Effect of Acupuncture on the Notch Signaling Pathway in Rats with Brain Injury / 中国结合医学杂志

<p><b>OBJECTIVE</b>To observe the effect of acupuncture on the Notch signaling pathway in rats with traumatic brain injury and to explore the pathogenesis of acupuncture intervention on traumatic brain injury.</p><p><b>METHODS</b>Feeney's freefall epidural impact method was used to establish a traumatic brain injury model in rats; the rats were randomly divided into a normal group, sham operation group, model group and acupuncture group. Acupuncture was performed in the Baihui (DU 20), Shuigou (DU 26), Fengfu (DU 16), Yamen (DU 15) and Hegu (LI 4) acupoints in the rat, and Yamen was punctured via Fengfu. Then, the rats in each group were randomly divided into three subgroups, namely the day 3 subgroup, day 7 subgroup and day 14 subgroup according to treatment duration. The modified neurological severity scores (mNss) method was used to perform neurobehavioral scoring for evaluating the degree of injury in the rats. The hematoxylin-eosin (HE) staining method was used to observe the pathological change in the brain tissue of rats in each group. Real-time fluorescent quantitative polymerase chain reaction (Q-PCR) technology was used to detect changes in the Notch1, Hes1 and Hes5 gene expression levels in the cortex on the injured side. Western blot was used to detect the protein expression changes.</p><p><b>RESULTS</b>One day after modeling, the mNss scores in the model group and in the acupuncture group were significantly higher than those in the normal and sham operation groups (P<0.01) ; there was no statistically significant difference between the normal group and the sham operation group. The scores decreased with increased treatment time, and the scores in the acupuncture group decreased more significantly than those in the model group (P<0.01). The pathological examination by the HE staining method demonstrated that the brain tissue of the rats in the acupuncture and model groups relatively significantly changed. The Notch1 gene expression level in the acupuncture group was significantly higher than the level in all of the other groups (P<0.01) ; the Hes1 and Hes5 gene expression levels were also higher in the acupuncture group. The expression changes of the Notch1 and Hes1 protein were consistent with that of mRNA. In each experimental group, the mNss score and the pathological results by the HE staining method were consistent with the mRNA results.</p><p><b>CONCLUSION</b>Acupuncture could significantly promote high expression levels of Notch1, Hes1 and Hes5 in the brain tissue of traumatic brain injury rats. Therefore, acupuncture might be an important intervention for inducing endogenous stem cell proliferation and for promoting nerve repair.</p>

Effects ofDanlong Xingnao Formula on Proliferation of Neural Stem Cells in Sub Ventricular Zone and Expressions of Hes1 and Hes5 in Cerebral Ischemia-Reperfusion Injury Model Rats / 中国中医药信息杂志

Objective To study effects ofDanlong Xingnao Formula (DLXNF) on proliferation of neural stem cells (NSCs) and the expressions of Hes1 and Hes5 in sub ventricular zone (SVZ) in cerebral ischemia-reperfusion injury model rats; To explore the mechanism of promoting the proliferation of NSCsMethods Eighty male SD rats were randomly divided into sham-operation group, model group, edaravone group andDLXNF group. The focal cerebral ischemia reperfusion injury models were prepared by suture method, and 7 d after reperfusion, the SVZ brain tissue of ischemia side was taken. The proliferation of cells was detected by Brdu labeling fluorescence immunocytochemistry; Hes1, Hes5 mRNA and protein expressions were detected by fluorescence real-time quantitative PCR and Western blot method in each group.Results Compared with the sham-operation group, Brdu positive cell rate in other groups increased more obviously, and the expressions of Hes1, Hes5 mRNA and protein also increased significantly (P<0.01). Compared with the model group, Brdu positive cell rate increased significantly in edaravone group and DLXNF group, and the expressions of Hes1, Hes5 mRNA and protein increased significantly (P<0.01). The expression of Hes1 mRNA in DLXNF group was superior to that in edaravone group (P<0.01), and other indexes had no significant difference.Conclusion DLXNF can promote the proliferation of NSCs in SVZ in cerebral ischemia-reperfusion injury model rats, and up-regulate the expressions of Hes1 and Hes5, whose mechanism may be related to the activation of Notch signaling pathway.

PROTEOMIC ANALYSIS OF CELL DIFFERENTLATION IN THE SUBVENTRICULAR ZONE OF Hes5 KNOCKOUT MICE / 解剖学报

Objective Hes5 is one of the critical effectors of the Notch pathway and its expression Call inhibit neuronal differentiation.Which protein expression is influenced in cell differentiation by HeS5 remains to be determined.Methods Experiments were designed to study changes in protein expression in the subventricular zone(SVZ)of the HesS knockout mice at the postnatal day 8 using the new method of proteomic isobaric tags for relative and absolute quantification(ITRAQ),and confirmed expression of the up-gnlated and down-regulated proteins by Western blotting.Results Nineteen proteins showed differences in expression levels in the SVZ of Hes5 knockout mice.When compared tO the wild type mice,expressions of 11 proteins in the knockout mice were up-regulated,and the differentially expressed proteins showed ratios of knockout/widtype between 1.20 and 1.32.Expressions of 9 proteins were down-regulated,with their ratios were between 0.77 and 0.83.The upregulated proteins have been shown to be involved in neurogenesis,cell adhesion,migration and signal transduction.The downregulated proteins have been mainly shown to be involved in growth inhibition and cytoskeleton.Westem bloning analysis further confirmed the expression of proteins from iTRAQ.Conclusion Our findings suggest that Hes5 may influence cell differentiation by regulating expression of some proteins involved in cell growth and migration,and its cytoskeleton.

Proliferation effects of astragaloside on neural atem cells in vitro / 中国药理学通报

Aim To explore the effects of astragaloside(AS)on the proliferation of neural stem cells(NSCs).Methods The NSCs of embryonic day(E)16 SD rat were cultured in vitro.The NSCs were identified by the immunocytochemical(ICC)staining of Nestin.The ICC staining of BrdU was adopted to characterize the proliferation of NSCs.The number of neurospheres and the ICC staining of BrdU were performed to identify their proliferation properties.Real time RT-PCR technique was used to investigate the proliferation mechanisms of the AS on the NSCs.Results Characteristic protein(Nestin)of NSCs labeling,BrdU labeling were positive showed by immunocytochemical staining.The ICC of BrdU labeling results showed that different dosages of AS could promote the proliferation of NSCs in vitro.The proliferation rate of NSCs was increased extremely significant(P

PROTEOMIC ANALYSIS OF CELL DIFFERENTIATION IN THE SUBVENTRICULAR ZONE OF Hes5 KNOCKOUT MICE / 解剖学报

Objective Hes5 is one of the critical effectors of the Notch pathway and its expression can inhibit neuronal differentiation.Which protein expression is influenced in cell differentiation by Hes5 remains to be determined. Methods Experiments were designed to study changes in protein expression in the subventricular zone(SVZ) of the Hes5 knockout mice at the postnatal day 8 using the new method of proteomic isobaric tags for relative and absolute quantification(iTRAQ),and confirmed expression of the up-gulated and down-regulated proteins by Western blotting. Results Nineteen proteins showed differences in expression levels in the SVZ of Hes5 knockout mice.When compared to the wild type mice,expressions of 11 proteins in the knockout mice were up-regulated,and the differentially expressed proteins showed ratios of knockout/widtype between 1.20 and 1.32.Expressions of 9 proteins were down-regulated,with their ratios were between 0.77 and 0.83.The up-regulated proteins have been shown to be involved in neurogenesis,cell adhesion,migration and signal transduction.The down-regulated proteins have been mainly shown to be involved in growth inhibition and cytoskeleton.Western blotting analysis further confirmed the expression of proteins from iTRAQ.Conclusion Our findings suggest that Hes5 may influence cell differentiation by regulating expression of some proteins involved in cell growth and migration,and its cytoskeleton.