血府逐瘀汤下调Caveolin-1抑制自噬改善大鼠脑出血损伤的效应及机制研究  

Study on the Effect and Mechanism of Xuefu Zhuyu Decoction(血府逐瘀汤)Down-regulating Caveolin-1Inhibiting Autophagy to Improve Cerebral Hemorrhage Injury in Rats

作　　者：蔡利 兰蕾 徐胜波 陈湘闻 杨鸿 黄东振 CAI Li;LAN Lei;XU Shengbo;CHEN Xiangwen;YANG Hong;HUANG Dongzhen(Guangxi International Zhuang Medical Hospital,Nanning,53001,China)

机构地区：[1]广西国际壮医医院,广西南宁530001

出　　处：《中医药导报》2023年第2期19-23,42,共6页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：广西壮族自治区中医药管理局自筹经费科研课题(GXZYZ20210042)。

摘　　要：目的:探究血府逐瘀汤下调窖蛋白-1(Caveolin-1)抑制自噬改善大鼠脑出血(ICH)损伤的效应及机制。方法:将60只大鼠按随机数字表法分为空白组、假手术组、模型组、低剂量组、中剂量组、高剂量组,每组10只。采用自体股动脉血注入脑尾状核建立ICH大鼠模型。各给药组大鼠分别灌胃给予相应药物,空白组、模型组、假手术组大鼠灌胃等体积生理盐水,1次/d,连续给药3周。观察各组大鼠状态,并行神经功能缺损评分。采集各组大鼠脑组织,HE染色观察脑组织结构的改变情况,免疫组织化学法检测LC3Ⅱ阳性表达,qPCR法检测Caveolin-1 mRNA相对表达量;Western blotting检测Caveolin-1、p-PI3K/PI3K、p-Akt/Akt、p-mTOR/mTOR、Caspase-3、Bcl-2/Bax、Beclin-1蛋白表达情况。结果:假手术组大鼠上述指标与空白组比较,差异均无统计学意义(P>0.05);与空白组比较,模型组大鼠神经功能缺损评分明显降低(P<0.05),脑组织血肿区域明显,神经元胞浆减少,核固缩,脑组织中Caveolin-1、Caspase-3、Beclin-1、LC3Ⅱ蛋白相对表达量及Caveolin-1 mRNA相对表达量均明显升高(P<0.05),p-PI3K/PI3K、p-Akt/Akt、p-mTOR/mTOR、Bcl-2/Bax均明显降低(P<0.05);与模型组比较,低、中、高剂量组大鼠神经功能缺损评分均明显升高(P<0.05),脑组织血肿区域缩小、坏死神经元数目减少,脑组织中Caveolin-1、Caspase-3、Beclin-1、LC3Ⅱ蛋白相对表达量及Caveolin-1 mRNA相对表达量均明显降低(P<0.05),p-PI3K/PI3K、p-Akt/Akt、p-mTOR/mTOR、Bcl-2/Bax均明显升高(P<0.05);中、高剂量组上述指标均优于低剂量组(P<0.05)。结论:血府逐瘀汤可下调Caveolin-1激活PI3K/Akt/mTOR通路抑制自噬,保护神经元而改善大鼠ICH损伤。Objective:To explore the effect and mechanism of Xuefu Zhuyu Decoction on inhibiting autophagy by down-regulating Caveolin-1 and improving the damage of cerebral hemorrhage(ICH)in rats.Methods:60 SD rats were randomly divided into blank group,sham operation group,model group,Xuefu Zhuyu Decoction low-dose,middle-dose and high-dose group,10 rats in each group.The ICH rat model was established by injecting autologous blood from femoral artery into caudate nucleus.Rats in each treatment group were given corresponding drugs by gavage,and rats in the blank group,model group and sham operation group were given the same volume of normal saline by gavage once a day for 3 weeks.Observe the state of rats in each group and score the neurological deficit.Collect the brain tissue of rats in each group,observe the changes of brain tissue structure by HE staining,detect the positive expression of LC3Ⅱby immunohistochemistry,and detect the relative expression of caveolin-1 mRNA by qPCR.The expression levels of Caveolin-1,p-PI3K/PI3K,p-Akt/Akt,p-mTOR/mTOR,Caspase-3,Bcl-2/Bax,and Beclin-1 were detected by Western blotting.Results:Compared with the blank group,the above indexes of rats in the sham operation group had no significant difference(P>0.05).Compared with the blank group,the neurological deficit score of rats in the model group was significantly decreased(P<0.05),the hematoma area of brain tissue was significantly reduced,the neuron cytoplasm was reduced,the nucleus was pyknotic,the relative expression of Caveolin-1,Caspase-3,Beclin-1,LC3 II protein and Caveolin-1 mRNA in brain tissue were significantly increased(P<0.05),and the relative expression of p-PI3K/PI3K,p-Akt/Akt,p-mTOR/mTOR,Bcl-2/Bax were significantly decreased(P<0.05).Compared with the model group,the neurological deficit score of rats in the low,middle and high dose groups was significantly increased(P<0.05),the hematoma area of brain tissue was reduced,the number of necrotic neurons was reduced,the relative expression of Caveolin-1,Caspase-3,Beclin-1,LC3 I

关 键 词：脑出血 血府逐瘀汤 自噬 窖蛋白-1 大鼠 

分 类 号：R285.5[医药卫生—中药学]