松龄血脉康胶囊对大鼠脑缺血再灌注损伤内质网应激凋亡相关蛋白的影响

Effects of Songling Xuemaikang Capsule on Endoplasmic Reticulum Stress Related Apoptosis Proteins in Rats with Cerebral Ischemia Reperfusion Injury

目的探讨松龄血脉康胶囊对大鼠脑缺血再灌注损伤的保护作用及其对内质网应激细胞凋亡的影响。方法从50只雄性SD大鼠中随机选取11只为假手术组,其余大鼠采用大脑中动脉栓塞法制备脑缺血再灌注损伤模型,造模成功的33只大鼠随机分为模型组、松龄血脉康组及依达拉奉组,每组11只,分别予蒸馏水[10 mL/(kg·d)]、松龄血脉康胶囊溶液[10 mL/(kg·d)]及依达拉奉注射液[3.15 mL/(kg·d)]干预5 d。神经功能评分检测造模后24 h及给药5 d后的神经功能;错步实验检测错步次数;HE染色观察大鼠大脑缺血区形态结构;TUNEL染色法观察细胞凋亡;Western blot法检测Cleaved Caspase-3及内质网应激促凋亡蛋白CHOP、Caspase-12的表达。结果神经功能评分显示,造模24 h后,与假手术组相比,造模组评分显著降低(P<0.01),表示造模成功。造模5 d后,模型组神经功能评分仍低于假手术组(P<0.01);松龄血脉康组与依达拉奉组神经功能评分显著高于模型组(P<0.05),且分别显著高于本组造模后24 h评分(P<0.01或P<0.05)。错步实验检测发现,模型组的错步次数显著高于假手术组(P<0.01);松龄血脉康组及依达拉奉组错步次数显著低于模型组(P<0.01)。HE染色结果发现,与模型组相比,松龄血脉康组及依达拉奉组缺血区脑组织形态结构显著改善。TUNEL染色结果显示,模型组阳性细胞百分比较假手术组显著增加(P<0.01);松龄血脉康组及依达拉奉组阳性细胞百分比较模型组显著减少(P<0.01)。Western blot检测结果显示,模型组Cleaved Caspase-3与CHOP、Caspase-12相对蛋白表达量较假手术组显著增高(P<0.01);松龄血脉康组及依达拉奉组Cleaved Caspase-3及CHOP、Caspase-12相对蛋白表达量较模型组明显减少(P<0.05或P<0.05)。结论松龄血脉康胶囊可能是通过降低内质网应激促凋亡蛋白CHOP、Caspase-12的表达减少细胞凋亡,减轻脑缺血再灌注损伤,发挥神经保护作用。

Objective To investigate the protective effect of Songling Xuemaikang Capsule on cerebral ischemia reperfusion injury in rats and its effect on the apoptosis of endoplasmic reticulum stress cells.Methods A total of 11 male SD rats were randomly selected from 50 male SD rats as the sham group.The other rats were established by right middle cerebral artery embolization to prepare cerebral ischemia reperfusion injury model.33 rats were successfully modeled,and were randomly divided into model group,Songling Xuemaikang group and edaravone group,with 11 rats in each group.And they were respectively given distilled water[10 mL/(kg·d)],Songling Xuemaikang Capsules solution[10 mL/(kg·d)]and edaravone injection[3.15 mL/(kg·d)].They were all treated for 5 days.Neurological function score was used to detect the neurological function 24 hours after modeling and 5 days after administration.Stagger-step experiment was used to detect the number of stagger-step.HE staining was used to observe the morphology and structure of cerebral ischemia in rats.Cell apoptosis was observed by TUNEL staining.The expression of Cleaved Caspase-3 and endoplasmic reticulum stress pro-apoptotic proteins CHOP and Caspase-12 were detected by Western blot.Results Neurological function score showed that,24 hours after modeling,the score of the modeling group was significantly lower than that of the sham group(P<0.01),indicating that the modeling was successful.After 5 days of modeling,the neurological function score of model group was still lower than that of sham group(P<0.01).The neurological function score in Songling Xuemaikang group and edaravone group was significantly higher than that in model group(P<0.05),and was significantly higher than that in self group 24 hours after modeling(P<0.05,P<0.01)respectively.Stagger-step experiment showed the number of wrong steps in the model group was significantly higher than that in the sham group(P<0.01).The number of wrong steps in Songling Xuemaikang group and edaravone group was significantly lower than that in model group(P<0.01).The results of HE staining showed that,compared with the model group,the morphological structure of ischemic brain tissue in Songling Xuemaikang group and edaravone group was significantly improved.TUNEL staining showed that the percentage of positive cells in the model group was significantly higher than that in the sham group(P<0.01).The percentage of positive cells in Songling Xuemaikang group and edaravone group was significantly lower than that in model group(P<0.01).Western blot results showed that the relative protein expression levels of Cleaved Caspase-3,CHOP and Caspase-12 in model group were significantly higher than those in sham group(P<0.01).The relative protein expression levels of Cleaved Caspase-3,CHOP and Caspase-12 in Songling Xuemaikang group and edaravone group were significantly decreased compared with model group(P<0.05 or P<0.01).Conclusion Songling Xuemaikang Capsule may reduce cell apoptosis by reducing the expression of endoplasmic reticulum stress pro-apoptotic proteins CHOP and Caspase-12,alleviate cerebral ischemia reperfusion injury and play a neuroprotective role.