Evaluation of the antifibrotic potency by knocking down SPARC, CCR2 and SMAD3

Ding, Weifeng; Pu, Weilin; Jiang, Shuai; Ma, Yanyun; Liu, Qingmei; Wu, Wenyu; Chu, Haiyan; Zou, Hejian; Jin, Li; Wang, Jiucun; Zhou, Xiaodong

doi:10.1101/376459

Cited by 5 publications

(6 citation statements)

References 46 publications

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“…The main significance of our study is the use of USF cells that are directly taken from patient stricture region and considering that these USF cells might be critical factor in the development of fibrosis. Conventionally, the antifibrotic potency assays have been conducted using cell lines [37,38].…”

Section: Discussionmentioning

confidence: 99%

The effect of human amniotic epithelial cells on urethral stricture fibroblasts

2019

J Clin Transl Res

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Background: Urethral stricture disease (USD) is effectively managed by buccal mucosa (BM) urethroplasty. Lack of adequate healthy BM has led to the use of autologous tissue-engineered BM grafts. Such grafts are costly, not easily scalable and recurrence of the stricture is still a problem. Hence, there is a requirement for cost-effective, scalable cells with innate antifibrotic properties which seem to be fulfilled by human amniotic epithelial cells (HAMECs). The effect of HAMECs on USD is unknown. Aim: To study the effect of HAMECs-CM on human urethral stricture fibroblast (USF) cells by using in-vitro migration assay and molecular techniques. Materials and Methods: USF cells were derived from six patients undergoing urethroplasty. HAMECs were derived from one placenta after delivery. The effect of HAMECs-CM on USF cell migration was observed using a standard in vitro scratch assay over a period of 3 days. The effect of HAMECs-CM on the expression levels of markers alpha-smooth muscle actin (α-SMA) and tissue inhibitor of metalloproteinases (TIMP-1) in USF cells was also examined. Results: The HAMECs-CM suppressed the migration of USF cells in in vitro scratch assay. The HAMECs-CM consistently downregulated α-SMA, but not TIMP-1. Conclusions: HAMECs have shown antifibrotic activity on USF cells in this in vitro study. Relevance for Patients: HAMECs could serve as an alternative cell source for tissue-engineered urethroplasty.

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Section: Discussionmentioning

confidence: 99%

The effect of human amniotic epithelial cells on urethral stricture fibroblasts

2019

J Clin Transl Res

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“…All patients were aged 10-15, three of them were girls, two were boys. After being washed, the tissues were cultured for preparation of HTFs, as described previously 11 . The HTFs were cultivated in Dulbecco's Modi ed Eagle's medium (DMEM) containing 10% of fetal bovine serum (FBS) at 37 C in a humidi ed atmosphere of 5% CO 2 .The cells were passaged every 3-5 days.…”

Section: Primary Cell Culture and Quality Controlmentioning

confidence: 99%

Secreted protein, acidic and rich in cysteine is crucial for the vascular endothelial growth factor-stimulated fibrotic process in human Tenon’s fibroblasts

Xiang

Luo

Chen

et al. 2020

Preprint

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Background Aberrant post-surgical scarring is responsible for failure of glaucoma filtration surgery (GFS) and is attributed to strong fibrotic process of human Tenon’s fibroblasts (HTFs). Vascular endothelial growth factor (VEGF) and secreted protein, acidic and rich in cysteine (SPARC) contribute to angiogenesis and fibrosis. However, whether SPARC can regulate the VEGF-mediated fibrotic process in HTFs has not been clarified. This study aimed to examine how SPARC and VEGF crosstalk to regulate the expression of Collagen-I and matrix metalloproteinase 9 (MMP9) as well as the ERK signalling in HTFs. Methods Human Tenon's capsule tissues were cultured for preparation of HTFs, which were characterized by immunofluorescence. The effects of VEGF treatment on SPARC, Collagen-I and MMP9 expression and ERK phosphorylation were determined by Western blot, quantitative RT-PCR and immunofluorescence. The proliferation and wound healing induced by VEGF were examined in HTFs and SPARC-silenced HTFs. Results Following successful passages, immunofluorescent assays indicated that HTFs at passages 3-9 displayed unique characters of fibroblasts with Vimentin, but not keratin, expression. Treatment with VEGF significantly up-regulated SPARC, Collagen-I and MMP9 expression and ERK phosphorylation, and promoted the proliferation and wound healing of HTFs. The stimulatory effects of VEGF were significantly mitigated by SPARC silencing in HTFs. Conclusions Our data provided novel evidence that SPARC was crucial for the VEGF-stimulated fibrotic process in HTFs and may be a novel target for anti-fibrotic therapies post GFS.

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“…SPARC can regulate ECM remodelling by modulating collagen expression and MMP activity 9,10 . To understand the regulatory role of SPARC in VEGF-induced fibrotic process in HTFs, we generated stable SPARC-silenced HTFs by lentivirus-based shRNA technology.…”

Section: Sparc Silencing Mitigates the Vegf-induced Fibrotic Process mentioning

confidence: 99%

“…Secreted protein, acidic and rich in cysteine (SPARC) is one of the matricellular proteins that are crucial for scar formation and tissue fibrosis [9][10] . SPARC can regulate the expression of Collagens and matrix metalloproteinases (MMPs), as well as the extracellular matrix (ECM) remodeling.…”

Section: Introductionmentioning

confidence: 99%

Secreted protein, acidic and rich in cysteine is crucial for the vascular endothelial growth factor-stimulated fibrotic process in human Tenon’s fibroblasts

Xiang¹,

Luo²,

Chen³

et al. 2019

Preprint

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Background Aberrant post-surgical scarring is responsible for failure of glaucoma filtration surgery (GFS) and is attributed to strong fibrotic process of human Tenon’s fibroblasts (HTFs). Vascular endothelial growth factor (VEGF) and secreted protein, acidic and rich in cysteine (SPARC) contribute to angiogenesis and fibrosis. However, whether SPARC can regulate the VEGF-mediated fibrotic process in HTFs has not been clarified. This study aimed to examine how SPARC and VEGF crosstalk to regulate the expression of Collagen-I and matrix metalloproteinase 9 (MMP9) as well as the ERK signalling in HTFs.Methods Human Tenon's capsule tissues were cultured for preparation of HTFs, which were characterized by immunofluorescence. The effects of VEGF treatment on SPARC, Collagen-I and MMP9 expression and ERK phosphorylation were determined by Western blot, quantitative RT-PCR and immunofluorescence. The proliferation and wound healing induced by VEGF were examined in HTFs and SPARC-silenced HTFs.Results Following successful passages, immunofluorescent assays indicated that HTFs at passages 3-9 displayed unique characters of fibroblasts with Vimentin, but not keratin, expression. Treatment with VEGF significantly up-regulated SPARC, Collagen-I and MMP9 expression and ERK phosphorylation, and promoted the proliferation and wound healing of HTFs. The stimulatory effects of VEGF were significantly mitigated by SPARC silencing in HTFs.Conclusions Our data provided novel evidence that SPARC was crucial for the VEGF-stimulated fibrotic process in HTFs and may be a novel target for anti-fibrotic therapies post GFS.

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Evaluation of the antifibrotic potency by knocking down SPARC, CCR2 and SMAD3

Cited by 5 publications

References 46 publications

The effect of human amniotic epithelial cells on urethral stricture fibroblasts

The effect of human amniotic epithelial cells on urethral stricture fibroblasts

Secreted protein, acidic and rich in cysteine is crucial for the vascular endothelial growth factor-stimulated fibrotic process in human Tenon’s fibroblasts

Secreted protein, acidic and rich in cysteine is crucial for the vascular endothelial growth factor-stimulated fibrotic process in human Tenon’s fibroblasts

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