Effect Of Traditional Chinese Medicine Decoction Of Activating Blood And Removing Stasis And Supplementing Essence And Marrow On Keap1-Nrf2-ARE Signaling Pathway In Mice With Experimental Cerebral Hemorrhage

Objective:The model of cerebral hemorrhage in experimental mice was established in this study.To investigate the effect of the KEAP1-NRF2-ARE signaling pathway in mice with experimental intracerebral hemorrhage(ICH)by using the decoction of activating blood and removing stasis and supplementing essence and marrow as the treatment.To provide theoretical basis for improving secondary injury after cerebral hemorrhage in mice by activating blood and removing stasis and supplementing essence and marrow.Methods:1.The selection of wild type C57 male mice with the gene knockout male mice,22 g ～25 g,were randomly divided into wild type control,gene knock out control,wild type model group,knockout model group,wild type formula,gene knock out prescription group,a total of 156,reference Rynkowski make experimental cerebral hemorrhage model building method.2.The neurobehavioral score,dry-wet weight method and HE staining method were used to observe the intervention effects of breaking blood to remove blood stasis and filling essence and filling marrow on the neurological function deficit,brain water content and the pathological brain tissue around hematoma in mice with experimental cerebral hemorrhage.3.Western blot was used to detect the expression of Nrf2 signaling pathway related proteins,including Nrf2,upstream KEAP1,and downstream HO-1 and NQO1.4.RT-q PCR was used to detect the expression of Nrf2 signaling pathway related genes,including Nrf2,upstream KEAP1,and downstream HO-1 and NQO1,and correlation analysis was conducted for each indicator.Results:1.The Longa score of the sham operation group was zero.Compared with the model group,the wild-type prescription group and the control group could significantly reduce the neurological deficit score(P<0.05),and the function recovery in the prescription group was the most obvious,which was significantly different from that in the gene knockout group(P<0.05).2.The water content of brain in the model group was significantly higher than that in the sham group(P<0.05),the water content of brain in the prescription group and control group was significantly lower than that in the model group,and the decrease in the prescription group was more significant than that in the gene knockout group(P<0.05).3.He staining of the model group,the prescription group and the control group showed obvious pathological changes in the brain tissue,but the damage of the brain tissue in the prescription group and the control group was less than that in the model group,and there was a significant difference compared with the gene knockout group(P<0.05).4.The contents of Nrf2,KEAP1,HO-1 and NQO1 in the brain tissues around hematoma were detected by Western blot at 3 days.The contents of Nrf2,KEAP1,HO-1and NQO1 in the treatment group and the control group were lower than those in the model group,with significant differences(P<0.05),and there were significant differences in the ability to decrease Nrf2,Keap1,HO-1 and NQO1 in the treatment group compared with the knockout group(P<0.05).5.The m RNA expression of Nrf2,KEAP1,HO-1 and NQO1 was consistent by RT-q PCR.Compared with the sham operation group,the expression levels of Nrf2,KEAP1,HO-1 and NQO1 in the model group were significantly increased(P<0.01);Compared with model group,the expression of wild prescription group was significantly decreased(P <0.05),and there was significant difference between knockout formula group and knockout formula group(P<0.05).Conclusion:1.The traditional Chinese medicine decoction can improve the degree of neurological dysfunction and brain edema after cerebral hemorrhage in mice,and can effectively relieve the secondary injury of cerebral hemorrhage.2.The traditional Chinese medicine decoction can up-regulate the expression of KEAP1,Nrf2,HO-1 and NQO1 in mice after intracerebral hemorrhage,and the possible mechanism of its neuroprotective effect may be related to the anti-oxidation effect of activating KEAP1-Nrf2-ARE pathway.3.Nrf2 gene knockout aggravates neuronal death,neurological function and oxidative stress injury after intracerebral hemorrhage in mice.Traditional Chinese medicine decoction may treat secondary injury of intracerebral hemorrhage by regulating Nrf2.