| Objective: By constructing a rat MCAO/R(Middle cerebral artery occlusion/reperfusion)model,in vivo intervention was carried out with Blood Rongluo Fang decoction and edaravone dexturol injection,the mechanism of inhibition of cell pyrosis and angiogenesis after cerebral infarction in rats was studied.Methods:Thirty SD rats were randomly divided into sham group,model group,HXRLF group,edaravone dextrol(YDLFYKC)group,and live blood rong party + edaravone dextrol(HXRLF+YDLFYKC)group,with 6 rats in each group.Only the common carotid artery was isolated in the Sham group,and the MCAO/R model was prepared by the tetheric method in the remaining groups.The HXRLF+YDLFYKC group used a drug solution with a concentration of 1.17g/ml,gavage at a dose of 1ml/100 g,and intraperitoneal injection of edaravone dexturol injection(0.29ml/100g);The HXRLF group used crude drug concentration of 1.17g/ml solution,gavage at a dose of 1ml/100 g,and intraperitoneal injection of the corresponding dose of normal saline;The YDLFYKC group was administered intraperitoneally with edaravone dextronicol injection(0.29ml/100g),and the corresponding dose of distilled water was gavage;The rats in the Sham group and Model group had a corresponding dose of distilled water for gavage and intraperitoneal injection of the corresponding dose of normal saline,which was administered once a day for 7 days.2 h after modeling,rats were scored for neurological deficits;After 7 days of drug intervention,abdominal aorta phlebotomy was taken from the brain,the brain tissue was isolated on ice,and the brain tissue was sectioned,and HE(Hematoxylin-eosins staining)staining was stained to observe the pathological morphological changes of hippocampal tissue;Western Blotting detected the protein content of VEGF,VEGFR2,CDC42,RAC1,GSDMD in hippocampal tissues.The colocalization and expression changes of angioneoprotein VEGF and pyrozoprotein GSDMD were detected by immunofluorescence doublestain co-labeling.The Pearson correlation coefficient method was used to analyze the correlation between VEGF protein and GSDMD protein.Results:1.Neurological deficit score: Compared with the Sham group,the score of neurological deficit in the Model group was significantly increased(P<0.01),and compared with the Model group,the score of neurological deficit in the HXRLF group,YDLFYKC group and HXRLF+YDLFYKC group was significantly reduced(P<0.01),among which the HXRLF+YDLFYKC group decreased most significantly(P<0.01).2.Observation of brain histopathological morphological changes by HE staining: through the observation of the histopathological morphology of rat hippocampus,it was found that the hippocampal tissue structure of the Model group was obviously abnormal,the neuronal damage was serious,the number of cells was reduced,the arrangement was disordered,the soma deformation was deformed,the nucleus was solidified,and the nucleoli were cleavaged.The other drug groups were able to reduce nerve cell damage in the MCAO/R model to varying degrees.3.Western Blotting: Compared with the Sham group,the expression of VEGF,VEGFR2,CDC42,and RAC1 proteins was increased(P<0.01)and the expression of GSDMD proteins(P<0.01)was reduced in rat hippocampal tissues in the Model group.Compared with the Model group,the expression of VEGF,VEGFR2,CDC42 and RAC1 proteins was increased in rat hippocampal tissues in each drug group(P<0.01)and the expression of GSDMD protein was decreased(P<0.01).Compared with HXRLF+YDLFYKC group,the expression of VEGF,VEGFR2,CDC42 and RAC1 proteins in rat hippocampal tissues of HXRLF+YDLFYKC group was lower than that in HXRLF+YDLFYKC group(P<0.01),and the expression of GSDMD protein was higher than that in HXRLF+YDLFYKC group(P<0.01),while VEGF,VEGFR2,CDC42,and There was no significant difference in the expression of RAC1 and GSDMD proteins(P>0.05).4.Immunofluorescence double stain and Pearson correlation analysis:Compared with the Sham group,the fluorescence expression of VEGF in rat hippocampal tissue in the Model group was elevated(P<0.01)and the fluorescence expression of GSDMD was decreased(P<0.01).Compared with the Model group,the fluorescence expression of VEGF(P<0.01)was increased to different degrees and the fluorescence expression of GSDMD(P<0.01)was decreased to different degrees.Compared with HXRLF+YDLFYKC group,the fluorescence expression of VEGF and GSDMD proteins in hippocampal tissues of HXRLF group and YDLFYKC group was lower than that in HXRLF+YDLFYKC group(P<0.01),and the fluorescence expression of GSDMD was higher than that in HXRLF+YDLFYKC group(P<0.01),while there was no significant difference in fluorescence expression of VEGF and GSDMD proteins in hippocampal tissues between HXRLF group and YDLFYKC group(P>0.05).The Pearson correlation coefficient method showed a high negative correlation between VEGF protein and GSDMD protein in rat hippocampal tissue.Conclusion:1.Combining edaravone dexenol can improve MCAO/R rat neurological deficits,which may be by inhibiting the expression of cytopyrozotic protein GSDMD,reducing the inflammatory cascade,upregulating the expression of angiogenesis-related proteins VEGF,VEGFR2,CDC42,RAC1,promoting angiogenesis,and exerting neuroprotective effects.2.Inhibit cell pyrosis,reduce inflammatory response,promote angiogenesis,and the efficacy of blood revitalization is comparable to that of edaravone dextrol,and the effect can be enhanced when used in combination. |
PDF Full Text Request