| Obojective In order to improve the clinical treatment rate of post-stroke depression,the mechanism of traditional Chinese medicine compound Jieyuningshen decoction on PSD rats with stagnation of fire and spleen deficiency is especially discussed to provide objective laboratory data support for clinical treatment.Method Experiment 1:140 SD male rats(200±20 g)were divided into blank group(Control),stroke group(MCAO),model group(PSD),fluoxetine group(Flu),Jieyu Ningshen Decoction high dose group(JYNS-H),Jieyu Ningshen Decoction medium dose group(JYNS-M),and Jieyu Ningshen Decoction low dose group(JYNS-L).Blank groups cannot be processed.MCAO was performed only in stroke group.The model group,fluoxetine group and Jieyu Ningshen decoction(high,middle and low)dose group received 4 week unpredictable mild stimulation(CUMS)after MCAO,including:(1)Fasting and water deprivation for 24 hours;(2)Day and night upside down for 24 hours;(3)Shake cage(200 Hz 5 min);(4)Wet cage 100 m L of water sprinkled on the cage inside for 24 hours;(5)Tail clamping for 2 min;(6)Electric shock to sole of foot(0.9m A,15s×4 times);(7)Forced swimming test(4℃ for 5 min).After the treatment,fluoxetine or Jieyu Ningshen Decoction was given 4 week in high,medium and low doses.At the 4th and 8th weeks of the experiment,urine xylose excretion rate,body weight measurement,forced swimming experiment,sugar water consumption experiment,novelty inhibition feeding experiment and behavioral evaluation of open box experiment were performed.Experiment 2: Behavioral evaluation and HE staining of cerebral cortex were performed after gastric lavage.Experiment 3: Western blot was used to detect protein expression of ERK,p-ERK,CREB,p-CREB and BDNF in rat hippocampus and cortex,and ELisa was used to detect BDNF content in rat hippocampus and cortex.Experiment 4:Sixty SD male rats(200±20 g)were randomly divided into three groups(n=20).Middle cerebral artery occlusion(MCAO)was performed for 30 min,60 min and 90 min respectively,and neurological deficit score was performed 24 hours after operation. TTC staining was performed on 3 brain tissues randomly selected from each group to evaluate cerebral infarction volume,and the remaining rats were evaluated for 14-day survival rate.Results Experiment 1: Behavioral Results(1)Compared with the blank group,the urine xylose excretion rate of rats in the model group,fluoxetine group and Jieyuningshen decoction group decreased significantly(P<0.05)in the 4th week experiment.At the same time,the body mass,immobility time of forced swimming,sugar water consumption,novel inhibition of food intake latent time,open-box experiment score results showed significant differences(P<0.01)compared with the blank group,and the modeling was in accordance with PSD rats of depression of fire and spleen deficiency.(2)In the 8th week of the experiment,compared with the blank group,the behavioral indexes of rats in the model group were significantly different(P<0.01).Compared with the model group,the behavioral tests in fluoxetine group and the high-dose group of Jieyuningshen decoction improved significantly(P<0.01).The behavioral changes in the middle-dose group of Jieyuningshen decoction were different(P<0.05).The low-dose group of Jieyuningshen decoction showed an improvement trend,but there was no statistical difference(P>0.05)Experiment 2: Histomorphology After HE staining,cortical cells in PSD group were loose in arrangement,with more empty motor cell and nuclear shrinkage and migration.Cortical cells in fluoxetine group and high and medium dose groups of Jieyu Ningshen decoction were closely arranged,more orderly,and the number of nuclei increased.Experiment 3: Western-blot detection of p-ERK/ERK,p-CREB/CREB and BDNF: Compared with the model group,the high-dose group(P<0.01)and the medium-dose group(P < 0.05)of Jieyuningshen decoction can up-regulate the expression of p-ERK/ERK,p-CREB/CREB and BDNF protein.Fluoxetine group can improve the expression of p-ERK/ERK and BDNF protein(p < 0.05),but has no effect on the expression of p-CREB/CREB protein.Elisa showed that BDNF content in hippocampus and cortex increased in fluoxetine group and Jieyu Ningshen decoction high and medium dose groups(P<0.05).Experiment 4: Improvement of Post Stroke Depression Model Rats The survival rate of rats in the 30 min group was 80%,but the neurological deficit score and cerebral infarction volume could not reach the standard established by stroke model.Compared with the 30 min group,the neurological deficit scores in the 60 min and 90min groups were all greater than 1 point,and there was statistical difference in cerebral infarction volume(P<0.05).The survival rate of rats in 90 min group was only 30%,which could not provide guarantee for subsequent experiments.Therefore,60 min is the best time point for cerebral ischemia reperfusion in PSD model rats.Conclusion 1.The high and medium dose of Jieyu Ningshen Decoction can obviously improve the depressive behavior of rats,which is equivalent to that of fluoxetine group.Jieyu Ningshen Decoction promotes neuron survival,which may play an antidepressant role through ERK-CREB-BDNF signaling pathway.2.The optimal time point for cerebral ischemia-reperfusion in the establishment of post-stroke depression model is 60 min,which can not only simulate stable pathological changes of cerebral infarction,improve survival rate,but also provide guarantee for superposition of subsequent depression models. |
PDF Full Text Request