黄芩甙对慢性脑低灌注大鼠认知功能及其对海马内LC-3 Ⅱ和Beclin-1表达的影响
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摘要
目的观察黄芩甙对慢性脑低灌注大鼠学习记忆功能及海马区自噬相关蛋白LC-3Ⅱ和Beclin-1表达的影响。方法将50只SD大鼠按随机数字表法分为对照组、模型组和黄芩甙低、中、高剂量组,每组各10只。采用永久性双侧颈总动脉结扎建立慢性脑低灌注大鼠模型,黄芩甙组腹腔注射3种不同剂量的黄芩甙(60、120、240 mg·kg~(-1)·d~(-1)),对照组和模型组给予等量0. 9%(质量分数)氯化钠注射液腹腔注射。4周后采用Morris水迷宫法进行行为学检测,Western blotting法检测自噬相关蛋白LC-3Ⅱ和Beclin-1的表达。结果与对照组相比,模型组Morris水迷宫平均逃避潜伏期和游泳距离均明显延长(P <0. 01),空间探索实验中在原平台所在象限游泳时间明显缩短(P <0. 01),海马区自噬相关蛋白LC-3Ⅱ和Beclin-1表达明显增加(P <0. 01)。与模型组比较,黄芩甙干预后,大鼠学习记忆能力显著改善(P <0. 01或P <0. 05),海马内LC-3Ⅱ和Beclin-1表达均显著低于模型组(P <0. 01或P <0. 05),差异有统计学意义。结论黄芩甙能够改善慢性脑低灌注大鼠的学习记忆功能,并抑制自噬相关蛋白LC-3Ⅱ和Beclin-1的表达。
Objective To investigate the effects of baicalin on the cognitive ability and the expression of LC-3Ⅱand Beclin-1 proteins in hippocampus of the rats with chronic cerebral hypoperfusion. Methods A total of 50 rats were randomly divided into five groups: control group,model group,low,medium,and high dose of baicalin groups with 10 rats in each group. The chronic cerebral hypoperfusion rat model was established by permanent bilateral common carotid artery occlusion. Baicalin groups were treated with intraperitoneal injection of baicalin in three different dosages( 60,120,240 mg·kg~(-1)·d~(-1)),while those in the other two groups were treated with the same volume of saline.After four weeks,the spatial learning and memory ability were observed by Morris water maze test,and the expression of LC-3 Ⅱand Beclin-1 in the hippocampus were revealed with Western blotting. Results Both escape latency and swimming distance of the chronic cerebral hypoperfusion rats were significantly prolonged in comparison of those in normal control( P < 0. 01),and swimming time in the platform of previous quadrant was significantly shorter in model group( P < 0. 01). Furthermore,the expression of LC-3 Ⅱ and Beclin-1 protein increased in model group compared with the control group( P < 0. 01). Four weeks of treatment with baicalin remarkably improved the learning and memory ability of chronic cerebral hypoperfusion rats( P < 0. 01 or P < 0. 05),decreased the expression of LC-3 Ⅱand Beclin-1 in hippocampus of the model rats( P < 0. 01 or P < 0. 05). Conclusion Baicalin can improve cognitive ability of the chronic cerebral hypoperfusion rats,and inhibit autophagy by decreasing the expression of LC-3 Ⅱ and Beclin-1 in the hippocampus.
Objective To investigate the effects of baicalin on the cognitive ability and the expression of LC-3Ⅱand Beclin-1 proteins in hippocampus of the rats with chronic cerebral hypoperfusion. Methods A total of 50 rats were randomly divided into five groups: control group,model group,low,medium,and high dose of baicalin groups with 10 rats in each group. The chronic cerebral hypoperfusion rat model was established by permanent bilateral common carotid artery occlusion. Baicalin groups were treated with intraperitoneal injection of baicalin in three different dosages( 60,120,240 mg·kg~(-1)·d~(-1)),while those in the other two groups were treated with the same volume of saline.After four weeks,the spatial learning and memory ability were observed by Morris water maze test,and the expression of LC-3 Ⅱand Beclin-1 in the hippocampus were revealed with Western blotting. Results Both escape latency and swimming distance of the chronic cerebral hypoperfusion rats were significantly prolonged in comparison of those in normal control( P < 0. 01),and swimming time in the platform of previous quadrant was significantly shorter in model group( P < 0. 01). Furthermore,the expression of LC-3 Ⅱ and Beclin-1 protein increased in model group compared with the control group( P < 0. 01). Four weeks of treatment with baicalin remarkably improved the learning and memory ability of chronic cerebral hypoperfusion rats( P < 0. 01 or P < 0. 05),decreased the expression of LC-3 Ⅱand Beclin-1 in hippocampus of the model rats( P < 0. 01 or P < 0. 05). Conclusion Baicalin can improve cognitive ability of the chronic cerebral hypoperfusion rats,and inhibit autophagy by decreasing the expression of LC-3 Ⅱ and Beclin-1 in the hippocampus.
引文
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[20] Chen C,Li X H,Gao P L,et al. Baicalin attenuates Alzheimer-like pathological changes and memory deficits induced by amyloidβ1-42 protein[J]. M etab Brain Dis,2015,30(2):537-544.
[21] Zhao J W,Lu S,Yu H,et al. Baicalin and ginsenoside Rb1 promote the proliferation and differentiation of neural stem cells in Alzheimer's disease model rats[J]. Brain Res,2018,1678:187-194.
[2] Hamano T,Hayashi K,Shirafuji N,et al. The implications of autophagy in alzheimer's disease[J]. Curr Alzheimer Res,2018,15(14):1283-1296.
[3] Liang W,Huang X,Chen W. The effects of baicalin and baicalein on cerebral ischemia:a review[J]. Aging Dis,2017,8(6):850-867.
[4] Chen C,Li X,Gao P,et al. Baicalin attenuates alzheimER-like pathological changes and memory deficits induced by amyloidβ1-42 protein[J]. M etab Brain Dis,2015,30(2):537-544.
[5] Wang Z,Fan J,Wang J,et al,Chronic cerebral hypoperfusion induces long-lasting cognitive deficits accompanied by long-term hippocampal silent synapses increase in rats[J].Behav Brain Res,2016,301:243-252.
[6] Morris R. Developments of a water maze procedure for studying spatial learning in the rat[J]. J Neurosci M ethods,1984,11(1):47-60.
[7] Iadecola C. The pathobiology of vascular dementia[J].Neuron,2013,80(4):844-866.
[8] Zhao Y,Gong C X. From chronic cerebral hypoperfusion to Alzheimer-like brain pathology and neurodegeneration[J]. Cell Mol Neurobiol,2015,35(1):101-110.
[9] Kim S,Kang I H,Nam J B,et al. Ameliorating the effect of astragalosideⅣon learning and memory deficit after chronic cerebral hypoperfusion in rats[J]. M olecules,2015,20(2):1904-1921.
[10] Jing Z,Shi C,Zhu L,et al. Chronic cerebral hypoperfusion induces vascular plasticity and hemodynamics but also neuronal degeneration and cognitive impairment[J]. J Cereb Blood Flow M etab,2015,35(8):1249-1259.
[11] Wang D,Kiesinger P R. Autophagy,neuron-specific degradation and neurodegeneration[J]. Autophagy,2012,8(4):711-713.
[12] Kang R,Zeh H J,Lotze M T,et al. The beclin 1 netw ork regulates autophagy and apoptosis[J]. Cell Death Differ,2011,18(4):571-580.
[13] Klionsky D J,Abdalla F C,Abeliovich H,et al. Guidelines for the use and interpretation of assays for monitoring autophagy[J]. Autophagy,2012,8(4):445-544.
[14] Ghavami S,Cunnington R H,Yeganeh B,et al. Autophagy regulates trans fatty acid-mediated apoptosis in primary cardiac my fibroblasts[J]. Biochim Biophys Acta,2012,1823(12):2274-2286.
[15] Gong C,Bauvy C,Tonelli G,et al. Beclin 1 and autophagy are required for the tumorigenicity of breast cancer stem-like/progenitor cells[J]. Oncogene,2013,32(18):2261-2272.
[16] Dou Z,Xu C,Donahue G,et al. Autophagy mediates degradation of nuclear lamina[J]. Nature,2015,527(7576):105-109.
[17]邹文颖,李玉梅,付剑亮.慢性脑低灌注所致认知障碍大鼠海马区自噬机制的初步研究[J].中国临床神经科学,2016,24(1):33-41.
[18] Ohta K,Mizuno A,Ueda M,et al. Autophagy impairment stimulates PS1 expression and gamma-secretase activity[J]. Autophagy,2010,6(3):345-352.
[19]徐昕,颜小华.黄芩苷治疗脑损伤的研究进展[J].实用临床医学,2012,13(8):131-136.
[20] Chen C,Li X H,Gao P L,et al. Baicalin attenuates Alzheimer-like pathological changes and memory deficits induced by amyloidβ1-42 protein[J]. M etab Brain Dis,2015,30(2):537-544.
[21] Zhao J W,Lu S,Yu H,et al. Baicalin and ginsenoside Rb1 promote the proliferation and differentiation of neural stem cells in Alzheimer's disease model rats[J]. Brain Res,2018,1678:187-194.