Network Pharmacology Analysis And Mechanism Of Action Of Guizhi Xinjia Decoction In The Treatment Of Cervical Spondylosi

Objective: exploring the mechanism of Guizhi Xinjia Decoction(GZXJD)in the treatment of cervical spondylosis and effectively relieve pain,to provide a basis for clinical application.Methods: Part 1,through the network pharmacology and molecular docking technology to explore the theoretical basis of GZXJD in the treatment of cervical spondylosis,to guide the direction of animal experiment.In Part2,the cervical disc degeneration model was constructed by the dynamic and static imbalance method,and the mechanical withdrawal threshold(MWT)of each group was detected by Von Frey fibers(preoperative,postoperative12 w,treatment 2w,4w,8w).Cervical degeneration was evaluated by anterior and lateral cervical X-ray and MRI images(12W after modeling and 8W after treatment).HE staining was used to detect the morphological changes of cervical intervertebral disc.The contents of substance P(SP),prostaglandin E2(PGE2)and tumor necrosis factor α(TNF-α)in peripheral serum were detected by enzyme linked immunosorbent assay(ELISA),and the expression levels of TNF-α in intervertebral disc and transient receptor potential cation channel,subfamily V,member 1(TRPV1)in spinal cord tissue were detected by immunohistochemistry(IHC)(12W after modeling,2W,4W and 8W after treatment).Part 3,GZXJD,contrast positive drug-celecoxib capsule(Celebrex)was used to treat cervical intervertebral disc degeneration model rats by intragastric administration for 4W.The contents of SP,PGE2,TNF-α in peripheral serum and the expression level of TNF-α in cervical intervertebral disc were compared,and all data were statistically analyzed and compared.Results:1.The possible active ingredients of GZXJD in relieving pain of cervical spondylosis are Betulin,Stigmasterol,Quercetin,Zingiberone,and so on,which are related to TNF,IL-6,IL-1 β,PIK3 CA,STAT3 and other inflammatory related targets.Design the expression of MAPK,PI3K-AKT and other signal pathways.Molecular docking confirmed that the binding energy of Betulin with ESR1,Betulin,Quercetin,Shyobunone,Stigmasterol and FGF23,Betulin with IL-1 β target was high,suggesting that it may be the key active components to exert the effect.2.The contents of SP and TNF-α in peripheral serum in GZXJD2,4 and 8W groups were significantly lower than those in Model at the same time groups(P<0.01);And the contents of PGE2 in GZXJD 4W and 8W groups were significantly lower than those in Model groups at the same time groups(P<0.01).The levels of serum PGE2 and TNF-α in GZXJD group were slightly higher than those in Celebrex group,but there was no significant difference between GZXJD group and GZXJD group(P>0.05).HE staining of cervical intervertebral disc in GZXJD group showed less infiltration of inflammatory cells than in Model group at the same time period.3.TRPV1 receptor expression in spinal cord in GZXJD 4W and 8W groups was4.significantly lower than that in Model 4W and 8W groups(P<0.01),and there was no significant difference between GZXJD 4w group and 8w group(P>0.05).The expression of TNF-α in cervical intervertebral disc in GZXJD 8W group was significantly lower than that in Model 8W group(P<0.05),and the expression level of TNF-α in GZXJD group was lower than that in Celebrex group,but the difference was not significant(P>0.05).Conclusion:1.The results of network pharmacology analysis indicated that GZXJD may act on multiple inflammatory targets such as TNF,IL-6,IL-1β and other inflammatory targets in the treatment of cervical spondylosis.2.The method of dynamic and static imbalance can effectively accelerate the degeneration of cervical intervertebral disc in rats and successfully establish the rat model of cervical intervertebral disc degeneration.GZXJD can delay intervertebral disc degeneration in varying degrees.3.GZXJD can reduce the expression of SP,PGE2,TNF-α in serum and TNF-α in cervical intervertebral disc in cervical disc degeneration model rats,reduce the expression of TRPV1 ion channel in spinal cord,increase the pain threshold of mechanical stimulation,and play a role in relieving pain.effect;4.There was no significant difference between GZXJD and celecoxib in regulating systemic and local inflammatory response in cervical disc degeneration rats.