Open Access

P21‑activated kinase 1 mediates angiotensin II‑induced differentiation of human atrial fibroblasts via the JNK/c‑Jun pathway

  • Authors:
    • Yafei Zhou
    • Ying Xie
    • Tao Li
    • Peng Zhang
    • Tangting Chen
    • Zhongcai Fan
    • Xiaoqiu Tan
  • View Affiliations

  • Published online on: January 18, 2021     https://doi.org/10.3892/mmr.2021.11846
  • Article Number: 207
  • Copyright: © Zhou et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Cardiac fibrosis is a common pathophysiological condition involved in numerous types of cardiovascular disease. The renin‑angiotensin system, particularly angiotensin II (AngII), serves an important role in cardiac fibrosis and remodeling. Furthermore, p21‑activated kinase 1 (PAK1) is a highly conserved serine/threonine protein kinase, which is abundantly expressed in all regions of the heart. However, the role of PAK1 in AngII‑mediated activation of cardiac fibroblasts remains unknown. Therefore, the present study aimed to investigate the role of PAK1 in cardiac fibroblasts and its underlying mechanisms. Human cardiac fibroblasts (HCFs) were cultured and treated with PAK1 inhibitor IPA‑3 or transduced with PAK1 short hairpin (sh)RNA by lentiviral particles to silence PAK1 expression levels. Subsequently, the cell proliferation and migration abilities of the HCFs were determined. Western blot analysis was used to detect the phosphorylation status of Janus kinase (JNK) and c‑Jun. A Cell Counting Kit‑8 assay showed that PAK1 inhibition following treatment of HCFs with 5 µM IPA‑3 or PAK1‑shRNA, significantly attenuated AngII‑induced proliferation of fibroblasts. In addition, wound healing and Transwell migration assays demonstrated that inhibition of PAK1 significantly inhibited AngII‑induced cell migration. Finally, decreased PAK1 expression levels downregulated AngII‑mediated upregulation of α‑smooth muscle actin (α‑SMA), collagen I, phosphorylated (p)‑JNK and p‑c‑Jun, a downstream molecule of JNK signaling. These findings indicate that PAK1 contributes to AngII‑induced proliferation, migration and transdifferentiation of HCFs via the JNK/c‑Jun pathway.

Introduction

Cardiac fibrosis is a common pathophysiological condition and a key step in the occurrence and maintenance of the majority of cardiovascular diseases, including atrial fibrillation (AF), hypertension and heart failure (1,2). Fibrosis promotes systolic and diastolic dysfunction, as well as heart rhythm disturbances. Therefore, suppression of cardiac fibrosis may impede cardiac remodeling progression and may be a therapeutic target for AF and other types of cardiovascular disease (3).

Cardiac fibrosis is a complicated process associated with myocardial infarction, pressure and/or volume overload, mechanical stress, metabolic dysfunction and aging (46). One of the most important cellular and molecular mechanisms of cardiac fibrosis is differentiation of fibroblasts into myofibroblasts, which is characterized by the expression of secretory and contractile proteins, such as α-smooth muscle actin (α-SMA) and matrix metalloproteinases (MMPs), and production of large amounts of extracellular matrix (ECM) components, such as collagen I and III (1,7). Activated myofibroblasts regulate matrix and cardiomyocyte remodeling. It has been reported that activation of the renin-angiotensin system (RAS) is involved in the pathogenesis of cardiac fibrosis remodeling (2,8,9). Furthermore, studies have shown that angiotensin II (AngII) increases cardiac fibroblast proliferation and migration, as well as collagen expression levels (10,11). Previous studies have suggested that a variety of cell signaling pathways, such as TGF-β and mitogen-activated protein kinases (MAPKs) pathways, are involved in the differentiation, proliferation and migration of cardiac fibroblasts (1,1214). TGF-β-activated kinase and downstream p38 MAPK, ERK1/2 and Janus kinase (JNK) pathways are associated with AngII- or TGF-β-mediated activation of proliferation and migration of cardiac fibroblasts (15). Therefore, the inhibition of AngII-mediated activation of myofibroblasts may be an important strategy for treating cardiac fibrosis.

P21-activated kinases (PAKs) are a group of serine/threonine protein kinases that are activated by cell division cycle 42 and Rac family small GTPase 1. The PAK family consists of six members, namely PAK1-6 (1618). PAK1, 2 and 3 are abundantly expressed in the human heart and PAK1 is involved in actin-based cytoskeletal remodeling (19). Previous studies have shown that PAK1 contributes to TGF-β-induced cancer cell proliferation, migration and invasion (2023); these effects have also been reported in cardiomyocytes. Therefore, it has been demonstrated that PAK1 serves a key role in protection against cardiac hypertrophy (24). However, the role of PAK1 in cardiac fibroblasts has not been fully investigated. Therefore, the present study aimed to investigate the role of PAK1 in cardiac fibroblasts and the underlying mechanisms.

Materials and methods

Chemicals and reagents

IPA-3, a direct non-ATP-competitive PAK1 inhibitor, was purchased from MedChemExpress (cat. no. HY-15663). AngII was obtained from Sigma-Aldrich (cat. no. 4474-91-3; Merck KGaA).

RNA interference

The short hairpin (sh)RNA lentiviral particle package for PAK1 interference (PAK1-shRNA) was purchased from Shanghai Jikai Gene Chemical Technology Co., Ltd. Human cardiac fibroblasts (HCFs) were cultured to 60–70% confluence and then transduced with shRNA package (1×106 cells transduced with 5 µl lentivirus) according to the manufacturer's instructions. Following transduction for 48–72 h, the cells were used for subsequent experiments. The knocked-down mRNA and protein levels of PAK1 were determined using reverse transcription-quantitative PCR (RT-qPCR) and western blot analysis, respectively. The target sequence of PAK1-shRNA was 5′-CCGCTTGCTTCAAACATCAAA-3′. The sequence of negative control (NC)-shRNA (scrambled control) was 5′-TTCTCCGAACGTGTCACGT-3′.

Cell culture

Adult HCFs and fibroblast medium-2 were purchased from ScienCell Research Laboratories, Inc. (cat. no. 6320; sciencellonline.com/human-cardiac-fibroblasts-juvenile-atrial.html). Cell passage did not exceed 7–10 generations.

Cell migration assay

Fibroblast migration ability was measured using wound healing and Transwell migration assays. Briefly, fibroblasts were cultured to 60–70% confluence in 3.5-cm diameter dishes. Cells were starved of serum for 12 h, then divided into the following groups: i) Control; ii) AngII; iii) IPA-3; and iv) AngII + IPA-3 or i) NC; ii) AngII; iii) PAK1-shRNA; and iv) AngII + PAK1-shRNA for wound healing and Transwell migration assay. The multiplicity of infection of PAK1-shRNA package by lentiviral particles in the present study was difficult to determine because human adult cardiac fibroblasts can proliferate. In order to ensure the efficiency of virus interference, confluence of the cells was decreased for PAK1-shRNA silencing. In order to determine the wound area, a thin scratch (wound) was made in the central area of the dish using 1-ml pipette tips. Following washing with fibroblast medium-2 to remove detached cells, fibroblasts were cultured with fresh medium with 5% fetal bovine serum. Images of the scratch wounds were captured with a light microscope (magnification, ×4) at 0 and 24 h, and the wound width was calculated using the following formula: (T0 - T24/T0) × 100%, where T0 and T24 indicated the width of the wound area at the start time (0 h) and end point (24 h), respectively. For the Transwell migration assay, fibroblasts (density, 3×104) were rehydrated in 100 µl fibroblast medium-2 supplemented with 0.1% bovine serum albumin. Following digestion and resuspension in serum-free fibroblast medium-2, fibroblasts were placed into a Costar Transwell chamber with 8.0-µm pore size membrane (Corning, Inc.). Fibroblasts were seeded into 6-well plates containing 200 µl fibroblast medium-2 supplemented with 5% fetal bovine serum at 37°C with 5% CO2 for 12 h. Subsequently, fibroblasts on the inner side of the membrane were removed with a cotton bud; migrated cells on the outer side of the membrane were stained with 0.5% crystal violet for 10 min at room temperature, then observed with a light microscope (magnification, ×4) and counted manually.

Cell proliferation assay

CCK-8 kit was used to analyze cell proliferation in the present study (25). HCFs (density, 1×104 cells/well) were cultured in 96-well plates and grown to 50% confluence at 37°C with 5% CO2. Subsequently, the cells were cultured in serum-free fibroblast medium-2 for 12 h followed by treatment with 1 µM AngII in the presence or absence of 5 µM IPA-3 for 24 h at 37°C with 5% CO2. Following incubation for 24 h, the cells were treated with 10 µl CCK-8 solution (Nanjing Jiangcheng Bioengineering Institute) at 37°C with 5% CO2 for 4 h according to the manufacturer's instructions. Finally, the optical density of each well at a wavelength of 450 nm was measured using a Spectramax M5 microplate reader (Molecular Devices, LLC).

RT-qPCR

Total RNA from cells was isolated with TRIzol (Qiagen China Co., Ltd.) and reverse transcribed into cDNA using the ReverTra Ace qPCR RT Master Mix (Toyobo Life Science) as follows: 37°C for 15 min; 50°C for 5 min; and 98°C for 5 min. Subsequently, the expression levels of PAK1 in PAK1-shRNA-treated cells were quantified by qPCR using SYBR Green (Qiagen China Co., Ltd.) as follows: 95°C for 10 sec; 52°C for 10 sec; and 72°C for 10 sec for 40 cycles. The following primer pairs were used: PAK1: Forward, 5′-TCCGCCAGATGCTTTGACCC-3′ and reverse, 5′-AGCCTCCAGCCAAGTATTCC-3′; and GAPDH: Forward, 5′-GTGGACCTGACCTGCCGTCT-3′ and reverse, 5′-GGAGGAGTGGGTGTCGCTGT-3′. The expression levels of PAK1 were normalized to those of GAPDH and calculated using the 2−ΔΔCq method (26).

Western blot analysis

In order to determine the protein expression levels, cells were lysed with ice-cold RIPA buffer (Beyotime Institute of Biotechnology) supplemented with protease and phosphatase inhibitor cocktail (Thermo Fisher Scientific, Inc.), and the protein concentration was measured via the bicinchoninic acid method. Cell lysate (30 µg) was resolved by 5% SDS-PAGE and transferred onto PVDF membranes. Following blocking for 2 h with 5% non-fat dry milk in TBS-T [20.00 mM Tris (pH 8.0), 150.00 mM NaCl, 0.05% Tween-20] at room temperature, membranes were incubated with primary antibodies against α-SMA, collagen І (both 1:2,000; cat. nos. GB11044 and GB11022, respectively; both Servicebio, Inc.); JNK, phosphorylated (p)-JNK; ERK, p-ERK; p38, p-p38 (all 1:1,000; cat. nos. 9252, 9255, 4696, 4370, 8690 and 4511, respectively; all Cell Signaling Technology, Inc.) and GAPDH (1:5,000; cat. no. G9295; Sigma-Aldrich; Merck KGaA) at 4°C overnight. The PVDF membrane was then incubated with horseradish peroxidase (HRP)-conjugated anti-mouse and anti-rabbit IgG and secondary antibodies (both 1:2,000; cat. nos. 7076 and 7074, respectively; both Cell Signaling Technology, Inc.) at room temperature for 1 h and treated with Chemiluminescent HRP Substrate (cat. no. WBKLS0500; EMD Millipore). Images were captured and immunoreactive bands were quantified using Quantity One software v4.6.6 (Bio-Rad Laboratories, Inc.).

Statistical analysis

Data are expressed as the mean ± SEM. Results were analyzed by one-way ANOVA followed by Tukey's posthoc comparison. P<0.05 was considered to indicate a statistically significant difference.

Results

IPA-3-mediated inhibition of PAK1 attenuates AngII-induced migration and transdifferentiation of HCFs

The effect of PAK1 on AngII-induced migration of HCFs was first investigated. Growth and migration of HCFs were decreased using a PAK1 inhibitor, IPA-3 (concentration, 5 µM) or PAK1-shRNA, respectively (Figs. 1 and 2). Exposure of HCFs to 1 µM AngII for 24 h increased their migration ability, as indicated by Transwell and wound healing migration assays (Fig. 1A and C). Compared with the control group, treatment of HCFs with AngII significantly increased wound closure (P<0.05; n=5) and cell migration rate (P<0.01; n=3; Fig. 1B and D). Furthermore, co-treatment of AngII-treated HCFs with 5 µM IPA-3 decreased wound closure (P<0.01, n=5) and relative cell migration rate (P<0.01; n=3) compared with the AngII-treated group (Fig. 1B and D).

Collagen I and α-SMA were used as markers of transdifferentiation of fibroblasts into myofibroblasts. Treatment of HCFs with AngII significantly upregulated α-SMA (P<0.05, n=4) and collagen I (P<0.01, n=3) expression levels, whereas co-treatment with IPA-3 downregulated AngII-mediated upregulation of both proteins (P<0.01; Fig. 3A-C). AngII also significantly increased the expression levels of PAK1 in fibroblasts (Fig. 3D and E). Finally, CCK-8 assay revealed that IPA-3 significantly inhibited AngII-induced cell proliferation (P<0.05, n=8; Fig. 3F).

PAK1 knockdown attenuates AngII-induced migration and transdifferentiation of HCFs

Subsequently, the effect of PAK1-knockdown (PAK1-shRNA) on AngII-induced migration and transdifferentiation of HCFs was investigated. Interference rate of PAK1-shRNA is presented in Fig. 4A-C. PAK1-shRNA significantly decreased the mRNA (Fig. 4A) and protein levels (Fig. 4B and C) of PAK1 in HCFs. Similar results were observed between HCFs exposed to IPA-3 and HCFs transduced with PAK1-shRNA. Transwell and wound healing migration assays and western blot analysis demonstrated that PAK1 knockdown significantly inhibited AngII-induced wound closure (Fig. 2A and B) and cell migration (Fig. 2C and D) and PAK1 knockdown significantly (Fig. 4A-C) decreased the expression of collagen I and α-SMA (Fig. 4D-F), respectively.

JNK/c-Jun pathway is involved in the effects of PAK1 on HCFs

Furthermore, the underlying signaling pathway associated with the effect of PAK1 on fibroblasts was investigated. The results showed that the JNK/c-Jun pathway is involved in the inhibitory effect of PAK1 on AngII-induced migration and transdifferentiation of HCFs (Fig. 5). Treatment of HCFs with AngII increased phosphorylation of JNK and its downstream molecule c-Jun, whereas PAK1 inhibition with IPA-3 (Fig. 5D-F) or PAK1-shRNA (Fig. 5A-C) significantly attenuated the effect of AngII. Knockdown of PAK1 with PAK1-shRNA significantly decreased AngII-induced phosphorylation of JNK and c-Jun (Fig. 5A-C). Similarly, IPA-3 significantly decreased AngII-induced phosphorylation of JNK and c-Jun (Fig. 5D-F).

Discussion

Activation of myofibroblasts is a key step in cardiac fibrosis (1,6,27). The present study showed that p21-activated kinase 1 (PAK1) contributed to angiotensin II (AngII)-mediated activation of myofibroblasts, whereas its inhibition significantly attenuated the effects of AngII on human cardiac fibroblasts (HCFs) (Fig. 5G).

Activation of the renin-angiotensin system (RAS), particularly AngII, is involved in numerous types of cardiovascular disease by promoting cardiac fibroblast differentiation into myofibroblasts (28). Therefore, in the present study AngII-activated myofibroblasts were used to investigate the underlying mechanism of PAK1 in HCFs.

The activation of myofibroblasts is associated with overexpression of α-smooth muscle actin (α-SMA) and collagen I and II (29). Consistent with previous studies, the present study demonstrated that treatment of HCFs with AngII significantly upregulated α-SMA and collagen I expression levels (8,30). Furthermore, inhibition/downregulation of PAK1 activity using IPA-3 inhibitor or shRNA interference, respectively, attenuated AngII-mediated overexpression of α-SMA and collagen I. These findings indicate that PAK1 may be involved in AngII-mediated differentiation of HCFs. The present study also found that treatment with IPA-3 or PAK1-shRNA in the absence of AngII inhibited the migration and proliferation of fibroblasts. These data indicated that PAK1 was involved in the basic properties of fibroblasts, such as proliferation and migration.

The activation of myofibroblasts is characterized by increased cell migration and proliferation ability. Activation of RAS or upregulation of AngII increase cardiac fibroblast migration and proliferation (31). Herein, Transwell and wound healing migration assays also confirmed the effect of AngII on the migration ability of HCFs. Additionally, inhibition or downregulation of PAK1 significantly decreased AngII-induced migration and proliferation of HCFs.

It has been documented that numerous signaling pathways are involved in AngII-mediated activation of myofibroblasts, such as the TGFβ1/Smad and MAPK pathways (14,32). However, the mechanism underlying the effect of PAK1 in AngII-activated HCFs remains unknown. c-Jun is an important downstream molecule of the JNK pathway and promotes the proliferation of fibroblasts (33). The results of the present study demonstrated that the JNK/c-Jun pathway mediated the effects of PAK1 on AngII-induced myofibroblasts.

To the best of our knowledge, the present study is the first to demonstrate that PAK1 contributes to AngII-mediated activation of myofibroblasts via the JNK/c-Jun pathway (Fig. 5G). The effect of PAK1 on fibroblasts was different from that observed in cardiomyocytes in previous study (20,34). Therefore, it was hypothesized that PAK1 downregulation may decrease the AngII-induced differentiation, proliferation and migration of myofibroblasts, thus providing a novel approach in preventing cardiac fibrosis. In the present study, the role of PAK1 was investigated only in vitro using an HCF cell line; therefore, in vivo studies using a PAK1-knockout mouse model should be performed to further investigate the role of PAK1 in myofibroblasts.

Acknowledgements

Not applicable.

Funding

The present study was supported by the National Natural Science Foundation of China (grant nos. 31300948 and 81670310) and the Outstanding Youth Foundation of Sichuan province of China (grant no. 2020JDJQ0047).

Availability of data and materials

The datasets generated and/or analyzed during the current study are available from the corresponding author on reasonable request.

Authors' contributions

XT, YZ and ZF designed the experiment. YZ, YX, TL, and PZ performed the experiments. XT, YZ, YX and TC performed the data analysis. XT, YZ and ZF wrote the manuscript. All authors read and approved the final manuscript.

Ethics approval and consent to participate

Not applicable.

Patient consent for publication

Not applicable.

Competing interests

The authors declare that they have no competing interests.

References

1 

Frangogiannis NG: Cardiac fibrosis: Cell biological mechanisms, molecular pathways and therapeutic opportunities. Mol Aspects Med. 65:70–99. 2019. View Article : Google Scholar : PubMed/NCBI

2 

Prabhu SD and Frangogiannis NG: The Biological Basis for Cardiac Repair After Myocardial Infarction: From Inflammation to Fibrosis. Circ Res. 119:91–112. 2016. View Article : Google Scholar : PubMed/NCBI

3 

Ma ZG, Yuan YP, Wu HM, Zhang X and Tang QZ: Cardiac fibrosis: New insights into the pathogenesis. Int J Biol Sci. 14:1645–1657. 2018. View Article : Google Scholar : PubMed/NCBI

4 

Humeres C and Frangogiannis NG: Fibroblasts in the Infarcted, Remodeling, and Failing Heart. JACC Basic Transl Sci. 4:449–467. 2019. View Article : Google Scholar : PubMed/NCBI

5 

Ji Y, Qiu M, Shen Y, Gao L, Wang Y, Sun W, Li X, Lu Y and Kong X: MicroRNA-327 regulates cardiac hypertrophy and fibrosis induced by pressure overload. Int J Mol Med. 41:1909–1916. 2018.PubMed/NCBI

6 

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

7 

Weber KT: Fibrosis in hypertensive heart disease: Focus on cardiac fibroblasts. J Hypertens. 22:47–50. 2004. View Article : Google Scholar : PubMed/NCBI

8 

Frangogiannis NG: Cardiac fibrosis: Cell biological mechanisms, molecular pathways and therapeutic opportunities. Mol Aspects Med. 65:70–99. 2019. View Article : Google Scholar : PubMed/NCBI

9 

Ma H, Kong J, Wang YL, Li JL, Hei NH, Cao XR, Yang JJ, Yan WJ, Liang WJ, Dai HY and Dong B: Angiotensin-converting enzyme 2 overexpression protects against doxorubicin-induced cardiomyopathy by multiple mechanisms in rats. Oncotarget. 8:24548–24563. 2017. View Article : Google Scholar : PubMed/NCBI

10 

Chen T, Li M, Fan X, Cheng J and Wang L: Sodium Tanshinone IIA Sulfonate Prevents Angiotensin II–Induced Differentiation of Human Atrial Fibroblasts into Myofibroblasts. Oxid Med Cell Longev. 2018:67125852018. View Article : Google Scholar : PubMed/NCBI

11 

Dong X, Yu S, Wang Y, Yang M, Xiong J, Hei N, Dong B, Su Q and Chen J: (Pro)renin receptor-mediated myocardial injury, apoptosis, and inflammatory response in rats with diabetic cardiomyopathy. J Biol Chem. 294:8218–8226. 2019. View Article : Google Scholar : PubMed/NCBI

12 

Li R, Xiao J, Qing X, Xing J, Xia Y, Qi J, Liu X, Zhang S, Sheng X, Zhang X, et al: Sp1 Mediates a Therapeutic Role of MiR-7a/b in Angiotensin II–Induced Cardiac Fibrosis via Mechanism Involving the TGF-β and MAPKs Pathways in Cardiac Fibroblasts. PLoS One. 10:e01255132015. View Article : Google Scholar : PubMed/NCBI

13 

Działo E, Tkacz K and Błyszczuk P: Crosstalk between the TGF-β and WNT signalling pathways during cardiac fibrogenesis. Acta Biochim Pol. 65:341–349. 2018. View Article : Google Scholar : PubMed/NCBI

14 

Gu J, Liu X, Wang QX, Tan HW, Guo M, Jiang WF and Zhou L: Angiotensin II increases CTGF expression via MAPKs/TGF-β1/TRAF6 pathway in atrial fibroblasts. Exp Cell Res. 318:2105–2115. 2012. View Article : Google Scholar : PubMed/NCBI

15 

Trial J and Cieslik KA: Changes in cardiac resident fibroblast physiology and phenotype in aging. Am J Physiol Heart Circ Physiol. 315:H745–H755. 2018. View Article : Google Scholar : PubMed/NCBI

16 

Rane CK and Minden A: P21 activated kinase signaling in cancer. Semin Cancer Biol. 54:40–49. 2019. View Article : Google Scholar : PubMed/NCBI

17 

Sun X, Su VL and Calderwood DA: The subcellular localization of type I p21-activated kinases is controlled by the disordered variable region and polybasic sequences. J Biol Chem. 294:14319–14332. 2019. View Article : Google Scholar : PubMed/NCBI

18 

Rudolph J, Crawford JJ, Hoeflich KP and Wang W: Inhibitors of p21-activated kinases (PAKs). J Med Chem. 58:111–29. 2015. View Article : Google Scholar : PubMed/NCBI

19 

Wang Y, Wang S, Lei M, Boyett M, Tsui H, Liu W and Wang X: The p21-activated kinase 1 (Pak1) signalling pathway in cardiac disease: From mechanistic study to therapeutic exploration. Br J Pharmacol. 175:1362–1374. 2018. View Article : Google Scholar : PubMed/NCBI

20 

Al-Azayzih A, Gao F and Somanath PR: P21 activated kinase-1 mediates transforming growth factor β1-induced prostate cancer cell epithelial to mesenchymal transition. Biochim Biophys Acta. 1853:1229–1239. 2015. View Article : Google Scholar : PubMed/NCBI

21 

Chen QY, Xu LQ, Jiao DM, Yao QH, Wang YY, Hu HZ, Wu YQ, Song J, Yan J and Wu LJ: Silencing of Rac1 modifies lung cancer cell migration, invasion and actin cytoskeleton rearrangements and enhances chemosensitivity to antitumor drugs. Int J Mol Med. 28:769–776. 2011.PubMed/NCBI

22 

Yang Y, Du J, Hu Z, Liu J, Tian Y, Zhu Y, Wang L and Gu L: Activation of Rac1-PI3K/Akt is required for epidermal growth factor-induced PAK1 activation and cell migration in MDA-MB-231 breast cancer cells. J Biomed Res. 25:237–245. 2011. View Article : Google Scholar : PubMed/NCBI

23 

Zhang C, Lee HJ, Shrivastava A, Wang R, McQuiston TJ, Challberg SS, Pollok BA and Wang T: Long-Term In Vitro Expansion of Epithelial Stem Cells Enabled by Pharmacological Inhibition of PAK1-ROCK-Myosin II and TGF-β Signaling. Cell Rep. 25:598–610.e5. 2018. View Article : Google Scholar : PubMed/NCBI

24 

Liu W, Zi M, Naumann R, Ulm S, Jin J, Taglieri DM, Prehar S, Gui J, Tsui H, Xiao RP, et al: Pak1 as a novel therapeutic target for antihypertrophic treatment in the heart. Circulation. 124:2702–2715. 2011. View Article : Google Scholar : PubMed/NCBI

25 

Buttke TM, McCubrey JA and Owen TC: Use of an aqueous soluble tetrazolium/formazan assay to measure viability and proliferation of lymphokine-dependent cell lines. J Immunol Methods. 157:233–240. 1993. View Article : Google Scholar : PubMed/NCBI

26 

Mazzei M, Vascellari M, Zanardello C, Melchiotti E, Vannini S, Forzan M, Marchetti V, Albanese F and Abramo F: Quantitative real time polymerase chain reaction (qRT-PCR) and RNAscope in situ hybridization (RNA-ISH) as effective tools to diagnose feline herpesvirus-1-associated dermatitis. Vet Dermatol. 30:491–e147. 2019. View Article : Google Scholar : PubMed/NCBI

27 

Talman V and Ruskoaho H: Cardiac fibrosis in myocardial infarction-from repair and remodeling to regeneration. Cell Tissue Res. 365:563–581. 2016. View Article : Google Scholar : PubMed/NCBI

28 

Holtz J: Pathophysiology of heart failure and the renin-angiotensin-system. Basic Res Cardiol. 88 (Suppl 1):183–201. 1993.PubMed/NCBI

29 

Petrov VV, Fagard RH and Lijnen PJ: Stimulation of collagen production by transforming growth factor-beta1 during differentiation of cardiac fibroblasts to myofibroblasts. Hypertension. 39:258–263. 2002. View Article : Google Scholar : PubMed/NCBI

30 

Harada M, Luo X, Qi XY, Tadevosyan A, Maguy A, Ordog B, Ledoux J, Kato T, Naud P, Voigt N, et al: Transient receptor potential canonical-3 channel-dependent fibroblast regulation in atrial fibrillation. Circulation. 126:2051–2064. 2012. View Article : Google Scholar : PubMed/NCBI

31 

Alex L and Frangogiannis NG: The Cellular Origin of Activated Fibroblasts in the Infarcted and Remodeling Myocardium. Circ Res. 122:540–542. 2018. View Article : Google Scholar : PubMed/NCBI

32 

Hirsch AT, Pinto YM, Schunkert H and Dzau VJ: Potential role of the tissue renin-angiotensin system in the pathophysiology of congestive heart failure. Am J Cardiol. 66:D22–D332. 1990. View Article : Google Scholar

33 

Sabapathy K, Hochedlinger K, Nam SY, Bauer A, Karin M and Wagner EF: Distinct roles for JNK1 and JNK2 in regulating JNK activity and c-Jun-dependent cell proliferation. Mol Cell. 15:713–725. 2004. View Article : Google Scholar : PubMed/NCBI

34 

Harms FL, Kloth K, Bley A, Denecke J, Santer R, Lessel D, Hempel M and Kutsche K: Activating Mutations in PAK1, Encoding p21-Activated Kinase 1, Cause a Neurodevelopmental Disorder. Am J Hum Genet. 103:579–591. 2018. View Article : Google Scholar : PubMed/NCBI

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Spandidos Publications style
Zhou Y, Xie Y, Li T, Zhang P, Chen T, Fan Z and Tan X: P21‑activated kinase 1 mediates angiotensin II‑induced differentiation of human atrial fibroblasts via the JNK/c‑Jun pathway. Mol Med Rep 23: 207, 2021
APA
Zhou, Y., Xie, Y., Li, T., Zhang, P., Chen, T., Fan, Z., & Tan, X. (2021). P21‑activated kinase 1 mediates angiotensin II‑induced differentiation of human atrial fibroblasts via the JNK/c‑Jun pathway. Molecular Medicine Reports, 23, 207. https://doi.org/10.3892/mmr.2021.11846
MLA
Zhou, Y., Xie, Y., Li, T., Zhang, P., Chen, T., Fan, Z., Tan, X."P21‑activated kinase 1 mediates angiotensin II‑induced differentiation of human atrial fibroblasts via the JNK/c‑Jun pathway". Molecular Medicine Reports 23.3 (2021): 207.
Chicago
Zhou, Y., Xie, Y., Li, T., Zhang, P., Chen, T., Fan, Z., Tan, X."P21‑activated kinase 1 mediates angiotensin II‑induced differentiation of human atrial fibroblasts via the JNK/c‑Jun pathway". Molecular Medicine Reports 23, no. 3 (2021): 207. https://doi.org/10.3892/mmr.2021.11846