乌司他丁对脑出血大鼠局部黏着斑激酶/细胞外信号调节激酶信号通路及神经元凋亡的影响  

Effects of ulinastatin on FAK/ERK signaling pathway and neuronal apoptosis in rats with intracerebral hemorrhage

作　　者：苗志凯 夏清岫[1] 王艳[1] MIAO Zhikai;XIA Qingxiu;WANG Yan(Department of Neurosurgery,Cangzhou Hospital of Integrated Traditional Chinese and Western Medicine,Cangzhou,Hebei 061000,China)

机构地区：[1]河北省沧州中西医结合医院神经外科,河北沧州061000

出　　处：《安徽医药》2022年第11期2217-2222,I0002,共7页Anhui Medical and Pharmaceutical Journal

摘　　要：目的探讨乌司他丁(UTI)对脑出血(ICH)大鼠局部黏着斑激酶(FAK)/细胞外信号调节激酶(ERK)信号通路及神经元凋亡的影响。方法建立ICH大鼠模型,使用随机数字表法将造模成功的大鼠随机分为模型组(尾静脉注射0.9%氯化钠溶液)、抑制剂组(FAK抑制剂PF562271,50 mg/kg灌胃)、UTI组(尾静脉注射10万U/kg UTI)、UTI+抑制剂组(FAK抑制剂PF562271,50 mg/kg灌胃同时尾静脉注射10万U/kg UTI),每组20只,另设20只为假手术组(尾静脉注射0.9%氯化钠溶液)作为对照,所有处理均每天1次,连续7 d。干湿比重法测定各组大鼠脑组织含水率;酶联免疫吸附(ELISA)法各组大鼠血清中炎性因子含量;原位末端标记(TUNEL)法检测各组大鼠海马组织神经元凋亡;免疫组织化学分析法检测大鼠海马组织抗凋亡因子B细胞淋巴瘤-2(Bcl-2)和促凋亡因子Bcl相关X(Bax)、胱天蛋白酶3(caspase-3)表达;尼氏染色法检测各组大鼠海马组织中神经元存活情况;蛋白质印迹法(Western blotting)法检测大鼠海马组织中FAK蛋白表达和ERK1/2磷酸化水平。结果与假手术组相比,模型组大鼠脑组织含水率(84.51±7.42)%、炎性因子含量、神经元凋亡指数(44.51±3.77)%以及促凋亡因子Bax(3.05±0.41)、caspase-3(3.27±0.46)表达和ERK1/2磷酸化水平升高,神经元存活数目、抑凋亡因子Bcl-2(1.23±0.21)表达以及FAK(0.29±0.07)蛋白表达水平降低(P<0.05)。与模型组相比,UTI组大鼠脑组织含水率(71.43±6.11)%、炎性因子含量、神经元凋亡指数(15.34±0.76)%以及促凋亡因子Bax(2.03±0.15)、caspcase-3(2.27±0.61)表达和ERK1/2磷酸化水平降低,神经元存活数目、抑凋亡因子Bcl-2(2.67±0.27)表达以及FAK(0.58±0.09)蛋白表达升高(P<0.05);抑制剂组大鼠脑组织含水率(92.83±7.56)%、炎性因子含量、神经元凋亡指数(51.99±5.65)%以及促凋亡因子Bax(3.73±0.37)、caspcase-3(3.99±0.26)表达和ERK1/2磷酸化水平升高,神经元存活数目、抑凋亡因Objective To investigate the effects of ulinastatin(UTI)on focal adhesion kinase(FAK)/extracellular signal regulated ki-nase(ERK)signal pathway and neuronal apoptosis in rats with intracerebral hemorrhage.Methods ICH rat models were establishedand randomly divided into model group(tail vein injection of 0.9%sodium chloride solution),inhibitor group(FAK inhibitor PF562271,50 mg/kg by gavage),UTI group(tail vein injection of 100000 U/kg UTI),UTI+inhibitor group(gavage of FAK inhibitor pf562271 and50 mg/kg,meanwhile,tail vein injection of 100000 U/kg UTI),with 20 rats in each group,another 20 rats were divided into sham opera-tion group(tail vein injection of 0.9%sodium chloride solution)as control,all treatments were once a day for 7 consecutive days.Thewater content of brain tissue was measured by dry wet specific gravity method;the content of inflammatory factors in serum of rats ineach group was detected by enzyme-linked immunosorbent assay(ELISA);TUNEL method was used to detect neuronal apoptosis in hip-pocampus;the expressions of anti apoptotic factor Bcl-2 and pro apoptotic factor Bax,caspase-3 were detected by immunohistochemistry;the survival of neurons in hippocampal tissues of rats were detected by Nysch staining;and Western blotting was used to detect FAKprotein expression and ERK1/2 phosphorylation level in hippocampus.Results Compared with those in the sham operation group,thewater content of brain tissue(84.51±7.42)%,inflammatory factor content,neuronal apoptosis index(44.51±3.77)%,Bax(3.05±0.41)andcaspase-3(3.27±0.46)expressions,ERK1/2 phosphorylation level in model group were higher,while the expressions of Bcl-2(1.23±0.21)and FAK(0.29±0.07)protein was lower(P<0.05).Compared with those in the model group,the water content of brain tissue(71.43±6.11)%,inflammatory factor content,neuronal apoptosis index(15.34±0.76)%,Bax(2.03±0.15)and caspase-3(2.27±0.61)expres-sions,ERK1/2 phosphorylation level in UTI group were lower,while the number of neurons alive,the expressions of Bcl-2(2.67±0.27)and

关 键 词：乌司他丁 脑出血 局部黏着斑激酶/细胞外信号调节激酶 神经元凋亡 

分 类 号：R743.34[医药卫生—神经病学与精神病学]