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目的观察并探讨络瘀通胶囊对大鼠脑缺血-再灌注后神经功能的影响及其保护机制。方法采用改良Longa法制备大鼠大脑中动脉阻塞(MCAO)模型,缺血1.5 h后进行再灌注。选取10只大鼠作为假手术组,将40只造模成功的雄性SD大鼠按照随机数字表法分为4组,每组10只:模型组、络瘀通中剂量组(LYTM组)、络瘀通高剂量组(LYTH组)及胞磷胆碱钠组(CS组)。分别于造模后3 d及7 d采用12分评分及前肢踩空实验评价大鼠神经功能,应用蛋白免疫印迹法(Western blot)检测造模3 d后大鼠缺血侧及假手术组同侧脑组织中脑源性神经营养因子(BDNF)、碱性成纤维生长因子(b-FGF)及磷酸化蛋白激酶/蛋白激酶(p-AKT/AKT)的表达,及造模后7 d同侧脑组织中Caspase-12的表达并进行组间比较。结果 (1)神经功能评分:造模后3 d各组12分评分及前肢踩空实验评分差异均无统计学意义(均P>0.05);造模后7 d,LYTM组、LYTH组、CS组较模型组均明显改善(均P<0.05)。(2)Western blot结果显示:1模型组BDNF及b-FGF表达明显少于假手术组(均P<0.05);与模型组比较,LYTM组、LYTH组、CS组均可上调BDNF和b-FGF表达,以LYTH组作用最明显(P<0.01)。2与假手术组比较,模型组p-AKT/AKT表达明显减少(P<0.05);与模型组比较,LYTM组、LYTH组、CS组p-AKT/AKT表达增加,以LYTH组和CS组明显,差异均有统计学意义(均P<0.05)。3在Caspase-12的表达中,模型组裂解后Caspase-12表达较假手术组明显升高,差异有统计学意义(P<0.05)。与模型组比较,LYTM组、LYTH组Caspase-12前体(pro Caspase-12)和裂解后Caspase-12表达有减少趋势,但差异无统计学意义(均P>0.05);CS组Caspase-12前体和裂解后Caspase-12表达水平明显低于模型组(P<0.05)。结论络瘀通胶囊可通过促进神经元存活及再生对大鼠脑缺血-再灌注性损伤起到保护作用,且该保护作用也可能与抑制神经细胞凋亡有关。
Objective To observe and explore the effect of Luo Yu Tong Capsule on neurological function after cerebral ischemia reperfusion in rats and its protective mechanism. Methods Rat model of middle cerebral artery occlusion (MCAO) was established by modified Longa method. After ischemia for 1.5 h, reperfusion was performed. Ten rats were selected as the sham operation group. Forty successful male SD rats were randomly divided into 4 groups (n = 10): model group, LYTM group, Tonglu Tong high dose group (LYTH group) and Citicoline sodium group (CS group). The neurological function of rats was assessed by score of 12 and foregut test at 3 days and 7 days after modeling respectively. Western blotting was used to detect the neuronal function in the ischemic and sham-operated rats after 3 days The expression of brain derived neurotrophic factor (BDNF), basic fibroblast growth factor (b-FGF) and phosphorylated protein kinase / protein kinase (p-AKT / AKT) The expression of Caspase-12 in lateral brain tissues was compared between groups. Results (1) Neurological score: There were no significant differences in scores of 12 points and foregut trawl test scores on the 3rd day after modeling (all P> 0.05). On the 7th day after modeling, LYTM group, LYTH group, CS Group than the model group were significantly improved (all P <0.05). (2) Western blot results showed that the expression of BDNF and b-FGF in model group was significantly less than that in sham operation group (all P <0.05). Compared with model group, LYTM group, LYTH group and CS group could up- FGF expression, the role of LYTH group most obvious (P <0.01). Compared with sham group, the expression of p-AKT / AKT in model group was significantly decreased (P <0.05). Compared with model group, the expression of p-AKT / AKT increased in LYTM group, LYTH group and CS group. Significant differences were statistically significant (P <0.05). The expression of Caspase-12 in model group was significantly higher than that in sham operation group after the cleavage of Caspase-12, the difference was statistically significant (P <0.05) .3. Compared with model group, the expressions of Caspase-12 and Caspase-12 in LYTM group and LYTH group showed a decreasing trend, but the difference was not statistically significant (all P> 0.05); Caspase-12 Caspase-12 expression level in precursor and lysis group was significantly lower than that in model group (P <0.05). Conclusion Luo Yu Tong capsule can protect neurons from cerebral ischemia-reperfusion injury by promoting the survival and regeneration of neurons, and the protective effect may be related to the inhibition of neuronal apoptosis.