丹参酮ⅡA通过PI3K/AKT信号通路调节脊髓神经元凋亡的实验研究  

Experimental study on effect of tanshinone IIA in spinal cord compression through PI3K/AKT signaling pathway

作　　者：刘伟[1] 赵宝辉[1] 王建忠[1] 刘晓峰[1] 马晓勇[1] 雷鸣[1] Liu Wei;Zhao Baohui;Wang Jianzhong;Liu Xiaofeng;Ma Xiaoyong;Lei Ming(Department of Orthopaedic,the Affiliated Hospital of Hebei University,Baoding 071000,China)

机构地区：[1]河北大学附属医院骨科,保定071000

出　　处：《中华骨科杂志》2019年第10期622-629,共8页Chinese Journal of Orthopaedics

摘　　要：目的通过建立脊髓压迫模型探讨丹参酮ⅡA对受压迫的脊髓组织中神经元凋亡的影响及其分子机制。方法 24只体重为250~300 g的普通级SD大鼠作为实验动物简单随机化方法分为4组,每组6只:假手术组、脊髓压迫损伤组(spinal cord injury,SCI组)、丹参酮ⅡA组、LY2904002组(LY组)。采用动脉钳夹闭脊髓的方式对后3组建立脊髓压迫模型。假手术组给予生理盐水腹腔注射;SCI组给予生理盐水腹腔注射;丹参酮ⅡA组予30 mg/kg丹参酮ⅡA腹腔注射;LY组予0.3 mg/kg LY294002腹腔注射,5 min后予30 mg/kg丹参酮ⅡA腹腔注射。干预后3 d,通过斜板角度和Basso-Beattie-Bresnahan(BBB)脊髓损伤的行为学评分来评估大鼠的运动功能,并通过qPCR、组织染色、蛋白质免疫印迹等分子生物学实验手段检测受压脊髓组织中凋亡基因、PI3K/AKT信号分子的表达量。结果 SCI组BBB评分(3.31±0.45)分、斜板角度9.31°±1.02°及受压脊髓组织中GSK-3β 0.35±0.06、CyclinD1 0.25±0.06、Bcl-2 0.38±0.06、p-PI3K 0.32±0.05、p-AKT 0.29±0.07的相对表达量均低于假手术组,受压脊髓组织中Caspase-9 3.27±0.54 、Caspase-3 2.73±0.35的蛋白相对表达量高于假手术组;丹参酮ⅡA组大鼠的BBB评分(9.31±1.02)分、斜板角度24.95°±3.52°以及受压脊髓组织中GSK-3β 0.74±0.09、CyclinD1 0.69±0.11、Bcl-2 0.83±0.13、p-PI3K 0.77±0.11、p-AKT 0.69±0.08的蛋白相对表达量均高于SCI组,受压脊髓组织中Caspase-9 1.78±0.22、Caspase-3 1.64±0.2的蛋白表达量低于SCI组;LY组大鼠的BBB评分(0.43±0.07)分、斜板角度0.38°±0.06°以及受压脊髓组织中GSK-3β 0.43±0.07、CyclinD1 0.38±0.06、Bcl-2 0.49±0.09的蛋白相对表达量均低于丹参酮ⅡA组,受压脊髓组织中Caspase-9 2.54±0.38、Caspase-3 2.25±0.37的蛋白相对表达量高于丹参酮ⅡA组。结论丹参酮ⅡA可通过激活PI3K/AKT信号通路来抑制受压脊髓组织中的细胞凋亡、减轻脊髓压迫大鼠模型的�Objective To explore the effect of tanshinone IIA on neuronal apoptosis in the compressed spinal cord and its molecular mechanism by establishing a spinal cord compression model. Methods Twenty-four SD rats weighing 250-300 g were the experimental animals. The spinal cord compression model was established by clamping the spinal cord with arterial clamp. Six rats in each group were randomly selected and randomly divided into sham group (Sham group, given intraperitoneal injection of normal saline at the same dose once a day) and spinal cord compression injury group (SCI group, given normal salts). Water intraperitoneal injection, once a day), tanshinone IIA group (TAN group, 30 mg/kg tanshinone IIA intraperitoneal injection, once a day), LY2904002 group (LY group, 0.3 mg/kg LY294002 intraperitoneal injection, 5 min after 30 mg/kg tanshinone IIA intraperitoneal injection, once 1 d). After 3 d of intervention, the motor function of rats were evaluated by inclined plane test and BBB score. The expression of apoptotic genes and PI3K/AKT signaling molecules in the compressed spinal cord were detected by qPCR, immunohistochemistry and Western blot. Results The BBB score 3.31±0.45 points, inclined angle 9.31°±1.02°, GSK-3β 0.35±0.06, CyclinD1 0.25±0.06, Bcl-2 0.38±0.06, p-PI3K 0.32±0.05, p-AKT 0.29±0.07 protein expression in SCI group were lower than those in Sham group. The protein expression of Caspase-9 3.27±0.54 and Caspase-3 2.73±0.35 in SCI group was higher than that in Sham group. The BBB score 9.31±1.02 points, inclined angle 24.95°±3.52°, GSK-3β 0.74±0.09, CyclinD1 0.69±0.11, Bcl-2 0.83±0.13, p-PI3K 0.77±0.11, p-AKT 0.69±0.08 in TAN group were higher than those in SCI group BBB score 3.31 ±0.45 points, inclined angle 9.31°±1.02°, GSK-3β 0.35±0.06, CyclinD1 0.25±0.06, Bcl-2 0.38±0.06, p-PI3K 0.32±0.05, p-AKT 0.29±0.07. The protein expression levels of Caspase-9 1.78±0.22 and Caspase-3 1.64±0.2 in TAN group were lower than those in SCI group 3.27±0.54 and Caspase-3 2.73±0.35. Th

关 键 词：脊髓压迫症 丹参酮 磷酸肌醇3-激酶类 细胞凋亡 

分 类 号：R651.2[医药卫生—外科学]