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

 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300 μg·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300 μg·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（300 μg·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300 μg·mL-1能抑制RhoA、ROCK-Ⅰ、CTGF的mRNA与蛋白表达；而且可通过RhoA/ROCK-Ⅰ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。

Effects of Asiaticoside on Dermal Fibroblasts in Hypertrophic Scar

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 (P＜0.05). 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 (P＜0.05). 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.