目的 探讨疏血通注射液对缺血性脑卒中大鼠神经功能恢复的影响，并探索其作用机制。方法 采用大脑中动脉闭塞（MCAO）法建立大鼠缺血性脑卒中模型，将40只SD大鼠随机分为假手术组、模型组及疏血通注射液低、高剂量（1、2 mL/kg）组，每组10只。在缺血性脑卒中后3 h及3、7、14 d评估神经功能评分，并测量脑梗死体积；利用苏木素-伊红（HE）染色观察海马缺血区域病理形态学变化；采用免疫荧光染色检测BrdU⁺/Nestin⁺和BrdU⁺/DCX⁺阳性细胞数量，Western blotting分析突触后致密区95（PSD-95）和突触小泡蛋白（SYP）蛋白相对表达水平。构建体外氧糖剥夺/复氧（OGD/R）细胞模型，采用细胞计数试剂盒-8（CCK-8）法检测0～80 μmol/mL疏血通注射液对PC12细胞的增殖情况，并采用Western blotting疏血通注射液对磷脂酰肌醇3激酶（PI3K）-蛋白激酶B（Akt）通路的影响。结果 与模型组相比，在第3、7、14天，疏血通注射液低、高剂量组神经功能缺损评分降低（P＜0.01）。缺血后14 d，疏血通注射液低、高剂量组脑梗死体积降低，BrdU⁺/Nestin⁺和BrdU⁺/DCX⁺阳性细胞数量增加，SYP和PSD-95蛋白表达水平显著升高（P＜0.01）。与模型组相比，10 μmol/mL疏血通注射液组p-PI3K/PI3K和p-Akt/Akt蛋白水平显著升高（P＜0.01），而这些作用可被Akt抑制剂MK-2206和PI3K抑制剂BKM120减弱。结论 疏血通注射液可通过激活PI3K/Akt通路改善缺血性脑卒中大鼠的神经功能。

Objective To explore the effect of Shuxuetong Injection on the recovery of neurological function in rats with ischemic stroke, and to explore its mechanism. Methods The rat model of ischemic stroke was established by middle cerebral artery occlusion (MCAO). Forty SD rats were randomly divided into sham operation group, model group, Shuxuetong Injection low- and high-dose (1, 2 mL/kg) groups, with 10 rats in each group. The neurological function score was evaluated at 3 h, 3, 7 and 14 d after ischemic stroke, and the volume of cerebral infarction was measured. Hematoxylin-eosin (HE) staining was used to observe the pathomorphological changes of hippocampal ischemic area. The number of BrdU⁺/Nestin⁺ and BrdU⁺/DCX⁺ positive cells was detected by immunofluorescence staining. The relative expression levels of postsynaptic density area 95 (PSD-95) and synaptophysin (SYP) protein were analyzed by Western blotting. The in vitro oxygen-glucose deprivation/reoxygenation (OGD/R) cell model was constructed. The proliferation of PC12 cells treated with 0 — 80 μmol/mL Shuxuetong Injection was detected by cell counting kit-8 (CCK-8) method, and the effect of Shuxuetong Injection on phosphatidylinositol 3 kinase (PI3K)-protein kinase B (Akt) pathway was detected by Western blotting. Results Compared with the model group, the neurological deficit scores of the Shuxuetong Injection low and high dose groups were decreased on the 3rd, 7th and 14th days (P < 0.01). On the 14th day after ischemia, the cerebral infarction volume was decreased, the number of BrdU⁺/Nestin⁺ and BrdU⁺/DCX⁺ positive cells was increased, and the expression levels of SYP and PSD-95 protein were significantly increased in Shuxuetong Injection low-dose and high-dose groups (P < 0.01). Compared with the model group, the levels of p-PI3K/PI3K and p-Akt/Akt proteins in the 10 μmol/mL Shuxuetong Injection group were significantly increased (P < 0.01), and these effects could be attenuated by Akt inhibitor MK-2206 and PI3K inhibitor BKM120. Conclusion Shuxuetong Injection can improve the neurological function of ischemic stroke rats by activating the PI3K/Akt pathway.

R285.5

河南省科技攻关计划项目（242102311261）；河南省高等学校重点科研项目计划（23A360018）