The Influence Of Herbal Pair Of Astragali Radix And Salviae Miltiorrhizae On Neuronal Apoptosis Of HIBD Rats

Background:Neonatal hypoxic-ischemic encephalopathy(HIE)which is one of the most common diseases and can cause to neonatal death and permanent nervous system injury,brought a huge burden to family and society. Nowadays, treatment of HIE is still bases on the "three support treatment and three symptomatic treatment ",but it can't improve prognosis effectively, so we need to find more safe and effective treatment to HIE. As a treasure of our country,traditional Chinese medicine has a good effect in HIE and provides a mew prospect to it.Our experiment aims to explore the mechanism that herbal pair of Astragali Radix and Salviae Miltiorrhizae Radix et Rhizoma treats HIE bases on the theory of supplementing Qi and activating blood circulation.Objective:Hypoxic-ischemic brain damage (HEBD) rats were given a gavage of herbal pair of Astragali Radix and Salviae Miltiorrhizae Radix et Rhizoma in our experiment. Exploring the mechanism of herbal pair in the treatment of HIE by observing the change of cell apoptosis related gene and protease and controlling apoptosis pathway to provide the basis for clinical treatment.Methods:Groups:A total of 77 SD rats in clean level ages 7d, with the birth weight of 15-20g were randomly divided into different three groups:sham operation group (n=28)?HIBD model group (n=28) and traditional Chinese medicine treatment group(n=21). Then sham operation group and HIBD model group were randomly divided into four subgroups (n=7 in each subgroups) based on different time points at 2h?1d?3d?7d after HIBD.And traditional Chinese medicine treatment group was randomly divided into three subgroups(n=7 in each subgroups) based on different time points at 1d?3d? 7d after HIBD.The preparation of HIBD model:HEBD model rats were prepared by clamping their right common carotid artery and keeping them into hypoxic environment of 8% O2 and 37? concentration for 2 hours. In the sham operation group, the right common carotid artery was exposed but wasn't ligated or exposed hypoxia.Interventions:The treatment group:Matching Astragali Radix and Salviae militorrhizae by 1:1 and gavage was given by 0.3ml/10g once a day and continued seven days.HE8D model group was given a gavage of normal saline with 0.3ml/10g. Sham operation group rats were fed by their mother.Observing brain tissue:Rats were sacrificed in different time points and collected their brain tissue. Then observed the morphologic changes of brain tissue under light microscope,and the expression of Caspase-3^ Bcl-2 and Bax in right brain were detected with immunohistochemistry.Results:(l)Observation of behavior:Neonatal SD rats have a lot of abnormal behaviors after HIBD. (2) We could observe various morphologic abnormality in rats brain in HIBD model group at different time, but morphology of brain tissue of rats in treatment group hadn't obvious changes. (3)HE staining:Cellular morphous and arrangement were normal in brain tissue in sham operation group;Cell of brain tissue of rats in HIBD group were scatteredly arranged, vacuolization, necrosis,nuclear condensation and nuclear fragmentation were observed;Compared with HIBD model group,,the pathological changes of brain tissue lessened significantly and cell arranged regularly in treatment group.(4)Immunohistochemical results:The expressions of Caspase-3 and Bax in brain tissue in HIBD group were higher than which in sham operation group in different time points. There was statistical significant difference between them (P<0.05). Compared with sham operation group.the expression of Bcl-2 in HIBD group was increased in 2 hours,there was statistical significant difference between them (p<0.05);The expression of Bcl-2 in HIBD group was higher than that of sham operation group in Id and 3d,but there were no statistical significant changes(p>0.05);The expression of Bcl-2 in HIBD group was lower compared with that of sham operation group in 7d. The expression of Caspase-3 and Bax in treatment group was lower compared with HIBD group and the expression of Bcl-2 in treatment was slightly higher than that of HIBD, there were statistical significant changes at different time points (P<0.05).Conclusions:1> By clamping right common carotid artery and keeping rats into hypoxic environment of 8% O2 concentration for 2 hours making HIBD model is successful.2?The herbal pair of Astragali Radix and Salviae Miltiorrhizae Radix et Rhizoma could make the expression of caspase-3 in brain tissue descend obviously.3?The herbal pair of Astragali Radix and Salviae Miltiorrhizae Radix et Rhizoma could descend the expression of Bax and increase the expression of Bcl-2 in brain tissue obviously.4?The herbal pair of Astragali Radix and Salviae Miltiorrhizae Radix et Rhizoma could alleviate pathological injuries coursed by HIBD. It could inhibit neuronal apoptosis and provide protective effects for hypoxia-ischemia-induced neuronal injury of neonatal rats.