积雪草苷对增生性瘢痕成纤维细胞结缔组织生长因子及RhoA/ROCK-Ⅰ调控信号的影响  

Effects of Asiaticoside on Dermal Fibroblasts in Hypertrophic Scar

作　　者：戴丽冰[1,2] 潘姝[1,2] 沈雁[1,2] 黄粤[1,2] 梁蓉[1,2] 康宁[1,2] 李叶扬[1,2] 李孝建[1,2] 谢有富[1,2] 李罡[1,2] 张琰[1,2] 

机构地区：[1]暨南大学医学院第四附属医院 [2]广州市红十字会医院广州市创伤外科研究所,广州510220

出　　处：《中国药学杂志》2010年第14期1067-1072,共6页Chinese Pharmaceutical Journal

基　　金：广东省中医药局科研项目(2007401);广州市中医药;中西医结合科研项目(2007A16);广州市科技局项目(2008J1-C121);广东省自然科学基金(9152800001000009);广州市医药卫生科技重点项目(2005-ZDi-03;2006-ZDi-08;2008-ZDi-08;2009-ZDi-05;2009-ZDi-12)

摘　　要：目的研究积雪草苷对增生性瘢痕成纤维细胞结缔组织生长因子(CTGF)的影响及RhoA/ROCK-Ⅰ信号通路在其中的作用。方法分离培养人增生性瘢痕组织来源的成纤维细胞,应用积雪草苷300μg.mL-1及RhoA的下游效应器Rho激酶的特异抑制剂Y-27632对实验细胞进行干扰实验。分为对照组、积雪草苷组、Y-27632组、积雪草苷+Y-27632组。运用荧光定量PCR(SYBR Green)及免疫荧光细胞化学的方法检测各组瘢痕成纤维细胞中RhA、ROCK-Ⅰ、CTGF mRNA与蛋白表达。结果积雪草苷300μg.mL-1能抑制RhoA、ROCK-Ⅰ、CTGF的mRNA与蛋白表达,与对照组比较具有统计学意义(P<0.05);Y-27632能阻碍积雪草苷的作用,积雪草苷(300μg.mL-1)+Y-27632组,CTGF的mRNA与蛋白表达明显高于单纯积雪草苷组(P<0.05),与对照组及Y-27632组比较无显著性差异(P>0.05);Y-27632组与对照组比较RhoA、ROCK-Ⅰ、CTGF的mRNA与蛋白表达没有统计学意义。结论积雪草苷300μg.mL-1能抑制RhoA、ROCKⅠ-、CTGF的mRNA与蛋白表达;而且可通过RhoA/ROCKⅠ-信号通路抑制瘢痕成纤维细胞中CTGF mRNA与蛋白表达。这可能是积雪草苷治疗瘢痕的作用机制之一。OBJECTIVE To study the effects of asiaticoside on connective tissue growth factor（CTGF） and RhoA/ROCK signal pathway in fibroblasts obtained from hypertrophic scar.METHODS Fibroblasts were obtained from human hypertrophic scars and cultured in the mediums,which were divided into 4 groups： the control group,the asiaticoside group,the Y-27632 group and the asiaticoside＋Y-27632 group.Fluorescent quantitative PCR assay（FQ-PCR） and immunofluorescence cytochemistry were used to quantify the expression and secretion of RhocA,ROCK-Ⅰ and CTGF in the hypertrophic scar fibroblasts of each group.RESULTS To compared with the control group,the expression and secretion of RhocA,ROCK-Ⅰ and CTGF were significantly inhibited in asiaticoside group（P0.05）.But the effects of asiaticoside were inhibited by Y-27632 group.To compared with the asiaticoside group,the expression and secretion of ROCK-Ⅰ and CTGF in the asiaticoside＋Y-27632 group were up regulated（P0.05）.But among the asiaticoside＋Y-27632 group,the Y-27632 group and the control group,no significant difference was found.Between the Y-27632 group and the control group no difference was found neither.CONCLUSION The expression and secretion of RhocA,ROCK-Ⅰ and CTGF in hypertrophic scar fibroblasts through RhoA/ROCK-Ⅰ signal pathway were inhibited by asiaticoside,which may be one of the molecular mechanism of asiaticoside in hypertrophic scar formation.

关 键 词：增生性瘢痕 成纤维细胞 积雪草苷 RHOA ROCK-Ⅰ 结缔组织生长因子 

分 类 号：R965[医药卫生—药理学]