高压氧对血管性痴呆大鼠认知功能障碍的影响及抗凋亡机制  

Effects of hyperbaric oxygen therapy on vascular dementia and its anti-apoptosis mechanism

作　　者：赵慧 安娜 陈文艳 Zhao Hui;An Na;Chen Wenyan(Department of General Medicine,Shanxi Bethune Hospital,Shanxi Academy of Medical Sciences,Tongji Shanxi Hospital,Third Hospital of Shanxi Medical University,Taiyuan 030032,China)

机构地区：[1]山西白求恩医院(山西医学科学院同济山西医院)综合医疗科,山西医科大学第三医院,太原030032

出　　处：《中华实验外科杂志》2023年第9期1729-1732,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(81971094);山西白求恩医院人才引进项目(2022RC09)。

摘　　要：目的观察高压氧(HBO)对血管性痴呆(VD)大鼠认知功能障碍的保护性作用,并探讨相关抗凋亡机制。方法采用随机数字表法将39只雄性SD大鼠分为假手术组、模型组、HBO组,每组13只。HBO组于造模后6 h进行治疗,HBO治疗压力为0.2 MPa,稳压吸氧60 min,1次/d。假手术组与模型组呼吸常压空气。28 d后,采用Morris水迷宫实验评价各组空间学习与记忆能力。断头取海马组织,苏木精-伊红(HE)染色观察神经元病理学变化,蛋白免疫印迹法(Western blot)检测B细胞淋巴瘤/白血病-2(bcl-2)、B细胞淋巴瘤/白血病-2相关X蛋白(bax)、半胱氨酸天冬氨酸特异性蛋白酶(Caspase)-3、磷脂酰激酶3(PI3K)、磷酸化蛋白激酶B(p-Akt)的相对表达量,原位末端转移酶标记法(TUNEL)计算神经细胞凋亡率。多组间比较采用单因素方差分析,两组间比较采用LSD法。结果HBO治疗可减轻VD大鼠认知功能障碍,HBO组逃避潜伏期显著低于模型组[(28.96±4.67)s比(46.81±6.75)s,t=5.909,P<0.01],平台象限时间比、穿越平台次数显著高于模型组[(30.31±6.45)%、(2.00±0.85)次比(21.57±5.26)%、(1.08±0.67)次,t=3.640、2.930,P<0.01],海马区细胞排列紊乱、胞核皱缩、炎性细胞浸润的程度减低,正常神经元计数显著高于模型组[(100.58±7.99)个/高倍镜视野比(91.25±7.62)个/高倍镜视野,t=2.928,P<0.01]。bcl-2、PI3K、p-Akt蛋白相对表达显著高于模型组(1.32±0.20、0.84±0.07、1.01±0.05比0.87±0.13、0.42±0.09、0.56±0.07,t=3.314、6.712、9.409,P<0.05),bax蛋白、Caspase-3蛋白相对表达显著低于模型组(0.84±0.03、0.75±0.08比1.26±0.06、1.65±0.45,t=3.404、10.537,P<0.05),细胞凋亡率无明显变化[(25.00±3.67)%比(29.40±3.65)%,t=1.901,P>0.05]。结论HBO可能通过上调PI3K/Akt信号通路,减轻神经细胞损伤,调节细胞凋亡,保护VD大鼠认知功能。Objective To observe the protective effect of hyperbaric oxygen(HBO)on cognitive impairment in rats with vascular dementia(VD)and its related anti-apoptosis mechanism.Methods A total of 39 male Sprague-Dawley(SD)rats were randomly assigned into Sham,VD and HBO groups(n=13 each).The HBO group was given HBO therapy 6 h after the modeling,and the sham and VD groups inhaled air at normal pressure.At 28th day after the modeling,the changes in spatial learning and memory ability were measured by Morris water maze test.Histological changes of hippocampal brain slices were examined by hematozylin-eosin(HE)staining.The expression levels of B cell lymphoma/lewkmia-2(bcl-2),bcl-2 associated X protein(bax),cysteinyl aspartate specific proteinase-3(Caspase-3),phosphatidylinositol 3 kinase(PI3K)and phosphorylated protein kinase B(p-Akt)in hippocampal brain slices were detected by Western blotting.The apoptosis rate was measured by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling(TUNEL).Results The HBO therapy could ameliorate the cognitive injury in VD.The average escape latency in the HBO group was significantly shorter than that in the VD group[(28.96±4.67)s vs.(46.81±6.75)s,t=5.909,P<0.01].Platform quadrant time ratios and the number of times crossing the platform area were significantly higher than those in the VD group[(30.31±6.45)%,(2.00±0.85)vs.(21.57±5.26)%,(1.08±0.67),t=3.640,2.930,P<0.01].The arrangement of hippocampal cells was less disordered,nucleus pyknosis and the infiltration of inflammatory cells were lower and the number of normal neurons was greater in the HBO group than those in the VD group[(100.58±7.99)vs.(91.25±7.62),t=2.928,P<0.01].The expression of bcl-2,PI3K and p-Akt was significantly higher[(1.32±0.20),(0.84±0.07),(1.01±0.05)vs.(0.87±0.13),(0.42±0.09),(0.56±0.07),t=3.314,6.712,9.409,P<0.05],and that of bax and Caspase-3 was significantly lower[(0.84±0.03),(0.75±0.08)vs.(1.26±0.06),(1.65±0.45),t=3.404,10.537,P<0.05]in the HBO group than those in the VD gr

关 键 词：高压氧 血管性痴呆 神经细胞损伤 凋亡 磷脂酰激酶3/蛋白激酶B 

分 类 号：R749.13[医药卫生—神经病学与精神病学]