PAR4 Effects The Visceral High Sensitivity Of Rat Abdominal And Impacts On The Mast Cell P2X7 And NOS Expressed

Purpose:Now the study of pain,P2 receptors have been attended by the broad masses of scientists.P2 X and P2 Y are in the P2 receptors.P2X7 receptor is a kind of the P2 X receptors.P1 receptors and protease activated receptor PARs,are all in the G protein coupled receptor(G-protein coupled receptor,GPCR).According to the new study,scientists have found that PARs includes four subtypes.The four subtypes are PAR1,PAR2,PAR3,PAR4.In the study of pain and inflammation of signal conditioning,protease can make GPCR N-terminal cracking,and let the receptor to form a new N-terminal.New N-end receptors after the formation of connection and activate itself,which cast protease activated receptor itself to adjust the action of pain and inflammation process.Nitric oxide synthase(NOS),which belongs to a kind of isozyme,is in endothelial cells,macrophages and nerve cells.NOS can be divided into nerves induced nitric oxide synthase and nitric oxide synthase,endothelial nitric oxide synthase.NOS play a role in many physiological process when the inflammatory stimulation after the organization.But for PAR4 activation after adjusting mechanism of the body's inflammatory process,that is not clear.Especially for PARs,P2 X,NOS receptor in the resent study is not clear.This course is now a hotspot of scientific research personnel.Scientists are provided a new treatment of pain and inflammation in a new research field.This topics,which through the use of high visceral sensitivity in rats to make experimental model,are adjust to the PAR4 receptor activation in the organism.The function of gastrointestinal factors such as NOS,PAR4,P2X7 expressed in intervention.Experimental results provide more effective treatments for patients in irritable bowel syndrome(IBS).Material:1.In CRD modeling,adult SD rats use straight colon expansion technology.Founding experiment of IBS rat model,the animal model are in AWR scores.We select qualified animal as the animal model of IBS.For qualified animal models,we use the function of animal experimental instrument detection.By using instrument BL420-F system,we evaluate the abdominal muscle electromyography in the abdominal muscles and observe muscle mobility.2.IBS group animal models,which given PAR4-AP in anus perfusion,make liquid into rats rectum tissue.These rats are AWR score again using semi-quantitative method.Then,experimental animals use BL420-F system,which electromyogram of abdominal muscle testing,to observe the muscle mobility.3.When building the animal model,I extract the rat intestinal tissue.Using immunohistochemical method.I combining with toluidine blue staining technique,which observation of mast cells in IBS group model of rats and normal rats model and three groups into PAR4-AP IBS rats model in the change of intestinal mucosa,and take the effects on NOS,P2X7,PAR4.4.Using immunohistochemical technique,I am combined to use of laser confocal microscopy method,which Tryptase(Tryptase,AA4,is specific marker of mast cells)and PAR4,P2X7,NOS in experimental animals intestinal tissue structure.5.To extract the normal group,the IBS group,PAR4 agonist,three groups of SD rat dorsal root ganglion,I use IHC method to observe the MC.Further observe PAR4 receptor is changed in nerve tissue.1?PAR4 receptors on IBS rat model of abdominal pain after activation.(1)the CRD technology expansion rats is used for the experimental rats after who run AWR standards,by dividing into rats.In compared with normal group rats,I found that IBS group in SD rats,who run AWR are obviously on the high side.After dealing with the rectum expansion,the visceral high sensitive to rise.PAR4 excited by SD rats in the group,which runs AWR score results IBS group is low,and the rat model after joining PAR4 agonist is visceral high sensitivity when be artificially low.(2)The rat model by abdominal muscle electromyography is detect.Results show that in IBS group rats model,PAR4 agonist can significantly reduce.Because rectum expansion is caused by average peak,when reduce the curve of peak area.2?After activation,PAR4 receptor on IBS group rats model and the influence of P2X7 and NOS positive mast cells.(1)For PAR4 positive cells expression,IBS group of cecum tissue slices.PAR4 positive cells number significantly higher than the PAR4 excited group and normal group.PAR4 excited showed positive reaction of mast cells in the group,that mast cell numbers are higher than normal group.(2)The influence of PAR4 activate of mast cells.In IBS group within the cecum tissue section,I found that the number of mast cells AA4 is positive,which than the normal group and PAR4 excited group,and number of positive cells are increased significantly.(3)PAR4 activation effect on NOS positive cells.When cecum in IBS group within the organization,NOS shows positive cell count compared with the normal group,by increased cell count.After dealing with the PAR4 agonist of the experimental group,NOS positive mast cell count agreed to count more than the normal group.In the group of IBS and PAR4 excited the number of NOS positive cells have the same number.(4)PAR4 activated the expression of P2X7 positive cell numbers changed.In the normal group and PAR4 group,P2X7 positive cell count was lower than that in group IBS,and the normal group is lower than the PAR4 excited.3?In IBS group rats model,PAR4,P2X7 and NOS expression are changed in mast cells.Using immunohistochemical method,I combined to use of laser confocal microscope technology.Under the microscope I found that by bone marrow stem cells are induced by nurturing of mast cells and rats intestinal mucosal mast cells are in IBS group,PAR4,P2X7 and NOS expressed.PAR4,P2X7 NOS are exist with AA4.4?To extract the normal group,IBS group and PAR4 agonist,I dorsal root ganglion of three groups of SD rats.Extracted the ganglion of dyeing,I use immunohistochemical method to found the NOS and P2X7 staining positive expression cells.IBS and PAR4 excited group can increased obviously,which positive number of mast cells and the IBS group,significant higher than the PAR4 excited group.Conclusion:1.IBS group rats who runs AWR score higher,which visceral high sensitivity is high.Adding a dose of PAR4 agonist to rats,the viscera can be highly sensitive.2.The normal group and PAR4 excited of mast cells(MC)are compared with IBS group,which can reduced cell number.The number of mast cells in the normal group are lower than PAR4 excited number within the group.3.In the process of the regulation of visceral highly sensitive abdominal pain,P2X7 can mediated by mast cells,PAR4 factor and NOS factor.