摘要
目的:观察槲皮素对双环己酮草酰二腙(cuprizone,CPZ)诱导脱髓鞘模型小鼠髓鞘再生的影响。方法:将50只C57BL/6J小鼠按随机数字表法分为正常组,模型组,槲皮素低、中、高剂量组,每组10只。除正常组外,其余各组小鼠给予0.2%CPZ饲料进行喂养以诱导小鼠脱髓鞘模型。槲皮素低、中、高剂量组灌胃槲皮素溶液25、50、100 mg/kg,正常对照组和模型组灌胃等体积蒸馏水,1次/d。连续灌胃5周,每周记录小鼠体重,并进行转棒实验。5周后取材,采用快蓝染色及透射电镜观察小鼠胼胝体髓鞘病理学改变,并计算G-ratios值;采用免疫荧光染色法测定小鼠胼胝体区髓磷脂碱性蛋白(myelin basic protein,MBP)和少突胶质细胞转录因子(oligodendrocyte transcription factor 2,Olig2)蛋白表达;采用Western blot法测定小鼠脑组织MBP与环核苷酸-3'磷酸水解酶(cyclic nucleotide-3'phosphate hydrolase,CNPase)蛋白表达。结果:第2~5周,与模型组比较,槲皮素中、高剂量组小鼠体重升高(P<0.01),转棒维持时间延长(P<0.01);与模型组比较,槲皮素低、中、高剂量组胼胝体脱髓鞘评分[(2.23±0.25)分、(1.50±0.15)分、(1.14±0.97)分比(2.83±0.18)分]降低(P<0.01)、G-ratio值[(0.75±0.05)、(0.75±0.08)、(0.73±0.08)比(0.87±0.05)]降低(P<0.01);槲皮素中、高剂量组小鼠胼胝体区MBP[(37.40±2.41)、(37.40±1.14)比(24.40±3.65)]、Olig2[(7.40±1.14)、(4.60±1.14)比(2.80±0.84)]阳性表达升高(P<0.05),脑组织MBP蛋白[(1.32±0.12)、(0.80±0.34)比(0.21±0.07)]、CNPase蛋白[(0.72±0.13)、(1.06±0.36)比(0.36±0.21)]表达升高(P<0.05)。结论:50、100 mg/kg剂量的槲皮素可减轻CPZ诱导的脱髓鞘模型小鼠的髓鞘损伤,促进髓鞘再生,有一定的神经保护作用。
Objective To observe the effect of quercetin on myelin regeneration in desyelinating mice induced by cuprizone(CPZ).Methods A total of 50 C57BL/6J mice were randomly divided into normal control group,model group,low,medium and high dose quercetin groups,10 mice in each group.In addition to the normal control group,mice demyelinating model was induced by feeding with 0.2%CPZ rodent feed.Quercetin solution was administered to the low,medium and high dose quercetin groups at 25,50 and 100 mg/kg,and to the normal control group and the model group at 1 time/d.After 5 weeks of continuous gavage,the weight of mice was recorded every week,and the experiment of rotating bar was carried out.After 5 weeks,the changes of the myelin sheath in the corpus callosum of mice were observed by luxol fast blue(LFB)and transmission electron microscope(TEM).Immunofluorescence method was used to determine the protein expressions of myelin basic protein(MBP)and oligodendrocyte transcription 2actor(Olig2)in mouse brain tissue.The expression of MBP and cyclic nucleotide-3'phosphate hydrolase(CNPase)in mouse corpus callosum was determined by Western blot.Results After 2-5 weeks,compared with the model group,the body mass of the medium and high dose quercetin groups significantly increased(P<0.01),and the score of turning rod significantly increased(P<0.01).The LFB observation showed that the demyelination score of corpus callosum in low,medium and high dose quercetin groups(2.23±0.25,1.50±0.15,1.14±0.97 vs.2.83±0.18)significantly decreased(P<0.01).TEM observation showed that the G-ratio value in low,medium and high dose quercetin groups(0.75±0.05,0.75±0.08,0.73±0.08 vs.0.87±0.05)significantly reduced(P<0.01).Immunofluorescence observation showed that the positive expression of MBP(37.40±2.41,37.40±1.14 vs.24.40±3.65)and Olig2(7.40±1.14,4.60±1.14 vs.2.80±0.84)in the medium and high dose quercetin groups significantly increased(P<0.05).WB showed that the expression of MBP protein(1.32±0.12,0.80±0.34 vs.0.21±0.07)and CNPase protein(0.72±0.13,1.06±0.36 vs.0.36±0.21)in the medium and high dose quercetin groups significantly increased(P<0.05).Conclusions Quercetin(50-100 mg/kg)can reduce myelin injury and promote myelin regeneration in CPZ mice,as a neuroprotective effect on CPZ mice.
作者
于佳
孙雅琴
赵晖
李康宁
王蕾
Yu Jia;Sun Yaqin;Zhao Hui;Li Kangning;Wang Lei(School of Traditional Chinese Medicine,Beijing Key Lab of TCM Collateral Disease Theory Research,Capital Medical University,Beijing 100069,China;Department of Traditional Chinese Medicine,Beijing Tiantan Hospital,Capital Medical University,Beijing 100070,China)
出处
《国际中医中药杂志》
2020年第1期39-45,共7页
International Journal of Traditional Chinese Medicine
基金
国家自然科学基金(81873252、81573898)
北京市自然科学基金(7182020、7174298)。
