Heat injured stromal cells-derived exosomal EGFR enhances prostatic wound healing after thulium laser resection through EMT and NF-κB signaling
Heat injured stromal cells-derived exosomal EGFR enhances prostatic wound healing after thulium laser resection through EMT and NF-κB signaling
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DOI：
10.1002/pros.23827
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发表时间：
2019-08-01
期刊：
影响因子：
2.8
通讯作者：
Han, Bang-Min
中科院分区：
文献类型：
--
作者：
Shi, Fei;Deng, Zheng;Han, Bang-Min
本研究探讨了前列腺基质成纤维细胞和上皮细胞的浅热损伤及其相互作用，以调节伤口愈合和潜在的分子事件。方法将前列腺间质成纤维细胞和上皮细胞单独培养或共培养，经浅热损伤后，检测细胞增殖、迁移、凋亡、细胞周期分布及基因表达。收集热损伤的WPMY-1细胞的上清液用于外泌体提取和评估。此外，比格犬接受了前列腺的铥激光切除术（TmLRP），并随机分为吉非替尼、GW 4869和对照治疗组，用于组织学分析、组织再上皮化和前列腺伤口表面上的表皮生长因子受体（EGFR）表达。免疫荧光法检测p63阳性基底祖细胞转分化和巨噬细胞极化; ELISA法检测尿中细胞因子水平。结果浅度热损伤使细胞进入应激状态，串扰增强。前列腺间质成纤维细胞产生和分泌更多的exosomal-EGFR和其他细胞因子和趋化因子后，浅热损伤，导致前列腺上皮细胞的增殖和迁移在伤口愈合过程中增加。在TmLRP程序后，吉非替尼和GW 4869治疗组中犬前列腺尿道的伤口愈合比动物对照组中慢。免疫荧光和ELISA显示，EGFR表达减少中断巨噬细胞极化，但增加炎症反应。结论浅热损伤可促进前列腺基质细胞与前列腺上皮细胞的相互作用，促进创面愈合。间质源性外泌体-EGFR在巨噬细胞极化和前列腺伤口愈合的平衡中起着至关重要的作用。
Background This study investigated shallow heat injury to prostate stromal fibroblasts and epithelial cells and their interaction to regulate the wound healing and the underlying molecular events. Methods Prostate stromal fibroblasts and epithelial cells were cultured individually or cocultured and subjected to shallow heat injury for assessments of cell proliferation, migration, apoptosis, cell cycle distribution, and gene expression. The supernatant of heat-injured WPMY-1 cells was collected for exosome extraction and assessments. Furthermore, beagle dogs received thulium laser resection of the prostate (TmLRP) and randomly divided into Gefitinib, GW4869, and control treatment for the histological analysis, tissue re-epithelialization, and epidermal growth factor receptor (EGFR) expression on the prostatic wound surface. Immunofluorescence was to evaluate p63-positive basal progenitor cell trans-differentiation and macrophage polarization and ELISA was to detect cytokine levels in beagles' urine. Results Shallow heat injury caused these cells to enter a stressed state and enhanced their crosstalk. The prostate stromal fibroblasts produced and secreted more exosomal-EGFR and other cytokines and chemokines after shallow heat injury, resulting in increased proliferation and migration of prostate epithelial cells during wound healing. The wound healing of the canine prostatic urethra following the TmLRP procedure was slower in the Gefitinib and GW4869 treatment group than in the control group of animals. Immunofluorescence and ELISA showed that reduced EGFR expression interrupted macrophage polarization but increased the inflammatory response. Conclusions Shallow heat injury was able to promote the interaction of prostate stromal cells with prostate epithelial cells to enhance wound healing. Stromal-derived exosomal-EGFR plays a crucial role in the balance of the macrophage polarization and prostatic wound healing.