破血化瘀填精补髓中药汤剂对核因子E 2相关因子2基因敲除脑出血小鼠氧化应激的影响  

Effect of blood stasis removing, essence replenishing decoction on oxidative stress in Nrf2 gene knocked-out intracerebral hemorrhage mice

作　　者：丁子惠 袁月 宋双 李萍[1] Ding Zihui;Yuan Yue;Song Shuang;Li Ping(School of Preclinical Medicine,Changchun University of Traditional Chinese Medicine,Changchun 130117,Jilin Province,China)

机构地区：[1]长春中医药大学基础医学院,130117 [2]江苏省中医药研究院

出　　处：《中华老年心脑血管病杂志》2022年第5期536-540,共5页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

基　　金：国家自然科学基金(81603557);2021年度长春中医药大学研究生“橘井杯”学术科研创新及创业项目(YK202101)。

摘　　要：目的 观察以破血化瘀、填精补髓法组方的中药汤剂对核因子E 2相关因子2(Nrf2)基因敲除脑出血模型小鼠氧化应激水平的影响。方法 将60只Nrf2基因敲除小鼠(Nrf2;)和60只野生型小鼠随机分为6组:假手术组、Nrf2;假手术组、模型组、Nrf2;模型组、方剂组、Nrf2;方剂组,每组20只。采用基底核区注射自体血方法制作小鼠脑出血模型,评估小鼠神经功能状态,小鼠麻醉清醒后开始给药干预,检测脑出血小鼠脑组织病理表达,应用Western blot法和免疫组织化学法检测超氧化物歧化酶(SOD)2和过氧化物还原酶3(PRDX3)的表达,应用试剂盒检测SOD、谷胱甘肽过氧化物酶(GPx)、过氧化氢酶(CAT)活力及丙二醛、一氧化氮(NO)的含量,探讨破血化瘀、填精补髓中药汤剂对其的影响。结果 模型组神经细胞坏死、数目减少、排列紊乱,脑组织疏松、水肿明显。Nrf2;模型组大量红细胞浸润,脑组织结构破坏,神经细胞坏死严重。方剂组和Nrf2;方剂组少量红细胞浸润,间质水肿较轻。治疗3 d后与模型组和Nrf2;模型组比较,方剂组和Nrf2;方剂组的Longa评分显著降低[(0.75±0.55)分、(1.25±0.64)分vs(1.70±0.66)分、(1.85±0.49)分]。脑出血后3 d,与假手术组和Nrf2;假手术组比较,模型组和Nrf2;模型组的SOD、GPx、CAT活力显著降低,MDA和NO的含量均显著升高(P<0.05);与Nrf2;方剂组比较,方剂组的SOD[(89.41±6.59)U/mg vs(74.73±4.03)U/mg]、GPx[(17.33±1.20)U/mg vs(14.44±0.96)U/mg]、SOD2(0.499±0.045 vs 0.443±0.057)、PRDX3(0.345±0.021 vs 0.297±0.013)显著增高,MDA[(1.83±0.14)nmol/mg vs(2.28±0.19)nmol/mg]显著降低,差异有统计学意义(P<0.05)。结论 破血化瘀、填精补髓中药汤剂可通过调节Nrf2减轻脑出血后氧化应激水平,从而保护脑出血后继发损伤的脑组织。Objective To study the effect of blood stasis removing, essence replenishing decoction on oxidative stress in Nrf2 gene knocked-out mice.Methods Sixty Nrf2 gene knocked-out mice and 60 wild-type mice were randomly divided into sham operation group, Nrf2/sham operation group, model group, Nrf2/model group, formula group and Nrf2/formula group(20 in each group).The cerebral hemorrhge model of mice was established by injecting autologous blood into the basal ganglia.The neurological function of mice was evaluated.The drug intervention was started after awake under anesthesia.The pathological expression in brain tissue of Nrf2 gene knocked-out intracerebral hemorrhage mice was detected.The expressions of superoxide dismutase 2(SOD2) and peroxidase reductase 3(PRDX3) in Nrf2 gene knocked-out intracerebral hemorrhage mice were detected by Western blot and immunohistochemistry respectively.The activities of SOD and glutathione peroxidase(GPx),catalase(CAT) in Nrf2 gene knocked-out intracerebral hemorrhage mice were tested and the serum levels of malondialde-hyde(MDA) and nitric oxide(NO) in Nrf2 gene knocked-out intracerebral hemorrhage mice were measured with agent kits.The effect of blood stasis removing, essence replenishing and marrow reinforcing Chinese traditional medicinal decoction on oxidative stress in Nrf2 gene knocked-out intracerebral hemorrhage mice was analyzed.Results The nerve cells were significantly necrosed, the number of nerve cells was significantly reduced, the arrangement of nerve cells was significantly disturbed, and the brain tissue was significantly loose with edema in model group.A large number of red blood cells was infiltrated, the structure of brain tissue was destroyed and the nerve cells were severely necrosed in Nrf2/model group.A small number of red blood cells was infiltrated and the interstitial edema was milder in formula group and Nrf2/formula group.The Longa score was significantly lower in formula group and Nrf2/formula group than in model group and Nrf2/model group after 3 days

关 键 词：活血祛瘀剂 汤剂 小鼠 基因敲除 脑出血 氧化性应激 超氧化物歧化酶 

分 类 号：R285.5[医药卫生—中药学]