血清MMP-9、蛋白激酶B和Cyt-C联合检测在脑出血大鼠模型中的诊断价值
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摘要
目的分析血清基质金属蛋白酶9(MMP-9)、蛋白激酶B(AKT)和细胞色素C(Cyt-C)联合检测在脑出血大鼠模型中的诊断价值。方法 2017年1—6月在西电集团医院进行实验,选取健康雄性SD大鼠30只,随机数字表法分为空白组、假手术组和模型组各10只。空白组大鼠不做任何处理,假手术组大鼠开颅但不注血,模型组大鼠进行脑出血模型建造,在模型建造成功后的1 d、3 d和5 d对3组大鼠的行为学和神经功能分比较,并提取尾部静脉血比较1 d、3 d和5 d时间的MMP-9、AKT和Cyt-C水平,5 d后断头处死采集组织进行HE染色,并进行病理学检查,最后对MMP-9、AKT和Cyt-C与疾病严重程度的相关性和ROC曲线下面积分析比较。结果术后1 d、3 d、5 d,模型组大鼠平衡木评分、Longa评分和神经功能缺损程度评分明显高于空白组和假手术组大鼠(F_(平衡木)=4.518、11.216、13.588,P均=0.000;F_(longa)=5.286、9.812、13.925,P均=0.000;F_(神经功能)=48.891、55.165、51.599,P均=0.000);模型组大鼠MMP-9、Cyt-C表达水平在不同时间段均显著高于空白组、假手术组大鼠(P均=0.000),AKT水平低于空白组和假手术组大鼠(P均=0.000),且升高和降低趋势随着时间的延长而逐渐明显;MMP-9及Cyt-C与脑出血严重程度呈正相关(r/P=-0.496/0.024,0.507/0.020),AKT与脑出血严重程度呈负相关(r/P=0.582/0.018);联合检测的ROC曲线下面积明显大于单项MMP-9、AKT和Cyt-C检测的ROC曲线下面积,特异度、敏感度、阳性预测值、阴性预测值以及Youden指数均高于单项指标检测。结论 MMP-9、AKT和Cyt-C在脑出血的发病中均具有特异性表达,3项指标联合检测的诊断效能较高,能够为临床脑出血的治疗及预后提供重要的理论依据。
Objective To analyze the diagnostic value of combined detection of serum matrix metalloproteinase 9(MMP-9), protein kinase B(AKT) and cytochrome C(Cyt-C) in rats with cerebral hemorrhage.Methods From January to June 2017, 30 healthy male SD rats were selected and randomly divided into blank group, sham-operated group and model group with 10 rats each. Rats in the blank group did not do any treatment, rats in the sham-operated group were craniotomy without blood injection. Rats in the model group underwent cerebral hemorrhage model construction. Behavioral and neurological functions of the three groups were compared on the 1 st, 3 rd and 5 th day after the model was successfully constructed. The levels of MMP-9, AKT and Cyt-C in the caudal vein blood were extracted and compared on the 1 st, 3 rd and 5 th day. Tissues were decapitated and killed 5 days later for HE staining. Pathological examination was carried out. Finally, the correlation between MMP-9, AKT and Cyt-C and the severity of disease and the area under ROC curve were analyzed and compared. Results At 1 d, 3 d, and 5 d after operation, the balance beam score, Longa score, and neurological deficit score of the model group were significantly higher than those of the blank group and the sham operation group(F_(balance beam) = 4.518, 11.216, 13.588,P= 0.000; F_(longa) = 5.286, 9.812, 13.925,P=0.000; F_(neurological function) = 48.891, 55.165, 51.599,P=0.000); MMP-9 and Cyt-C expression levels in the model group were significantly higher in different time periods. In the blank group and sham-operated group(P=0.000), the AKT level was lower than that in the blank group and the sham-operated group(P=0.000), and the increase and decrease trend gradually became obvious with the prolongation of time; MMP-9 and Cyt-C were positively correlated with the severity of cerebral hemorrhage(r/P=-0.496/0.024, r/P=0.507/0.020), and AKT was negatively correlated with the severity of cerebral hemorrhage(r/P=0.582/0.018). The area under the ROC curve of the combined detection was significantly larger than the area under the ROC curve of single MMP-9, AKT and Cyt-C detection. The specificity, sensitivity, positive predictive value, negative predictive value and Youden index were higher than the single index test. Conclusion MMP-9, AKT and Cyt-C have specific expression in the pathogenesis of cerebral hemorrhage. The combined detection of three indicators has high diagnostic efficiency, which can provide important theoretical basis for the treatment and prognosis of clinical cerebral hemorrhage.
Objective To analyze the diagnostic value of combined detection of serum matrix metalloproteinase 9(MMP-9), protein kinase B(AKT) and cytochrome C(Cyt-C) in rats with cerebral hemorrhage.Methods From January to June 2017, 30 healthy male SD rats were selected and randomly divided into blank group, sham-operated group and model group with 10 rats each. Rats in the blank group did not do any treatment, rats in the sham-operated group were craniotomy without blood injection. Rats in the model group underwent cerebral hemorrhage model construction. Behavioral and neurological functions of the three groups were compared on the 1 st, 3 rd and 5 th day after the model was successfully constructed. The levels of MMP-9, AKT and Cyt-C in the caudal vein blood were extracted and compared on the 1 st, 3 rd and 5 th day. Tissues were decapitated and killed 5 days later for HE staining. Pathological examination was carried out. Finally, the correlation between MMP-9, AKT and Cyt-C and the severity of disease and the area under ROC curve were analyzed and compared. Results At 1 d, 3 d, and 5 d after operation, the balance beam score, Longa score, and neurological deficit score of the model group were significantly higher than those of the blank group and the sham operation group(F_(balance beam) = 4.518, 11.216, 13.588,P= 0.000; F_(longa) = 5.286, 9.812, 13.925,P=0.000; F_(neurological function) = 48.891, 55.165, 51.599,P=0.000); MMP-9 and Cyt-C expression levels in the model group were significantly higher in different time periods. In the blank group and sham-operated group(P=0.000), the AKT level was lower than that in the blank group and the sham-operated group(P=0.000), and the increase and decrease trend gradually became obvious with the prolongation of time; MMP-9 and Cyt-C were positively correlated with the severity of cerebral hemorrhage(r/P=-0.496/0.024, r/P=0.507/0.020), and AKT was negatively correlated with the severity of cerebral hemorrhage(r/P=0.582/0.018). The area under the ROC curve of the combined detection was significantly larger than the area under the ROC curve of single MMP-9, AKT and Cyt-C detection. The specificity, sensitivity, positive predictive value, negative predictive value and Youden index were higher than the single index test. Conclusion MMP-9, AKT and Cyt-C have specific expression in the pathogenesis of cerebral hemorrhage. The combined detection of three indicators has high diagnostic efficiency, which can provide important theoretical basis for the treatment and prognosis of clinical cerebral hemorrhage.
引文
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[2] Kamei H, Wakimoto Y, Murakami Y, et al. A Case of a pregnant woman with thrombosis in an artificial aortic valve resulting in severe cerebral hemorrhage in the newborn [J]. Case Rep Obstet Gynecol, 2018,2018:6154382. DOI: 10.1155/2018/6154382.
[3] 张极星,张艳国,赵亮,等.急性期脑出血患者血清ICAM-1、MMP-9、TNF-α、IL-6水平和出血灶周围水肿脑组织体积的相关性[J].山东医药,2017,57(27):58-60.DOI:10.3969/j.issn.1002-266X.2017.27.017.
[4] 徐磊,闫兴洲,李震宇等.醒脑开窍针法结合康复训练对缺血性脑卒中大鼠神经修复和梗死灶周围皮质神经生长相关蛋白-43表达的影响[J].针刺研究,2017,42(3):223-228.DOI:10.13702/j.1000-0607.2017.03.006.
[5] 俞天虹,潘华阳.补阳还五汤及其拆方对脑缺血大鼠神经功能及血管生成的影响[J].中国脑血管病杂志,2017,14(2):87-93.DOI:10.3969/j.issn.1672-5921.2017.02.006.
[6] 李丽敏,于芳苹,张黎,等.盐酸法舒地尔对脑出血大鼠炎性反应及继发性脑损伤的影响研究[J].中华神经医学杂志,2017,16(11):1081-1090.DOI:10.3760/cma.j.issn.1671-8925.2017.11.001.
[7] 李治华,陈曦,臧卫东,等.Wnt/β-catenin通路对大鼠脑出血的保护作用[J].郑州大学学报:医学版,2015,50(6):749-752,753.DOI:10.13705/j.issn.1671-6825.2015.06.005.
[8] Haegens NM, Gathier CS, Horn J, et al. Induced Hypertension in Preventing Cerebral Infarction in Delayed Cerebral Ischemia After Subarachnoid Hemorrhage [J]. Stroke, 2018,49(11):2630-2636. DOI:10.1161/STROKEAHA.118.022310.
[9] Zhao L, Arbel-Ornath M, Wang X, et al. Matrix metalloproteinase 9-mediated intracerebral hemorrhage induced by cerebral amyloid angiopathy [J]. Neurobiol Aging, 2015,36(11):2963-2971. DOI:10.1016/j.neurobiolaging.2015.07.016.
[10] 张秀峰,刘宏,王静辉,等.瑞舒伐他汀对脑出血大鼠血清IL-6、脑组织AQP-4与MMP-9表达水平和神经功能的影响[J].河北医药,2018,40(18):2815-2818.DOI:10.3969/j.issn.1002-7386.2018.18.023.
[11] 宁培云,赵洪勇,赵志超,等.丹红注射液对实验性脑出血大鼠脑水肿及血浆MMP-9、TIMP-1、NF-κB水平的影响[J].疑难病杂志,2017,16(8):821-824.DOI:10.3969/j.issn.1671-6450.2017.08.017.
[12] Xi T, Jin F, Zhu Y, et al. MicroRNA-126-3p attenuates blood-brain barrier disruption, cerebral edema and neuronal injury following intracerebral hemorrhage by regulating PIK3R2 and Akt[J]. Biochem Biophys Res Commun,2017,494(1-2):144-151. DOI:10.1016/j.bbrc.2017.10.064.
[13] 陈状,温锦崇,江澈,等.乌司他丁联合微创血肿清除术对高血压脑出血患者神经功能、氧化应激反应的影响[J].疑难病杂志,2018,17(9):884-887.DOI:10.3969/j.issn.1671-6450.2018.09.005
[14] 赵明媚,袁少勇,陈瑶刚,等.小骨窗微创血肿清除术联合甘露醇对高血压脑出血患者炎性因子、血清铁蛋白和血清P物质的影响[J].疑难病杂志,2018,17(8):776-780.DOI:10.3969/j.issn.1671-6450.2018.08.005
[15] 向勇,朱建萍,刘丹荣,等.rhEPO通过调节PI3K/AKT信号通路改善大鼠脑出血后神经元损伤[J].中南医学科学杂志,2017,45(5):454-458.DOI:10.15972/j.cnki.43-1509/r.2017.05.007.
[16] 刘绍晨,吴晓光,杨岚,等.补阳还五汤对脑出血大鼠脑组织水通道蛋白4表达及血-脑脊液屏障通透性的影响研究[J].中国全科医学,2016,19(21):2557-2561.DOI:10.3969/j.issn.1007-9572.2016.21.013.
[17] 陈朝晖,洪溪屏,兰频,等.百里醌对大鼠蛛网膜下腔出血后脑细胞凋亡的影响[J].医学研究杂志,2018,47(3):89-93.DOI:10.11969/j.issn.1673-548X.2018.03.023.
[18] 王跃华,林贵军,饶芝国,等.阿托伐他汀对大鼠脑出血后血肿周围组织细胞凋亡和细胞色素c表达的影响[J].国际脑血管病杂志,2013,21(3):191-196.DOI:10.3760/cma.j.issn.1673-4165.2013.03.006.
[19] 肖兴花.粒细胞集落刺激因子对实验性脑出血大鼠血肿周围组织中Bax及Bcl-2蛋白表达的影响[J].中国老年学杂志,2015,(11):2927-2928,2929.DOI:10.3969/j.issn.1005-9202.2015.11.020.
[20] 殷旭华,李冬梅,于慧玲,等.不同时间点脑出血大鼠脑组织中缺氧诱导因子1α和线粒体细胞色素C的表达[J].中国老年学杂志,2014,18(20):5790-5791.DOI:10.3969/j.issn.1005-9202.2014.20.079.