益气活血化浊解毒方对脑缺血再灌注损伤大鼠Bcl-2、Bax和Caspase-3表达的影响  

Effect of Yiqi Huoxue Huazhuo Jiedu Decoction on the Expression of Bcl-2,Bax and Caspase-3 Proteins in Rats with Cerebral Ischemia Reperfusion Injury

作　　者：孙玲玲 谢占维 孙亚萍 石瑞 崔怡娴 田军彪[2] SUN Lingling;XIE Zhanwei;SUN Yaping(Hebei College of Chinese Medicine,Shijiazhuang Hebei 050091,China)

机构地区：[1]河北中医学院研究生学院,河北石家庄050091 [2]河北中医学院第一附属医院,河北石家庄050011

出　　处：《四川中医》2023年第8期70-75,共6页Journal of Sichuan of Traditional Chinese Medicine

基　　金：河北省省级科技计划资助(重点研发计划项目)(编号:20377710D);河北省中医药管理局科研计划重点项目(编号:Z2022008);河北中医学院科研能力提升重点项目(编号:KTZ2019028);河北中医学院科研能力提升一般项目(编号:KTZ2019013);河北中医学院研究生创新资助项目(编号:XCXZZBS2022008)。

摘　　要：目的:探讨益气活血化浊解毒方是否可通过调控Bcl-2、Bax和Caspase-3的表达减轻大鼠脑缺血再灌注损伤。方法:72只SPF级雄性SD大鼠,按随机数字表法分为假手术组(Sham)、模型组(MCAO)、益气活血化浊解毒方低剂量组(TCM-L)、中剂量组(TCM-M)、高剂量组(TCM-H)、尼莫地平组(Nim),每组12只大鼠。建立大鼠大脑中动脉闭塞模型,缺血2h后再灌注。造模24h后,各用药组分别予相应浓度的药物灌胃,其余各组等容积生理盐水灌胃,连续治疗3d。采用改良神经功能缺损评分评估大鼠神经功能损伤情况;盐酸2,3,5-三苯基四氮(TTC)染色观察脑梗死体积;TUNEL染色检测神经细胞凋亡情况;Western blot检测Bcl-2、Bax和Caspase-3蛋白表达的情况;荧光定量PCR法检测Caspase-3mRNA表达的情况。结果:(1)与Sham组比较,MCAO组的大鼠神经功能缺损症状显著加重(P<0.05);脑梗死体积占比增大(P<0.05);缺血区细胞凋亡率增加(P<0.05);Western blot检测显示大鼠脑组织Bax、Caspase-3蛋白表达明显升高,Bcl-2蛋白表达明显下降(P<0.05);PCR检测显示Caspase-3mRNA表达显著升高(P<0.05)。(2)与MCAO组比较,TCM组和Nim组的神经功能缺损评分均明显降低(P<0.05);脑梗死体积百分比减少(P<0.05);缺血区细胞凋亡率降低(P<0.05);Western blot检测显示大鼠脑组织Bax、Caspase-3蛋白表达明显降低,Bcl-2蛋白表达显著升高(P<0.05);PCR检测显示Caspase-3mRNA表达显著降低(P<0.05)。结论:益气活血化浊解毒方可通过上调Bcl-2蛋白表达,下调Bax、Caspase-3蛋白及Caspase-3mRNA表达抑制神经细胞凋亡,减轻脑缺血再灌注损伤,发挥神经保护作用。Objective To investigate whether Yiqi Huoxue Huazhuo Jiedu Decoction can alleviate cerebral ischemia reperfusion injury in rats by regulating the expression of Bcl-2,Bax and Caspase-3.Methods 72 SPF grade male SD rats were randomly divided into sham operation group(Sham),model group(MCAO),low dose group(TCM-L),medium dose group(TCM-M),high dose group(TCM-H),and nimodipine group(Nim),with 12 rats in each group.The middle cerebral artery occlusion model of rats was established and reperfusion was performed after 2 hours of ischemia.After 24 hours of modeling,each drug group was given the corresponding concentration of drug by gavage,and the other groups were given the same volume of normal saline by gavage,and the treatment lasted for 3 days.The nerve function damage of rats was evaluated by the modified nerve function defect score;The volume of cerebral infarction was observed by 2,3,5-triphenyl tetrazolium(TTC)staining;TUNEL staining was used to detect the apoptosis of nerve cells;Western blot was used to detect the expression of Bcl-2,Bax and Caspase-3 proteins;The expression of caspase-3 mRNA was detected by fluorescence quantitative PCR.Results⑴Compared with Sham group,the symptoms of nerve function defect in MCAO group were significantly increased(P<0.05);The proportion of cerebral infarction volume increased(P<0.05);The rate of apoptosis in ischemic area increased(P<0.05);Western blot showed that the expression of Bax and Caspase-3 protein in rat brain tissue increased significantly,while the expression of Bcl-2 protein decreased significantly(P<0.05);PCR detection showed that the expression of caspase-3 mRNA was significantly increased(P<0.05).⑵Compared with MCAO group,the scores of neurological deficits in TCM group and Nim group were significantly lower(P<0.05);The volume percentage of cerebral infarction decreased(P<0.05);The rate of apoptosis in ischemic area decreased(P<0.05);Western blot showed that the expression of Bax and Caspase-3 protein in rat brain tissue was significantly decreased,while t

关 键 词：益气活血化浊解毒方 脑缺血再灌注损伤 细胞凋亡 Bcl-2 BAX Caspase-3 

分 类 号：R285.5[医药卫生—中药学]