Abstract

PAEONOL AND QUERCITRIN, ACTIVE COMPONENTS OF XIANGQING ANODYNE SPRAY, SYNERGISTICALLY IMPLEMENT ANTI-INFLAMMATORY EFFECTS BY SUPPRESSING P38, ERK, AND AKT SIGNALING PATHWAYS

Xin Tong, Shudong Wang, Ru Cheng, Shanshan Hou, Guangli Zhang, Christian Hanson, Guangming Gong, Xuechun Shan, Wenya Liu, Wenhui Qian and Jihua Fu*

ABSTRACT

Background: Previously, we revealed that Xiangqing anodyne spray (XQAS), a traditional Chinese medicine formulation, significantly decrease acute soft-tissue injury by suppressing p38 and AKT activation. Here, we clarified the effects of XQAS were exerted by its active components which inhibit MAPKs and AKT signaling pathways. Methods: Carrageenan-induced paw edema with acute inflammation in rats were topically applied with XQAS, and THP-1 cell-derived macrophages exposed to TNF-α were treated with ethanol extracts of Xiangqing (EEXQ), Cynanchum Paniculatum (EECP), and Illicium Henryi (EEIH), two herbs composing of XQAS, or with paeonol and quercitrin, two key active components contained in EECP and EEIH, respectively. Protein expression and levels of inflammation-related factors were determined via western blotting and quantitative assay kits, respectively. Results: XQAS could dose-dependently inhibit Carrageenan-induced paw edema with acute inflammation by suppressing the activation of NF-κB. In TNF-α-treated macrophages, the effects of EEXQ were resulted from suppressing TNF-α-induced phosphorylation of ERK1/2, p38, and AKT in a dose-dependent manner. However, the inhibitory effects of EECP and paeonol were mainly occurred on p38, while those of EEIH and quercitrin were primarily pointed to AKT. Moreover, they exhibited similar and moderate inhibitory effect on the phosphorylation of ERK1/2 but fail to influence the phosphorylation of JNK. When composing EECP and EEIH into the EEXQ, it results in a great inhibitory effect on the activation of NF-κB in a synergistic manner. Conclusion: the anti-inflammatory effects of XQAS are attributed to its key active components paeonol and quercitrin by multi-target effects in a synergistic manner.

Keywords: Xiangqing anodyne spray (XQAS); active component; Anti-inflammatory effect; Mitogen-activated protein kinases (MAPKs); Protein kinase B (PKB or AKT).