Department of Dermatology, Air Force General Hospital, Chinese PLA, Beijing, China.
Photodermatol Photoimmunol Photomed. 2011 Apr;27(2):90-6. doi: 10.1111/j.1600-0781.2011.00577.x.
Clinical studies have demonstrated that photodynamic therapy (PDT) for hyperplastic dermatosis results in a beneficial outcome. Hypertrophic scar (HS) is a pathological process characterized by fibroblastic hyperproliferation. However, it is unclear whether photochemical interactions between PDT and fibroblasts contribute to a beneficial outcome. To investigate the primary photochemical effects of PDT, we studied the efficacy of 630 nm PDT on human fibroblasts from HS using hematoporphyrin monomethyl ether (HMME) as a photosensitizer.
Fibroblasts were cultured from nontreated HSs, and cells at passage 4-6 were used for the experiments. Morphological and biochemical changes in fibroblasts were assessed by Hoechst 33258 staining and annexin V-FITC/PI flow cytometry (FCM). Caspase-3 activity assay and immunofluorescence staining were performed to investigate caspase-3 expression in fibroblasts.
The morphological features of cell apoptosis were viewed under a fluorescent microscope by Hoechst 33258 staining. FCM indicated that the apoptotic rate was significantly increased after HMME-PDT, and caspase-3 activity was observed.
Low-level exposure to 630 nm PDT mediated by HMME appears to induce fibroblast apoptosis and stimulate caspase-3 activation. However, the effect of HMME-PDT on fibroblasts needs further investigation to determine its therapeutic potential for HS.
临床研究表明,光动力疗法(PDT)治疗增生性皮肤病可获得良好的效果。增生性瘢痕(HS)是一种以成纤维细胞过度增殖为特征的病理过程。然而,目前尚不清楚 PDT 与成纤维细胞之间的光化学反应是否有助于获得良好的效果。为了研究 PDT 的主要光化学效应,我们使用血卟啉单甲醚(HMME)作为光敏剂,研究了 630nm PDT 对 HS 人成纤维细胞的疗效。
从未治疗的 HS 中培养成纤维细胞,并使用第 4-6 代细胞进行实验。通过 Hoechst 33258 染色和膜联蛋白 V-FITC/PI 流式细胞术(FCM)评估成纤维细胞的形态和生化变化。通过 caspase-3 活性测定和免疫荧光染色研究成纤维细胞中 caspase-3 的表达。
通过 Hoechst 33258 染色,荧光显微镜下观察到细胞凋亡的形态特征。FCM 表明,HMME-PDT 后细胞凋亡率显著增加,且观察到 caspase-3 活性。
低水平的 630nm PDT 介导的 HMME 似乎诱导成纤维细胞凋亡并刺激 caspase-3 激活。然而,HMME-PDT 对成纤维细胞的影响需要进一步研究,以确定其对 HS 的治疗潜力。