文章摘要
俞蕾媛,盛少琴,潘弘毅,等.三七重楼生化汤对成纤维细胞功能的影响及机制研究[J].浙江中医药大学学报,2020,44(12):1215-1221.
三七重楼生化汤对成纤维细胞功能的影响及机制研究
Effects of Sanqi Chonglou Shenghua Decoction on Biological Function of Fibroblasts and Its Mechanism
投稿时间:2020-06-14  
DOI:10.16466/j.issn1005-5509.2020.12.015
中文关键词: 三七重楼生化汤  成纤维细胞  VEGF  TGF-β1  ERK信号通路  增殖  迁移  黏附
英文关键词: Sanqi Chonglou Shenghua Decoction  fibroblasts  VEGF  TGF-β1  ERK signaling pathway  proliferation  migration  adhesion
基金项目:浙江省中医药科技计划重点资助项目(2018ZZ013);浙江省医药卫生科技计划项目(2019KY477)
作者单位E-mail
俞蕾媛 浙江中医药大学第二临床医学院 杭州 310053  
盛少琴 浙江中医药大学附属第二医院 shshaoqin@sina.com 
潘弘毅 浙江中医药大学第三临床医学院  
刘佳俐 浙江中医药大学附属第二医院  
范梦梦 浙江中医药大学第二临床医学院 杭州 310053  
潘鸿燕 云和县中医院  
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中文摘要:
      [目的]观察三七重楼生化汤(Sanqi Chonglou Shenghua Decoction,SCD)对L929成纤维细胞增殖、黏附、迁移及愈合相关蛋白表达的影响,并探讨其机制。[方法]常规培养L929成纤维细胞,随机分为SCD低、中、高浓度组及对照组,SCD各浓度组予不同浓度SCD(1.25、2.5、5mg·mL-1),对照组予等体积培养基,以实时细胞分析技术(real time cellular analysis,RTCA)、细胞增殖检测试剂盒(cell counting kit-8,CCK-8)分别检测细胞增殖、黏附能力,划痕实验检测细胞迁移情况,Western blot检测血管内皮细胞生长因子(vascular endothelial growth factor,VEGF)、转化生长因子-β1(transforming growth factor-β1,TGF-β1)、细胞外调节蛋白激酶1/2(extracellular regulated protein kinases 1/2,ERK1/2)、磷酸化细胞外调节蛋白激酶1/2(phospho-extracellular regulated protein kinases 1/2,p-ERK1/2)蛋白表达。[结果]SCD促进成纤维细胞增殖,且呈动态变化。RTCA示:SCD干预12~24h,中、高浓度促进细胞增殖(P<0.01),但组间比较,差异无统计学意义(P>0.05)。CCK-8结果示:SCD干预24h,低、中、高浓度组均促进细胞增殖(P<0.01)。与低浓度组比较,中、高浓度组促增殖作用更显著 (P<0.01)。与中浓度组比较,高浓度组促增殖作用更显著 (P<0.05)。SCD促进成纤维细胞迁移。干预24h,中、高浓度组促进细胞迁移(P<0.05,P<0.01),且高浓度组促进作用更显著(P<0.01)。干预36h,低、中、高浓度SCD均具有促进迁移的作用(P<0.05,P<0.01)。与低浓度组比较,中、高浓度促进作用更显著(P<0.05,P<0.01)。与中浓度组比较,高浓度组促进迁移作用更显著(P<0.01)。SCD促进成纤维细胞黏附(P<0.01),但不同浓度间差异无统计学意义(P>0.05)。SCD高浓度组可上调成纤维细胞VEGF、p-ERK1/2的表达(P<0.01),但SCD不影响TGF-β1表达(P>0.05)。[结论]SCD可通过促进成纤维细胞增殖、黏附和迁移,并促进VEGF蛋白表达,从而发挥促进剖宫产瘢痕愈合的作用,其机制可能与ERK信号通路激活有关。
英文摘要:
      [Objective] To observe the effects of Sanqi Chonglou Shenghua Decoction(SCD) on proliferation, adhesion, migration and expression of healing related proteins of L929 fibroblasts and explore its related mechanism. [Methods] L929 fibroblasts were cultured and randomly divided into SCD low dose group, SCD medium dose group, SCD high dose group and control group. The drug intervention groups were treated with SCD at different concentrations (1.25,2.5,5mg·mL-1), while control group was treated with medium of same volume. Real time cellular analysis(RTCA) and cell counting kit-8(CCK-8) assay were used to detect the proliferation and adhesion of L929. The scratch test was used to detect the migration of L929. Western blot was used to detect the protein expressions of vascular endothelial growth factor(VEGF), transforming growth factor-β1(TGF-β1), extracellular regulated protein kinases 1/2(ERK1/2), phospho-extracellular regulated protein kinases 1/2(p-ERK1/2). [Results] SCD promoted fibroblast proliferation dynamically. RTCA showed medium and high dose groups promoted proliferation after 12~24h hours' intervention(P<0.01), but there was no significant difference between two groups(P>0.05). CCK-8 assay showed SCD promoted cell proliferation in low, medium and high dose groups(P<0.01). Compared with the low dose group, the promoting effect in medium and high dose groups was more significant(P<0.01). Compared with the medium dose group, the promoting effect in high dose group was more significant(P<0.05). SCD promoted fibroblast migration. After 24 hours' intervention, the medium and high dose groups promoted migration(P<0.05,P<0.01), the promoting effect in high dose group was more significantly(P<0.01). After 36 hours' intervention, the low, medium and high dose groups promoted migration(P<0.05,P<0.01). Compared with low dose group, the promotion effect of medium and high dose group was more significant(P<0.05,P<0.01). Compared with the medium dose group, the promoting effect in high dose group was more significant(P<0.01). Compared with control group, SCD promoted fibroblast adhesion(P<0.01), but there was no significant difference between different concentrations (P>0.05). SCD high dose group could upregulate the expression of VEGF and p-ERK1/2(P<0.01), but it did not affect the expression of TGF-β1(P>0.05). [Conclusion] SCD can promote the proliferation, adhesion and migration of fibroblasts and increase the protein expression of VEGF, so as to promote the healing of cesarean section scars, and it may be related to the activation of ERK signaling pathway.
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