脑泰通组方对局灶性脑缺血再灌注大鼠模型的影响及保护机制

Protective Effect and Mechanism of Naotaitong Formula in the Rat Model of Focal Cerebral Ischemia-reperfusion Injury

目的:观察脑泰通组方对局灶性脑缺血再灌注大鼠模型的影响及保护机制。方法:构建局灶性脑缺血再灌注大鼠模型,成模后随机分为模型对照组、阳性药物组、中药组、联合用药组,另选取健康斯泼累格·多雷(SD)大鼠作为假手术组。观察并评估各组大鼠的美国国立卫生研究院卒中量表(NIHSS)评分、脑组织含水量、脑梗死体积百分比,免疫组织化学法检测脑组织Bcl-2相关X蛋白(Bax)、B细胞淋巴瘤-2(Bcl-2)的平均光密度值,蛋白质免疫印迹法、实时定量PCR(QT-PCR)检测脑组织叉头框转录因子3a(FoxO3a)基因、低氧诱导因子-1α(HIF-1α)、核因子κB(NF-κB)、脑源性神经营养因子(BDNF)蛋白及mRNA的表达水平。结果:与假手术组比较,模型对照组NIHSS评分,脑组织含水量,脑组织缺血体积百分比,脑组织FoxO3a、HIF-1α、NF-κB、BDNF蛋白和mRNA的表达水平,Bax、Bcl-2平均光密度值以及Bax/Bcl-2值均显著升高(均P<0.05);与模型对照组比较,药物干预各组的NIHSS评分、脑组织含水量、脑梗死体积百分比、Bax平均光密度值、Bax/Bcl-2值、脑组织NF-κB蛋白和mRNA的表达水平显著降低(均P<0.05),其中联合用药组的降低程度最大(均P<0.05),Bcl-2平均光密度值,脑组织FoxO3a、HIF-1α、BDNF蛋白和mRNA的表达水平显著升高(均P<0.05),其中联合用药组的升高程度最大(均P<0.05)。结论:脑泰通组方能有效保护局灶性脑缺血再灌注大鼠模型脑组织损伤,改善局部脑缺血,其作用机制可能与脑泰通组方调控FoxO3a/HIF-1α/NF-κB信号通路相关。

Objective:To study the effect and protective mechanism of Naotaitong Formula in the rat model of focal cerebral ischemia-reperfusion injury.Methods:Rats were modeled for focal cerebral ischemia-reperfusion injury and then randomized into model control,positive control,Naotaitong,and drug combination groups.Healthy SD rats were selected as the sham operation group.The National Institute of Health stroke scale(NIHSS)score and the water content and infarct volume percentage in the brain tissue were evaluated and determined.The immunohistochemical method was employed to measure the average optical density values of B-cell lymphoma-2(Bcl-2)and Bcl-2-associated X protein(Bax).Western blotting and RT-PCR were employed to determine the protein and mRNA levels of forkhead box O3a(FoxO3a),hypoxia-inducible factor 1 alpha(HIF-1α),nuclear factor-kappa B(NF-κB),brain-derived neurotrophic factor(BDNF)in the brain tissue.Results:Compared with the sham operation group,the modeling increased the NIHSS score,water content and infarct volume percentage in the brain tissue,the protein and mRNA levels of FoxO3a,HIF-1α,NF-κB,and BDNF in the brain tissue,the average optical density values of Bax and Bcl-2,and the Bax/Bcl-2 ratio(P<0.05).Compared with the model control group,drug interventions reduced the NIHSS score,water content and infarct volume percentage in the brain tissue,the average optical density of Bax,the Bax/Bcl-2 ratio,and the protein and mRNA levels of NF-κB in the brain tissue(P<0.05),and the drug combination group showed the greatest reduction(P<0.05).In addition,the average optical density of Bcl-2,the protein and mRNA levels of FoxO3a,HIF-1α,and BDNF were increased(P<0.05),and the drug combination group had the greatest increase(P<0.05).Conclusion:Naotaitong Formula can protect the brain tissue from focal cerebral ischemia injury in the rat model by regulating the FoxO3a/HIF-1α/NF-κB signaling pathway.