莪术二酮对增生性瘢痕成纤维细胞的作用及机制探讨

Pharmacological analysis of curdione on hypertrophic scar fibroblast

目的探讨莪术二酮调节增生性瘢痕成纤维细胞增殖、转化、胶原分泌的作用及机制。方法采用人源增生性瘢痕成纤维(HSF)细胞进行培养,加入不同浓度的莪术二酮进行干预。MTT法检测细胞增殖抑制率,ELISA法检测细胞上清中I型胶原蛋白(COL-I)、III型胶原蛋白(COL-III)、基质金属蛋白酶抑制剂-1(TIMP-1)及基质金属蛋白酶1(MMP-1)水平,免疫组化法分析细胞α-平滑肌肌动蛋白(α-SMA)表达情况,Western blotting法检测细胞内PI3K/Akt/m TOR通路及转化生长因子β1(TGF-β1)/Smads通路相关分子表达情况。结果莪术二酮可有效抑制HSF细胞增殖;与对照组比较,不同浓度莪术二酮可明显减少细胞TIMP-1、COL-I及COL-III的分泌(P<0.05、0.01),促进MMP-1合成(P<0.05、0.01),同时莪术二酮能明显抑制PI3K、Akt、m TOR、Smad3的磷酸化及TGF-β1、α平滑肌肌动蛋白(α-SMA)的表达(P<0.05、0.01),且呈浓度依赖性。结论莪术二酮可通过下调PI3K/Akt/m TOR及TGF-β1/Smads信号传导通路,同时抑制HSF细胞增殖、转化,减少胶原分泌,促进胶原酶解,继而有效改善增生性瘢痕的发生与发展。

Objective To investigate the effects of curdione on the HSF proliferation, transformation, and collagen secretion. Methods After the human HSF was treated with curdione, the proliferation inhibition ratio was measured using MTT method. Meanwhile, the TIMP-1, MMP-1, COL-I, and COL-III were detected by ELISA method, the α-SMA was analyzed by IHC technology, and the PI3 K/Akt/m TOR and TGF-β1/Smads related molecular were evaluated by Western blotting. Results Curdione could reduce the proliferation inhibition ratio. Compared with control group, the TIMP-1, COL-I, and COL-III secretion were inhibited by curdione significantly（P 〈 0.05, P 〈 0.01）, while the MMP-1 levels was significantly increased by curdione（P 〈 0.05, P 〈 0.01）. The results also indicated that the expression levels of p-PI3 K, p-Akt, p-m TOR, p-Smad3, TGF-β1, and α-SMA were significantly suppressed by curdione with concentration dependence（P 〈 0.05, P 〈 0.01）. Conclusion Curdione could effectively improve the hypertrophic scar by inhibiting the HSF proliferation, transformation, and collagen secretion, and accelerating the collagen enzymolysis via PI3 K/Akt/m TOR and TGF-β1/Smads pathways.