The Effect Of ZD On Some Anti-inflammatory Factors Of Non Open Soft Tissue Injury

ZD preparation is a classic prescription for the treatment of limb and foot non open soft tissue damage in horse animals.It consists of 12 flavors of traditional Chinese Medicine.Earlier experiments showed that ZD could inhibit the content of IL-1 beta,IL-6 and IL-8 in the cell inflammation model in vitro,but the influence on other inflammatory factors was not yet studied.In this study,by using lipopolysaccharide(LPS)to stimulate the cell inflammation model in vitro,the effect of ZD on the related inflammatory factors was observed,and the molecular mechanism of the anti-inflammatory effect of ZD preparation was preliminarily studied.(1)MTT assay and ELISA assay were used to detect the effect of ZD preparations on the content of TNF-? in the cell inflammatory model and the effect of TNF-? on L929 cell death induced by TNF-?.The results showed that ZD preparation could inhibit the content of TNF-? in the cell inflammation model,and also inhibit the toxic effect of TNF-? on L929 cells.(2)The effects of ZD preparations on COX-1 and COX-2mRNA,protein expression and activity of macrophages in mice were detected by RT-PCR,Western Blot and ELISA.The results showed that the expression of COX-2 mRNA,the expression of COX-2 protein and the content of COX-2 were inhibited by ZD,but there was no obvious effect on the expression and activity of COX-1mRNA and protein.(3)the influence of ZD preparation on the ratio of cAMP,cGMP,PDE and cAMP/cGMP in cell inflammatory model was detected by ELISA.The results showed that ZD preparation could increase the content of cAMP and cAMP/cGMP ratio,and inhibit the content of PDE,but had no significant effect on cGMP.Conclusion:(1)through the toxicity test of ZD preparation to mouse peritoneal macrophage Raw264.7 and mouse fibroblast L929,it is found that the toxicity of ZD preparation is very small and can be safely applied to the cell under the experimental concentration.(2)ZD preparations at the onset of inflammation can effectively inhibit the content of TNF-? in inflammatory cells,and the inflammatory development period can be achieved by effectively reducing the activity of TNF-? in cells.(3)ZD preparations can achieve the aim of anti-inflammatory by effectively inhibiting the expression of COX-2 mRNA and protein in the inflammatory phase.(4)ZD preparation has no inhibitory effect on the nonspecific inhibitory target COX-1,and has no obvious effect on the expression and content of COX-1 mRNA and protein.(5)ZD preparation can effectively inhibit the content of PDE in inflammatory cells at the onset of inflammation,and can effectively improve the content of cAMP and the ratio of cAMP/cGMP to play an anti-inflammatory effect,but have no obvious effect on the content of cGMP.