CAJ | 학술논문

目的分析不同幅度牵张力对正常皮肤成纤维细胞(NSFB)向增生性瘢痕成纤维细胞(HSFB)转化的诱导作用.方法体外培养增生性瘢痕患者的NSFB和HSFB.加载牵张力的NSFB作为实验组,分N10％组、N15％组和N20％组3组,分别加载幅度为10％、15％、20％的周期性牵张力24 h,不加载牵张力的NSFB和HSFB作为对照组(N0组和H0组).通过观察细胞形态,以细胞计数试剂盒(CCK-8)检测细胞增殖,实时定量荧光PCR法检测整合素β1、P130Cas、转化生长因子-β1(TGF-β1)、Ⅰ型胶原a1链(COL1A1)、Ⅲ型胶原a1链(COL3A1)的基因表达,酶联免疫吸附(ELISA)法检测Ⅰ、Ⅲ型胶原和TGF-β1蛋白表达,分析NSFB的转化程度.结果机械牵张力刺激NSFB增殖,N10％组细胞增殖水平显著高于N20％组(0.91±0.09比0.8±0.07,P＜0.05).牵张力作用下各个基因表达上调,N10％组整合素β1、P130Cas、TGF-β1、COL1A1、COL3A1的mRNA表达水平均显著高于N20％组(3.14±0.24、5.01±0.49、1.55±0.08、2.38±0.08、1.37±0.11比1.78±0.32、3.07±0.41、1.18±0.05、1.97±0.05、1.26±0.19,均P＜0.05).机械牵张力引起TGF-β1蛋白表达上调,N10％组TGF-Jβ1的蛋白表达水平显著高于N20％组[(459±12.96)比(372±10.49) pg/ml,P＜0.05].实验组Ⅰ、Ⅲ型胶原蛋白表达水平均显著高于N0组,但低于H0组(均P＜0.05);N10％组Ⅰ型胶原蛋白表达水平显著高于N20％组[(12.66±1.16)比(11.23±0.75)ng/ml,P＜0.05].结论 10％幅度牵张力对NSFB有明显的诱导转化作用,能够使NSFB表现出HSFB的部分生化特征.
목적 분석불동폭도견장력대정상피부성섬유세포(NSFB)향증생성반흔성섬유세포(HSFB)전화적유도작용.방법 체외배양증생성반흔환자적NSFB화HSFB.가재견장력적NSFB작위실험조,분N10％조、N15％조화N20％조3조,분별가재폭도위10％、15％、20％적주기성견장력24 h,불가재견장력적NSFB화HSFB작위대조조(N0조화H0조).통과관찰세포형태,이세포계수시제합(CCK-8)검측세포증식,실시정량형광PCR법검측정합소β1、P130Cas、전화생장인자-β1(TGF-β1)、Ⅰ형효원a1련(COL1A1)、Ⅲ형효원a1련(COL3A1)적기인표체,매련면역흡부(ELISA)법검측Ⅰ、Ⅲ형효원화TGF-β1단백표체,분석NSFB적전화정도.결과 궤계견장력자격NSFB증식,N10％조세포증식수평현저고우N20％조(0.91±0.09비0.8±0.07,P＜0.05).견장력작용하각개기인표체상조,N10％조정합소β1、P130Cas、TGF-β1、COL1A1、COL3A1적mRNA표체수평균현저고우N20％조(3.14±0.24、5.01±0.49、1.55±0.08、2.38±0.08、1.37±0.11비1.78±0.32、3.07±0.41、1.18±0.05、1.97±0.05、1.26±0.19,균P＜0.05).궤계견장력인기TGF-β1단백표체상조,N10％조TGF-Jβ1적단백표체수평현저고우N20％조[(459±12.96)비(372±10.49) pg/ml,P＜0.05].실험조Ⅰ、Ⅲ형효원단백표체수평균현저고우N0조,단저우H0조(균P＜0.05);N10％조Ⅰ형효원단백표체수평현저고우N20％조[(12.66±1.16)비(11.23±0.75)ng/ml,P＜0.05].결론 10％폭도견장력대NSFB유명현적유도전화작용,능구사NSFB표현출HSFB적부분생화특정.
Objective To explore the role of cyclic stretch with different amplitudes on the transformation of normal skin fibroblasts (NSFB) into hypertrophic scar fibroblasts (HSFB).Methods NSFB and HSFB were cultured in vitro.Cyclic stretch with 10％,15％ and 20％ amplitude was applied for 24 h on NSFB of experimental groups (Group N10％,Group N15％ and Group N20％).And control groups (groups N0 and H0),NSFB and HSFB were cultured without stretch.Cell Counting Kit-8 method was used to detect the proliferation of fibroblasts.The gene expressions of integrin β1,P130Cas,TGF-β1,COL1A1 and COL3 A1 were detected by real-time polymerase chain reaction (PCR).And the protein expressions of TGF-β1,collagen types Ⅰ and Ⅲ were detected by enzyme-linked immunosorbent assay.Results Cyclic stretch had stimulatory effect on cell proliferation.The proliferation capacity of group N10％ was higher than that of group N20％ (0.91 ± 0.09 vs 0.8 ± 0.07,P ＜ 0.05).Cyclic stretch could induce increased levels of gene expression.Gene expression levels of integrin β1,P130Cas,TGF-β1,COL1A1 and COL3 A1 in group N10％ was higher than those in group N20％ (3.14 ± 0.24,5.01 ± 0.49,1.55 ± 0.08,2.38 ± 0.08,1.37 ±0.11 vs 1.78±0.32,3.07 ±0.41,1.18 ±0.05,1.97 ±0.05,1.26 ±0.19; all P ＜0.05).Cyclic stretch could stimulate NSFB to secrete TGF-β1.The protein level of TGF-β1 in group N10％ was higher than that in group N20％ ((459 ± 12.96) vs (372 ± 10.49) pg/ml,P ＜0.05).The protein levels of collagen types Ⅰ and Ⅲ in experiment groups were higher than those in group N0 and lower than those in group H0 (all P ＜ 0.05).The protein level of collagen type Ⅰ in group N10％ was higher than that in group N20％ ((12.66±1.16) vs (11.23 ±0.75) ng/ml,P ＜0.05).Conclusions NSFB loaded cyclic stretch with 10％ amplitude could exhibit some biological characteristics of HSFB.And cyclic stretch with optimal amplitude might transform NSFB into HSFB.