黄角颗粒对脑缺血再灌注损伤大鼠炎性因子及凋亡相关蛋白的影响

Effect of Huangjiao Granule on Inflammatory Factors and Apoptosis-related Proteins in Rats with Cerebral Ischemia-reperfusion Injury

目的探讨黄角颗粒对脑缺血再灌注损伤大鼠炎性因子及凋亡相关蛋白的影响。方法 SPF级Sprague-Dawley健康雄性大鼠40只随机分为假手术组、模型组和黄角颗粒组,每组10只,余10只备用。采用线栓法构建大鼠脑缺血2 h再灌注模型,其中假手术组和模型组在假手术前或缺血再灌注前30 min予生理盐水10 ml/kg灌胃,黄角颗粒组在缺血再灌注前30 min予黄角颗粒溶液10 ml/kg(生药含量1 g/ml)灌胃。再灌注24 h后,Longa评分法对各组大鼠进行神经功能评分,TTC染色检测各组脑梗死体积百分比,HE染色观察各组脑组织病理形态,TUNEL检测各组脑细胞凋亡,ELISA检测各组血清中白细胞介素(IL)-1β、IL-6和肿瘤坏死因子α(TNF-α)的含量,Western blotting检测各组大鼠脑组织中cleaved-caspase 3、cleaved-caspase 9、Bax和Bcl-2蛋白的表达。结果与模型组相比,黄角颗粒组神经功能评分显著降低(P<0.001),脑梗死体积百分比和脑细胞凋亡率降低(P<0.05);血清中IL-1β、IL-6和TNF-α的表达水平降低(P<0.05);脑组织中cleaved-caspase 3、cleaved-caspase 9和Bax的表达均降低(P<0.05),Bcl-2的表达升高(P<0.05)。结论黄角颗粒对脑缺血再灌注损伤大鼠的保护作用与抑制炎症反应和减少细胞凋亡相关。

Objective To investigate the effect of Huangjiao granule on inflammatory factors and apoptosis-related proteins in rats with cerebral ischemia-reperfusion injury.Methods A total of 40 Sprague-Dawley rats were divided into sham operation group,model group and Huangjiao granule group,with ten rats in each group,and ten rats standby.The cerebral ischemia for two hours and reperfusion model was established by suture method.The sham operation group and the model group were given saline 10 ml/kg intragastrically 30 minutes before operation.The Huangjiao granule group was given Huangjiao granule solution 10 ml/kg(the content of crude drug was 1 g/ml)intragastrically 30 minutes before ischemia-reperfusion.Longa scoring method was used to evaluate the neurological function score 24 hours after reperfusion,while the percentage of cerebral infarction volume was detected by TTC staining,the pathological morphology of brain tissue was observed by HE staining,the cell apoptosis of brain tissue was detected with TUNEL,the levels of interleukin(IL)-1β,IL-6 and tumor necrosis factorα(TNF-α)in serum were detected by ELISA,the expression of cleaved-caspase 3,cleaved-caspase 9,Bax and Bcl-2 proteins of brain tissue was evaluated byWestern blotting.Results Compared with the model group,the neurological score decreased,the percentage of cerebral infarction volume and the apoptosis rate of the brain tissue decreased in Huangjiao granule group(P<0.05).The levels of IL-1β,IL-6 and TNF-αin serum and the expression of cleaved-caspase 3,cleaved-caspase 9 and Bax proteins of brain tissues significantly decreased(P<0.05),and the expression of Bcl-2 protein of brain tissues inreased in Huangjiao granule group(P<0.05).Conclusion The protective effect of Huangjiao granule on rats with cerebral ischemia-reperfusion injury may be related to the inhibition of inflammatory response and the reduction of apoptosis.