舒血宁注射液对急性脑梗死大鼠脑组织的保护作用及其机制

doi:10.13481/j.1671-587x.20200321

摘要/Abstract

摘要： 目的：探讨舒血宁注射液对急性脑梗死大鼠脑组织的保护作用，并阐明其作用机制。方法：50只大鼠随机分为对照组、假手术组、模型组、尼莫地平组和舒血宁注射液组（n=10）。模型组、尼莫地平组和舒血宁注射液组采用颈内动脉线栓法建立急性脑梗死大鼠模型。假手术组大鼠分离颈总动脉、颈外动脉和颈内动脉后，只结扎颈外动脉；对照组大鼠不作手术处理。舒血宁注射液组大鼠造模成功后给予舒血宁注射液，尼莫地平组大鼠造模后给予尼莫地平，对照组、模型组和假手术组大鼠给予等体积生理盐水。检测各组大鼠神经功能缺损评分、脑组织含水量和相对脑梗死面积，HE染色观察各组大鼠脑组织形态表现，免疫组织化学染色检测各组大鼠脑组织中生长分化因子15（GDF-15）和C反应蛋白（CRP）表达水平，ELISA法检测各组大鼠脑组织中肿瘤坏死因子α（TNF-α）、白细胞介素6（IL-6）和白细胞介素1β（IL-1β）水平。结果：对照组和假手术组大鼠脑皮层结构完整，细胞排列整齐；模型组大鼠神经细胞胞浆和胞核皱缩，神经细胞与毛细管周围间质疏松；舒血宁注射液组大鼠脑皮质神经细胞、神经胶质细胞肿胀程度及间质疏松程度均减轻，病理组织形态表现与尼莫地平组相近。与对照组和假手术组比较，模型组大鼠脑组织含水量升高（P<0.05），脑组织中CRP表达水平和TNF-α、IL-6和IL-1β水平升高（P<0.05），GDF-15表达水平明显降低（P<0.05）。与模型组比较，尼莫地平组和舒血宁注射液组大鼠神经功能缺损评分降低（P<0.05），脑组织含水量明显降低（P<0.05），相对脑梗死面积减小（P<0.05），脑组织中CRP表达水平和TNF-α、IL-6及IL-1β水平均明显降低（P<0.05），GDF-15表达水平明显升高（P<0.05）。与尼莫地平组比较，舒血宁组大鼠神经功能缺损评分，脑组织含水量，相对脑梗死面积，脑组织中CRP和GDF-15表达水平及TNF-α、IL-6、IL-1β水平比较差异均无统计学意义（P>0.05）。结论：舒血宁注射液可上调急性脑梗死大鼠脑组织GDF-15表达，抑制CRP表达，降低细胞炎性因子水平，改善神经功能，对急性脑梗死起保护作用。

关键词: 脑梗死, 舒血宁注射液, 生长分化因子15, C反应蛋白

Abstract: Objective: To investigate the protective effect of Shuxuening Injection on the brain tissue of the rats with acute cerebral infarction,and to elucidate its mechanism. Methods: Fifty rats were randomly divided into control group, sham operation group, model group, nimodipine group and Shuxuening Injection group(n=10).The rat models of acute cerebral infarction were made by internal carotid artery suture method in model group, nimodipine group and Shuxuening Injection group. The common carotid arteries, the external carotid arteries and the internal carotid arteries of the rats in sham operation group were separated,and only the external carotid arteries were ligated; the rats in control group were not treated with operation; the rats in Shuxuening Injection group were given Shuxuening Injection, the rats in nimodipine group were given nimodipine,and the rats in control group, model group and sham operation group were given normal saline at the same volume.The score of neurological impairment, water contents in brain tissue and relative cerebral infarction areas of the rats in various groups were measured.HE staining was used to observe the morphology of brain tissue of the rats in various groups,and immumohistochemical staining was used to detect the expression levels of growth differentiation factor -15(GDF-15) and C-reactive protein (CRP) in brain tissue of the rats in various groups. ELISA method was used to detect the levels of tumor necrosis factor-α(TNF-α), interleukin- 6(IL-6)and interleukin-1β(IL-1β)in brain tissue of the rats in various groups. Results: The cortical structures of cortex of the rats in control group and sham operation group were complete and the cells were arranged neatly;the cytoplasm and nucleus of the nerve cells of the rats in model group were wrinkled, and the interstitium between nerve cells and capillaries were loose; the swelling degrees of cerebral cortical nerve cells and glial cells and the loose degrees of stroma of the rats in Shuxuening Injection group were reduced, and the histopathological manifestations were similar to those in nimodipine group. Compared with control group and sham operation group, the water content in brain tissue of the rats in model group was significantly increased(P<0.05), and the expression levels of CRP,the levels of TNF-α, IL-6 and IL-1β in brain tissue of the rats in model group were significantly increased(P<0.05), while the expression level of GDF-15 was significantly decreased(P<0.05).Compared with model group, the scores of neurological impairment of the rats in nimodipine group and Shuxuening Injection group were significantly decreased (P<0.05), and the relative cerebral infarction areas(P<0.05),the water contents in brain tissue(P<0.05),the expression levels of CRP,the levels of TNF-α, IL-6, and IL-1β in brain tissue of the rats in nimodipine group and Shuxuening Injection group were significantly decreased(P<0.05), while the expression levels of GDF-15 were significantly increased(P<0.05).Compared with nimodipin group, the score of neurological impairment,the relative cerebral infarction area, the water content in brain tissue,the expression levels of CRP and GDF-15 and the levels of TNF-α, IL-6 and IL-1β in brain tissue of the rats in Shuxuening Injection group had no statistical significances (P>0.05). Conclusion: Shuxuening Injection can protect the acute cerebral infarction by up-regulating the expression of GDF-15 and inhibiting the expression of CRP in brain tissue of the rats with acute cerebral infarction, reducing the levels of inflammatory cytokines and improving the neurological function.

Key words: cerebral infarction, Shuxuening Injection, brain tissue, growth differentiation factor 15, C-reactive protein

中图分类号:

R743.33

焦光美, 单海雷, 张晓璇, 赵亮, 窦志杰, 康玲伶, 马征, 孙艳军, 杨宁. 舒血宁注射液对急性脑梗死大鼠脑组织的保护作用及其机制[J]. 吉林大学学报(医学版), 2020, 46(03): 557-562.

JIAO Guangmei, SHAN Hailei, ZHANG Xiaoxuan, ZHAO Liang, DOU Zhijie, KANG Lingling, MA Zheng, SUN Yanjun, YANG Ning. Protective effect of Shuxuening Injection on brain tissue of rats with acute cerebral infarction and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2020, 46(03): 557-562.

参考文献

[1] 王硕,何俗非,翟静波,等.丹红注射液药理作用及临床应用研究进展[J].中国中医药信息杂志,2014,21(3):128-131.
[2] 牛国忠.急性脑梗死血管内治疗新进展[C]//浙江省科学技术协会.2015年浙江省神经病学学术年会论文汇编.杭州:浙江省科学技术协会,2015:15-17.
[3] 朱云波,李佳佳,马征,等.阿替普酶对急性脑梗死大鼠血管内皮细胞中Claudin-1和Claudin-5蛋白表达的影响[J].吉林大学学报(医学版),2017,43(6):1137-1141.
[4] LI T H, ZHANG Y Y, TIAN J W, et al. Ginkgo biloba pretreatment attenuates myocardial ischemia-reperfusion injury via mitoBKCa[J]. Am J Chin Med, 2019, 47(5):1057-1073..
[5] 张建平, 石军, 孔延夺. 银杏提取物舒血宁注射液治疗慢性脑供血不足患者的临床疗效[J]. 临床医学研究与实践, 2017,14(30):119-120.
[6] JIE L I, MIN X U. Effect of adjuvant Shuxuening injection therapy on the inflammatory response mediated by TLRs in patients with acute exacerbation of COPD[J]. J Hainan Med Univ, 2017, 23(4):44-47.
[7] AGO T, SADOSHIMA J. GDF15, a cardioprotective TGF-β superfamily protein[J]. Circ Res, 2006, 98(3):294-297.
[8] WOLLERT K C, KEMPF T, PETER T,et al. Prognostic value of growth-differentiation factor-15 in patients with non-ST-elevation acute coronary syndrome[J]. Circulation, 2007, 115(8):962-971.
[9] 刘文,张欢,钱琳.丹红注射液对大鼠脑缺血再灌注后GDF-15表达的影响研究[J].中南药学,2014,12(9):876-879.
[10] LONGA E Z, WEINSTEIN P R, CARLSON S, et al. Reversible middle cerebral artery occlusion without craniectomy in rats[J]. Stroke, 1989,20(1):84-91.
[11] 姜丹, 李彬, 马军, 等. 舒血宁注射液对脑缺血再灌注大鼠缺血半暗带水通道蛋白1和水通道蛋白9表达的影响[J]. 中风与神经疾病杂志, 2010, 27(11):1018-1021.
[12] 贾真, 李晓艳. 舒血宁注射液对大鼠心肌缺血再灌注损伤的保护作用研究[J]. 国医论坛, 2010, 25(2):41-42.
[13] 中华医学会神经病学分会脑血管病学组急性缺血性脑卒中诊治指南撰写组.中国急性缺血性脑卒中诊治指南2010[J].中国临床医生,2011,39(3):67-73.
[14] 任超,刘小芳,解丰帆.急性脑梗死治疗进展[J].中华医学实践杂志, 2008, 7(8):680-682.
[15] 廖金明, 余梦黎, 秦莎莎, 等. 羟基红花黄色素A、芍药苷单用及联合对脑缺血再灌注损伤大鼠脑组织NF-κB及炎症因子的影响[J]. 中药药理与临床, 2018, 34(3):51-55.
[16] 唐永鑫, 李慧琴. 白果内酯对脑缺血损伤保护作用的研究进展[J]. 药物评价研究, 2018, 41(7):1203-1209.
[17] HUA J, YIN N, YANG B, et al. Ginkgolide B and bilobalide ameliorate neural cell apoptosis in α-synuclein aggregates[J]. Biomed Pharmacother, 2017, 96:792-797.
[18] 沈亚雯, 赵燕燕, 王文安. 长春西汀与舒血宁对脑缺血性疾病的血流动力学影响[J]. 实用临床医药杂志, 2014, 18(23):8-10.
[19] 代岩, 金博, 许学宗. 舒血宁注射液联合孟鲁司特钠对AECOPD患者肺功能及血清4-HNE、bFGF、VCAM-1、TGF-β1水平变化的影响[J]. 陕西中医, 2018, 39(8):1037-1039.
[20] 田国忠, 魏罡, 李梅秀, 等.葛瑞林对心肌梗死致心力衰竭大鼠保护作用及生长分化因子-15的影响[J].解剖学报,2013,44(2):274-278.
[21] 唐淑俊, 梅凤君. 炎性细胞因子和C-反应蛋白在急性脑梗死中的变化及相关性研究[J]. 河北医科大学学报, 2010, 31(11):1347-1350.
[22] DONG X, LIU W J, YANG P K. Joint detection of serum Cys-C, IL-6 and VEGF levels in patients with hypertensive acute cerebral infarction[J]. Int J Clin Exp Med, 2007, 17(9):600-607.
[23] LORENZO P. Involvement of IL-1beta in acute stress-induced worsening of cerebral ischaemia in rats[J]. Eur Neuropsychopharmacol, 2007, 17(9):600-607.
[24] 滕海英, 曹丽霞, 赵丽娟. TNF-α hs-CRP LDL-c与老年急性脑梗死的相关性研究[J]. 中国实用神经疾病杂志, 2016(24):61-63.
[25] 姚杰鹏,刘昕,李志安,等.同型半胱氨酸对急性高血压性脑梗死颈动脉斑块稳定性及D-二聚体影响临床研究[J].中国实用内科杂志,2019,39(1):93-94.