Wang YW,Zhang H,Cao P,et al.Influences and mechanism of extracellular vesicles from dermal papilla cells of mice on human hypertrophic scar fibroblasts[J].Chin J Burns Wounds,2024,40(3):258-265.DOI: 10.3760/cma.j.cn501225-20231107-00185.
Citation: Wang YW,Zhang H,Cao P,et al.Influences and mechanism of extracellular vesicles from dermal papilla cells of mice on human hypertrophic scar fibroblasts[J].Chin J Burns Wounds,2024,40(3):258-265.DOI: 10.3760/cma.j.cn501225-20231107-00185.

Influences and mechanism of extracellular vesicles from dermal papilla cells of mice on human hypertrophic scar fibroblasts

doi: 10.3760/cma.j.cn501225-20231107-00185
Funds:

General Program of National Natural Science Foundation of China 82272268

Industry-University-Research Innovation Fund of Ministry of Education of China 2021JH030

More Information
  •   Objective   To investigate the influences and mechanism of extracellular vesicles from dermal papilla cells (DPC-EVs) of mice on human hypertrophic scar fibroblasts (HSFs).   Methods   The study was an experimental research. The primary dermal papilla cells (DPCs) of whiskers were extracted from 10 6-week-old male C57BL/6J mice and identified successfully. The DPC-EVs were extracted from the 3 rd to 5 th passage DPCs by ultracentrifugation, and the morphology was observed through transmission electron microscope and the particle diameter was detected by nanoparticle tracking analyzer ( n=3) at 24 h after culture. The 3 rd passage of HSFs were divided into DPC-EV group and phosphate buffer solution (PBS) group, which were cultured with DPC-EVs and PBS, respectively. The cell scratch test was performed and cell migration rate at 24 h after scratching was calculated ( n=5). The cell proliferation levels at 0 (after 12 h of starvation treatment and before adding DPC-EVs or PBS), 24, 48, 72, and 96 h after culture were detected by using cell counting kit 8 ( n=4). The protein expressions of α-smooth muscle actin (α-SMA) and collagen typeⅠ (ColⅠ) in cells at 24 h after culture were detected by immunofluorescence method and Western blotting, and the protein expression of Krüppel-like factor 4 (KLF4) in cells at 24 h after culture was detected by Western blotting. After the 3 rd passage of HSFs were cultured with DPC-EVs for 24 h, the cells were divided into blank control group, KLF4 knockdown group, and KLF4 overexpression group according to the random number table. The cells in blank control group were only routinely cultured for 48 h. The cells in KLF4 knockdown group and KLF4 overexpression group were incubated with KLF4 knockdown virus for 24 h, then the cells in KLF4 knockdown group were routinely cultured for 24 h while the cells in KLF4 overexpression group were incubated with KLF4 overexpression virus for 24 h. The protein expressions of KLF4, α-SMA, and ColⅠ in cells were detected by Western blotting at 48 h after culture.   Results   At 24 h after culture, the extracted DPC-EVs showed vesicular structure with an average particle diameter of 108.8 nm. At 24 h after scratching, the migration rate of HSFs in PBS group was (54±10)%, which was significantly higher than (29±8)% in DPC-EV group ( t=4.37, P<0.05). At 48, 72, and 96 h after culture, the proliferation levels of HSFs in DPC-EV group were significantly lower than those in PBS group (with t values of 4.06, 5.76, and 6.41, respectively, P<0.05). At 24 h after culture, the protein expressions of α-SMA and ColⅠ of HSFs in DPC-EV group were significantly lower than those in PBS group, while the protein expression of KLF4 was significantly higher than that in PBS group. At 48 h after culture, compared with those in blank control group, the protein expression of KLF4 of HSFs in KLF4 knockdown group was down-regulated, while the protein expressions of α-SMA and ColⅠ were both up-regulated; compared with those in KLF4 knockdown group, the protein expression of KLF4 of HSFs in KLF4 overexpression group was up-regulated, while the protein expressions of ColⅠ and α-SMA were down-regulated.   Conclusions   The DPC-EVs of mice can inhibit the proliferation and migration of human HSFs and significantly inhibit the expressions of fibrosis markers α-SMA and ColⅠ in human HSFs by activating KLF4.

     

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    Created with Highcharts 5.0.7Chart context menuAccess Area Distribution其他: 8.3 %其他: 8.3 %其他: 0.1 %其他: 0.1 %Falls Church: 0.1 %Falls Church: 0.1 %三明: 0.3 %三明: 0.3 %三门峡: 0.3 %三门峡: 0.3 %上海: 0.6 %上海: 0.6 %东营: 0.1 %东营: 0.1 %丽水: 0.9 %丽水: 0.9 %佛山: 0.6 %佛山: 0.6 %六安: 0.4 %六安: 0.4 %兰州: 0.1 %兰州: 0.1 %内江: 0.1 %内江: 0.1 %包头: 0.4 %包头: 0.4 %北京: 1.1 %北京: 1.1 %南京: 1.3 %南京: 1.3 %南宁: 0.3 %南宁: 0.3 %南平: 0.6 %南平: 0.6 %南昌: 0.3 %南昌: 0.3 %南通: 0.1 %南通: 0.1 %厦门: 0.5 %厦门: 0.5 %台州: 1.5 %台州: 1.5 %吉林: 0.5 %吉林: 0.5 %咸阳: 2.3 %咸阳: 2.3 %哥伦布: 0.3 %哥伦布: 0.3 %商洛: 0.1 %商洛: 0.1 %嘉兴: 0.7 %嘉兴: 0.7 %圣安东尼奥: 0.1 %圣安东尼奥: 0.1 %大连: 1.6 %大连: 1.6 %天津: 0.2 %天津: 0.2 %宁德: 0.9 %宁德: 0.9 %宁波: 0.9 %宁波: 0.9 %宜春: 0.7 %宜春: 0.7 %宣城: 0.1 %宣城: 0.1 %宿迁: 1.6 %宿迁: 1.6 %常州: 0.1 %常州: 0.1 %常德: 0.7 %常德: 0.7 %广元: 0.1 %广元: 0.1 %广安: 0.2 %广安: 0.2 %广州: 2.0 %广州: 2.0 %廊坊: 0.1 %廊坊: 0.1 %张家口: 1.0 %张家口: 1.0 %徐州: 0.9 %徐州: 0.9 %成都: 0.1 %成都: 0.1 %扬州: 0.8 %扬州: 0.8 %抚州: 1.8 %抚州: 1.8 %新乡: 0.1 %新乡: 0.1 %无锡: 1.2 %无锡: 1.2 %日照: 0.3 %日照: 0.3 %昆明: 0.5 %昆明: 0.5 %朝阳: 1.0 %朝阳: 1.0 %杭州: 1.7 %杭州: 1.7 %枣庄: 0.4 %枣庄: 0.4 %柳州: 0.2 %柳州: 0.2 %梧州: 0.2 %梧州: 0.2 %榆林: 1.7 %榆林: 1.7 %武汉: 0.5 %武汉: 0.5 %汕头: 0.6 %汕头: 0.6 %江门: 1.4 %江门: 1.4 %池州: 0.6 %池州: 0.6 %沈阳: 0.9 %沈阳: 0.9 %泉州: 0.2 %泉州: 0.2 %法兰克福: 0.1 %法兰克福: 0.1 %泰州: 0.7 %泰州: 0.7 %泸州: 0.4 %泸州: 0.4 %济南: 1.4 %济南: 1.4 %济源: 0.2 %济源: 0.2 %海口: 0.2 %海口: 0.2 %海得拉巴: 0.3 %海得拉巴: 0.3 %淮北: 0.4 %淮北: 0.4 %淮安: 0.5 %淮安: 0.5 %深圳: 0.3 %深圳: 0.3 %温州: 0.5 %温州: 0.5 %渭南: 1.0 %渭南: 1.0 %湖州: 1.0 %湖州: 1.0 %湘潭: 0.3 %湘潭: 0.3 %湘西: 0.3 %湘西: 0.3 %漯河: 0.3 %漯河: 0.3 %漳州: 1.5 %漳州: 1.5 %珠海: 0.2 %珠海: 0.2 %盐城: 0.5 %盐城: 0.5 %盘锦: 0.2 %盘锦: 0.2 %石家庄: 0.6 %石家庄: 0.6 %福州: 0.9 %福州: 0.9 %秦皇岛: 0.7 %秦皇岛: 0.7 %纽约: 0.1 %纽约: 0.1 %绍兴: 0.9 %绍兴: 0.9 %绵阳: 0.4 %绵阳: 0.4 %自贡: 0.2 %自贡: 0.2 %舟山: 0.7 %舟山: 0.7 %芒廷维尤: 9.0 %芒廷维尤: 9.0 %芜湖: 0.1 %芜湖: 0.1 %芝加哥: 0.2 %芝加哥: 0.2 %苏州: 0.2 %苏州: 0.2 %荆州: 0.3 %荆州: 0.3 %荆门: 0.8 %荆门: 0.8 %莆田: 0.3 %莆田: 0.3 %营口: 0.9 %营口: 0.9 %葫芦岛: 0.6 %葫芦岛: 0.6 %衢州: 0.1 %衢州: 0.1 %襄阳: 0.2 %襄阳: 0.2 %西宁: 1.9 %西宁: 1.9 %西安: 1.2 %西安: 1.2 %辽阳: 0.8 %辽阳: 0.8 %运城: 0.1 %运城: 0.1 %连云港: 2.5 %连云港: 2.5 %邯郸: 0.1 %邯郸: 0.1 %邵阳: 0.3 %邵阳: 0.3 %郑州: 0.2 %郑州: 0.2 %郴州: 0.7 %郴州: 0.7 %酒泉: 0.1 %酒泉: 0.1 %重庆: 5.3 %重庆: 5.3 %金华: 0.8 %金华: 0.8 %铁岭: 2.0 %铁岭: 2.0 %锦州: 0.2 %锦州: 0.2 %长春: 0.4 %长春: 0.4 %长沙: 0.4 %长沙: 0.4 %随州: 0.4 %随州: 0.4 %雅安: 0.7 %雅安: 0.7 %青岛: 0.2 %青岛: 0.2 %鞍山: 5.8 %鞍山: 5.8 %鹰潭: 0.4 %鹰潭: 0.4 %黄冈: 1.4 %黄冈: 1.4 %黄南: 0.3 %黄南: 0.3 %黄山: 0.1 %黄山: 0.1 %黄石: 0.4 %黄石: 0.4 %其他其他Falls Church三明三门峡上海东营丽水佛山六安兰州内江包头北京南京南宁南平南昌南通厦门台州吉林咸阳哥伦布商洛嘉兴圣安东尼奥大连天津宁德宁波宜春宣城宿迁常州常德广元广安广州廊坊张家口徐州成都扬州抚州新乡无锡日照昆明朝阳杭州枣庄柳州梧州榆林武汉汕头江门池州沈阳泉州法兰克福泰州泸州济南济源海口海得拉巴淮北淮安深圳温州渭南湖州湘潭湘西漯河漳州珠海盐城盘锦石家庄福州秦皇岛纽约绍兴绵阳自贡舟山芒廷维尤芜湖芝加哥苏州荆州荆门莆田营口葫芦岛衢州襄阳西宁西安辽阳运城连云港邯郸邵阳郑州郴州酒泉重庆金华铁岭锦州长春长沙随州雅安青岛鞍山鹰潭黄冈黄南黄山黄石

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