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[摘要]
目的 考察苦碟子注射液對(duì)大鼠腦缺血再灌注損傷的保護(hù)作用,并從炎癥角度探討其保護(hù)機(jī)制。方法 改良線栓法建立大鼠大腦中動(dòng)脈栓塞(MCAO)模型,激光多普勒血流儀(LDF)監(jiān)測(cè)大鼠腦血流變化。雄性SD大鼠隨機(jī)分為假手術(shù)、模型組及苦碟子注射液高、低劑量組。缺血2 h再灌注24 h后進(jìn)行神經(jīng)功能缺損評(píng)分;斷頭取鬧,稱腦濕質(zhì)量,計(jì)算腦指數(shù);TTC染色法檢測(cè)腦梗死面積;HE染色觀察腦組織病理形態(tài);并取血、腦組織,Elisa試劑盒檢測(cè)炎癥因子表達(dá),Western Blotting技術(shù)檢測(cè)TLR-4、NF-κB蛋白表達(dá)。結(jié)果 與假手術(shù)組比較,模型組神經(jīng)功能缺損較嚴(yán)重(P<0.01),腦指數(shù)和腦梗死面積也顯著升高;給予苦碟子注射液后能改善神經(jīng)功能缺損癥狀(P<0.01),顯著降低模型組的腦指數(shù)和腦梗死面積,減少神經(jīng)元壞死,同時(shí)苦碟子注射液還可顯著降低腦勻漿和血清TNF-α水平(P<0.05),增加局部腦組織IL-10水平(P<0.05)。Western Blotting結(jié)果顯示,苦碟子注射液可降低TLR-4、NF-κB蛋白的表達(dá)(P<0.05)。結(jié)論 苦碟子注射液對(duì)大鼠腦缺血再灌注損傷有明確的保護(hù)作用,具有降低腦缺血再灌注后TNF-α,并升高IL-10水平的作用,其作用機(jī)制可能與其下調(diào)TLR-4/NF-κB信號(hào)通路有關(guān)。
[Key word]
[Abstract]
Objective To investigate the protective effects of Kudiezi (KDZ) Injection on cerebral ischemia reperfusion injury in rats and to explore its protective mechanism.Methods The rat model of middle cerebral artery occlusion (MCAO) was established by modified suture method, and cerebral blood flow was monitored using laser Doppler flowmetry (LDF). Male SD rats were randomly divided into control group, model group, Kudiezi Injection high and low dose groups. After ischemia-reperfusion for 24 h, the neurological scores were evaluated. After anesthesia, the blood samples and brain tissues were collected, and the expression of inflammatory was detected by Elisa. Western blotting was used to detect the expression of TLR-4 and NF-κB protein.Results Behavioral scores showed that neural function defect was serious in model group compared with control group (P<0.01). In model group, cerebral index and cerebral infarction area were significantly higher than those of the control group; After KDZ intervention, the symptoms of neurological deficit was alleviated (P<0.01), the cerebral index and cerebral infarction area of mode were decreased, and the neuronal necrosis was reduced. Kudiezi Injection could significantly reduce the cerebral homogenate and serum levels of TNF-α (P<0.05) and increase IL-10 level (P<0.05). Western blotting showed that Kudiezi Injection could reduce the expression of TLR-4 and NF-κB protein (P<0.05).Conclusion Kudiezi Injection has protective effect on cerebral ischemia reperfusion rats. After ischemia-reperfusion, Kudiezi Injection could reduce the levels of TNF-α and raise IL-10. Its mechanism may be associated to the down-regulation of TLR-4/NF-κB signaling pathway.
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