沙棘总黄酮对人增生性瘢痕成纤维细胞增殖、迁移和凋亡的影响  

Effects of total flavonoids of sea buckthorn on proliferation,migration and apoptosis of human hypertrophic scar fibroblasts

作　　者：牛梓晗 余扬[1] 艾江 李文博 卜盼盼 马少林[1] Niu Zihan;Yu Yang;Ai Jiang;Li Wenbo;Bu Panpan;Ma Shaolin(Department of Plastic Surgery,the First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,China)

机构地区：[1]新疆医科大学第一附属医院整形外科,乌鲁木齐830054

出　　处：《中华实验外科杂志》2022年第12期2340-2343,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(81750345)。

摘　　要：目的:探究沙棘总黄酮(TFH)对人增生性瘢痕成纤维细胞(Fbs)增殖、迁移和凋亡的影响。方法:增生性瘢痕(HS)组织取自2019年9月至2020年3月新疆医科大学第一附属医院整形外科10例HS患者,组织块法培养Fbs,通过细胞计数实验(CCK-8)检测不同浓度TFH对Fbs增殖活力的影响,细胞划痕实验检测TFH对Fbs迁移能力的影响,流式细胞术检测TFH对Fbs细胞周期、凋亡的影响,分析细胞周期比例和凋亡率;蛋白质印迹法(Western blot)检测TFH对Fbs中Ⅰ型胶原(Col1)、Ⅲ型胶原(Col3)、α-平滑肌肌动蛋白(α-SMA)、B淋巴细胞瘤-2(bcl-2)、bcl-2相关X蛋白(bax)、半胱氨酰天冬氨酸特异性蛋白酶(Caspase)-3蛋白表达影响。组间比较采用单因素方差分析。结果:50、100、200、400、800μg/ml TFH呈浓度依赖性抑制Fbs细胞增殖[(16.35±2.41)%比(20.86±2.58)%比(40.89±0.33)%比(76.23±0.14)%比(85.04±0.58)%,F=75.63,P<0.05]。干预24 h半数抑制浓度(IC 50)=224.2μg/ml,分为对照组、150μg/ml TFH组、300μg/ml TFH组。随TFH浓度增大,伤口愈合面积减小(5.74±0.54比2.55±0.32比2.07±0.41,F=18.43,P<0.01),G_(0)/G_(1)期细胞比例减少[(86.24±1.71)%比(79.42±0.48)%比(74.64±1.92)%,F=44.72,P<0.01],S期和G_(2)/M期比例增多[(9.27±1.28)%比(12.57±0.47)%比(17.36±1.90)%,F=44.72,P<0.05,(4.49±0.44)%比8%比8%,F=190.6,P<0.01],Fbs凋亡率升高[(0.40±0.26)%比(19.43±0.91)%比(31.17±1.46)%,F=719.3,P<0.01],Fbs中Col1、Col3、α-SMA、bcl-2、Caspase-3蛋白表达减少(1.07±0.01比0.73±0.01比0.58±0.01,F=10840,P<0.01;1.14±0.03比0.87±0.01比0.70±0.01,F=446.50,P<0.01;1.14±0.02比0.87±0.01比0.69±0.01,F=1314,P<0.01;1.13±0.02比0.68±0.01比0.56±0.01,F=2756,P<0.01;0.92±0.01比0.89±0.01比0.61±0.01,F=1039,P<0.01],bax蛋白表达水平升高(0.91±0.01比1.07±0.01比1.03±0.01,F=192.90,P<0.05),作用呈浓度依赖性。结论:TFH可抑制人增生性瘢痕Fbs增殖、迁移能力,诱导细胞凋亡,调节胶原代谢。Objective To investigate the effects of total flavones of hippophae rhamnoides(TFH)on proliferation,migration and apoptosis of human hypertrophic scar fibroblasts(Fbs).Methods The hypertrophic scar(HS)samples were taken from 10 HS patients in the plastic surgery department of the First Affiliated Hospital of Xinjiang Medical University from September 2019 to March 2020.Fbs were cultured by tissue block method,and the effects of different concentrations of TFH on the proliferation activity of Fbs were detected by cell counting experiment,the effects of TFH on the migration ability of Fbs were detected by cell scratch experiment,and the effects of TFH on the cell cycle and apoptosis of Fbs were detected by flow cytometry,and the cell cycle proportion and apoptosis rate of each group were analyzed;Western blotting was used to detect the effects of TFH on the expression of typeⅠcollagen(Col1),typeⅢcollagen(Col3),α-smooth muscle actin(α-SMA),B cell lymphoma/leukemia-2(bcl-2),bcl-2 associated X protein(bax),and cysteinyl aspartate-specific protease(Caspase)-3 proteins in Fbs.One-way ANOVA was used for between-group comparisons.Results 50,100,200,400,800μg/ml TFH inhibited Fbs proliferation in concentration-dependent manner[(16.35±2.41)%vs.(20.86±2.58)%vs.(40.89±0.33)%vs.(76.23±0.14)%vs.(85.04±0.58)%,F=75.63,P<0.05].24 h half maximal inhibitory concentration(IC50)=224.2μg/ml.Divided into control group,150μg/ml TFH group,300μg/ml TFH group.With the increase of TFH concentration,the proportion of cells in the G_(0)/G_(1)phase decreased[(86.24±1.71)%vs.(79.42±0.48)%vs.(74.64±1.92)%,F=44.72,P<0.01],S phase and G_(2)/M phase increased[(9.27±1.28)%vs.(12.57±0.47)%vs.(17.36±1.90)%,F=44.72,P<0.05,(4.49±0.44)%vs.8%vs.8%,F=190.6,P<0.01],and the apoptosis rate increased[(0.40±0.26)%vs.(19.43±0.91)%vs.(31.17±1.46)%,F=719.3,P<0.01].The expression of Col1,Col3,α-SMA,bcl-2 and Caspase-3 proteins in Fbs was reduced with an increase in TFH concentration(1.07±0.01 vs.0.73±0.01 vs.0.58±0.01,F=10840,P<0.01;1.14

关 键 词：沙棘总黄酮 成纤维细胞 增殖 迁移 凋亡 

分 类 号：R285[医药卫生—中药学]