S mad交互蛋白1在增生性瘢痕组织中的表达及其对纤维化相关因子的影响
S mad교호단백1재증생성반흔조직중적표체급기대섬유화상관인자적영향
Expression of Smad interacting protein 1 in hyperplastic scar tissue and effects on fibrosis-related factors
  • 万方数据
    中华损伤与修复杂志（电子版） 중화손상여수복잡지（전자판）
    Chinese Journal of Injury Repair and Wound Healing
    2014年 2期 144-150 ,共7页

    方小兵%胡晓龙%石继红%蔡维霞%白晓智%贾文斌%刘佳琦%韩夫%胡大海

    방소병%호효룡%석계홍%채유하%백효지%가문빈%류가기%한부%호대해

    Smad交互蛋白1%增生性瘢痕成纤维细胞%转化生长因子β1%α平滑肌肌动蛋白%结缔组织生长因子 Smad交互蛋白1%增生性瘢痕成纖維細胞%轉化生長因子β1%α平滑肌肌動蛋白%結締組織生長因子
    Smad교호단백1%증생성반흔성섬유세포%전화생장인자β1%α평활기기동단백%결체조직생장인자
    Smad interacting protein 1%Hypertrophic scars fibroblasts%Transforming growthfactor beta1%α-smooth muscle actin%Connective tissue growth factor
    目的：研究Smad 交互蛋白1（SIP1）在人增生性瘢痕组织中的表达及其对纤维化相关因子的影响。方法收集临床整形手术切取的9例患者增生性瘢痕标本及其自体正常皮肤标本。分离培养原代人增生性瘢痕成纤维细胞（HSFBs），取第3～5代细胞用于实验。（1）免疫组织化学法检测增生性瘢痕组织标本及其自体正常皮肤组织标本中SIP1的表达情况。（2）真核表达质粒pcDNA 3．1（＋）/SIP1转染HSFBs。按照随机数字表法将细胞分为6组：对照组；pcDNA3．1（＋）组（空载体组）；pcDNA3．1（＋）/SIP1组；对照＋TGF-β1组；pcDNA3．1（＋）＋TGF-β1组；pcDNA3．1（＋）/SIP1＋TGF-β1组。于转染后第48 h和72 h分别提取细胞总RNA和细胞总蛋白。应用实时荧光定量PCR法检测各组细胞α平滑肌肌动蛋白（α-SMA）及结缔组织生长因子（CTGF）的mRNA表达水平；采用Western-blotting检测α-SMA的蛋白表达水平。结果（1）免疫组织化学染色结果显示：增生性瘢痕组织中SIP1表达明显低于自体正常皮肤组织。（2）pcDNA3．1（＋）/SIP1转染HSFBs后纤维化相关因子的表达：①α-SMA的mRNA和蛋白表达水平：与对照组和pcDNA3．1（＋）组相比，pcDNA3．1（＋）/SIP1组在 mRNA和蛋白表达水平均显著下调，差异有统计学意义（P＜0．05）。当细胞给予TGF-β1刺激后，各组α-SMA表达均上调，但pcDNA3．1（＋）/SIP1＋TGF-β1组mRNA和蛋白表达水仍低于对照＋TGF-β1组，差异有统计学意义（P＜0．05）。② CTGF的mRNA表达水平：与对照组和pcDNA3．1（＋）组相比，pcDNA3．1（＋）/SIP1组在mRNA表达水平显著下调，差异有统计学意义（P＜0．05）。当细胞给予TGF-β1刺激后，各组CTGF表达均显著上调，但pcDNA3．1（＋）/SIP1＋TGF-β1组mRNA表达水仍低于对照＋TGF-β1组，差异有统计学意义（P＜0．05）。结论与正常皮肤组织相比较，SIP1在增生性瘢痕组织中低表达。SIP1基因转染 HSFBs 可降低纤维化相关因子α-SMA 和CTGF的表达，有利于抑制瘢痕形成和减轻挛缩。
    목적：연구Smad 교호단백1（SIP1）재인증생성반흔조직중적표체급기대섬유화상관인자적영향。방법수집림상정형수술절취적9례환자증생성반흔표본급기자체정상피부표본。분리배양원대인증생성반흔성섬유세포（HSFBs），취제3～5대세포용우실험。（1）면역조직화학법검측증생성반흔조직표본급기자체정상피부조직표본중SIP1적표체정황。（2）진핵표체질립pcDNA 3．1（＋）/SIP1전염HSFBs。안조수궤수자표법장세포분위6조：대조조；pcDNA3．1（＋）조（공재체조）；pcDNA3．1（＋）/SIP1조；대조＋TGF-β1조；pcDNA3．1（＋）＋TGF-β1조；pcDNA3．1（＋）/SIP1＋TGF-β1조。우전염후제48 h화72 h분별제취세포총RNA화세포총단백。응용실시형광정량PCR법검측각조세포α평활기기동단백（α-SMA）급결체조직생장인자（CTGF）적mRNA표체수평；채용Western-blotting검측α-SMA적단백표체수평。결과（1）면역조직화학염색결과현시：증생성반흔조직중SIP1표체명현저우자체정상피부조직。（2）pcDNA3．1（＋）/SIP1전염HSFBs후섬유화상관인자적표체：①α-SMA적mRNA화단백표체수평：여대조조화pcDNA3．1（＋）조상비，pcDNA3．1（＋）/SIP1조재 mRNA화단백표체수평균현저하조，차이유통계학의의（P＜0．05）。당세포급여TGF-β1자격후，각조α-SMA표체균상조，단pcDNA3．1（＋）/SIP1＋TGF-β1조mRNA화단백표체수잉저우대조＋TGF-β1조，차이유통계학의의（P＜0．05）。② CTGF적mRNA표체수평：여대조조화pcDNA3．1（＋）조상비，pcDNA3．1（＋）/SIP1조재mRNA표체수평현저하조，차이유통계학의의（P＜0．05）。당세포급여TGF-β1자격후，각조CTGF표체균현저상조，단pcDNA3．1（＋）/SIP1＋TGF-β1조mRNA표체수잉저우대조＋TGF-β1조，차이유통계학의의（P＜0．05）。결론여정상피부조직상비교，SIP1재증생성반흔조직중저표체。SIP1기인전염 HSFBs 가강저섬유화상관인자α-SMA 화CTGF적표체，유리우억제반흔형성화감경련축。
    Objective To study the expression of Smad interacting protein 1 (SIP1)in hyperplasticscar tissue and its effects on fibrosis-related factors.Methods Collection of clinical specimens that were remove from 9 patients of hypertrophic scars and autologous normal skin specimens. Primary human hypertrophic scars fibroblasts (HSFBs)were isolated and cultured in vitro.The 3rd to 5th generation cells were used in the experiments.(1 )The expression levels of SIP1 in hypertrophic scars and autologous normal skin specimens were detected by immunohistochemistry.(2)pcDNA3.1 (+)/SIP1 plasmid transfection HSFBs.HSFBs were divided into 6 groups according to a random number table cells.① control group;②pcDNA3.1 (+)group(empty vector group);③ pcDNA3.1 (+)/SIP1 group;④ control +TGF-β1 group;⑤ pcDNA3.1 (+) +TGF-β1 group;⑥ pcDNA3.1 (+)/SIP1 +TGF-β1 group.After transfection for 48 h and 72 h,total RNA and proteins were extracted from the cells,respectively.The mRNA expression levels of α-SMA and CTGF were detected by real-time quantitative PCR.The protein expression levels ofα-SMA were detected by Western blotting.Results (1 )Immunohistochemical staining showed that expression of SIP1 in hypertrophic scar tissue was significantly reduced compared with autologous normal skin tissue.(2)The expression of fibrosis-related factors after pcDNA3.1 (+)/SIP1 transfected into HSFBs.① mRNA and protein expression levels ofα-SMA showed that compared with the control group and pcDNA3.1 (+)group,in pcDNA3.1 (+)/SIP1 group,the α-SMA expression leves both at mRNA and protein were significantly reduced,the differences were statistically significant (P<0.05 ).After cells stimulate with TGF-β1 ,α-SMA expression in each group were raised,but the expression levels both mRNA and protein in pcDNA3.1 (+)/SIP1 +TGF-β1 group remained lower than the control +TGF-β1 group and the differences were statistically significant (P<0.05 ).② CTGF mRNA expression levels showed that compared with the control group and pcDNA3.1 (+)group,in pcDNA3.1 (+)/SIP1 group,the mRNA expression levels of CTGF were significantly reduced,the differences were statistically significant (P <0.05).After cells stimulate with TGF-β1,CTGF expression in each group were raised,but the expression levels of mRNA in pcDNA3.1 (+)/SIP1 +TGF-β1 group remained lower than the control +TGF-β1 groupandthedifferenceswerestatisticallysignificant(P<0.05).Conclusions Comparedtonormalskin tissue,SIP1 expression in hypertrophic scar tissue were decreased.HSFBs transfected SIP1 gene can reduce the expression of fibrosis-related factors (α-SMA and CTGF ) that may be inhibit scars formation and attenuate their contracture.
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