异搏定对增殖瘢痕成纤维细胞凋亡及细胞信号传导作用  

The Effect of Verapamil on Cellular Apoptosis in Fibroblasts Derived from Keloid and Hypertrophic Scars

作　　者：滕建英[1] 徐少骏[2] 谢菁[2] 吴艾竞[2] 沈明强[2] 

机构地区：[1]杭州师范大学临床医学院,浙江杭州310036 [2]杭州师范大学附属医院整形外科,浙江杭州310015

出　　处：《健康研究》2011年第2期86-90,F0002,F0003,共7页Health Research

摘　　要：目的目的探讨异搏定对瘢痕疙瘩和增生性瘢痕成纤维细胞凋亡及相关的细胞信号传导通路的影响,并与类固醇的作用进行比较。方法对6例瘢痕疙瘩、增生性瘢痕及6例皮肤标本进行细胞培养,对不同成纤维细胞在不同异搏定浓度的培养基中的生长情况进行观察。采用免疫组织化学、凝胶电泳及FACE ELISA方法,通过检测Bax和Bcl-2蛋白表达、特异性DNA梯状条带以及激活(磷酸化)的ERK1/2和JNK的OD值,对不同成纤维细胞在异搏定(0.1 mg/ml)作用后的细胞凋亡及相关细胞信号传导通路进行了研究,同时,以地塞米松(0.1 mg/ml)作用后的结果为对比。结果①当异搏定浓度为0.1 mg/ml,可诱导瘢痕疙瘩、增生性瘢痕成纤维细胞的凋亡;②异搏定可引起瘢痕疙瘩和增生性瘢痕成纤维细胞Bax/Bcl-2比值明显升高,DNA经电泳后可见梯形条带;但不引起皮肤成纤维细胞Bax/Bcl-2表达的改变且未见明显梯形条带;③瘢痕疙瘩成纤维细胞在异搏定作用后JNK细胞传导通路被激活,增生性瘢痕成纤维细胞在异搏定作用后ERK1/2细胞传导通路被抑制而JNK细胞传导通路被激活,皮肤成纤维细胞在异搏定作用后ERK1/2细胞传导通路被激活而JNK细胞传导通路无明显改变。④地塞米松可通过激活ERK1/2和JNK细胞传导通路诱导三类不同来源成纤维细胞发生细胞凋亡。结论异搏定和地塞米松均可诱导增生性瘢痕、瘢痕疙瘩成纤维细胞凋亡,但细胞传导通路不尽相同。异搏定和地塞米松对皮肤成纤维细胞作用不同。Objective To investigate the effect of verapamil on cellular apoptosis and related signal pathways in fibroblasts derived from keloid and hypertrophic scar and to compare its function with steroid.Methods 6 samples of keloid,hypertrophic scars and 6 samples of normal skin tissues were collected respectively and primary fibroblasts were cultured.Fibroblasts were treated with different concentrations of verapamil（1,0.5,0.25,0.1 and 0.01 mg/ml） and the effects of verapamil and dexamethasone were compared.The expression of Bax and Bcl-2 in situ and DNA ladders in fibroblasts were detected by histochemical staining and gel electrophoresis respectively.FACE ELISA was performed for detecting ERK1/2 and JNK pathways.Results ① 0.1mg/ml verapamil induced the apoptosis of fibroblasts from keloid and hypertrophic scar.② The ratios of Bax/Bcl-2 increased in fibroblast from keloid and hypertrophic scar,compared to those from normal skin tissues.③ DNA ladders were detected in fibroblasts from keloida and hypertrophic scars treated with verapamil,whereas,no obvious DNA ladder was detected in the control group.④ Verapamil activated JNK pathway in fibroblasts of keloids and hypertrophic scars,but only prohibited ERK1/2 pathway in fibroblasts of hypertrophic scar;In fibroblasts of skin Verapamil activated ERK1/2 pathway but didn’t affect JNK pathway.⑤ Dexamethasone induced apoptosis in all three types of fibroblasts through activating ERK1/2 and JNK pathway.Conclusion Verapamil and dexamethasone induce apoptosis in fibroblasts from keloid and hypertrophic scar through different pathways.The effects of verapamil and dexamethasone on fibroblasts from skin and their relative pathways were different

关 键 词：瘢痕疙瘩 增生性瘢痕 异搏定 细胞凋亡 细胞信号传导 

分 类 号：R622[医药卫生—整形外科]