This study aimed to further explore the relevant mechanism of action by network pharmacology integrated with animal experimental verification based on previous proven effective treatment of vertebral artery type of
cervical spondylosis(CSA) by
Panlongqi Tablets. Bionetwork analysis was performed to establish
drug-disease interaction network, and it was found that the key candidate targets of
Panlongqi Tablets were enriched in multiple signaling pathways related to CSA pathological links, among which
phosphatidylinositol 3-kinase(PI3 K)/
serine-threonine kinase(AKT/PKB) signaling pathway was the most significant. Further, mixed modeling method was used to build the CSA rat model, and the rats were divided into normal, model,
Panlongqi Tablets low-, medium-and high-dose(0.16, 0.32, 0.64 g·kg~(-1)) and Jingfukang Granules(positive
drug, 1.35 g·kg~(-1)) groups. After successful modeling, the rats were administered for 8 consecutive weeks. Pathological changes of rat cervical muscle tissues were detected by
hematoxylin-
eosin(HE) staining, and the content of interleukin-1β(IL-1β),
tumor necrosis factor-α(TNF-α), vascular endothelial cell
growth factor(
VEGF) and
chemokine(C-C motif) ligand 2(CCL2) in rat serum and/or cervical tissues was determined by
enzyme-linked
immunosorbent assay(ELISA). Western blot was employed to detect the
protein expression levels of
chemokine(C-C motif) receptor 2(CCR2), PI3 K, AKT, phosphorylated AKT(p-AKT),
I-kappa-B-kinase beta(
IKK-beta/IKKβ), nuclear factor kappa B(NF-κB P65) and phosphorylated nuclear factor kappa B(NF-κB p-P65) in rat cervical tissues, and positive expression of p-NF-κB P65 in rat cervical muscle tissues was detected by immunofluorescence. The results showed that
Panlongqi Tablets at different doses improved the degree of muscle
fibrosis and
inflammation in cervical muscle tissues of CSA rats, and reduced the content of inflammatory factors IL-1β, TNF-α,
VEGF, CCL2 and CCR2 in serum and/or cervical tissues. The
protein expression levels of PI3 K, p-AKT, IKKβ and p-NF-κB P65 as well as the nuclear entry of p-NF-κB P65 in cervical tissues were down-regulated. These findings suggest that
Panlongqi Tablets can significantly inhibit the inflammatory response of CSA rats, and the mechanism of action may be related to the down-regulation of the activation of PI3 K/AKT signaling pathway.