| Objective:To observe the effect of "baihui""Qubin"on the expression of Nrf2,ho-1 and NQO1 in acute cerebral hemorrhage in rats with cerebral hemorrhage,To explore the protective effect of acupuncture on nrf2-are signaling pathway and its downstream target protein on peripheral nerve cells in rats with cerebral hemorrhage,and to provide a basis for clinical treatment of acute intracerebral hemorrhage.Methods:The model of cerebral hemorrhage was prepared by injecting autologous blood in stereotactic brain.A total of 54 healthy male Wistar rats were selected,which were randomly divided into sham operation group,model group and acupuncture group(model + acupuncture group).Each group was divided into 3 subgroups according to Id,3d and 7d at different time points,and 6 rats in each subgroup.Longa scoring method was used to evaluate the neurological deficit of rats in each group.HE staining was used to observe the morphological changes of brain tissue in rats.Immunohistochemistry,Western-blot and rt-pcr method were used to detect the expression of Nrf2,ho-1,NQO1 protein and mRNA in rats of rats at different time points,and the correlation analysis was carried out.Results:1.Neurobehavioral scoring results:no neurological deficits were observed in the sham operation group.The rats in the model group showed different levels of neurological defects,and the postoperative 3d nerve function was the most severe,with 7d reduction after surgery.The score of the acupuncture group at each time point was lower than that of the same time point model group.It is indicated that acupuncture can significantly reduce the degree of neurological defects in rats,improve the symptoms of neurological defects in rats,and have significant differences with the model group(P<0.05).2.HE staining results:no hemorrhage was seen in the brain tissue of rats in the sham operation group,and the brain structure was intact.In the model group,the brain tissue of the rats showed obvious hemorrhage in the light microscope,tissue edema was observed around the lesion,the glial cells proliferated,inflammatory cells invaded,and the change was most obvious at 3 d,and decreased at 7 d.Compared with the time point model group,the damage degree of brain tissue was reduced.3.Immunohistochemical test results showed that there was a small amount of Nrf2,ho-1,NQO1 positive cells in rats at different time points in the sham operation group,and there was no significant difference in time points(P>0.05).Model group rats all time points are obvious Nrf2,HO-1,NQO1 positive cells expressing,postoperative 1 d began to accumulate,postoperative 3 d peak,to postoperative 7 d expression began to decline,compared with control group,the difference was statistically significant(P<0.01).Comparison between acupuncture group and model group showed no significant difference in 1d time after surgery(P>0.05).The expression of Nrf2,ho-1 and NQO1 positive cells in the acupuncture group in 3d and 7d time was higher than that in the same time point model group,and the difference was statistically significant(P<0.05).4.Results of Western blot method:Nrf2,ho-1 and NQO1 protein were expressed in a small number of time points in the sham operation group,and there was no significant change at all times(P>0.05).Nrf2 model group rats,HO-1,the amount of NQO1 protein expression in postoperative 1 d has obviously increased,postoperative 3 d protein expression in most,has receded in the 7 d after the rush hour,the expression is still higher than control group(P<0.01).The expression levels of Nrf2,ho-1 and NQO1 protein in the acupuncture group were significantly higher than that in the model group,and the difference was statistically significant(P<0.01).5.Results of rt-pcr method:there were a small amount of Nrf2,ho-1 and NQO1 mRNA expression in each time point of the sham operation group,with no significant difference at each time point(P>0.05).Postoperative 1d,the rats in the model group had a significant amount of Nrf2,ho-1,NQO1 mRNA expression,and postoperative 3d reached a peak,and the postoperative 7d was lower than the peak,but still higher than the sham operation group(P<0.01).The expression of Nrf2,ho-1 and NQO1 mRNA in the acupuncture group was significantly higher than that in the model group,and the difference was statistically significant(P<0.01).6.Correlation analysis showed that the expression of Nrf2 and ho-1 was positively correlated and the correlation coefficient was 0.862.Nrf2 was positively correlated with NQO1 expression and the correlation coefficient was 0.883.The expression of NQO1 and ho-1 was positively correlated with the correlation coefficient of 0.988.Conclusions:1.Baihui Qubin scalp acupuncture therapy can significantly reduce the score of brain tissue defect in rats with cerebral hemorrhage and improve the symptoms of nerve defect in rats.2.Baihui Qubin scalp acupuncture therapy can reduce the infiltration of inflammatory cells,reduce the degree of cerebral edema,and improve the damage of nerve cells around the hematoma.3.Baihui Qubin scalp acupuncture therapy can promote the expression of Nrf2,ho-1 and NQO1,and thus play a role in brain protection against oxidative stress.4.Baihui Qubin scalp acupuncture therapy can regulate the expression of ho-1 and NQO1 by activating Nrf2 signaling pathway to play an anti-oxidative protective effect. |
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