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目的:探究丹参酮ⅡA磺酸钠(STS)对人增生性瘢痕(HS)的成纤维细胞(Fb)超微结构及自由基代谢平衡的影响,从抗氧化途径寻找一种新的治疗HS的有效方法。方法:收集术中7例HS组织进行Fb体外培养,取第3～6代传代细胞进行实验观察,设置实验组和对照组,实验组设置不同浓度组STS(0.05,0.075,0.10,0.125,0.15,0.20mg/ml)干预培养,对照组加入等量不含STS的DMEM培养液。①干预培养48h后应用倒置相差显微镜和透射电镜观察细胞形态及超微结构改变;②分别于培养12h,24h,48h后,应用化学比色法检测细胞内丙二醛(MDA)含量、黄嘌呤氧化酶(XOD)活力、总超氧化物歧化酶(T-SOD)活力及谷胱甘肽过氧化物酶(GSH-Px)活力变化。结果:①透射电镜见实验组细胞内线粒体肿胀及空泡样变性,粗面内质网扩张、囊泡变及脱颗粒,胶原纤维束减少,细胞坏死等现象;②与对照组比较,实验组MDA含量及XOD活力明显降低(P<0.05或P<0.01),且存在浓度-时间效应。与对照组比较,实验组T-SOD和GSH-Px活力水平有不同程度升高,其升高程度受药物浓度和作用时间影响。与对照组比较,药物作用12h时,高浓度组(0.15mg/ml和0.20mg/ml)T-SOD活力水平升高明显(P<0.05);24h时,0.125～0.20mg/ml组显著升高(P<0.05或P<0.01);48h时,0.05mg/ml～0.10mg/ml组显著升高(P<0.05或P<0.01),0.15mg/ml和0.20mg/ml组显著降低(P<0.01)。与对照组比较,药物作用12h时,0.075～0.15mg/ml组GSH-Px活力水平明显升高(P<0.05或P<0.01),24h和48h时0.05mg/ml和0.075mg/ml组明显升高(P<0.01),48h时0.20mg/ml组GSH-Px活力水平明显降低(P<0.01)。结论:STS可通过降低自由基产生及增强抗氧化酶活性调节瘢痕成纤维细胞自由基代谢平衡和生物学功能。
Objective: To investigate the effect of sodium tanshinone Ⅱ A sulfonate (STS) on the ultrastructure of fibroblasts (Fb) and the balance of free radical metabolism in human hypertrophic scars (HS), and to find out a new effective way to treat HS from antioxidant pathway method. Methods: Seven cases of HS tissues were collected for Fb culture in vitro. The passage cells from passage 3 to passage 6 were collected for experimental observation. Experimental and control groups were set up. The experimental groups were given different concentrations of STS (0.05,0.075,0.10,0.125,0.15 , 0.20mg / ml) intervention culture, the control group was added an equal amount of STEM-free DMEM culture medium. ① The morphological and ultrastructural changes of cells were observed by inverted phase contrast microscope and transmission electron microscope after 48 h intervention; ② The contents of malondialdehyde (MDA), xanthine Oxidase (XOD) activity, total superoxide dismutase (T-SOD) activity and glutathione peroxidase (GSH-Px) activity changes. Results: (1) The phenomenon of mitochondrial swelling and vacuolar degeneration, rough endoplasmic reticulum dilation, vesicle change and degranulation, collagen fiber bundles reduction and cell necrosis were observed in the experimental group by transmission electron microscopy. ②Compared with the control group, MDA content and XOD activity decreased significantly (P <0.05 or P <0.01), and there was concentration-time effect. Compared with the control group, the experimental group T-SOD and GSH-Px activity levels increased to varying degrees, the degree of increase by the drug concentration and duration of action. Compared with the control group, the activity of T-SOD in high concentration group (0.15mg / ml and 0.20mg / ml) increased significantly at 12h (P <0.05); at 24h, 0.125 ~ 0.20mg / (P <0.05 or P <0.01). At 48 hours, 0.05mg / ml ~ 0.10mg / ml group was significantly increased (P <0.05 or P <0.01), 0.15mg / ml and 0.20mg / ml group was significantly lower P <0.01). Compared with the control group, the activity of GSH-Px in the 0.075-0.15mg / ml group was significantly increased at 12h (P <0.05 or P <0.01), and significantly increased at 0.05mg / ml and 0.075mg / ml at 24h and 48h (P <0.01). At 48h, the activity of GSH-Px in 0.20mg / ml group was significantly decreased (P <0.01). Conclusion: STS can regulate free radical metabolism and biological function of scar fibroblasts by reducing the production of free radicals and enhancing the activity of antioxidant enzymes.