The Relative Molecular Mechanism Of COX-2 In Pelvic Inflammation,Scar And Stroke

Background:Inflammation was considered as one of the major pathogenic factors,which was involved in many chronic diseases.Local and continuous inflammation may develop into systemic and eventually induce cancer.During the process the formation of chronic diseases,COX-2 was present as an inflammatory mediator.By catalyzing the synthesis of prostaglandins,it exerted an inflammatory response and caused cells to be infiltrated by inflammatory cytokines and chemokines that destroyed the balance of internal environment.Nowdays,anti-inflammatory drugs by targeting COX-2 had poor efficacy and side effects during the treatment of chronic diseases.Based above,we investigated the molecular mechanism of COX-2 in chronic diseases to slove this problem.Aim:In this paper,we explored the function and relative cellular signals of COX-2 in the chronic deseases including pelvic inflammatory disease,scars and stroke.It provided a theoretical basis for the development of new drugs targeting COX-2 in chronic diseases.Method:1.A chronic rat pelvic inflammatory disease model was established by co-exposure of bacteria and hormones.Gram-stained strains were used to confirm the estrus cycle.In vitro,the inflammatroy effect of bacterial infection was evaluated by those events:a general view of the accessory organs?ovaries,uterus?,a detection of SOD and MDA,the HE staining of accessory organs.In vivo,COX-2 expression was detected by immunohistochemical staining to compare the ability of different strains to cause inflammation.2.LPS was used to stimulate NIH3T3 cells to establish a inflammatory cell model during scar formation.The protein and mRNA expression levels of COX-2 and?-catenin were detected by WB,PCR and immunofluorescence staining to initially determine the relationship between COX-2 and?-catenin;the co-transfection technology was used to explore the role of?-catenin in the transcription of COX-2;With the treatment of PGE₂ and LPS,and the expression and location of?-catenin was detected by immunofluorescence staining.So that we can make it clear that how LPS stimulate the expression of?-catenin.3.In vivo,primary brain cells were stimulated by LPS to simulate the environment when brain was damaged.The protein expression of the autophagy-related factors?LC3,p62 and lamp-1?,and the inflammatory factors?COX-2 and?-catenin?were detected by Western blot.The result determined the association between COX-2/?-catenin and autophagy.In vitro,a rat cerebral hemorrhage model was established by collagenase.The thermal pain threshold,cold pain threshold,and mechanical pain threshold of the rats were used to evaluate the animal behavioral after injection of collagenase.TEM observation method was used to evaluate the degree of neurons damage.Electroacupuncture was applied to the injured rats,and analgesic effect of electroacupuncture was evaluated by detecting the thermal pain threshold,cold pain threshold,and mechanical pain threshold of rats;The morphology of neurons was observed to evaluate therapeutic effect of electroacupuncture.Protein expressions of LC3,p62 and lamp-1 were detected by Western blot to determine the regulation of autophagy by electroacupuncture treatment.Results:1.A rats model of Chronic pelvic inflammatory disease was established successfully:?1?The uterus and ovaries of rats showed obvious swelling and erosion in the model group.?2?The SOD activity decreased and the MDA content increased in the serum.?3?The uterus in model groups showed inflammatory infiltrates and structural disorder?4?Overexpression of COX-2 appeaered in the accessory tissues2.?1?Both LPS and TWS119 can up-regulate the expression of COX-2 and?-catenin.With the treatment of corresponding inhibitor,the expression of both proteins would decrease.?2?The results of luciferase report showed that the overexpression of?-catenin increased the protein expression of COX-2.This phenomenon is more pronounced with the treatment of TWS119.?3?PGE₂?downstream of COX-2?induces the expression and nuclear metastasis of?-catenin in dependent dose.Meanwhile,AH6908 can suppress that phenomenon.Similarly,it also happened when LPS influence the expression and location of?-catenin.3.?1?In vivo,LPS induced the expression of LC3II/I and lamp-1 in primary brain cells.Meanwhile,the expression of p62 decreased.So that cell autophagy occured.?2?With the treatment of inhibitors of COX-2 and?-catenin,the expression of the above protein was reversed.?3?the rats showed hyperalgesia after the injection of collagenase.Electron microscopy showed swelling of neurons and atrohy of nuclei.Histopathology showed that the Nissl body of the model group was light and its number reduced.?4?Electroacupuncture can relieve pain of rats and repair organelles of neuronal cells and injured brain by effectively upregulating the protein expression of LC3 and p62 as well as downregulating expression of lamp-1 protein.In conclusion:1.COX-2 is associated with chronic pelvic inflammatory disease,scars and stroke when inflammation occurs.2.During wound repair,activated COX-2 reglated the nuclear metastasis of?-catenin through the PGE₂/EP2 pathway.In vivo,?-catenin has a transcriptional effects on COX-2.As a result,the"?-catenin and COX-2"regulatory loop could potentially be a therapeutic target for wound healing.3.Electroacupuncture may reglate the expression of autophagy by inhibiting COX-2 to protect and repair nerve cells and thereby it exerts an analgesic effect.