Anti-inflammatory And Antinociceptive Activities And Mechanisms Of Danqiao Perfusate

Endometritis is one of the common diseases occurred in postpartum dairy cows, which is the inflammation caused by the infection of endometrium, resulting in infertility. Endometritis caused high economic loss due to prolonged intercalving intervals, decreased conception rate and pregnancy rate, reduced milk production, increased treatment costs and cull rate. Currently, antibiotic and hormone were widely used to treat endometritis in clinic. However, with the high-dose application such as antibiotics, hormone, residues in milk and drug-resistant were emergence, which caused great harm to the health of human bodies. Traditional chinese medicine(TCM) could improve of immune function, resist diseases of bodies, and there is no drug residues and toxic side effects, so it was actively explored in the treatment of endometritis.The preparation of Danqiao perfusate was according to the traditional Chinese medicine(TCM) knowledge and the pathogenesis of endometritis,which was used to prevent and treat endometritis. The anti-inflammatory effect of Danqiao perfusate on Endometritis and its possible mechanism is not clear. Therefore this paper is composed of three sections to study the anti-inflammatory, antinociceptive effect and mechanism of Danqiao perfusion, which could provide theoretical basis and data for the clinical trials and further development of anti-inflammatory using TCM.1. The anti-inflammatory activity of Danqiao Perfusate was studied by xylene-induced ear swelling, acetic acid-induced Evans blue leakage test and cotton pellet-induced granuloma tissue formation in mice. The results showed that Danqiao perfusate(40 g/kg) could inhibit xylene-induced ear swelling and Danqiao perfusate(40 g/kg, 20 g/kg) suppress acetic acid-induced increasing of blood capillary permeability and granulomatous tissue formation, indicating more than 40 g/kg Danqiao perfusate has effect on acute and chronic inflammation.2. The analgesic activity of Danqiao Perfusate was studied by acetic acid-induced writhing response and formalin tests in mice. The results showed that Danqiao perfusate(40 g/kg, 20 g/kg) could decrease acetic acid-induced writhing numbers and formalin-induced licking foot times in second phase, indicating Danqiao perfusate involved in peripheral inflammation.3. We tried to further investigate anti-inflammatory and the possible underlying mechanisms of Danqiao perfusate in LPS-stimulated RAW264.7 cells. In this study, the effect of non-toxic doses of Danqiao perfusate on the production and expression of pro-inflammatory cytokines and inflammatory mediator, including NO, PGE2,TNF-α, and IL-6 in LPS-induced RAW264.7 cells was evaluated by ELISA, Real-time PCR and Western blot. The result showed that Danqiao perfusate(600 μg/m L, 300 μg/m L, 100 μg/m L) dose-dependently suppressed the secretion of NO, gene expression of TNF-α, IL-6, i NOS, COX-2 and protein expression of NF-κB p65 in cells induced by LPS. Moreover, Danqiao perfusate(600 μg/m L) extremely significantly inhibit the secretion of PGE2、TNF-α and IL-6. In conclusion, This study preliminarily proves the protective effect of Danqiao perfusate on LPS-induced RAW264.7 macrophages. The molecular mechanism was associated with inhibition of NF-κB signal pathway and the expressions and secretion of inflammatory mediators and inflammatory cytokines. However, Danqiao perfusate treatment mildly inhibited the gene expression of IL-6, i NOS, COX-2 in RAW 264.7 cells compared to the control group, indicating Danqiao perfusate can’t completely blocking inflammation.