目的 探討疏血通注射液對缺血性腦卒中大鼠神經(jīng)功能恢復的影響，并探索其作用機制。方法 采用大腦中動脈閉塞（MCAO）法建立大鼠缺血性腦卒中模型，將40只SD大鼠隨機分為假手術組、模型組及疏血通注射液低、高劑量（1、2 mL/kg）組，每組10只。在缺血性腦卒中后3 h及3、7、14 d評估神經(jīng)功能評分，并測量腦梗死體積；利用蘇木素-伊紅（HE）染色觀察海馬缺血區(qū)域病理形態(tài)學變化；采用免疫熒光染色檢測BrdU⁺/Nestin⁺和BrdU⁺/DCX⁺陽性細胞數(shù)量，Western blotting分析突觸后致密區(qū)95（PSD-95）和突觸小泡蛋白（SYP）蛋白相對表達水平。構建體外氧糖剝奪/復氧（OGD/R）細胞模型，采用細胞計數(shù)試劑盒-8（CCK-8）法檢測0～80 μmol/mL疏血通注射液對PC12細胞的增殖情況，并采用Western blotting疏血通注射液對磷脂酰肌醇3激酶（PI3K）-蛋白激酶B（Akt）通路的影響。結果 與模型組相比，在第3、7、14天，疏血通注射液低、高劑量組神經(jīng)功能缺損評分降低（P＜0.01）。缺血后14 d，疏血通注射液低、高劑量組腦梗死體積降低，BrdU⁺/Nestin⁺和BrdU⁺/DCX⁺陽性細胞數(shù)量增加，SYP和PSD-95蛋白表達水平顯著升高（P＜0.01）。與模型組相比，10 μmol/mL疏血通注射液組p-PI3K/PI3K和p-Akt/Akt蛋白水平顯著升高（P＜0.01），而這些作用可被Akt抑制劑MK-2206和PI3K抑制劑BKM120減弱。結論 疏血通注射液可通過激活PI3K/Akt通路改善缺血性腦卒中大鼠的神經(jīng)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

河南省科技攻關計劃項目（242102311261）；河南省高等學校重點科研項目計劃（23A360018）