厚朴酚通过抑制TGF-β1抑制增生性瘢痕成纤维细胞中的炎症因子

Magnolol inhibits TGF-β1 by inhibiting the expression of TGF-β1 inhibits inflammatory factors in hypertrophic scar fibroblasts

目的 探讨厚朴酚是否能够通过影响TGF-β1影响增生性瘢痕成纤维细胞的炎症反应。方法 自2019年9月至2020年9月,铁岭市第二人民医院外科从10例增生性瘢痕患者身上采集了10份增生性瘢痕组织样本,10例皮瓣移植患者术后采集10份正常皮肤组织样本。通过ELISA检测增生性瘢痕组织和正常皮肤组织中白细胞介素(interleukin, IL)-1β和肿瘤坏死因子-α(tumor necrosis factor-α, TNF-α)的表达情况。获取增生性瘢痕成纤维细胞后,分别采用0、10、20和40μmol/L厚朴酚处理增生性瘢痕成纤维细胞和正常成纤维细胞,在处理后24 h,采用ELISA和实时定量PCR检测检测细胞中转化生长因子-β1(transforming growth factor-β1, TGF-β1)、IL-1β和TNF-α的表达情况,Western blot实验检测TGF-β1的表达情况。分别在细胞中转染对照和TGF-β1后,采用0μmol/L和40μmol/L厚朴酚处理细胞,在处理后24 h时,采用ELISA和实时定量PCR检测细胞中IL-1β、TNF-α和TGF-β1的表达情况,Western blot实验检测TGF-β1的表达情况。结果 ELISA实验检测结果表明,增生性瘢痕组织中IL-1β和TNF-α表达水平明显高于正常组织(P<0.05)。与0μmol/L相比10、20和40μmol/L厚朴酚处理能显著抑制增生性瘢痕成纤维细胞中TGF-β1、IL-1β和TNF-α的表达(P<0.05)。Western blot、ELISA和实时定量PCR实验检测结果显示,不同浓度的厚朴酚作用能够下调增生性瘢痕成纤维细胞中TGF-β1、IL-1β和TNF-α的表达(P<0.05)。TGF-β1沉默后40μmol/L厚朴酚与0μmol/L的比较,增生性瘢痕成纤维细胞中炎症因子和TGF-β1的表达差异无统计学意义(P>0.05)。沉默TGF-β1和40μmol/L厚朴酚均能够显著下调炎症因子和TGF-β1的表达水平。结论 厚朴酚能够通过抑制TGF-β1后抑制增生性瘢痕成纤维细胞中IL-1β1和TNF-α的表达。

Objective To explore whether magnolol can affect the inflammatory response of hypertrophic scar fibroblasts by inhibiting the expression of TGF-β1. Methods From September 2019 to September 2020, 10 hypertrophic scar samples and 10 normal tissues were collected. The expression of IL-1β and TNF-α in hypertrophic scar and normal tissue was detected by ELISA. The hypertrophic scar fibroblasts and normal fibroblasts were respectively treated by magnolol of 0 μmol/L, 10 μmol/L, 20 μmol/L and 40 μmol/L.After treatment for 24 h, the expression of TGF-β1, IL-1β and TNF-α was detected by ELISA, real-time PCR and Western blot. After transfection with si-NC and si-TGF-β1, the fibroblasts were treated with magnolol of 0 μmol/L and 40 μmol/L for 24 h. The expression of IL-1β, TNF-α and TGF-β1 was detected by ELISA, real-time PCR and Western blot. Results The results of ELISA showed that the expression of IL-1β and TNF-α in hypertrophic scar was significantly higher than that in normal tissue. Magnolol of 10 μmol/L, 20 μmol/L and 40 μmol/L could significantly inhibit the expression of TGF-β1, IL-1β and TNF-α in hypertrophic scar fibroblasts(P<0.05). There were no significant differences in inflammatory factors and TGF-β1 in hypertrophic scar fibroblasts between magnolol of 40 μmol/L and0 μmol/L after TGF-β1 silencing(P>0.05). Both TGF-β1 silencing and magnolol of 40 μmol/L could down regulated the expression of inflammatory factors and TGF-β1. Conclusion Magnolol can inhibit the expression of IL-1β and TNF-α in hypertrophic scar fibroblasts by inhibiting TGF-β1.