远志汤对慢性脑低灌注大鼠认知功能及海马凋亡蛋白的影响

doi:10.3969/j.issn.1006-7795.2019.03.001

摘要/Abstract

摘要： 目的观察远志汤对慢性脑低灌注大鼠空间学习记忆能力及海马凋亡蛋白的影响。方法采用永久结扎大鼠双侧颈总动脉法造模。将42只SD大鼠采用数字表法随机分为正常对照组，模型组，假手术组，远志汤高、中、低剂量组[20、10、5 g/（kg·d）]及阳性药物对照组[盐酸多奈哌齐0.52 mg/（kg·d）]，每组各6只，分别灌胃给药，1次/d，给药4周。通过Morris水迷宫定位航行实验及空间探索实验，观察远志汤对慢性脑低灌注大鼠空间学习记忆能力的影响；通过免疫荧光染色法检测远志汤对慢性脑低灌注大鼠海马凋亡相关蛋白B细胞淋巴瘤-2（B-cell lymphoma-2，Bcl-2）、Bcl-2相关X蛋白抗体（Bcl-2 associated X protein，Bax）及半胱氨酸天冬氨酸特异性蛋白酶3（cysteinyl aspartate specific proteinase-3，Caspase-3）表达的影响。结果模型组大鼠较对照组和假手术组的逃避潜伏期和搜索距离明显延长（P<0.01），在原平台所在象限的搜索时间明显缩短（P<0.01）。而远志汤各剂量干预均可明显缩短大鼠的逃避潜伏期和搜索距离（P<0.01），并延长在原平台所在象限的搜索时间（P<0.01）。模型组大鼠海马Bax和Caspase-3的表达较对照组和假手术组增加，差异有统计学意义（P<0.05），Bcl-2则显著下降，差异有统计学意义（P<0.01）。而远志汤高、中剂量组干预均可明显减少Bax和Caspase-3的表达（P<0.05），增加Bcl-2的表达（P<0.05）。结论远志汤可能通过调节海马凋亡相关蛋白Bcl-2、Bax和Caspase-3的表达以改善慢性脑低灌注大鼠的认知功能。

关键词: 远志汤, 认知障碍, 凋亡, 慢性脑低灌注, 海马

Abstract: Objective To observe the effects of Yuanzhi decoction on spatial learning and memory ability and apoptosis-related protein in the hippocampus of rats with chronic cerebral hypoperfusion. Methods A rat model was developed with with ligating the bilateral common carotid arteries permanently.A total of 42 SD rats were divided into 7 groups according to the digital random method:control, model, sham, Yuanzhi decoction high[20 g/(kg·d)], medium[10 g/(kg·d)]and low[5 g/(kg·d)] dose intervention group, and the positive drug control group[donepezil hydrochloride 0.52 mg/(kg·d)] (n=6 in each group). The rats were treated with intragastric administration once a day for 4 weeks. The Morris water maze positioning navigation experiment and space exploration experiment were used to evaluate the spatial learning and memory ability in rats. Immunofluorescence staining was used to examine the expression of apoptosis-related proteins B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein (Bax) and cysteinyl aspartate specific proteinase-3 (Caspase-3) in the hippocampus of rats.Results Compared with the control group and the sham group, the escape latency and search distance of the model group were significantly longer (P<0.01), and the search time in the quadrant of the original platform was significantly shorter (P<0.01). The high, medium and low dose of Yuanzhi decoction intervention can significantly shorten the escape latency and search distance (P<0.01), and prolong the search time in the quadrant of the original platform (P<0.01). The expression of Bax and Caspase-3 in the hippocampus of the model group were significantly increased compared with the control group and the sham group (P<0.05 or P<0.01), and Bcl-2 was significantly decreased (P<0.01). The intervention of high and medium dose of Yuanzhi decoction can significantly reduce the expression of Bax and Caspase-3 (P<0.05) and increase the expression of Bcl-2 (P<0.05 or P<0.01). Conclusion Yuanzhi decoction could regulate the expression of apoptosis-related proteins Bcl-2, Bax and Caspase-3 in hippocampus and improve cognitive function of rats with chronic cerebral hypoperfusion.

Key words: Yuanzhi decoction, cognitive impairment, apoptosis, chronic cerebral hypoperfusion, hippocampus

中图分类号:

R243

刘妍, 陈玉静, 陈文强, 黄小波. 远志汤对慢性脑低灌注大鼠认知功能及海马凋亡蛋白的影响[J]. 首都医科大学学报, 2019, 40(3): 323-329.

Liu Yan, Chen Yujing, Chen Wenqiang, Huang Xiaobo. Effects of Yuanzhi decoction on cognitive function and apoptosis-related protein in hippocampus of rats with chronic cerebral hypoperfusion[J]. Journal of Capital Medical University, 2019, 40(3): 323-329.

参考文献

[1] Luo P, Lu Y, Li C, et al. Long-lasting spatial learning and memory impairments caused by chronic cerebral hypoperfusion associate with a dynamic change of HCN1/HCN2 expression in hippocampal CA1 region[J]. Neurobiol Learn Mem, 2015, 123:72-83.
[2] Li W, Yuan H, Yu Y, et al. Etidronate rescues cognitive deficits through improving synaptic transmission and suppressing apoptosis in 2-vessel occlusion model rats[J]. J Neurochem, 2017, 140(3):476-484.
[3] 黄小波, 陈文强, 陈玉静, 等. 远志-石菖蒲干预糖尿病认知障碍大鼠血清甲基乙二醛的研究[J]. 中国实验方剂学杂志, 2013, 19(24):223-226.
[4] 国家药典委员会. 中华人民共和国药典. 一部[M]. 北京:中国医药科技出版社, 2015:91-92, 156-157.
[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] Shih A Y, Blinder P, Tsai P S, et al. The smallest stroke:occlusion of one penetrating vessel leads to infarction and a cognitive deficit[J]. Nat Neurosci, 2013, 16(1):55-63.
[7] 陈士铎(清). 石室秘录. 中医临床必读丛书[M]. 北京:人民卫生出版社, 2006:319-320.
[8] 赵佶(宋). 圣济总录[M]. 北京:人民卫生出版社, 1982:1022.
[9] 王宁群, 黄小波, 魏翠柏, 等. 脑康Ⅱ号对脑梗死后血管性认知障碍患者的干预研究[J]. 中医药学报, 2014, 42(12):71-74.
[10] 陈玉静, 陈文强, 黄小波, 等. 远志皂苷元对甲基乙二醛神经毒性的抑制作用研究[J]. 北京中医药, 2016, 35(5):422-425.
[11] 王晓丽, 董妙先, 徐天娇, 等. β-细辛醚对缺糖缺氧再灌注损伤原代大鼠海马神经元的保护作用[J]. 中国病理生理杂志, 2014, 30(5):928-932.
[12] Huang X B, Chen Y J, Chen W Q, et al. Neuroprotective effects of tenuigenin on neurobehavior, oxidative stress, and tau hyperphosphorylation induced by intracerebroventricular streptozotocin in rats[J]. Brain Circ, 2018, 4(1):24-32.
[13] Deng M, Huang L, Ning B, et al. β-asarone improves learning and memory and reduces Acetyl Cholinesterase and Beta-amyloid 42 levels in APP/PS1 transgenic mice by regulating Beclin-1-dependent autophagy[J]. Brain Res, 2016, 1652:188-194.
[14] Jiang X M, Hu J H, Wang L L, et al. Ulinastatin alleviates neurological deficiencies evoked by transient cerebral ischemia via improving autophagy, Nrf-2-ARE and apoptosis signals in hippocampus[J]. Physiol Res, 2018, 67(4):637-646.
[15] Zhang Y, Yu S, Tuazon J P, et al. Neuroprotective effects of human bone marrow mesenchymal stem cells against cerebral ischemia are mediated in part by an anti-apoptotic mechanism[J]. Neural Regen Res, 2019, 14(4):597-604.
[16] Lei X, Lei L, Zhang Z, et al. Diazoxide inhibits of ER stress mediated apoptosis during oxygen glucose deprivation in vitro and cerebral ischemia reperfusion in vivo[J]. Mol Med Rep, 2018, 17(6):8039-8046.
[17] Srbulescu R F, Zupanc G K. Inhibition of caspase-3 mediated apoptosis improves spinal cord repair in a regeneration competent vertebrate system[J]. Neuroscience, 2010, 171(2):599-612.
[18] 冉丰,章歌雅,何治.超极化激活环核苷酸门控阳离子通道在脑缺血及其认知功能障碍中的作用[J].中国脑血管病杂志,2016,13(9):502-505.
[19] 曾晓莉,张文娟,赵博,等.慢性脑缺血神经保护机制及药物研究进展[J].中国脑血管病杂志,2017,14(11):607-611.
[20] 张羽乔,周俊山,张颖冬,等.不同程度颈动脉狭窄对大鼠认知功能及海马CA1区神经元凋亡的影响[J].中国脑血管病杂志,2016,13(10):529-534,557.