| ObjectiveTo investigate the effects of rhG-CSF on apoptosis and motor function in neonatal hypoxic-ischemic rats,so as to provide experimental basis for in-depth study on the cerebral protection of rhG-CSF.Method65 SPF Sprague-Dawley rats aged P7(Postnatal 7 days)was used in this study.The animals came from the laboratory animal center in Southern Medical University,both male and female,weight 13-18g.The animals were randomly divided into 2 groups,including acute-phase and long-term phase.Then,each group divide into 3 groups:(1)Sham(2)HI(3)HI+rhG-CSF,50 g/kg,intraperitoneal injection,1/day,animals for TTC and fluorescent staining were injected for 2 times and the Catwalk group were injected for 6 times totally.According to Rice-Vannucci method,the right carotid artery of the pups was ligated and then hypoxia was conducted in a closed chamber for 150 min.48hours later,the pups were tested with righting reflex,negative geotaxis and Front-limb suspension test.Each test was repeated 3 times and the time was recorded.Fluorescent staining was performed in P9.Five fields were randomly selected from each rat,and the red-green fluorescence ratio was calculated.TTC staining was also performed in P9,the brain of the pups was evenly cut into 5 pieces and the thickness of each piece was about 2mm.We calculate the infarct area ratio of each brain slice.At P21,the Catwalk XT system was used to analyze the gait of free-moving rats,and the parameters of each limb including standing time,step cycle,stride length,swing speed,max contact area and mean intensity were recorded.During the experiment,the rats were weighed once every 2 days in order to calculate the weight gain.Results1.Weight growth:compared with the Sham group,the weight growth of the rats in the HI group was significantly reduced(P<0.05).Compared with the HI group,the weight growth of the rats in the HI+rhG-CSF group increased significantly(P<0.05),in the five stages including P7 to P9,P9 to P11,P13 to P15,P15 to P17,and P19 to P21.Compared with the HI group,the weight growth of the rats in the HI+rhG-CSF group increased slightly(P>0.05),in the two stages including P11 to P13 and P17 to P19.2.Behavioral assessment results in the acute phase:compared with the Sham group,the neural behavior of the rats in the HI group significantly deteriorated(P<0.05).Compared with the HI group,the neural behavior of the rats in the HI+rhG-CSF group was significantly improved(P<0.05).3.TTC staining:compared with the Sham group,the cerebral infarction area of the rats in the HI group and the HI+rhG-CSF group increased significantly(P<0.05).Compared with the HI group,the cerebral infarction area of the HI+rhG-CSF group was significantly reduced(P<0.05).4.Fluorescence staining:compared with the Sham group,the red/green ratio of the rats in the HI group and the HI+rhG-CSF group were significantly reduced(P<0.05).Compared with the HI group,the red/green ratio of the rats in the HI+rhG-CSF group increased significantly.5.Catwalk gait analysis results:compared with the Sham group,the Stand and Step Cycle of the rats in the HI group were significantly increased(P<0.05),and the Stride Length,Swing Speed,Max Contact Area,Mean Intensity of the rats in the HI group were significantly reduced(P<0.05).Compared with the HI group,the Stand and Step Cycle of the rats in the HI+rhG-CSF group were significantly reduced(P<0.05),and the Stride Length,Swing Speed,Max Contact Area,Mean Intensity of the rats in the HI+rhG-CSF group were significantly increased(P<0.05).ConclusionCompared with Sham,hypoxic ischemia can increase the apoptosis of neonatal rats,reduce its weight growth and motor function.This kind of the damage to motor function in acute stage means slower movement in righting reflex,negative geotaxis and Front-limb suspension test.For long-term effect,it means slower action,disorder paw prints,reduced contact area with the ground.Using rhG-CSF shows an effect on the improvement of motor function in neonatal hypoxic-ischemic rats,reducing the slowness and muscle rigidity. |
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