独角莲根茎水提取物对瘢痕疙瘩成纤维细胞的抑制作用

摘要

Objective To study the inhibitory effect of Typhonium gigantewm Eng1.(AEoTGE) on the proliferation and apoptosis of KFB in vitro and to survey the death rate.Methods Samples of hypertrophic scars were collected and cultured.Only 4-8 passage cells were selected for experiment.Inverted microscope and transmission electron microscope were used to observe the morphogenesis and ultrastructure of KFB.The KFB cells were treated with AEoTGE in different concentrations(3.125,6.250,12.500,25.000,50.000,100.000 g/L) for 24 hours.The effect of AEoTGE on the proliferation and the IC50 of KFB was observed with MTT assay and EdU.The effect of AEoTGE on apoptosis of KFB was detected by flow cytometry.Results It showed that AEoTGE could inhibit the proliferation of KFB in an concentration-dependent style within the range of 3.125-100.000 g/L.The AEoTGE could obviously increase the apoptosis rate of the KFB compared with blank control group(P ＜ 0.05).The IC50 of AEoTGE was 35 g/L.FITC-Annexin V/PI showed that apoptosis rate of KFB in the AEoTGE group was (72.07 ± 0.70)％,while it was 23.5％ in blank control group (P ＜ 0.05).Conclusions AEoTGE could significantly inhibit the proliferating activity and induce apoptosis of KFB after co-culture for 24 hours.The IC50 is 35 g/L and the rate of apoptosis is (72.07 ±0.70)％.%目的 探讨独角莲根茎水提取物(AEoTGE)对瘢痕疙瘩成纤维细胞的抑制作用,测定其半数抑制浓度及凋亡率.方法 采用组织块法体外培养瘢痕疙瘩成纤维细胞,以透射电镜鉴定细胞超微结构.四甲基偶氮唑盐比色法(MTT法)检测不同浓度AEoTGE(3.125、6.250、12.500、25.000、50.000、100.000g/L)作用瘢痕疙瘩成纤维细胞24 h后的抑制率,计算其半数抑制浓度(IC50).采用5-乙炔基-2-脱氧尿苷(EdU)染色法测定瘢痕疙瘩成纤维细胞的增殖情况,流式细胞技术检测其凋亡率.结果 ①应用不同浓度(3.125～100.000 g/L) AEoTGE作用瘢痕疙瘩成纤维细胞24 h后,随AEoTGE浓度增加对细胞抑制作用增强,与对照组比较,细胞抑制率差异有统计学意义(P＜0.05)；②瘢痕疙瘩成纤维细胞经AEoTGE作用后的半数抑制浓度(IC50)为(35±0.27) g/L；③EdU染色示35 g/L AEoTGE组与对照组比较差异有统计学意义(P＜0.05)；④FITC-Annexin V/PI示AEoTGE组凋亡率为(72.07±0.70)％,对照组为(23.48±1.63)％(P＜0.05).结论 体外培养人瘢痕疙瘩成纤维细胞经AEoTGE作用24 h后,半数抑制浓度(IC50)为(35±0.27) g/L,并可明显诱导细胞凋亡,凋亡率为(72.07±0.70)％.