Ligustrazine mitigates chronic venous disease-induced pain hyperalgesia through desensitization of inflammation-associated TRPA1 activity in DRG

Abstract

Ethnopharmacological relevanceLigustrazine, an important active ingredient extracted from Ligusticum chuanxiong hort, has been widely used to cure cardiovascular diseases and exerts an analgesic effect. Aims of this studyThe aim of this study is to investigate whether ligustrazine mitigates chronic venous disease (CVeD)-induced pain and to explore its underlying mechanisms. Materials and methodsA mouse model of CVeD was established by vein ligature. Ligustrazine was administered intraperitoneally to CVeD mice for a single injection (20 mg/kg, 100 mg/kg, and 200 mg/kg) or once a day for three weeks (100 mg/kg and 200 mg/kg), and TRPA1 overexpressed HEK 293 cells were treated with ligustrazine (600 μM) in the presence of mustard oil (100 μM) for 2 min. Patch clamp and calcium imaging were used to measure the inhibitory response of ligustrazine on DRG neurons and TRPA1 transfected HEK293 cells. ResultsThe present results showed that mice receiving vein ligature surgery exhibited obvious pain hypersensitivity to mechanical, cold and thermal stimuli, whereas ligustrazine significantly reversed the pain hyperalgesia in CVeD mice. Furthermore, ligustrazine desensitized transient receptor potential ankyrin 1 (TRPA1) activity in the dorsal root ganglion (DRG) neurons, resulting in suppressing the DRG neuronal excitability in the CVeD mice. However, ligustrazine could not directly inhibit the response of TRPA1 transfected HEK293 cells to mustard oil. Strikingly, ligustrazine restricted the macrophage infiltration and decreased the mRNA levels of Interleukin-1β (IL-1β) and NOD-like receptor protein 3 (NLRP3) in the DRG neurons of the CVeD mice. ConclusionsThe present study provided evidence that ligustrazine alleviated pain hypersensitivity to mechanical, cold and thermal stimuli in CVeD mice. Ligustrazine could weaken the activity of TRPA1 in the DRG to mitigate CVeD-induced pain hyperalgesia mainly through inhibition of inflammation. Our findings identify that ligustrazine may be a new therapeutic agent for the treatment of CVeD-induced pain.