| Aims:To investigate the effects of ghrelin on TNF-??IL-6?NF-?B in serum and in vascular tissue,and observe the pathological changes of balloon vascular injury after carotid artery intima balloon injury in rats.Methods:A total of 72 healthy male SD rats?weighting about350-450g?,collected from the Experimental Animal Center of University of South China,were randomly divided into sham operation group?group A?,operation group?group B?and treatment group?group C?with 24 in each.Group A:general anesthesia and left common carotid artery of the rats isolated,but not into the balloon catheter;group B:the rats carotid artery injured by balloon catheter;group C:the rats given intraperitoneal injection of 100?g·kg-1·d-1 ghrelin after balloon injury;C group began to use drugs,A,B group were injected with physiological saline intraperitoneal injection,dose,frequency and termination time with the C group,rats in each group were fed with ordinary feed,drinking water without restriction.Respectively after 1 days,7 days,14 days and 28 days,the rats were killed,6 rats at each time point,then detect serum concentrations of TNF-alpha and IL-6 in serum-line by enzyme-linked immunosorbent assay?ELISA?;The left Carotid tissues of the three groups are harvested after 7?14 and 28 days after taking heart blood,tissues were used to HE staining,observed the intimal and the VSMC proliferation and the ratio of intimal and medial by optical microscope.seventh days after operation,detect the expression level of vascular tissue TNF-alpha IL-6 and NF-?B by real-time fluorescence quantitative PCR and Western Blot.Result:1.Tissue sections with HE staining,observe the changes of carotid artery intima and media area ratio,under the light microscope showed that the intima of rats in the A group the left carotid artery was monolayer endothelial cells,smooth as usual,the internal elastic membrane wavy,membrane into smooth muscle cells,connective membrane organization.Compared with A group,B group and C group after each time point of the balloon injury segment in vascular tissues,the1d vascular intimal damage,but no intima,intima/media ratio is 0;after the establishment of the model of 7d,14d and 28d were obvious segmental artery balloon injury and proliferation of cell wall visible,irregular arrangement of extracellular matrix increased,neointima formation,and gradually thickening,endothelium hyperplasia,wall thickening,intima after balloon operation that can cause neointimal formation;compared with B group,C group,intimal hyperplasia and tube wall thickening degree were significantly reduced?P<0.05?.2.The results of ELISA:each time point after surgery showed the serum inflammatory factor in A?B and C groups.TNF-alpha,and IL-6were all mild expression in A group;compared with group A,serum of TNF-alpha and IL-6 protein content were significantly higher in B and C groups?P<0.05?;compared with group B,the serum of TNF-alpha and IL-6 protein content was significantly decreased in C group?P<0.05?.3.The results of real-time PCR after surgery in the 7th day:The mRNA of TNF-alpha?IL-6 and NF-?B appeared moderate expression in A group;compared with A group,the mRNA levels of TNF-alpha?IL-6and NF-?B were significantly higher after balloon injury in B and C group?P<0.05?;compared with B group,the mRNA expression of TNF-alpha?IL-6 and NF-?B were significantly lower after balloon injury in C group?P<0.05?.4.The results of Western Blot after surgery in the 7th day:The TNF-alpha?IL-6 and NF-?B of vascular tissues appeared moderate protein expression in A group;compared with A group,the protein levels of TNF-alpha?IL-6 and NF-?B were significantly higher after balloon injury in B and C group?P<0.05?;compared with B group,the protein expression of TNF-alpha?IL-6 and NF-?B were significantly lower after balloon injury in C group?P<0.05?.Conclusion:1.Intimal hyperplasia is associated with vascular endothelial injury,and ghrelin can significantly inhibit intimal hyperplasia and protect vascular endothelium after carotid artery balloon injury in rats;2.Ghrelin may improve intimal hyperplasia by inhibitting the expression of inflammatory factors TNF-??IL-6?NF-?B in serum and vascular tissue after carotid artery intima balloon injury in rats. |
PDF Full Text Request