山萘酚通过抑制Akt磷酸化的水平抑制增生性瘢痕成纤维细胞的迁移和生长  

Kaempferol prevents hypertrophic scar fibroblasts from migrating and expanding through preventing the phosphorylation of Akt

作　　者：刘芮嫡 关莹 韩子阳 陶凯 王洪一 林枫 LIU Ruidi;GUAN Ying;HAN Ziyang;TAO Kai;WANG Hongyi;LIN Feng(Graduate School,Dalian Medical University,Dalian 116000,China)

机构地区：[1]大连医科大学研究生院,辽宁大连116000 [2]北部战区总医院烧伤整形科,辽宁沈阳110016

出　　处：《中国美容整形外科杂志》2023年第7期423-427,共5页Chinese Journal of Aesthetic and Plastic Surgery

基　　金：国家自然科学基金面上项目(81971845);国家自然基金青年基金(82002047)。

摘　　要：目的探讨山萘酚是否能通过抑制Akt的磷酸化水平抑制增生性瘢痕成纤维细胞(hypertrophic scar fibroblasts,HSFB)迁移的能力、阻滞其细胞周期、促进其凋亡,并抑制其增殖能力。方法选取自2021年2-12月,北部战区总医院烧伤整形科接受瘢痕切除术治疗的15例患者的增生性瘢痕(hypertrophic scars,HS)组织。获取HSFB后,将细胞分为3组,分别使用0、50、100μmol/L山萘酚对细胞进行处理,在处理后的第0、12、24、36、48 h采用MTT检测细胞的增殖情况,通过流式细胞技术检测不同细胞周期中的细胞占比;Hoechst 33258实验检测细胞的凋亡情况,Transwell实验检测细胞迁移能力;Western blot实验检测山萘酚对于蛋白激酶B(protein kinase B,Akt)、磷酸化蛋白激酶B(phosphorylated protein kinase B,p-Akt)表达的作用。将经过Akt si-RNA转染后的细胞分为4组,空白对照组、si-Akt组、50μmol/L山萘酚组、50μmol/L山萘酚+si-Akt组,MTT实验检测细胞增殖情况。结果在不同的时间点,50、100μmol/L山萘酚组与0μmol/L组比较,HSFB增殖受到显著的抑制(P<0.05);流式细胞技术显示,在山萘酚作用下G1期细胞的比例增加,并且S期的细胞比例下降(P<0.05)。Hoechst 33258实验显示,山萘酚组细胞的凋亡数明显高于0μmol/L组(P<0.05)。Transwell实验显示,山萘酚组的成纤维细胞被明显的抑制迁移的能力(P<0.05)。Western blot结果表明,山萘酚具有下调p-Akt表达的能力(P<0.05)。MTT实验显示,沉默Akt后再加山萘酚,HSFB并不能被抑制。结论山萘酚可以通过抑制Akt的磷酸化水平抑制HSFB迁移的能力、阻滞其细胞周期、促进其凋亡、并且抑制其增殖。Objective To investigate whether kaempferol can prevent hypertrophic scar fibroblasts(HSFB)from multiplying,prevent them from migrating,interrupt their cell cycle,or accelerat apoptosis by inhibiting the phosphorylation level of Akt.Methods Hypertrophic scar tissue was selected from 15 patients who received scar resection in the Department of Burn and Plastic Surgery of General Hospital of Northern Theater from February to December 2021.While collecting HSFB,the cells were split into three different groups,and treated with 0μmol/L,50μmol/L,100μmol/L kaempferol respectively.MTT was used to measure cell proliferation at 0 h,12 h,24 h,36 h,48 h post-treatment.Flow cytometry was used to measure the proportion of cells in various cell cycles,the Hoechst 33258 assay to quantify cell apoptosis,and the Transwell assay was used to test cell migratory capacity.By using a Western blot assay,the impact of kaempferol on the transcription of Akt and p-Akt was confirmed.The cells transfected with Akt si-RNA were divided into four groups:0μmol/L,si-Akt,Kaempferol,Kaempferol+si-Akt,and cell proliferation was detected by MTT assay.Results At various time intervals,it was seen that the 50 and 100μmol/L kaempferol groups significantly suppressed the multiplication of HSFB in comparison to the control group(P<0.05).Kaempferol dramatically down-regulated both the frequency of G1 phase cells and the amount of S phase cells,according to flow cytometry(P<0.05).The fibroblast apoptosis rate in the kaempferol group was substantially higher than that of the 0μmol/L,according to Hoechst 33258(P<0.05).The transwell analytics to demonstrate a substantial prevention of fibroblast migration in the kaempferol group(P<0.05).According to the results of a Western blot,kaempferol has the ability to suppress the expression of p-Akt(P<0.05).MTT assay showed that after silencing Akt and adding kaempferol,the HSFB could not be inhibited.Conclusion By preventing Akt phosphorylation,kaempferol prevents HSFB from migrating,hindering the cell cycle,sti

关 键 词：山萘酚 增生性瘢痕 迁移 增殖 

分 类 号：R622[医药卫生—整形外科]