scholarly journals CGRP protects bladder smooth muscle cells stimulated by high glucose through inhibiting p38 MAPK pathway in vitro

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Jun Xue ◽  
Yadong Liu ◽  
Sichong Zhang ◽  
Liucheng Ding ◽  
Baixin Shen ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Cell Viability ◽  
P38 Mapk ◽  
High Glucose ◽  
Muscle Cells ◽  
Control Group ◽  
Asiatic Acid ◽  
Bladder Smooth Muscle

AbstractThis study aimed to explore the effect of calcitonin gene-related peptide (CGRP) on bladder smooth muscle cells (BSMCs) under high glucose (HG) treatment in vitro. BSMCs from Sprague–Dawley rat bladders were cultured and passaged in vitro. The third-generation cells were cultured and divided into control group, HG group, HG + CGRP group, HG + CGRP + asiatic acid (AA, p-p38 activator) group, CGRP group, AA group, HG + CGRP + CGRP-8-37 (CGRP receptor antagonist) group and HG + LY2228820 (p38 MAPK inhibitor) group. The cell viability, apoptosis, malondialdehyde (MDA) and superoxide dismutase (SOD) levels of BSMCs were observed by the relevant detection kits. The expressions of α-SM-actin, p38 and p-p38 were detected by qRT-PCR or Western blot analysis. Compared with the control group, the cell viability, SOD and α-SM-actin levels of BSMCs were decreased and apoptotic cells, MDA and p-p38 levels were increased after HG treatment, while these changes could be partly reversed when BSMCs were treated with HG and CGRP or LY2228820 together. Moreover, AA or CGRP-8-37 could suppress the effect of CGRP on BSMCs under HG condition. Our data indicate that CGRP protects BSMCs from oxidative stress induced by HG in vitro, and inhibit the α-SM-actin expression decrease through inhibiting the intracellular p38 MAPK signaling pathway.

scholarly journals Protective Effects of Adiponectin against Cobalt Chloride-Induced Apoptosis of Smooth Muscle Cells via cAMP/PKA Pathway

2020 ◽  
Vol 2020 ◽  
pp. 1-11
Author(s):  
Jingjie Xiao ◽  
Yingying Zhang ◽  
Wei Zhang ◽  
Liang Zhang ◽  
Li Li ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Cell Viability ◽  
Cobalt Chloride ◽  
Muscle Cells ◽  
Camp Level ◽  
Control Group ◽  
Dependent Manner ◽  
Sod Activity

Adiponectin (APN) is an adipokine secreted from adipose tissue and exhibits biological functions such as microcirculation-regulating, hearing-protective, and antiapoptotic. However, the effect of APN on the apoptosis of spiral arterial smooth muscle cells (SMCs) under hypoxic conditions in vitro is not clear. We used cobalt chloride (CoCl2) to simulate chemical hypoxia in vitro, and the SMCs were pretreated with APN and then stimulated with CoCl2. The viability of cells and apoptosis were assessed by CCK-8 and flow cytometry, respectively. Superoxide dismutase (SOD) activity, malondialdehyde (MDA) levels, cAMP level, and the activity of PKA were detected by ELISA. Protein expression and localization were studied by Western blot and immunofluorescence analysis. In the present study, we found that APN exhibits antiapoptosis effects. CoCl2 exhibited decreased cell viability, increased apoptosis and MDA levels, and decreased SOD activity in a concentration-dependent manner, compared with the control group. Moreover, CoCl2 upregulated the expression levels of Bax and cleaved caspase-3 and then downregulated Bcl-2 levels in a time-dependent manner. Compared with the CoCl2 group, the group pretreated with APN had increased cell viability, SOD activity, PKA activity, cAMP level, and PKA expression, but decreased MDA levels and apoptosis. Lastly, the protective effect of APN was blocked by cAMP inhibitor SQ22536 and PKA inhibitor H 89. These results showed that APN protected SMCs against CoCl2-induced hypoxic injury via the cAMP/PKA signaling pathway.

Mechanical stretch regulates cell survival in human bladder smooth muscle cells in vitro

2002 ◽  
Vol 283 (6) ◽  
pp. F1192-F1199 ◽  
Author(s):  
David J. Galvin ◽  
R. William G. Watson ◽  
James I. Gillespie ◽  
Hugh Brady ◽  
John M. Fitzpatrick
Keyword(s):  
Smooth Muscle ◽  
Growth Factor ◽  
Smooth Muscle Cells ◽  
Muscle Cells ◽  
Mechanical Stretch ◽  
Bladder Smooth Muscle ◽  
Human Bladder ◽  
Cell Hypertrophy ◽  
Growth Factor Production

Our understanding of the pathophysiology of the overactive bladder is poor. It has been proposed that localized contractions result in the abnormal stretching of bladder smooth muscle. We hypothesize that stretch regulates the cellular processes that determine tissue size. The purpose of this study was to investigate the effect of stretch on apoptosis, proliferation, cell hypertrophy, and growth factor production in human bladder smooth muscle cells in vitro. Normal human detrusor muscle was obtained from patients undergoing radical cystectomy for invasive bladder cancer, and primary cultures were established. Cells were mechanically stretched on flexible plates at a range of pressures and times. Apoptosis was assessed by propidium iodide incorporation and flow cytometry. Radiolabeled thymidine and amino acid incorporation were used to assess proliferation and cell hypertrophy. ELISA and RT-PCR were used to assess growth factor production. Mechanical stretch inhibits apoptosis in a time- and dose-dependent manner and was associated with increases in the antiapoptotic proteins heat shock protein-70 and cIAP-1. Stretch also increases smooth muscle cell proliferation and hypertrophy, but hypertrophy is the more dominant response. These changes were associated with increases in IGF-1 and basic FGF and a decrease in transforming growth factor-β1. Mechanical stretch regulates apoptosis, proliferation, and cell hypertrophy in human bladder smooth muscle cells.

CHARACTERIZATION OF CULTURED BLADDER SMOOTH MUSCLE CELLS: ASSESSMENT OF IN VITRO CONTRACTILITY

1999 ◽  
Vol 162 (5) ◽  
pp. 1779-1784 ◽  
Author(s):  
BRADLEY P. KROPP ◽  
YUANYUAN ZHANG ◽  
JAMES J. TOMASEK ◽  
RICK COWAN ◽  
PETER D. FURNESS ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Muscle Cells ◽  
Bladder Smooth Muscle

scholarly journals RAGE Differentially Altered in vitro Responses in Vascular Smooth Muscle Cells and Adventitial Fibroblasts in Diabetes-Induced Vascular Calcification

2021 ◽  
Vol 12 ◽  
Author(s):  
Amber M. Kennon ◽  
James A. Stewart
Keyword(s):  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
P38 Mapk ◽  
Vascular Calcification ◽  
Vascular Smooth Muscle Cells ◽  
Muscle Cells ◽  
Adventitial Fibroblasts ◽  
Rage Expression

The Advanced Glycation End-Products (AGE)/Receptor for AGEs (RAGE) signaling pathway exacerbates diabetes-mediated vascular calcification (VC) in vascular smooth muscle cells (VSMCs). Other cell types are involved in VC, such as adventitial fibroblasts (AFBs). We hope to elucidate some of the mechanisms responsible for differential signaling in diabetes-mediated VC with this work. This work utilizes RAGE knockout animals and in vitro calcification to measure calcification and protein responses. Our calcification data revealed that VSMCs calcification was AGE/RAGE dependent, yet AFBs calcification was not an AGE-mediated RAGE response. Protein expression data showed VSMCs lost their phenotype marker, α-smooth muscle actin, and had a higher RAGE expression over non-diabetics. RAGE knockout (RKO) VSMCs did not show changes in phenotype markers. P38 MAPK, a downstream RAGE-associated signaling molecule, had significantly increased activation with calcification in both diabetic and diabetic RKO VSMCs. AFBs showed a loss in myofibroblast marker, α-SMA, due to calcification treatment. RAGE expression decreased in calcified diabetic AFBs, and P38 MAPK activation significantly increased in diabetic and diabetic RKO AFBs. These findings point to potentially an alternate receptor mediating the calcification response in the absence of RAGE. Overall, VSMCs and AFBs respond differently to calcification and the application of AGEs.

A Novel In-Vitro System for the Simultaneous Exposure of Bladder Smooth Muscle Cells to Mechanical Strain and Sustained Hydrostatic Pressure

2002 ◽  
Vol 124 (2) ◽  
pp. 208-213 ◽  
Author(s):  
Karen M. Haberstroh ◽  
Martin Kaefer ◽  
Natacha DePaola ◽  
Sarah A. Frommer ◽  
Rena Bizios
Keyword(s):  
Smooth Muscle ◽  
Hydrostatic Pressure ◽  
Smooth Muscle Cells ◽  
Collagen Type ◽  
Mechanical Strain ◽  
Muscle Cells ◽  
Collagen Type Iii ◽  
Bladder Smooth Muscle ◽  
Simultaneous Exposure

The novel hydrostrain system was designed in an effort to establish and maintain conditions that simulate the in-vivo mechanical environment of the bladder. In this laboratory system, ovine bladder smooth muscle cells on flexible, 10-cm-dia silastic membranes were exposed simultaneously to hydrostatic pressure (40 cm H2O, a pressure level currently associated with bladder pathologies) and mechanical strains (up to 25 percent) under standard cell culture conditions for 7 h. Under these conditions, Heparin Binding-Epidermal Growth Factor and Collagen Type III mRNA expression were significantly increased (p<0.01 and 0.1, respectively); however, no changes were observed in Collagen Type I mRNA expression. Decreases in the Collagen Type I:Type III ratio following simultaneous exposure of bladder smooth muscle cells to pathological levels of hydrostatic pressure and mechanical strain in vitro are in agreement with clinically observed increases in Collagen Type III with concomitant decreased human bladder compliance. The results of the present study, therefore, provide cellular/molecular level information relevant to bladder pathology that could have significant implications in the field of clinical urology.

Bladder Smooth Muscle Cells Interaction and Proliferation on PCL/PLLA Electrospun Nanofibrous Scaffold

2013 ◽  
Vol 36 (2) ◽  
pp. 113-120 ◽  
Author(s):  
Nasser Shakhssalim ◽  
Javad Rasouli ◽  
Reza Moghadasali ◽  
Farzaneh Sharifi Aghdas ◽  
Mohammad Naji ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Cell Viability ◽  
Muscle Cells ◽  
Nanofibrous Scaffold ◽  
In Vivo Studies ◽  
Bladder Smooth Muscle ◽  
Bladder Tissue ◽  
Nanofibrous Scaffolds

Purpose Numerous synthetic materials have been used for the bladder reconstruction; of which, nano-structured scaffolds are used as relevant implant to the bladder tissue-engineering. The aim of this study was to investigate the capacity of Poly ∊-caprolactone/poly-L-lactide acid (PCL/PLLA) nanofibrous scaffold, in supporting the maintenance and attachment of the human bladder smooth muscle cells (BdSMCs). Methods In this study, BdSMCs were isolated by enzymatic digestion method. Then, cells were seeded on PCL/PLLA nanofibrous scaffolds. Thereafter, cell attachment and expansion were analyzed by Hematoxylin and Eosin staining (H&E), immunohistochemistry, and scanning electron microscopy (SEM). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay confirmed that the nanostructured scaffold supported and maintained normal cell viability without inducing cytotoxic events. Results H&E staining, immunohistochemistry, and SEM showed that BdSMCs were attached and expanded on PCL/PLLA nanofibrous scaffolds after 14 days. Cell viability of BdSMCs on PCL/PLLA nanofibrous scaffolds increased during 14 days. Conclusion Our results showed that the novel porous nanofibrous electrospun scaffold is a biocompatible structure for attachment and adhesion of BdSMCs. However, there is not enough information on the stimulating effect of this nanofiber on the cells. Therefore, further in-vivo studies seem required to confirm such a nanofiber to be used in the bladder tissue-engineering.

scholarly journals Anti-Proliferative Effect of an Aqueous Extract ofPrunella vulgarisin Vascular Smooth Muscle Cells

2013 ◽  
Vol 2013 ◽  
pp. 1-10 ◽  
Author(s):  
Sun Mi Hwang ◽  
Yun Jung Lee ◽  
Yong Pyo Lee ◽  
Jung Joo Yoon ◽  
So Min Lee ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
P38 Mapk ◽  
Vascular Smooth Muscle Cells ◽  
Aqueous Extract ◽  
High Glucose ◽  
Muscle Cells ◽  
Prunella Vulgaris ◽  
Vsmc Proliferation

The abnormal proliferation of vascular smooth muscle cells (VSMCs) in arterial walls is an important pathogenic factor of vascular disorders such as diabetic atherosclerosis. We have reported the anti-inflammatory effect of an aqueous extract fromPrunella vulgaris(APV) in vascular endothelial cell. In the present study, APV exhibited inhibitory effects on high glucose-stimulated VSMC proliferation, migration, and invasion activities, inducing G1cell cycle arrest with downregulation of cyclins and CDKs and upregulation of the CKIs,p21waf1/cip1andp27kip1. Furthermore, APV dose dependently suppressed the high glucose-induced matrix metalloproteinase activity. High glucose-induced phosphorylation of ERK, p38 MAPK, was decreased by the pretreatment of APV. NF-κB activation by high glucose was attenuated by APV, as an antioxidant. APV attenuated the high glucose-induced decrease of nuclear factor E2-related factor-2 (Nrf2) translocation and heme oxygenase-1 (HO-1) expression. Intracellular cGMP level was also increased by APV treatment. These results demonstrate that APV may inhibit VSMC proliferation via downregulating ROS/NF-κB /ERK/p38 MAPK pathways. In addition, APV has a beneficial effect by the interaction of Nrf2-mediated NO/cGMP with HO-1, suggesting thatPrunella vulgarismay be useful in preventing diabetic atherosclerosis.

Sign in / Sign up

Export Citation Format

Share Document