The Effect And Mechanism Of Rhein On Inflammatory Response After Cerebral Ischemic Injury In Rats

Objective:By observing the effect of rhein on the inflammatory response after cerebral ischemic injury in rats and the effect of inflammatory factors TNF-?,ICAM-1,MMP-9,MMP-2 expression,we can study whether rhein can inhibit the inflammation Expression can play a protective role in the brain,provide theoretical basis for rhubarb preparations for clinical treatment of stroke,and provide experimental basis for the development of a new cerebrovascular protective drug.Method:In this study,144 healthy male SD rats were selected to establish a focal cerebral ischemia model of middle cerebral artery occlusion by a modified thread embolization method,and were randomly divided into sham operation group,model group,minocycline group,and rhein group 36 in each group,and each group was divided into three subgroups of 3d,7d and 14d according to the time of administration.After 2 hours of cerebral ischemia,drug intervention was given.The minocycline group(45mg/kg·d)and rhein group(3.46mg/kg·d)were intraperitoneally injected for 3d,7d,and 14d,respectively,model group and sham operation.Groups were injected intraperitoneally with equal volume of normal saline,once a day.Rats in each group were scored for neurological function 2 hours after cerebral ischemia and 1 hour after the last dose.TTC staining was used to observe the changes in cerebral infarction area of each group,and the treatment of cerebral ischemia with rhein was evaluated.Function;take ischemic brain tissue sections for immunofluorescence staining and Western blot detection to observe the effect of rhein on the expression of TNF-?,ICAM-1,MMP-9,MMP-2 and other inflammatory factors in ischemic brain tissue.Results:1.The model groups and each group rats showed varying degrees of nerve function defect3d,7d and 14d groups within each group:compared with control group,model group rats nerve function defect score were significantly higher,have significant difference(p<0.01);Compared with model group,rhein and minocycline group neural function defect scale are falling,But the difference is not statistically significant(p<0.05);Rhein and minocycline neural function defect scale difference between groups was not significant(p>0.05).Comparison between groups:3 d,7 d,14 d compared with model group,the neural function defect scale difference had no statistical significance(p>0.05);3 d,7 d,14 d rhein and minocycline group compared with the neural function defect scale with the treatment time is on the decline,but there was no statistically significant difference(p>0.05).2.Model group and treatment group rats all have different degree of cerebral infarction.3 d,7 d and 14 d groups within each area of cerebral infarction:compared with control group,model group,cerebral infarction area significantly increased,Differences are significant(P<0.01);Compared with model group,rhein and minocycline group cerebral infarction volume reduced obviously,have significant difference(p<0.01);Emodin and minocycline group between cerebral infarction volume difference was not significant.Comparison between groups:3 d,7 d,14 d rhein and compared the minocycline group,cerebral infarction area with the treatment time showed a trend of decrease,But the difference between the groups is not statistically significant(p>0.05).3.Ischemia half dark area TNF-?,ICAM-1,MMP-9,and the comparison of expression of MMP-2:3 d,7 d,14 d group comparison:compared with control group,model group rats after ischemia half dark area TNF-?,ICAM-1,MMP-9 and expression of MMP-2 were significantly increased,The difference was statistically significant(p<0.01);Compared with model group,rhein and minocycline group of TNF-?,ICAM-1,MMP-9,the expression of MMP-2 significantly decreased(P<0.01),rhein and minocycline group between the TNF-?,ICAM-1,MMP-9,the expression of MMP-2 had no significant difference(P>0.05).Comparison between groups:3 d,7 d,14 d rhein and minocycline group compared with the TNF-?,ICAM-1,MMP-9,the expression of MMP-2 with the treatment time is on the decline,but no significant difference between groups(P>0.05).Immunofluorescence analysis prove that rhein treatment for ischemic half dark stripe TNF alpha,ICAM-1,MMP9,the expression of MMP-2 has inhibitory effect.Western blot detection MMP-9 expression in ischemia half dark area prompt differences in model group and treatment group compared with control group with statistical significance(**P<0.05,***P<0.01).Conclusions:1.Rhubarb acid has a good protective effect on cerebral ischemia injury in rats,which can significantly improve the loss of nerve function and reduce the volume of cerebellar infarction.2.Rrheinic acid can inhibit the expression of TNF-?,icam-1,mmp-2 and mmp-9 inflammatory factors in the ischemic penumbra of rats after cerebral ischemia injury,so that the inflammatory response of MCAO model rats can play a protective role of rrheinic acid in the brain.3.The protective effect of rrheinic acid on the brain of rats with cerebral ischemia was time-dependent,and the expression of inflammatory factors in the ischemic penumbra was negatively correlated with the treatment time.