失荅剌知丸對(duì)大鼠局灶性腦缺血再灌注模型海馬區(qū)Notch1受體表達(dá)的影響
目的通過(guò)失荅剌知丸對(duì)大鼠局灶性腦缺血再灌注模型海馬區(qū)Notch1受體表達(dá)的影響,探討其修復(fù)和保護(hù)缺血性病灶及神經(jīng)元的機(jī)制。方法將102只SD大鼠隨機(jī)分為17組,每組6只,分別為空白組、假手術(shù)組,以及模型組、尼莫地平組、失荅剌知丸低劑量組、失荅剌知丸中劑量組和失荅剌知丸高劑量組的1d、3d、7d組。除空白組、假手術(shù)組外,各組均采用線栓法制備成局灶性腦缺血再灌注模型。其中空白組、假手術(shù)組、模型組用生理鹽水灌胃;尼莫地平組給予濃度為1.08g/L灌胃;失荅剌知丸高、中、低劑量組分別用濃度為4.5g/ml、3.0g/ml和1.5g/ml灌胃;以上各組均同時(shí)間灌胃,1d 2次。每組在缺血再灌注1d、3d、7d節(jié)點(diǎn)取材。用Garcia JH評(píng)分法對(duì)大鼠神經(jīng)功能缺損恢復(fù)程度進(jìn)行觀察;免疫熒光、Western-blot、RT-PCR檢測(cè)缺血側(cè)腦組織Notch1的表達(dá)水平。結(jié)果1.大鼠神經(jīng)功能評(píng)分的結(jié)果(1)空白組與假手術(shù)組對(duì)比各時(shí)間點(diǎn)無(wú)統(tǒng)計(jì)學(xué)意義(P>0.05);(2)模型組和各藥物組各時(shí)間點(diǎn)相較于空白組和假手術(shù)組均有統(tǒng)計(jì)學(xué)意義(P<0.05);(3)各藥物組各時(shí)間點(diǎn)相較于模型組均有統(tǒng)計(jì)學(xué)意義(P<0.05);(4)尼莫地平組與失荅剌知丸低劑量組對(duì)比各時(shí)間點(diǎn)有統(tǒng)計(jì)學(xué)意義(P<0.05),與失荅剌知丸中劑量組對(duì)比各時(shí)間點(diǎn)沒(méi)有統(tǒng)計(jì)學(xué)意義(p>0.05),與失荅剌知丸高劑量組對(duì)比各時(shí)間點(diǎn)差異性顯著(P<0.01);(5)失荅剌知丸低劑量組與失荅剌知丸中、高劑量組對(duì)比各時(shí)間點(diǎn)有統(tǒng)計(jì)學(xué)意義(P<0.05或P<0.01)。2.notch1免疫熒光的結(jié)果(1)空白組與假手術(shù)組對(duì)比各時(shí)間點(diǎn)無(wú)統(tǒng)計(jì)學(xué)意義(p>0.05);(2)模型組和各藥物組各時(shí)間點(diǎn)相較于空白組和假手術(shù)組均有統(tǒng)計(jì)學(xué)意義(p<0.05);(3)各藥物組各時(shí)間點(diǎn)相較于模型組均有統(tǒng)計(jì)學(xué)意義(p<0.05);(4)尼莫地平組與失荅剌知丸低劑量組對(duì)比各時(shí)間點(diǎn)有統(tǒng)計(jì)學(xué)意義(p<0.05),與失荅剌知丸中劑量組對(duì)比各時(shí)間點(diǎn)沒(méi)有統(tǒng)計(jì)學(xué)意義(p>0.05),與失荅剌知丸高劑量組對(duì)比各時(shí)間點(diǎn)差異性顯著(p<0.01);(5)失荅剌知丸低劑量組與失荅剌知丸中、高劑量組對(duì)比各時(shí)間點(diǎn)有統(tǒng)計(jì)學(xué)意義(p<0.05或p<0.01)。3.notch1蛋白的western-blot結(jié)果(1)空白組與假手術(shù)組對(duì)比各時(shí)間點(diǎn)無(wú)統(tǒng)計(jì)學(xué)意義(p>0.05);(2)模型組和各藥物組各時(shí)間點(diǎn)相較于空白組和假手術(shù)組均有統(tǒng)計(jì)學(xué)意義(p<0.05);(3)各藥物組各時(shí)間點(diǎn)相較于模型組均有統(tǒng)計(jì)學(xué)意義(p<0.05);(4)尼莫地平組與失荅剌知丸低劑量組對(duì)比各時(shí)間點(diǎn)有統(tǒng)計(jì)學(xué)意義(p<0.05),與失荅剌知丸中劑量組對(duì)比各時(shí)間點(diǎn)沒(méi)有統(tǒng)計(jì)學(xué)意義(p>0.05),與失荅剌知丸高劑量組對(duì)比各時(shí)間點(diǎn)差異性顯著(p<0.01);(5)失荅剌知丸低劑量組與失荅剌知丸中、高劑量組對(duì)比各時(shí)間點(diǎn)有統(tǒng)計(jì)學(xué)意義(p<0.05或p<0.01)。4.notch1的rt-pcr檢測(cè)結(jié)果(1)空白組與假手術(shù)組對(duì)比各時(shí)間點(diǎn)無(wú)統(tǒng)計(jì)學(xué)意義(p>0.05);(2)模型組和各藥物組各時(shí)間點(diǎn)相較于空白組和假手術(shù)組均有統(tǒng)計(jì)學(xué)意義(p<0.05);(3)各藥物組各時(shí)間點(diǎn)相較于模型組均有統(tǒng)計(jì)學(xué)意義(p<0.05);(4)尼莫地平組與失荅剌知丸低劑量組對(duì)比各時(shí)間點(diǎn)有統(tǒng)計(jì)學(xué)意義(p<0.05),與失荅剌知丸中劑量組對(duì)比各時(shí)間點(diǎn)沒(méi)有統(tǒng)計(jì)學(xué)意義(p>0.05),與失荅剌知丸高劑量組對(duì)比各時(shí)間點(diǎn)差異性顯著(p<0.01);(5)失荅剌知丸低劑量組與失荅剌知丸中、高劑量組對(duì)比各時(shí)間點(diǎn)有統(tǒng)計(jì)學(xué)意義(p<0.05或p<0.01)。結(jié)論1.失荅剌知丸對(duì)大鼠腦缺血再灌注損傷的神經(jīng)功能缺損具有明顯改善作用。2.失荅剌知丸有效干預(yù)腦缺血再灌注損傷的機(jī)制,可能與其激活notch信號(hào)轉(zhuǎn)導(dǎo)通路notch1受體發(fā)揮促進(jìn)或修復(fù)缺血區(qū)神經(jīng)元,進(jìn)而改善受損的腦神經(jīng)功能有關(guān)。
賈孟輝;
R29
免疫熒光法檢測(cè)各組大鼠腦組織Notch1表達(dá)（10×40）
各組腦組織Notch1蛋白含量表達(dá)β-actin
