E-selectin protein levels were analyzed after 2 h. (B) Cell ELISA of IL-1β (5 ng/ml, 90 min) stimulated HUVECs which were either left untreated (CTRL, black bar) or pretreated (30 min) with different doses (either 160, 130, 80, 60 μg/ml) of comfrey-RE (gray bars), comfrey-OP (20, 12, 10, 6 μg/ml dark gray bars) or the inhibitor PHA-408 (20, 10, 5, 2.5 μM light gray bars). Relative mRNA levels of E-selectin were normalized to GAPDH and expression levels are depicted as mean fold change compared to non-stimulated cells. (A) Real-time PCR of IL-1β (5 ng/ml, 90 min) stimulated HUVEC which were either left untreated (CTRL, black bar) or pretreated (30 min) with either 166, 125, 100 μg/ml of comfrey-RE (gray dotted bars) or with 20, 10, 5 μg/ml of comfrey-OP (black dotted bars). officinale root extract preparations affect E-selectin expression in a dose- and time-dependent manner. NF-κB Symphytum officinale comfrey endothelial cells inflammation transactivation transcription. These results provide a first mechanistic insight into the mode of action of a century-old popular herbal medicine. Furthermore, our biochemical studies provide evidence that comfrey inhibits NF-κB signaling at two stages: it dampens not only the activation of IKK1/2 and the subsequent IκBα degradation, but also interferes with NF-κB p65 nucleo-cytoplasmatic shuttling and transactivation. Both preparations inhibit the activation of NF-κB, a transcription factor of central importance for the expression of these and other pro-inflammatory genes. The extract, and especially its mucilage-depleted fraction, impair the interleukin-1 (IL-1) induced expression of pro-inflammatory markers including E-selectin, VCAM1, ICAM1, and COX-2. Here, we show that a hydroalcoholic extract of comfrey root impairs the development of a pro-inflammatory scenario in primary human endothelial cells in a dose-dependent manner. However, the molecular basis of its action remained elusive. Today, its topical use is based on its analgesic and anti-inflammatory effects, which have been substantiated by modern clinical trials. Symphytum officinale, commonly known as comfrey, constitutes a traditional medicinal plant with a long-standing therapeutic history, and preparations thereof have been widely used for the treatment of painful muscle and joint complaints, wound and bone healing, and inflammation.
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