摘要目的:基于转化生长因子-β1(TGF-β1)/Smads信号通路探讨肉苁蓉苯乙醇总苷(CPhGs)对人增生性瘢痕成纤维细胞(HSFbs)作用的机制。方法:人增生性瘢痕组织标本来自新疆医科大学第一附属医院2019年3月至6月收治的9例需要手术的增生性瘢痕患者，采用组织块培养法分离后传代培养人增生性瘢痕成纤维细胞，随机分为空白组、模型组(TGF-β1)、阳性对照组(TGF-β1＋5-氟尿嘧啶)、实验组(TGF-β1＋CPhGs)，通过细胞计数试剂盒(CCK-8)检测不同浓度CPhGs对HSFbs增殖的影响；实时荧光定量聚合酶链反应(RT-qPCR)检测各组中Smad2、Smad3、Smad7、Ⅰ型胶原(COL-1)和Ⅲ型胶原(COL-3)的mRNA的表达；蛋白质印迹法(Western blot)检测各组中Smad2、Smad3、磷酸化Smad2(p-Smad2)、磷酸化Smad3(p-Smad3)、Smad7、COL-1和COL-3蛋白的表达，多组间比较采用单因素方差分析。结果:60、90、120 mg/L CPhGs作用于成纤维细胞48 h后抑制率为[(36.87±0.06比43.52±0.04比78.11±0.03)%， F＝45.073， P<0.01]，CPhGs对细胞的抑制作用随干预时间的延长和药物浓度的增加而增强；RT-qPCR结果显示，实验组和阳性对照组COL-1、COL-3、Smad2、Smad3 mRNA表达低于模型组，Smad7 mRNA表达明显高于模型组，差异有统计学意义(0.21±0.04比0.3±0.25比1.56±0.01， F＝39.755， P<0.01)、(0.55±0.04比0.27±0.04比1.66±0.22， F＝61.997， P<0.01)、(0.55±0.03比0.78±0.76比1.06±0.15， F＝17.769， P<0.01)、(0.46±0.08比0.55±0.10比2.05±0.29， F＝56.796， P<0.01)、(1.77±0.59比0.11±0.02比0.04±0.04， F＝14.780， P<0.05)；Western blot结果显示：实验组和阳性对照组p-Smad2、p-Smad3、COL-1、COL-3蛋白表达低于模型组，Smad7蛋白表达明显高于模型组(0.16±0.03比0.11±0.02比0.06±0.00)、(0.12±0.02比0.07±0.01比0.04±0.01， F＝7.670， P<0.05)、(0.27±0.02比0.08±0.01比0.05±0.04， F＝34.291， P<0.01)、(0.33±0.02比0.16±0.02比0.17±0.01， F＝16.322， P<0.01)、(0.12±0.03比0.15±0.06比0.23±0.05， F＝3.936， P<0.05)，组间比较差异有统计学意义。 结论:CPhGs的可通过阻断TGF-β/Smad信号通路进而抑制HSFbs活化和增殖。

abstractsObjective:Based on the transforming growth factor-β1 (TGF-β1)/Smads signaling pathway to investigate the effects of phenylethanolglycosides from cistanchetubulosa (CPhGs) on human hypertrophic scar fibroblasts (HSFbs) and the possible mechanism.Methods:From March to June 2019, 9 patients with hypertrophic scar after burn were admitted. The fibroblasts were isolated from hypertrophic scar and cultured. Human hypertrophic scars were removed and fibroblasts were cultured in vitro. The blank group, model group, positive control group (TGF-β1 + 5-fluorouracil), and experimental group (TGF-β1 + CPhGs) were set up. The effects of CPhGs at different concentrations on the proliferation of HSFbs were detected by the cell counting kit-8(CCK-8) assay. The expression of Smad2, Smad3, Smad7, collagen Ⅰ (COL-1) and collagen Ⅲ (COL-3) mRNA was detected by real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) in each group. The expression of Smad2, Smad3, phosphorylated Smad2 (p-Smad2), phosphorylated Smad3 (p-Smad3), Smad7, COL-1 and COL-3 proteins was detected by Western blotting. Measurement data were expressed as mean±standard deviation ( Mean± SD). Multiple groups were compared using one-way ANOVA. Results:The inhibition rate of fibroblasts after treatment with CPhGs at 60, 90, and 120 mg/L for 48 h was (36.87±0.06)% vs. (43.52±0.04)% vs. (78.11±0.03)%, F value=45.070, P<0.01. The inhibitory effects of CPhGs on fibroblasts increased with the increase of drug concentration and the prolongation of action time. RT-qPCR results showed that as compared with the model group, the expression of COL-1, COL-3, Smad2, Smad3 mRNA was reduced, and the expression of Smad7 mRNA was increased in the experimental and positive control groups (0.21±0.04 vs. 0.3±0.25 vs. 1.56±0.01, F value=39.755, P<0.01), (0.55±0.04 vs. 0.27±0.04 vs. 1.66±0.22, F value=61.991, P<0.01), (0.55±0.03 vs. 0.78±0.76 vs. 1.06±0.15, F value=17.769, P<0.01), (0.46±0.08 vs. 0.55±0.10 vs. 2.05±0.29, F value=56.796, P<0.01), (1.77±0.59 vs. 0.11±0.02 vs. 0.04±0.04, F value=14.780, P<0.05); Western blotting results showed that the protein expression of p-Smad2 and p-Smad3, COL-1, COL-3, in the experimental group and positive control group was significantly lower than that in the model group, and the expression of Smad7 protein was significantly higher than that in the model group (0.16±0.03 vs. 0.11±0.02 vs. 0.06±0.00), (0.12±0.02 vs. 0.07±0.01 vs. 0.04±0.01, F value=7.670, P<0.05), (0.12±0.03 vs. 0.15±0.06 vs. 0.23±0.05, F value=3.936, P<0.05), (0.27±0.02 vs. 0.08±0.01 vs. 0.05±0.04, F value=34.291, P<0.01), (0.33±0.02 vs. 0.16±0.02 vs. 0.17±0.01 F value=16.322, P<0.01). Conclusion:CPhG inhibited hypertrophic scar fibroblasts proliferation and activation via blocking the TGF-β1/Smad signaling.