摘要
目的探讨黄芪甲苷Ⅳ(astragalosideⅣ,AS-Ⅳ)和丹参酮ⅡA(tanshinoneⅡA,Ta-ⅡA)联合应用对缺血性脑卒中大鼠神经保护作用及机制。方法建立大脑中动脉闭塞模型(线栓法),氧糖剥夺(oxyglycan deprivation,OGD)模拟体外缺氧缺糖情况。SD大鼠分为假手术组(Sham)、模型组(MCAO)和AS-Ⅳ+Ta-ⅡA(给药)组,每组20只。按照剂量给AS-Ⅳ和Ta-ⅡA后,评定3组大鼠神经功能、握杠时间、脑组织梗死及水肿情况;CCK-8实验筛选出最优联合给药浓度;酶联免疫吸附法检测炎性因子如肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-10(interleukin-10,IL-10)和白细胞介素-1β(interleukin-1β,IL-1β)水平;实时定量PCR(quantitative real time PCR,qRT-PCR)检测大鼠脑组织核因子相关因子2(Nrf2)和血红素氧合酶1(heme oxygenase enzyme 1,HO-1)mRNA水平;免疫荧光法检测脑组织神经元(cleaved-caspase 3,CC3)表达。结果模型组神经功能评分、脑梗死体积、脑水肿、丙二醛(malondialdehyde,MDA)和NO、TNF-α以及白细胞介素1β(IL-1β)、HO-1 mRNA、Nrf2 mRNA、CC3表达水平较假手术组明显增加(P<0.05);握力、IL-10、GSH和SOD水平明显降低(P<0.05)。与模型组相比,给药组神经功能评分、脑梗死体积、脑水肿、MDA和NO、TNF-α以及IL-1β、CC3表达水平降低(P<0.05);握力、IL-10、GSH及SOD、Nrf2 mRNA、HO-1mRNA水平明显增加(P<0.05)。CCK-8(cell counting kit-8,CCK-8)试剂盒结果显示最佳浓度为AS-Ⅳ15μg/mL、Ta-ⅡA 25μg/mL。体外实验中,与对照组相比,OGD组CC3表达增加;与OGD组相比,给药组降低。结论AS-Ⅳ联合Ta-ⅡA用药不仅能减轻缺血性脑卒中大鼠氧化应激和炎症反应,也能通过激活Nrf2/HO-1通路抑制神经元凋亡。
Objective To investigate the neuroprotective effect and mechanism of astragaloside IV(AS-Ⅳ)combined with tanshinone ⅡA(Ta-ⅡA)on ischemic stroke rats.Methods A model of middle cerebral artery occlusion(line plug method)was established,and oxygen glucose deprivation(OGD)method was used to simulate the hypoxia and glucose deficiency environment in vitro.SD rats were divided into three groups,sham group,model group,and AS-Ⅳ+Ta-ⅡA(20 in each group).After administration,neurological deficits were assessed by Zea-Longa score.The grip strength test was used to evaluate the grip time of rats.Infarct volume and brain tissue water content detection method were used to observe brain tissue infarction and edema;the Cell Counting Kit-8(CCK-8)experiment was used to screen out the optimal combined administration concentration;enzyme-linked immunosorbent assay was used to detect the levels of tumor necrosis factor-α(TNF-α),interleukin-10(IL-10)and interleukin(IL-1β);real-time quantitative PCR(qRT-PCR)was used to detect the changes of Nuclear factor erythroid 2-related factor 2(Nrf2)and Heme oxygenase 1(HO-1)in rat brain tissue;immunofluorescence was used to detect the expression level of Cleaved-Caspase 3(CC3)in brain tissue and neuronal cells.Results The neurological function score,cerebral infarction volume,cerebral edema,malondialdehyde(malondialdehyde,MDA),NO,TNF-α,and interleukin 1β(IL-1β),HO-1 mRNA,Nrf2 mRNA,and CC3 were significantly increased compared with the sham group(P<0.05);grip strength,IL-10,GSH,and SOD were significantly decreased(P<0.05).Compared with the model group,neurological function score,cerebral infarction volume,cerebral edema,MDA,and NO,TNF-αand IL-1β,CC3 expression levels decreased(P<0.05);grip strength,IL-10,GSH,SOD,and HO-1 mRNA levels increased significantly(P<0.05).The results of the CCK-8(cell counting kit-8,CCK-8)kit showed the optimal concentration of AS-Ⅳ 15μg/mL and Ta-ⅡA 25μg/mL.In vitro,CC3 expression was increased in the OGD group compared to the control group and decreased in the treated group compared to the OGD group.Conclusion AS-Ⅳ combined with Ta-ⅡA can not only reduce oxidative stress and inflammation in ischemic stroke rats,but also inhibit neuronal apoptosis by activating Nrf2/HO-1 pathway.
作者
石圆圆
傅德望
SHI Yuanyuan;FU Dewang(The First Affiliated Hospital of Jinzhou Medical University,Jinzhou 121000 China)
出处
《锦州医科大学学报》
2025年第1期39-44,共6页
Journal of Jinzhou Medical University
