β‐Adrenoceptor regulates contraction and inflammatory cytokine expression of human bladder smooth muscle cells via autophagy under pathological hydrostatic pressure

Abstract

Aims: Abnormal intravesical pressure created by partial bladder outlet obstruction (PBOO) triggered the progression from chronic inflammation to fibrosis, initiating structural and functional alterations of bladder. To elucidate the underlying mechanisms of contraction and inflammatory response, we investigated the isolated human bladder smooth muscle cells (hBSMC) under pathological hydrostatic pressure (HP) mimicking the in vivo PBOO condition. Methods: hBSMCs were subjected to HP of 200 cm H 2 O to explore the contraction and inflammatory cytokine expression of hBSMC treated with β-adrenoceptors (ADRBs) and/or autophagy signaling pathway agonists and/ or antagonists. Results: We showed that pathological HP induced the release of the proinflammatory cytokines, including monocyte chemotactic protein-1, regulated upon activation normal T cell expressed and secreted factor, and interleukin-6. HP downregulated ADRB2 and ADRB3 expression, which was consistent with the results of the PBOO rat model. ADRB2 or autophagy activation repressed pathological HP-induced proinflammatory cytokine production. ADRB2, ADRB3 or autophagy activation ameliorated the HP-enhanced contraction. The increased contraction and autophagy activity by ADRB2 agonist under HP conditions were reversed by pretreatment with antagonists of adenosine monophosphate-activated protein kinase (AMPK). Conclusion: The present study provides evidence that the ADRB3 agonist suppresses hBSMC contraction under pathological HP conditions. Moreover, the ADRB2 agonist negatively regulates the contraction and inflammatory response of hBSMCs through AMPK/mTOR-mediated autophagy under pathological HP. These findings provide a theoretical basis for potential therapeutic strategies for patients with PBOO.

RETRACTED: Survivin knockdown alleviates pathological hydrostatic pressure-induced bladder smooth muscle cell dysfunction and BOO-induced bladder remodeling via autophagy

“…Western blot (WB) procedures were as previously described (Chen et al, 2020b), the membranes were incubated overnight at 4 °C with the following antibodies: Microtubule-associated protein 1 light chain 3 beta (LC3B) (ab48394; 1:1,000; 17 kDa Frontiers in Cell and Developmental Biology frontiersin.org and 19 kDa), Beclin1 (ab207612; 1:2000; 52 kDa), Survivin (ab134170; 1:1,000; 16 kDa), GAPDH (ab9485; 1:2,500; 37 kDa),Col1 (ab96723; 1:1,000; 129 kDa), Fn(ab268020; 1: 1,000; 262 kDa), and PCNA(ab29; 1:1,000; 29 kDa) from Abcam (Cambridge, MA, United States). The membranes were then exposed to an immobilon chemiluminescent substrate.…”

“…Increasing evidence suggests that autophagy is closely related the bladder remodeling secondary to BOO due to its double-edged sword role in pro-cell survival and the pro-cell death properties ( Chen et al, 2021 ). Our previous study has demonstrated that autophagy regulated the biological function of BSMCs under HP ( Chen et al, 2020b ). The emerging data indicated that survivin negatively modulated autophagy levels in various diseases, which piqued our interest in exploring whether the survivin has a similar role in the bladder remodeling of BOO ( Ding et al, 2015 ; Pavel et al, 2018 ).…”

The preventive effects of colony‐stimulating factor 1 receptor (CSF‐1R) inhibition on bladder outlet obstruction induced remodeling

“…Next, we exposed human urothelial cells to hydrostatic pressure in our designed apparatus as previously described. 28 H 2 O (200 cm) was selected as pathological condition, whereas H 2 O (0 cm) was used for the control group. To investigate the stimulus for the CSF-1 during BOO, human urothelial cells were subjected to hydrostatic pressure for 1, 3, and 6 h. Western blot analysis showed that exposure to pathological hydrostatic pressure promoted CSF-1 production by human urothelial cells (data unpublished).…”

“…Therefore, we postulated that bladder urothelial cells were the main source for the endogenous CSF‐1. Next, we exposed human urothelial cells to hydrostatic pressure in our designed apparatus as previously described 28 . H 2 O (200 cm) was selected as pathological condition, whereas H 2 O (0 cm) was used for the control group.…”

Tim-1 alleviates lupus nephritis-induced podocyte injury via regulating autophagy

“…Autophagy stands for an ability of cells to prevent their own death [ 16 ], which could also protect cells against apoptosis [ 17 ]. In addition, some reports have proved that autophagy can be an important mediator in the immune system [ 18 , 19 ]. Meanwhile, autophagy is closely associated with progression of SLE/LN.…”