| Objective:To study the effects of scalp point cluster acupuncture combined with environmental stimulation on neurological deficit score and pathological changes of cerebral cortex in neonatal rats with hypoxic-ischemic brain damage,and to observe the effects of scalp cluster acupuncture combined with environmental stimulation on the expression of oxidative stress related indexes MDA,SOD and related proteins Nrf2 and HO-1 in cerebral cortex of neonatal rats with HIBD,so as to provide new experimental data and ideas for scalp point cluster acupuncture combined with environmental stimulation for the treatment of HIBD.Methods:90 seven-day-old neonatal Wistar rats were selected,and they were randomized into 5 groups: the sham operation group(group A),model group(group B),scalp point cluster acupuncture group(group C),environmental stimulation group(group D)and scalp point cluster acupuncture combined with environm-ental stimulation group(group E).Each group was further divided into three subgroups: 12 h,24h and 7d,with6 rats per group.HIBD model was made in each group using improved Rice-Vannucci.The rats in group A and group B were not treated.Group C was treated with cluster acupuncture of scalp points.Group D was treated with rich environmental stimulation.Group E was given rich environmental stimulation during acupuncture.After treatment at each time point,the neurological function of rats in each group was evaluated by modified Bederson;after decapitation,the pathological changes of cerebral cortex were observed by HE staining;the levels of MDA and SOD in cerebral cortex were detected by TBA colorimetry and xanthine oxidase method respectively;the expression of Nrf2 protein in brain tissue was detected by Western blot;the expression of HO-1 protein in brain tissue was detected by immunohistochemistry.Results:1.Bederson neurological deficit score: Compared with the group A,the neurological deficit scores of the other four groups at three time points were significantly different(P<0.01);compared with the group B,at 24 h and 7 d,the neurological deficit scores of C,D and E groups were significantly lower(P<0.01,P<0.05);compared with the group C and group D,there was no significant difference in neurological deficit score for the group E at 12 h and 24 h(P>0.05),but there was significant difference at 7 d(P<0.05).2.The results of HE staining of cerebral cortex: At three time points,the structure of cortical nerve cells in the group A was clear,neatly arranged,the shape was normal,and the nucleus was in the middle and the nucleolus was clear.In the group B,the arrangement of cells was disordered,the polarity was not clear,cells deformation and atrophy,nuclear pyknosis,cells injury and apoptosis.Compared with the group B,the form of nerve cells in the group C and the group D was relatively improved and the structure was relatively tight,but the boundary was still not clear and smooth,and the cells still showed swelling.At 24 h and 7 d,compared with groups C and D,the nerve cells in the group E were significantly improved,the shape of nerve cells was regular,the arrangement of nerve cells was neat,and the nucleolus was relatively clear.3.Levels of oxidative stress indexes MDA and SOD in cerebral cortex:(1)MDA content: Compared with the group A,except for the group E at 7 d,there were significant differences in MDA content among the other groups at three time points(P<0.01,P<0.05);compared with the group B,the contents of MDA in C,D and E groups were lower,and the results were significantly different(P<0.01,P<0.05);compared with group C and group D,there was significant difference in the content of MDA in the group E at24 h and 7 d(P <0.05).(2)SOD activity: Compared with the group A,there were extremely significant differences in SOD activity in brain tissue of the other four groups at three time points(P<0.01);compared with the group B,the activity of SOD in brain tissue of the group E at 12 h and group C,D and E at 24 h and 7 d was significantly higher,with significant differences(P<0.01,P<0.05);compared with group C and D,the activity of SOD in the group E was significantly higher at 24 h and 7 d,with significant differences(P<0.05).4.Nrf2 and HO-1 protein expression in cerebral cortex: Compared with the group A,the expression of Nrf2 and HO-1 protein in the cerebral cortex of group B,C,D and E gradually increased at three time points,and there was significant difference(P<0.01,P<0.05);compared with the group B,the expression of Nrf2 and HO-1 protein in group C,D and E was significantly higher at three time points,with extremely significant difference(P<0.01);compared with group C and D,there was extremely significant difference in the expression of Nrf2 protein in group E at three time points(P<0.01),and there was significant difference in the expression of HO-1 protein in group E at 24 h and 7 d(P<0.05).Conclusion:1.Scalp point cluster acupuncture combined with environmental stimula-tion can significantly improve the neurological deficit of neonatal rats with HIBD,and the effect is much better than that of single scalp point cluster acupuncture or environmental stimulation;2.Scalp point cluster acupuncture combined with environmental stimula-tion can reduce the damage of cerebral cortical neurons in HIBD neonatal rats,and the effect is much better than that of single scalp point cluster acupuncture or environmental stimulation;3.Scalp point cluster acupuncture combined with environmental stimulation can up-regulate the expression of Nrf2 and HO-1 protein in the cerebral cortex of HIBD neonatal rats,reduce the content of MDA in brain tissue,increase the activity of SOD in brain tissue,and reduce the oxidative stress injury of cerebral cortex of HIBD neonatal rats. |
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