摘要
目的探讨褪黑素对脑小血管病模型大鼠社交行为和脑白质损害的改善作用及其药理学机制。方法采用双侧颈总动脉结扎法制备慢性脑低灌注模型,44只SD大鼠随机分为假手术组、模型组、褪黑素5 mg/(kg·d)组和10 mg/(kg·d)组(每组各11只),分别以嗅探实验(嗅探时间和穿越次数)评价模型大鼠社交行为、倒置相差荧光显微镜观察胼胝体髓鞘碱性蛋白(MBP)表达水平、透射电子显微镜观察胼胝体白质髓鞘完整性(即G-ratio)、Western blotting法测定磷酸化哺乳动物雷帕霉素靶蛋白(pmTOR)/哺乳动物雷帕霉素靶蛋白(mTOR)比值。结果与假手术组相比,模型组大鼠嗅探时间(t=58.000,P=0.000)和穿越次数(t=20.000,P=0.000)减少,胼胝体MBP阳性细胞数目减少(t=20.400,P=0.000)、G-ratio表达水平升高(t=-9.800,P=0.025)、pmTOR/mTOR比值降低(t=20.336,P=0.000);予褪黑素[5或10 mg/(kg·d)]治疗后,大鼠嗅探时间(t=-12.600,P=0.001;t=-26.000,P=0.000)和穿越次数(t=-8.400,P=0.000;t=-10.200,P=0.000)、胼胝体pmTOR/mTOR比值(t=-6.022,P=0.014;t=-8.800,P=0.001)高于模型组但仍低于假手术组(t=45.000,P=0.000;t=32.000,P=0.000;t=11.600,P=0.000;t=9.800,P=0.000;t=14.314,P=0.000;t=11.536,P=0.000),同时胼胝体MBP阳性细胞数目亦增加(t=-16.800,P=0.001;t=-20.600,P=0.000)、G-ratio表达水平降低(t=8.600,P=0.041;t=9.200,P=0.030)但接近假手术组水平(均P> 0.05);褪黑素10 mg/(kg·d)组大鼠嗅探时间延长并且高于5 mg/(kg·d)组(t=-13.400,P=0.000)。结论褪黑素可以有效改善脑小血管病模型大鼠社交行为、减轻脑白质损害,其机制可能与提高mTOR磷酸化水平有关。
Objective To observe the protective function and mechanism of melatonin(MT)against white matter lesions in corpus callosum induced by chronic cerebral hypoperfusion(CCH)in cerebral small vessel disease(cSVD)rats.Methods Forty-four adult male Sprague-Dawley rats were divided into sham group,bilateral common carotid artery occlusion(BCCAO)group,MT1 group [BCCAO+melatonin5 mg/(kg·d)]and MT2 group[BCCAO+melatonin 10 mg/(kg·d)].Four weeks after operation,social activity(sniffing time and crossing number)was assessed to reflect apathetic behavior.Myelin basic protein(MBP)in corpus callosum was detected by immunofluorescence.Transmission electron microscope was used to compare the G-ratio.Western blotting test was used to detect phosphorylation level of mammalian target of rapamycin(mTOR).Results BCCAO group rats showed fewer sniffing time(t = 58.000,P = 0.000)and crossing number(t=20.000,P=0.000),lower level of MBP expression(t=20.400,P=0.000),higher level of G-ratio(t=-9.800,P=0.000),and lower pmTOR/mTOR ratio(t =20.336,P=0.000)than those of sham group; after treatment of melatonin[5 or 10 mg/(kg·d)],MT 1 group and MT2 group showed more sniffing time(t=-12.600,P=0.001; t=-26.000,P=0.000)and crossing number(t=-8.400,P=0.000; t=-10.200,P=0.000),higher pmTOR/mTOR ratio(t=-6.022,P=0.014; t=-8.800,P=0.001)than those of BCCAO group but still lower than those of sham group(t=45.400,P=0.000; t=32.000,P=0.000; t= 11.600,P=0.000; t=9.800,P=0.000; t=14.314,P=0.000; t= 11.536,P=0.000); while showed higher level of MBP expression(t=-16.800,P=0.001; t=-20.600,P=0.000),and lower level of G-ratio(t=8.600,P=0.041; t=9.200,P=0.030)than those of BCCAO group but similar with sham group(P>0.05,for all); MT2 group showed longer sniffing time than MT1 group(t =-13.400,P = 0.000).Conclusions Melatonin could ameliorate apathetic behavioral and white matter lesions in corpu callosums in rats with cSVD,which might be related to the up-regulate of phosphorylation level of mTOR.
引文
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