| 陈 勇,李文文,魏宽海,颜云峰,李思震,钟期富.P2RX3基因对创伤性脑损伤大鼠神经功能恢复的影响[J].现代生物医学进展英文版,2024,(17):3226-3232. |
| P2RX3基因对创伤性脑损伤大鼠神经功能恢复的影响 |
| Effect of P2RX3 Gene on Neurological Function Recovery in Rats with Traumatic Brain Injury |
| Received:January 31, 2024 Revised:February 26, 2024 |
| DOI：10.13241/j.cnki.pmb.2024.17.005 |
| 中文关键词: 创伤性脑损伤 嘌呤2X3（P2X3）受体（P2RX3） 神经功能恢复 BDNF/TrkB/Akt信号通路 |
| 英文关键词: Traumatic brain injury Purine 2X3 (P2RX3) Nerve function recovery BDNF/TrkB/Akt signal pathway |
| 基金项目:新疆维吾尔自治区科技支疆项目计划（指令性）项目（2022E02040）；广东省援疆农村科技（特派员）项目（KTPYJ2023010） |
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| 中文摘要: |
| 摘要 目的：探究嘌呤2X3（P2X3）受体（P2RX3）基因对创伤性脑损伤（TBI）大鼠神经功能恢复的影响。方法：通过颅脑损伤仪对大鼠进行TBI造模，造模后将54只TBI模型大鼠随机分为TBI组（n=10）、TBI+NC-sh组（n=11）、TBI+P2RX3-sh组（n=11）、TBI+NC-OE组（n=11）和TBI+P2RX3-OE组（n=11）将仅切开皮肤而未打击造模的大鼠作为Sham组（n=10）。TBI+NC-sh组、TBI+P2RX3-sh组、TBI+NC-OE组和TBI+P2RX3-OE组大鼠分别脑内注射NC-sh、P2RX3-sh、NC-OE组和P2RX3-OE。Sham组和TBI组大鼠脑内注射等量生理盐水。首次注射7 d后再次注射1次，14 d后进行取材。采用改良神经功能缺损评分法评价大鼠神经功能。采用Morris水迷宫实验评价认知功能。采用蔗糖偏好实验和旷场实验评价行为学。采用称重法测量脑组织含水量。采用TUNEL染色检测细胞凋亡。采用免疫荧光染色检测BDNF荧光强度。采用RT-qPCR检测P2RX3 mRNA水平。采用Western blot检测P2RX3、BDNF、TrkB、p-TrkB、蛋白激酶B（Akt）、p-Akt、Bcl-2和Bax蛋白水平。结果：与TBI组和TBI+NC-sh组比较，TBI+P2RX3-sh组大鼠的P2RX3 mRNA和蛋白相对水平降低（P<0.05），神经功能评分、逃避潜伏期和脑含水量均升高（P<0.05），60 s内的穿越平台次数、蔗糖偏好率和水平以及垂直活动分数均降低（P<0.05），TUNEL阳性率和Bax蛋白相对水平升高（P<0.05），BDNF相对荧光强度以及Bcl-2、BDNF、p-TrkB和p-Akt蛋白相对水平降低（P<0.05）。与TBI组和TBI+NC-OE组比较，TBI+P2RX3-OE组大鼠的P2RX3 mRNA和蛋白相对水平升高（P<0.05），神经功能评分、逃避潜伏期和脑含水量均降低（P<0.05），60 s内的穿越平台次数、蔗糖偏好率和水平以及垂直活动分数均升高（P<0.05），TUNEL阳性率和Bax蛋白相对水平降低（P<0.05），BDNF相对荧光强度以及Bcl-2、BDNF、p-TrkB和p-Akt蛋白相对水平升高（P<0.05）。结论：P2RX3在大鼠TBI后上调，且上调P2RX3具有促进神经功能恢复的作用，其机制与激活BDNF/TrkB/Akt信号通路有关。 |
| 英文摘要: |
| ABSTRACT Objective: To reveal the effect of purine 2X3 (P2X3) receptor (P2RX3) gene on neurological function recovery in rats with traumatic brain injury (TBI). Methods: The TBI model of rats was made by craniocerebral injury instrument. After the model establishment, 54 TBI model rats were randomly divided into TBI group (n=10), TBI+NC-sh group (n=11), TBI+P2RX3-sh group (n=11), TBI+NC-OE group (n=11) and TBI+P2RX3-OE group (n=11). Rats which only cut the skin without hitting were taken as Sham group (n=10). Rats in TBI+NC-sh group, TBI+P2RX3-sh group, TBI+NC-OE group and TBI+P2RX3-OE group were injected into the brain with NC-sh, P2RX3-sh, NC-OE and P2RX3-OE respectively. Normal saline was injected into the brain of rats in Sham group and TBI group. Rats were injected again 7 days after the first injection, and the samples were taken 14 days later. The modified neurological deficit score was used to evaluate the neurological function. Cognitive function was evaluated by Morris water maze test. Sucrose preference test and open field test were used to evaluate behavior. The water content of brain tissue was measured by weighing method. Apoptosis was detected by TUNEL staining. The fluorescence intensity of BDNF was detected by immunofluorescence staining. The level of P2RX3 mRNA was detected by RT-qPCR. The protein levels of P2RX3, BDNF, TrkB, p-TrkB, protein kinase B (Akt), p-Akt, Bcl-2 and Bax were detected by Western blot. Results: Compared with TBI group and TBI+NC-sh group, the relative levels of mRNA and protein of P2RX3 in TBI+P2RX3-sh group decreased (P<0.05), neurological function score, escape latency and brain water content were all increased (P<0.05), platform crossing times within 60 s, sucrose preference rate and horizontal and vertical activity scores all decreased (P<0.05), TUNEL positive rate and the relative protein level of Bax increased (P<0.05), the relative fluorescence intensity of BDNF and the relative protein levels of Bcl-2, BDNF, p-TrkB and p-Akt decreased (P<0.05). Compared with TBI group and TBI+NC-OE group, the relative levels of mRNA and protein of P2RX3 in TBI+P2RX3-sh group increased (P<0.05), neurological function score, escape latency and brain water content were all decreased (P<0.05), platform crossing times within 60 s, sucrose preference rate and horizontal and vertical activity scores all increased (P<0.05), TUNEL positive rate and the relative protein level of Bax decreased (P<0.05), the relative fluorescence intensity of BDNF and the relative protein levels of Bcl-2, BDNF, p-TrkB and p-Akt increased (P<0.05). Conclusion: P2RX3 is up-regulated after TBI in rats, and up-regulation of P2RX3 can promote the recovery of neurological function, and its mechanism is related to the activation of BDNF/TrkB/Akt signal pathway. |
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