目的 考察黃芪注射液對慢性腦缺血大鼠的影響及體外對缺血神經(jīng)細(xì)胞的保護(hù)作用。方法 采用永久性結(jié)扎雙側(cè)頸總動(dòng)脈建立慢性腦缺血大鼠模型，將造模成功大鼠隨機(jī)分為4組：模型組、假手術(shù)組、黃芪注射液組、尼莫地平陽性藥物組，每組10只，各組于術(shù)后3 d給藥治療，1次/d，連續(xù)60 d。給藥結(jié)束用開場實(shí)驗(yàn)和Morris水迷宮實(shí)驗(yàn)評價(jià)各組大鼠焦慮情緒與學(xué)習(xí)記憶能力，HE染色觀察海馬區(qū)細(xì)胞形態(tài)變化。采用連二亞硫酸鈉法誘導(dǎo)PC12細(xì)胞建立缺糖缺氧/復(fù)糖復(fù)氧（OGD/R）損傷模型，通過MTT法檢測細(xì)胞活力，流式細(xì)胞術(shù)檢測細(xì)胞凋亡率，評價(jià)黃芪注射液對體外缺血神經(jīng)細(xì)胞的影響。結(jié)果 曠場實(shí)驗(yàn)中，與模型組相比，黃芪注射液組大鼠第1～3天的站立次數(shù)顯著減少（P<0.05、0.01），第3天的運(yùn)動(dòng)距離明顯縮短（P<0.05）；水迷宮實(shí)驗(yàn)中，與模型組相比，黃芪注射液組大鼠定位航行實(shí)驗(yàn)第2天開始平均逃避潛伏期明顯縮短（P<0.05、0.01），空間探索實(shí)驗(yàn)的穿越平臺(tái)次數(shù)顯著增多（P<0.01）；HE染色結(jié)果顯示：模型組大鼠海馬較多錐體細(xì)胞固縮深染且細(xì)胞形狀不規(guī)則，黃芪注射液能有效改善海馬區(qū)神經(jīng)元細(xì)胞損傷。與對照組相比，模型組PC12細(xì)胞存活率顯著降低（P<0.01）；與模型組相比，40、200 mg/mL的黃芪注射液均可顯著提高OGD/R PC12細(xì)胞的存活率（P<0.01）；與模型組相比，200 mg/mL黃芪注射液可顯著降低PC12細(xì)胞凋亡率（P<0.01）。結(jié)論 黃芪注射液能在一定程度上緩解慢性腦缺血模型大鼠的認(rèn)知功能障礙，同時(shí)體外促進(jìn)缺血PC12細(xì)胞存活，其作用機(jī)制可能與調(diào)控細(xì)胞凋亡有關(guān)。

Objective To investigate the protective effect of Astragalus Injection on chronic cerebral ischemia rats and ischemic nerve cells in vitro. Methods In vivo experiment:Chronic cerebral ischemia rat model was established by permanent ligation of bilateral common carotid arteries. The rats were randomly divided into 4 groups:model group, sham operation group, Astragalus Injection group, and positive drug group, with 10 rats in each group. Each group was given medicine 3 days after operation, once daily for consecutive 60 days. After administration, the anxiety and learning and memory ability were evaluated by the opening experiment and Morris water maze test, and the morphological changes of cells in the hippocampus were observed by HE staining. In vitro experiment:PC12 cells were induced by sodium disulfite method to establish glucose deficiency hypoxia/glucose reoxygenation (OGD/R) injury model, cell viability was detected by MTT assay, cell apoptosis rate was detected by flow cytometry, and the effect of astragalus injection on ischemic nerve cells in vitro was evaluated. Results In the opening experiment, compared with the model group, the standing times of rats in the astragalus injection group were significantly reduced from day 1 to day 3 (P < 0.05, 0.01), and the movement distance on day 3 was significantly shortened (P < 0.05). In the water maze test, compared with the model group, the average escape latency of rats in Astragalus Injection group was significantly shortened from second day of positioning navigation test (P < 0.05, 0.01), and the number of times of crossing platform in space exploration test was significantly increased (P < 0.01). The results of HE staining showed that there were many pyramidal cells pyknosis and deep staining in the hippocampus of the model group, and the shape of the cells was irregular. Astragalus Injection group could effectively improve the damage of neurons in the hippocampus. Compared with the control group, the survival rate of PC12 cells in model group was statistically significant decreased (P < 0.01). Compared with model group, 40 mg/mL and 200 mg/mL Astragalus Injection could significantly improve the survival rate of PC12 cells (P < 0.01). Compared with model group, 200 mg/mL Astragalus Injection could significantly reduce the apoptosis rate of PC12 cells (P < 0.01). Conclusion Astragalus Injection can alleviate the cognitive impairment of rats with chronic cerebral ischemia to a certain extent, and promote the survival of PC12 cells after ischemia in vitro, and its mechanism may be related to the regulation of cell apoptosis.

R971

四川省科技廳項(xiàng)目（20ZDYF2365）；四川省大學(xué)生創(chuàng)新創(chuàng)業(yè)項(xiàng)目（202013705043）