破血化瘀、填精补髓方对脑出血模型小鼠脑组织TLR4/NF-κB/NLRP3信号通路的影响  

Effect of Blood-Breaking,Stasis-Resolving,Essence-Filling and Marrow Supplementing Formula on TLR4/NF-κB/NLRP3 Signaling Pathway of Brain Tissue in Cerebral Hemorrhage Model Mice

作　　者：乔颖 袁月 徐三鹏 宋双 李萍[1] QIAO Ying;YUAN Yue;XU Sanpeng;SONG Shuang;LI Ping(Changchun University of Chinese Medicine,Changchun 130117,China)

机构地区：[1]长春中医药大学,吉林长春130117

出　　处：《中医药学报》2023年第11期36-42,共7页Acta Chinese Medicine and Pharmacology

基　　金：国家自然科学基金项目(81603557)。

摘　　要：目的:探讨破血化瘀、填精补髓方治疗脑出血的相关作用机制。方法:野生型小鼠和Nrf2基因敲除小鼠各90只,采用小鼠尾动脉取血向脑内灌注的方法建立脑出血小鼠模型,造模成功后120只小鼠随机分为野生型模型组、野生型方剂组、Nrf2基因敲除模型组、Nrf2基因敲除方剂组,每组30只。另分别取30只野生型小鼠和Nrf2基因敲除小鼠脑内只插入微量注射针数分钟不注血作为野生型假手术组和Nrf2基因敲除假手术组。造模后对野生型方剂组及Nrf2基因敲除方剂组给予破血化瘀、填精补髓方10.41 g·kg^(-1)·d^(-1)灌胃,模型组及假手术组给予等量生理盐水。给药1 d、3 d后检测小鼠的行为学变化及脑含水量变化,3 d后HE染色观察脑组织病理形态学改变,用Western Blot以及免疫组化检测脑组织TLR4、NF-κB、NLRP3蛋白水平。结果:与野生型假手术组相比,野生型模型组小鼠的神经功能评分、脑含水量、HE染色病理细胞数、免疫组化阳性细胞数以及TLR4、NF-κB、NLRP3的蛋白表达水平均升高(P<0.05,P<0.01)。与野生型模型组比较,野生型方剂组小鼠上述各指标均明显改善(P<0.05)。相对于野生型模型组小鼠,Nrf2基因敲除型模型组小鼠上述各指标均有明显改善(P<0.05)。结论:破血化瘀、填精补髓方可以通过抑制TLR4/NF-κB/NLRP3信号通路抑制氧化反应及炎症反应,从而抑制脑出血后的神经细胞功能损伤,减少继发性脑损伤。Objective:To explore the related mechanism in the treatment of cerebral hemorrhage with the formula of breaking blood,resolving stasis,filling essence and supplementing marrow.Methods:90 wild-type mice and 90 Nrf2 gene knockout mice were used to establish mouse models of cerebral hemorrhage by taking blood from the mouse tail artery and injecting the blood into the brain.After successful modeling,120 mice were randomly divided into wild-type model group,wild-type formula group,Nrf2 gene knockout model group and Nrf2 gene knockout formula group,with 30 mice in each group.30 wild-type mice and 30 Nrf2 gene knockout mice were respectively taken as wild-type sham operation group and Nrf2 gene knockout sham operation group,and a microinjection needle was inserted into the brain of each of the mouse for a few minutes without blood injection.After modeling,wild-type formula group and Nrf2 gene knockout formula group were gavaged 10.41 g·kg^(-1)·d^(-1)of breaking blood,resolving stasis,filling essence and supplementing marrow formula,while no such treatment was given to model group and sham operation group.The behavioral changes and brain water quantity of the mice were detected 1 d and 3 d later,and the pathological shifts of brain tissue were surveyed by HE staining after 3 d.The protein levels of TLR4,NF-κB and NLRP3 in brain tissue were detected by Western Blot and immunohistochemistry.Results:Compared with sham operation group,the neural function score,brain water content,the number of HE stained pathological cells,the number of immunohistochemical positive cells,and the protein expression levels of TLR4,NF-κB and NLRP3 in model group were increased(P<0.05).Compared with the model group,the above indexes were significantly improved in the treatment groups of breaking blood,resolving stasis,filling essence,and supplementing marrow(P<0.05).Compared with wild-type mice,the above indexes of Nrf2 knockout mice were increased(P<0.05).Conclusion:Blood-breaking,stasis-resolving,essence-filling and marrow supplementing f

关 键 词：脑出血 破血化瘀、填精补髓方 基因敲除 TLR4/NF-κB/NLRP3信号通路 

分 类 号：R285.5[医药卫生—中药学]