洋葱提取物通过MAPK/ERK通路对病理性瘢痕成纤维细胞生长的影响  

Effects of onion extract on the growth of fibroblasts in pathological scar through MAPK/ERK pathway

作　　者：杨萍 李心怡[1] 李小静[1] YANG Ping;LI Xin-yi;LI Xiao-jing(Plastic Surgery Department,the First Affiliated Hospital of Anhui Medical University,Hefei 230000,Anhui Province,China)

机构地区：[1]安徽医科大学第一附属医院整形外科,安徽合肥230000

出　　处：《中国临床药理学杂志》2024年第21期3092-3096,共5页The Chinese Journal of Clinical Pharmacology

摘　　要：目的探讨洋葱提取物对病理性瘢痕成纤维细胞生长的影响,及其作用机制。方法将分离的成纤维细胞随机分为空白组(正常培养)、低剂量实验组(10μmol·L^(-1)洋葱提取物)、中剂量实验组(20μmol·L^(-1)洋葱提取物)、高剂量实验组(40μmol·L^(-1)洋葱提取物)、茴香霉素组[40μmol·L^(-1)洋葱提取物+10μmol·L^(-1)丝裂原活化蛋白激酶(MAPK)信号通路激活药茴香霉素]。用克隆形成实验检测细胞增殖情况,用流式细胞仪检测细胞凋亡情况,用划痕实验检测细胞迁移情况,用蛋白质印迹法检测相关蛋白的相对表达水平。结果空白组、低剂量实验组、中剂量实验组、高剂量实验组和茴香霉素组的细胞克隆形成数分别为(176.89±12.92)、(147.22±5.03)、(102.56±8.42)、(71.56±6.85)和(124.78±10.70)个,凋亡率分别为(4.19±0.29)%、(8.67±0.75)%、(17.59±1.32)%、(25.75±2.10)%和(10.88±1.06)%,迁移率分别为(0.60±0.05)%、(0.43±0.04)%、(0.31±0.02)%、(0.18±0.03)%和(0.51±0.04)%,磷酸化-MAPK(p-MAPK)蛋白相对表达水平分别为0.82±0.12、0.53±0.08、0.43±0.05、0.37±0.05和0.62±0.07,磷酸化-细胞外信号调节激酶1/2(p-ERK1/2)蛋白相对表达水平分别为0.93±0.10、0.73±0.07、0.60±0.07、0.47±0.07和0.76±0.07,alpha平滑肌肌动蛋白(α-SMA)蛋白相对表达水平分别为0.80±0.09、0.60±0.04、0.33±0.03、0.21±0.02和0.77±0.08,Ⅰ型胶原蛋白(CollagenⅠ)相对表达水平分别为0.82±0.08、0.63±0.07、0.43±0.03、0.31±0.02和0.69±0.06。低、中、高剂量实验组的上述指标分别与空白组比较,在统计学上差异均有统计学意义(均P<0.05);茴香霉素组的上述指标与高剂量实验组比较,在统计学上差异均有统计学意义(均P<0.05)。结论洋葱提取物可能通过抑制MAPK/ERK通路抑制成纤维细胞生长,并抑制细胞迁移、纤维化及细胞外基质沉积,诱导细胞凋亡,从而减轻病理瘢痕的形成。Objective To investigate the effect of onion extract on the growth of fibroblasts in pathological scar and its mechanism.Methods The isolated fibroblasts were randomly divided into blank group(normal cultured),experimental-L group(10μmol·L^(-1)onion extract),experimental-M group(20μmol·L^(-1)onion extract),experimental-H group(40μmol·L^(-1)onion extract),anisomycin group[40μmol·L^(-1)onion extract+10μmoL·L^(-1)mitogen activated protein kinase(MAPK)signaling pathway activator anisomycin].Cell proliferation was detected by clonogenesis assay,cell apoptosis was detected by flow cytometry,cell migration was detected by wound healing assay,and relative expression levels of related proteins was detected by Western blot assay.Results The colonies number of blank group,experimental-L group,experimental-M group,experimental-H group and anisomycin group was 176.89±12.92,147.22±5.03,102.56±8.42,71.56±6.85 and 124.78±10.70,respectively;the apoptosis rates were(4.19±0.29)%,(8.67±0.75)%,(17.59±1.32)%,(25.75±2.10)%and(10.88±1.06)%,respectively;the migration rates were(0.60±0.05)%,(0.43±0.04)%,(0.31±0.02)%,(0.18±0.03)%and(0.51±0.04)%,respectively;the relative expression levels of phosphorylation-MAPK(p-MAPK)protein were 0.82±0.12,0.53±0.08,0.43±0.05,0.37±0.05 and 0.62±0.07,respectively;the relative expression levels of phosphorylated extracellular signal-regulated kinase 1/2(p-ERK1/2)protein were 0.93±0.10,0.73±0.07,0.60±0.07,0.47±0.07 and 0.76±0.07,respectively;the relative expression levels of alpha smooth muscle actin(α-SMA)protein were 0.80±0.09,0.60±0.04,0.33±0.03,0.21±0.02 and 0.77±0.08,respectively;the relative expression levels of collagenⅠwere 0.82±0.08,0.63±0.07,0.43±0.03,0.31±0.02 and 0.69±0.06,respectively.The above indexes in experimental-L,-M,-H groups were compared with those in blank group,and the differences were statistically significant(all P<0.05);the above indexes in anisomycin group were significantly different from those in experimental-H group(all P<0.05).Co

关 键 词：洋葱提取物 病理性瘢痕 成纤维细胞 过度增殖 纤维化 丝裂原活化蛋白激酶/细胞外信号调节激酶通路 

分 类 号：R28[医药卫生—中药学]