Institut für Pharmazeutische Technologie, Technische Universität Braunschweig, Braunschweig, Germany; Center of Pharmaceutical Engineering, Technische Universität Braunschweig, Braunschweig, Germany.
Institut für Pharmazeutische Technologie, Technische Universität Braunschweig, Braunschweig, Germany; Center of Pharmaceutical Engineering, Technische Universität Braunschweig, Braunschweig, Germany.
Eur J Pharm Biopharm. 2018 May;126:108-114. doi: 10.1016/j.ejpb.2017.03.003. Epub 2017 Mar 11.
Hyperforin (HYP), one of the main bioactive compounds in extracts of Hypericum perforatum, is a potential drug candidate for the treatment of skin diseases. Since extracts have proven to support wound healing, in the present study effects of HYP on human dermal fibroblasts (HDF) were evaluated in 2D and 3D in vitro dermal constructs. Viability and cytotoxicity assays as well as a live-dead cell staining were performed to test at which concentration HYP reduces viability and/or shows cytotoxicity. Furthermore a differentiation between cytotoxic, anti-proliferative and anti-migratory effects was done. For the latter purpose a 2D migration assay was performed. HDF-induced contraction of a 3D artificial dermal (AD) construct was determined at given HYP concentration. Induction of apoptosis was examined by determination of caspase 3/7 activities. HYP reduced viability of HDF down to 70% at concentrations of 5-10µM. This decrease was not due to cytotoxicity but to a reduction in proliferation as shown from both the proliferation assay and the cytotoxicity assay as well as from live-dead cell staining. The 2D migration assay showed that HYP reduced migration activity of HDF cells at a concentration of 10µM. At this concentration HYP also reduced the HDF-induced contraction of collagen gels as 3D AD constructs. Apoptotic effects of HYP were excluded performing a caspase 3/7 activity detecting assay. The results show for the first time that HYP may be rather a potential candidate for treatment of hypertrophic scars than promoting effects which are understood as important in wound healing.
贯叶金丝桃素（HYP）是贯叶连翘提取物中的主要生物活性化合物之一，是治疗皮肤病的潜在候选药物。由于提取物已被证明能促进伤口愈合，因此本研究在 2D 和 3D 体外真皮模型中评估了 HYP 对人真皮成纤维细胞（HDF）的影响。通过进行细胞活力和细胞毒性测定以及死活细胞染色，检测 HYP 在何种浓度下降低细胞活力和/或表现出细胞毒性。此外，还区分了细胞毒性、抗增殖和抗迁移作用。为此目的，进行了 2D 迁移测定。在给定的 HYP 浓度下，测定 HDF 诱导的 3D 人工真皮（AD）构建体的收缩。通过测定半胱天冬酶 3/7 的活性来检测细胞凋亡的诱导。HYP 将 HDF 的活力降低至 5-10µM 时降低至 70%。这种减少不是由于细胞毒性，而是由于增殖减少，如增殖测定、细胞毒性测定和死活细胞染色所示。2D 迁移测定表明，HYP 在 10µM 浓度下降低了 HDF 细胞的迁移活性。在该浓度下，HYP 还降低了 HDF 诱导的胶原蛋白凝胶作为 3D AD 构建体的收缩。通过进行半胱天冬酶 3/7 活性检测测定排除了 HYP 的凋亡作用。结果首次表明，HYP 可能更适合治疗增生性瘢痕，而不是促进伤口愈合中理解的重要作用。
