摘要
目的探讨增生性瘢痕成纤维细胞（HSF）经过光敏剂血卟啉单甲醚（HMME）诱导，Smad3蛋白在光动力疗法（PDT）中的磷酸化效应。方法将原代培养的HSF分为对照组、PDT组、单纯光敏剂（PS）组和单纯（Laser）激光照射组，经Smad3-FITC染色后在荧光显微镜上观察各组细胞内Smad3的荧光强度；收集各组细胞经Western印迹法检测Smad3和磷酸化Smad3蛋白的含量。结果在荧光显微镜下观察到各组总体荧光强度相近，但与对照组核内荧光聚集较明显相比，PDT组HSF核内荧光微弱；HSF中磷酸化Smad3蛋白经PDT治疗后含量降低，与对照组相比，二者差异有统计学意义（0．92±0．15比0．20±0．02，P〈0．05），PS组和Laser组中磷酸化Smad3蛋白表达，差异无统计学意义（P〉0．05）。结论PDT能够降低Smad3蛋白磷酸化水平，从而抑制HSF增殖。
Objective To observe the phosphorylation of Smad3 in hyperplastic scar fibroblasts （HSFs） induced by hematoporphyrin monomerthyl ether （HMME） followed by photodynamic therapy （PDT）. Methods Fibroblasts were isolated from the hypertrophic scar tissues of 10 patients and subjected to culture in vitro. After 3-5 passages, the HSFs were divided into 4 groups： control group receiving no treatment, PDT group pretreated with HMME of 4 p^g/ml followed by PDT, HMME group induced by HMME alone, and laser group irradiated with laser alone. Fluorescence microscopy was used to observe the expression of Smad3 after immunofluorescent staining with anti-Smad3 antibody, and Western blot to detect the expression of Smad3 and phosphorylated Smad3 in these HSFs. Paired t test was conducted to compare the difference in Smad3 and phosphorylated Smad3 expression between these groups. Results The total fluorescence intensity of Smad3 was similar between these groups, but the intranuelear fluorescence signal was significantly weaker in the PDT group than in the control group. The level of phosphorylated Smad3 was statistically decreased in the PDT group compared with the control group （0.20 ± 0.02 vs. 0.92 ±0.15, P 〈 0.05）, but no significant difference was observed between the HMME group and laser group （P 〉 0.05）. Conclusion PDT may inhibit the proliferation of HSFs via attenuating the phosphorvlation of Smad3.
出处
《中华皮肤科杂志》
CAS
CSCD
北大核心
2013年第6期394-396,共3页
Chinese Journal of Dermatology
