BDNF對缺糖缺氧小鼠mPFC腦區神經元突觸形成的影響

喻曉路 傅慧

摘要：目的 ?探討腦源性神經生長因子（BDNF）在缺糖缺氧復灌條件下對mPFC腦區突觸前膜蛋白Synapsin-1的保護作用。方法 ?將18只小鼠隨機分為正常組、缺糖缺氧復灌組、缺糖缺氧復灌給藥組，每組6只。正常組mPFC腦區低糖DMEM培養基培養，缺糖缺氧復灌組（OGD/R）mPFC腦區EBSS培養基培養15 min后復灌1 h，缺糖缺氧復灌給藥組（OGD/R+BDNF）mPFC腦區EBSS培養基中加入BDNF培養15 min后復灌1 h，采用免疫印跡分析和RT-qPCR檢測分析突觸前膜蛋白Synapsin-1的表達量和mRNA表達量。結果 ?缺糖缺氧復灌組Synapsin-1蛋白和mRNA表達量低于正常組，差異有統計學意義（P<0.05）;缺糖缺氧復灌給藥組Synapsin-1蛋白表達量高于缺糖缺氧復灌組，差異有統計學意義（P<0.05）;缺糖缺氧復灌給藥組Synapsin-1蛋白和mRNA表達量與正常組比較，差異無統計學意義（P>0.05）。結論 ?缺糖缺氧對mPFC腦區神經元突觸有損傷作用，BDNF對神經元突觸起到保護作用。

關鍵詞：腦源性神經生長因子;mPFC;缺糖缺氧;突觸前膜蛋白

中圖分類號：R614 ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? 文獻標識碼：A ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? DOI：10.3969/j.issn.1006-1959.2020.17.018

文章編號：1006-1959（2020）17-0063-04

Abstract：Objective ?To investigate the protective effect of brain-derived nerve growth factor （BDNF） on the presynaptic membrane protein Synapsin-1 in mPFC brain under the condition of hypoglycemia and hypoxia reperfusion.Methods ?18 mice were randomly divided into normal group， hypoglycemia and hypoxia reperfusion group， hypoglycemia and hypoxia reperfusion administration group， 6 mice in each group. Normal group mPFC brain area was cultured in low-sugar DMEM medium， hypoglycemia hypoxia reperfusion group （OGD/R） mPFC brain area EBSS medium was cultured for 15 min and then reperfused for 1 h， hypoglycemia hypoxia reperfusion administration group （OGD/R+BDNF） mPFC brain area EBSS medium was cultured with BDNF for 15 min and reperfused for 1 h.Western blot analysis and RT-qPCR were used to analyze the expression and mRNA expression of the presynaptic membrane protein Synapsin-1.Results ?The expression of Synapsin-1 protein and mRNA in the hypoglycemia and hypoxia reperfusion group was lower than that of the normal group，the difference was statistically significant （P<0.05）;Synapsin-1 protein expression in the hypoglycemia and hypoxia reperfusion administration group was higher than that of the hypoglycemia hypoxia reperfusion group， the difference was statistically significant （P<0.05）; Synapsin-1 protein and Compared with the normal group， the mRNA expression level was not statistically different （P>0.05）.Conclusion ?Glucose and hypoxia could damage neuronal synapses in the mPFC brain region， and BDNF could protect neuronal synapses.

Key words：Brain-derived nerve growth factor;mPFC;Hypoglycemia and hypoxia;Presynaptic membrane protein

1.3統計學處理 ?采用統計學軟件SPSS 13.0進行處理，計量資料采用（x±s）表示，采用單因素方差分析，one-way ANOVE進行組間比較，P<0.05 為差異有統計學意義。

2結果

2.1三組小鼠mPFC腦區病理形態 ?鏡檢結果顯示，三組腦片中細胞核形態清晰、完整，提示模型制作成功，短時間的缺塘缺氧沒有造成大面積的組織壞死和細胞凋亡，完全符合后續實驗要求，見圖1。

2.2三組小鼠mPFC腦區Synapsin-1蛋白表達比較 ?WB檢測結果顯示，正常組Synapsin-1蛋白的相對表達量為（1.0573436±0.171485464），缺糖缺氧復灌組Synapsin-1蛋白的相對表達量為（0.702825767±0.03653467），缺糖缺氧復灌給藥組Synapsin-1蛋白的相對表達量為（0.845764±0.073608716）;缺糖缺氧復灌組Synapsin-1蛋白的表達量低于正常組、缺糖缺氧復灌給藥組，差異有統計學意義（P<0.05）;缺糖缺氧復灌給藥組與正常組Synapsin-1蛋白表達量比較，差異無統計學意義（P>0.05），見圖2。

2.3三組小鼠mPFC腦區Synapsin-1 mRNA相對表達量比較 ?正常組Synapsin-1 mRNA的相對表達量為（1.00000±1.00000），缺糖缺氧復灌組Synapsin-1 mRNA的相對表達量為（0.4126427±0.299804），缺糖缺氧復灌給藥組Synapsin-1 mRNA的相對表達量為（0.604788±0.271342）。……