Effect of Acupuncture on the Notch Signaling Pathway in Rats with Brain Injury.

OBJECTIVE: 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. METHODS: 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. RESULTS: 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. CONCLUSION: 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.

Effects of acupuncture on cortical expression of Wnt3a, ß-catenin and Sox2 in a rat model of traumatic brain injury.

OBJECTIVE: To observe the effects of acupuncture treatment on the expression of Wnt/ß-catenin signalling pathway-related genes (Wnt3a, ß-catenin and Sox2) in the injured cerebral cortex of rats with traumatic brain injury (TBI). METHODS: A controlled impact model of TBI was established using Feeney's free-drop method. Seventy-eight Sprague-Dawley rats were randomly divided into the following three groups: a normal group (n=18) that was left untreated; a model group (n=30) that received no treatment after TBI; and an acupuncture group (n=30) that received acupuncture (at LI4, GV20, GV26 and GV16) after TBI. Rats in each group were randomly and equally divided into 3-day, 7-day and 14-day subgroups according to the duration of therapy. Real-time fluorescence quantitative PCR (RT-qPCR) was used to measure mRNA expression of Wnt3a, ß-catenin and Sox2. Western blots were performed to determine the expression levels of WNT3a, ß-Catenin and SOX2. RESULTS: Wnt3a mRNA was upregulated in the 7-day and 14-day acupuncture subgroups compared with the corresponding model subgroups (p<0.05). ß-catenin expression was significantly increased in the 7-day and 14-day acupuncture subgroups compared with the corresponding model subgroups (p<0.01). In the 3-day and 7-day acupuncture subgroups, Sox2 expression was significantly higher than that in the normal and model groups (p<0.01 each). The levels of WNT3a, ß-catenin and SOX2 were generally consistent with the corresponding mRNA levels. CONCLUSIONS: Acupuncture exerts a regulatory effect on the Wnt/ß-catenin signalling pathway, which may in turn influence the proliferation and differentiation of endogenous neural stem cells.

[Clinical observation on electroacupuncture for arousing consciousness of comatose patients with severe trauma brain injury].

OBJECTIVE: To observe the arousal effect of electroacupuncture (EA) stimulation of Baihui (GV 20), Shuigou (GV 26), etc. on severe craniocerebral injury patients. METHODS: A total of 90 cases of severe craniocerebral injury were randomly allocated to routine medication, naloxone and EA groups, with 30 cases in each group. For patients of the routine medication group, mild hypothermia therapy, medicines for dehydration, hormonal therapy, vascular dilation, cerebral nutrition supporting, anti-inflammation, etc. were given. For patients of the naloxone group, intravenous drip of naloxone 0.4 mg/kg in the first 3 days, 0.2 mg/kg for 7 days and 0. 1 mg/kg afterwards. For patients of the EA group, EA (1 Hz/50 Hz) was given for 30 min once daily. All the treatments were conducted once a day for 14 days. The Glasgow Coma Scale (GCS) and Glasgow Outcome Scale (GOS) were used for assessing the therapeutic effect. RESULTS: In comparison with pre-treatment in each one of the routine medication, naloxone and EA groups, GCS scores were all obviously increased in the 3 groups following the treatment, and one month's follow-up (P<0. 05). The GCS scores of both naloxone and EA groups were significantly higher than those of the routine medication group (P<0.05). No significant difference was found between the naloxone group and EA group in GCS scores (P>0. 05). According to the GOS, one month's follow-up showed that of the three 30 cases in the routine medication, naloxone and EA groups, 6, 12 and 14 were improved, 8, 10 and 10 moderate handicap, 8, 3 and 2 severe handicap, 5, 3 and 2 vegetative state, and 3, 2 and 2 dead, with the arousal rates being 46. 66% , 73. 33% and 80. 00%, respectively. The therapeutic effects of both naloxone and EA groups were significantly superior to those of the routine medication group (P<0.05). CONCLUSION: EA intervention at early stage can promote the recovery of neurological function, accelerate the consciousness from coma and improve the outcomes of patients with severe traumatic brain injury.

Effect of acupuncture on proliferation and differentiation of neural stem cells in brain tissues of rats with traumatic brain injury.

OBJECTIVE: To observe the effect of acupuncture on proliferation and differentiation of neural stem cells in brain tissues of rats with traumatic brain injuny. METHODS: Thirty SD rats were randomly and equally allocated to the sham-operated, the model and the acupuncture groups. The traumatic brain injury model was established by the free drop method. For the rats in the acupuncture group, acupuncture was applied once a day for 7 days. Brain histotomy was carried out when treatments were completed. Immunohistochemical techniques were adopted to detect the cells that express nestin, neurofilament proteins (NF)-200 and glial fibrillary acidic proteins (GFAP), the markers of neural stem cells, neurons, astrocytes respectively. RESULTS: Compared to the sham-operated group, the number of nestin-positive cells and NF-200-positive cells in brain tissues was decreased significantly in the model group (P < 0.01), whereas the number of GFAP-positive cells was significantly increased P<0.01). Compared to the model group, the positive cells of nestin, NF-200, GFAP in brain tissues in the acupuncture group were increased obviously (P<0.01). CONCLUSIONS: Acupuncture can significantly increase the number of nestin-positive cells, NF-200-positive cells and GFAP-positive cells, indicating the significant increase of neural stem cells, neurons and astrocytes in number. Acupuncture can improve neuranagenesis by promoting the proliferation and differentiation of neural stem cells in brain tissues. This might be one of the mechanisms for acupuncture to treat traumatic brain injury and to promote the repair of nervous function.