High Hydrostatic Pressure Exacerbates Bladder Fibrosis through Activating Piezo1

Abstract

Objective

Bladder outlet obstruction (BOO) results in significant fibrosis in the chronic stage and elevated bladder pressure. Piezo1 is a type of mechanosensitive (MS) channel that directly responds to mechanical stimuli. To identify new targets for intervention in the treatment of BOO-induced fibrosis, this study investigated the impact of high hydrostatic pressure (HHP) on Piezo1 activity and the progression of bladder fibrosis.

Methods

Immunofluorescence staining was conducted to assess the protein abundance of Piezo1 in fibroblasts from obstructed rat bladders. Bladder fibroblasts were cultured under normal atmospheric conditions (0 cmH₂O) or exposed to HHP (50 cmH₂O or 100 cmH₂O). Agonists or inhibitors of Piezo1, YAP1, and ROCK1 were used to determine the underlying mechanism.

Results

The Piezo1 protein levels in fibroblasts from the obstructed bladder exhibited an elevation compared to the control group. HHP significantly promoted the expression of various pro-fibrotic factors and induced proliferation of fibroblasts. Additionally, the protein expression levels of Piezo1, YAP1, ROCK1 were elevated, and calcium influx was increased as the pressure increased. These effects were attenuated by the Piezo1 inhibitor Dooku1. The Piezo1 activator Yoda1 induced the expression of pro-fibrotic factors and the proliferation of fibroblasts, and elevated the protein levels of YAP1 and ROCK1 under normal atmospheric conditions in vitro. However, these effects could be partially inhibited by YAP1 or ROCK inhibitors.

Conclusion

The study suggests that HHP may exacerbate bladder fibrosis through activating Piezo1.

References

[1] DrakeMJ, LewisAL, YoungGJ, et al.. Diagnostic Assessment of Lower Urinary Tract Symptoms in Men Considering Prostate Surgery: A Noninferiority Randomised Controlled Trial of Urodynamics in 26 Hospitals. Eur Urol, 2020, 78(5): 701-710

[2] MetcalfePD, WangJ, JiaoH, HuangY, et al.. Bladder outlet obstruction: progression from inflammation to fibrosis. BJU Int, 2010, 106(11): 1686-1694

[3] AbramsPH, FarrarDJ, Turner-WarwickRT, et al.. The results of prostatectomy: a symptomatic and urodynamic analysis of 152 patients. J Urol, 1979, 121(5): 640-642

[4] ChenL, LvL, ZhangL, et al.. Metformin ameliorates bladder dysfunction in a rat model of partial bladder outlet obstruction. Am J Physiol Renal Physiol, 2021, 320(5): F838-F858

[5] ChenL, WeiTQ, WangY, et al.. Simulated bladder pressure stimulates human bladder smooth muscle cell proliferation via the PI3K/SGK1 signaling pathway. J Urol, 2012, 188(2): 661-667

[6] NguyenHT, AdamRM, BrideSH, et al.. Cyclic stretch activates p38 SAPK2-, ErbB2-, and AT1-dependent signaling in bladder smooth muscle cells. Am J Physiol Cell Physiol, 2000, 279(4): C1155-C1167

[7] WuYH, ChuehKS, ChuangSM, et al.. Bladder Hyperactivity Induced by Oxidative Stress and Bladder Ischemia: A Review of Treatment Strategies with Antioxidants. Int J Mol Sci, 2021, 22(11): 6014

[8] GaoX, WeiT, ChenJ, et al.. Cyclic hydrostatic pressure promotes uroplakin expression in human urothelial cells through activation of ERK1/2 signaling. Biochem Biophys Res Commun, 2018, 503(4): 2499-2503

[9] ZhaoM, WangL, WangM, et al.. Targeting fibrosis, mechanisms and cilinical trials. Signal Transduct Target Ther, 2022, 7(1): 206

[10] NiJ, WuQQ, LiaoHH, et al.. Bcl6 Suppresses Cardiac Fibroblast Activation and Function via Directly Binding to Smad4. Curr Med Sci, 2019, 39(4): 534-540

[11] SunW, MiH, HeDY, et al.. Liraglutide Suppresses Myocardial Fibrosis Progression by Inhibiting the Smad Signaling Pathway. Curr Med Sci, 2023, 43(5): 955-960

[12] ChenL, LvL, ZhangL, et al.. Metformin ameliorates bladder dysfunction in a rat model of partial bladder outlet obstruction. Am J Physiol Renal Physiol, 2021, 320(5): F838-F858

[13] ZhaoM, DingN, WangH, et al.. Activation of TRPA1 in Bladder Suburothelial Myofibroblasts Counteracts TGF-β1-Induced Fibrotic Changes. Int J Mol Sci, 2023, 24(11): 9501

[14] ChenG, GaoX, ChenJ, et al.. Actomyosin Activity and Piezo1 Activity Synergistically Drive Urinary System Fibroblast Activation. Adv Sci (Weinh), 2023, 10(33): e2303369

[15] JinP, JanLY, JanYN. Mechanosensitive Ion Channels: Structural Features Relevant to Mechanotransduction Mechanisms. Annu Rev Neurosci, 2020, 43: 207-229

[16] CosteB, MathurJ, SchmidtM, et al.. Piezo1 and Piezo2 are essential components of distinct mechanically activated cation channels. Science, 2010, 330(6000): 55-60

[17] LaiA, CoxCD, Chandra SekarN, et al.. Mechanosensing by Piezo1 and its implications for physiology and various pathologies. Biol Rev Camb Philos Soc, 2022, 97(2): 604-614

[18] LiX, HuJ, ZhaoX, et al.. Piezo channels in the urinary system. Exp Mol Med, 2022, 54(6): 697-710

[19] MichishitaM, YanoK, TomitaKI, et al.. Piezo1 expression increases in rat bladder after partial bladder outlet obstruction. Life Sci, 2016, 166: 1-7

[20] LluelP, DuquenneC, MartinD. Experimental bladder instability following bladder outlet obstruction in the female rat. J Urol, 1998, 160(6): 2253-2257 Pt 1

[21] DucheminAL, VignesH, VermotJ. Mechanically activated Piezo channels modulate outflow tract valve development through the Yap1 and Klf2-Notch signaling axis. Elife, 2019, 8: e44706

[22] ZhouT, GaoB, FanY, et al.. Piezo1/2 mediate mechanotransduction essential for bone formation through concerted activation of NFAT-YAP1-ß-catenin. Elife, 2020, 9: e52779

[23] JiangY, SongJ, XuY, et al.. Piezo1 regulates intestinal epithelial function by affecting the tight junction protein claudin-1 via the ROCK pathway. Life Sci, 2021, 275: 119254

[24] SpeichJE, SouthernJB, HendersonS, et al.. Adjustable passive stiffness in mouse bladder: regulated by Rho kinase and elevated following partial bladder outlet obstruction. Am J Physiol Renal Physiol, 2012, 302(8): F967-F976

[25] DeyA, VarelasX, GuanKL. Targeting the Hippo pathway in cancer, fibrosis, wound healing and regenerative medicine. Nat Rev Drug Discov, 2020, 19(7): 480-494

[26] ZhangF, IssahMA, FuHY, et al.. LATS1 Promotes B-ALL Tumorigenesis by Regulating YAP1 Phosphorylation and Subcellular Localization. Curr Med Sci, 2024, 44(1): 81-92

[27] FangY, LiQ, LiX, et al.. Piezo1 Participated in Decreased L-Type Calcium Current Induced by High Hydrostatic Pressure via. CaM/Src/Pitx2 Activation in Atrial Myocytes. Front Cardiovasc Med, 2022, 9: 842885