppress the TRP currents. Klotho-treated fibroblasts (100 pM, 48 h) had lower phosphorylated phospholipase C (PLC) (p-PLCβ3 Ser537) expression than the control. The PLC inhibitor, U73122 (1 µM, 48 h), reduced the migration, decreased SOCE and TRP currents, and lowered p-PLCβ3 in atrial fibroblasts, similar to Klotho. In the presence of the U73122 (1 µM, 48 h), Klotho (100 pM, 48 h) could not further modulate the migration and collagen synthesis nor suppress the TRP currents in human atrial fibroblasts. Conclusions: Klotho inhibited pro-fibrotic activities and SOCE by inhibiting the PLC signaling and suppressing the TRP currents, which may provide a novel insight into atrial fibrosis and arrhythmogenesis.
KW - atrial fibrillation
KW - fibroblast
KW - Klotho
KW - transient receptor potential channels
UR - http://www.scopus.com/inward/record.url?scp=85133705517&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85133705517&partnerID=8YFLogxK
U2 - 10.3390/biomedicines10071574
DO - 10.3390/biomedicines10071574
M3 - Article
AN - SCOPUS:85133705517
SN - 2227-9059
VL - 10
JO - Biomedicines
JF - Biomedicines
IS - 7
M1 - 1574
ER -