人参皂苷Rg1对阿尔茨海默症大鼠BDNF⁃TrkB信号通路的影响

doi:10.19586/j.2095-2341.2021.0183

生物技术进展 ›› 2022, Vol. 12 ›› Issue (3): 446-451.DOI: 10.19586/j.2095-2341.2021.0183

人参皂苷Rg1对阿尔茨海默症大鼠BDNF⁃TrkB信号通路的影响

陈晓光¹(), 潘晓峰¹, 王帆², 潘宋斌¹()

^1.武汉市第一医院（武汉市中西医结合医院）神经内科，武汉 430022
^2.武汉市第一医院（武汉市中西医结合医院）老年病科，武汉 430022

收稿日期:2021-11-15 接受日期:2021-12-16 出版日期:2022-05-25 发布日期:2022-05-26
通讯作者: 潘宋斌
作者简介:陈晓光 E-mail：a1980we12@163.com；
基金资助:
湖北省自然科学基金项目(2011CDB299);湖北省卫生和计划生育委员会项目(201742)

Effects of Ginsenoside Rg1 on BDNF-TrkB Signaling Pathways in Rats with Alzheimer′s Disease

Xiaoguang CHEN¹(), Xiaofeng PAN¹, Fan WANG², Songbin PAN¹()

^1.Department of Neurology，Wuhan No. 1 Hospital （Wuhan Hospital of Integrated Traditional Chinese Medicine and Western Medicine），Wuhan 430022，China
^2.Department of Geriatric，Wuhan No. 1 Hospital （Wuhan Hospital of Integrated Traditional Chinese Medicine and Western Medicine），Wuhan 430022，China

Received:2021-11-15 Accepted:2021-12-16 Online:2022-05-25 Published:2022-05-26
Contact: Songbin PAN

摘要/Abstract

摘要：

为了探究人参皂苷Rg1对阿尔茨海默症（Alzheimer's disease, AD）大鼠模型脑源性神经营养因子/酪氨酸激酶受体B（BDNF-TrkB）信号通路的影响，选取75只SD大鼠随机分为空白对照组、模型组、低剂量Rg1组、中剂量Rg1组及高剂量Rg1组，每组15只。取各组大鼠脑组织制备脑片，除空白对照组外，其他组加入Aβ1-42试剂制备AD模型，低剂量Rg1组、中剂量Rg1组和高剂量Rg1组分别使用60、120、240 μmol·L^-1 Rg1处理。采用HE染色观察脑组织病理损伤，TUNEL染色检测脑组织细胞凋亡，比色法测定脑片中乙酰胆碱（acetylcholine，Ach）、5-羟色胺（5-hydroxytryptamine，5-HT）水平和乙酰胆碱酯酶（acetylcholinesterase，TChE）活力，蛋白质印迹法检测各组脑片中切割后半胱氨酸蛋白酶-3（Cleaved Caspase-3）、B淋巴细胞瘤-2（B cell lymphoma-2，Bcl-2）、Bcl-2相关蛋白X（Bcl-2 associated X protein，Bax）及BDNF-TrkB信号通路相关蛋白表达情况。与空白对照组相比，模型组脑组织细胞凋亡数、Cleaved Caspase-3、Bax/Bcl-2及TChE水平显著增加，5-HT、Ach、BDNF及TrkB蛋白表达量显著降低（P<0.05）；与模型组相比，低剂量Rg1组、中剂量Rg1组和高剂量Rg1组脑组织细胞凋亡数、Cleaved Caspase-3、Bax/Bcl-2及TChE水平显著降低，5-HT、Ach、BDNF及TrkB蛋白表达量显著增加（P<0.05），且具有剂量依赖性。人参皂苷Rg1可有效保护阿尔茨海默症模型大鼠脑组织，抑制神经细胞凋亡，其作用机制可能与激活BDNF-TrkB信号通路相关。通过分析人参皂苷Rg1对AD大鼠模型的保护机制，以期为人参皂苷Rg1用于治疗AD奠定理论基础。

关键词: 阿尔茨海默症, 人参皂苷Rg1, 细胞凋亡, BDNF-TrkB信号通路

Abstract:

The aim of the study is to explore the effects of ginsenoside Rg1 on brain-derived neurotrophic factor/tyrosine kinase receptor B （BDNF-TrkB） signaling pathways in rats with Alzheimer's disease （AD）. A total of 75 SD rats were enrolled and randomly divided into blank control group， model group， low-dose Rg1 group， medium-dose Rg1 group and high-dose Rg1 group， with 15 rats in each group. The brain tissues in each group were collected to prepare brain slices. Except blank control group， Aβ1-42 reagent was added into each group to prepare AD models. The low-dose Rg1 group， medium-dose Rg1 group and high-dose Rg group were treated with 60， 120 and 240 μmol·L^-1 Rg1， respectively. The pathological damage of brain tissues was observed by HE staining. The apoptosis of brain tissues was detected by TUNEL staining. The levels of acetylcholine （Ach） and 5-hydroxytryptamine （5-HT）， and activity of acetylcholinesterase （TChE） in brain slices were detected by colorimetry. The expressions of cleaved cysteine proteinase-3 （Cleaved Caspase-3）， B cell lymphoma-2 （Bcl-2）， Bcl-2 associated X protein （Bax） and BDNF-TrkB signaling pathway related proteins in brain slices were detected by Western blot. Compared with blank control group， the number of brain tissues apoptosis， levels of Cleaved Caspase-3， Bax/Bcl-2 and TChE were significantly increased in model group， and 5-HT， Ach， BDNF and TrkB were significantly decreased in model group （P<0.05）. Compared with model group， the number of brain tissues apoptosis， levels of Cleaved Caspase-3， Bax/Bcl-2 and TChE were significantly decreased in low-dose group， medium-dose group and high-dose Rg1 group， and 5-HT， Ach， BDNF and TrkB were significantly increased in low-dose group， medium-dose group and high-dose Rg1 group （P<0.05）， which showing concentration-dependence. Ginsenoside Rg1 can effectively protect the brain tissue of AD rats and inhibit neuron apoptosis， which the mechanism of action may be related to activating BDNF-TrkB signaling pathways. By analyzing the protective mechanism of ginsenoside Rg1 on AD rat model， the paper was expected to lay a theoretical foundation for the use of ginsenoside Rg1 in the treatment of AD in this study.

Key words: Alzheimer's disease, ginsenoside Rg1, apoptosis, BDNF-TrkB signaling pathway

中图分类号:

R285.5

陈晓光, 潘晓峰, 王帆, 潘宋斌. 人参皂苷Rg1对阿尔茨海默症大鼠BDNF⁃TrkB信号通路的影响[J]. 生物技术进展, 2022, 12(3): 446-451.

Xiaoguang CHEN, Xiaofeng PAN, Fan WANG, Songbin PAN. Effects of Ginsenoside Rg1 on BDNF-TrkB Signaling Pathways in Rats with Alzheimer′s Disease[J]. Current Biotechnology, 2022, 12(3): 446-451.

图/表 7

图1 大鼠脑片HE染色结果注：放大倍数为200倍。

Fig.1 HE staining results of brain slices in rats

图2 大鼠脑片TUNEL染色结果注：放大倍数为400倍。

Fig.2 TUNEL staining results of brain slices in rats

图3 大鼠脑片中凋亡相关蛋白检测结果A：空白对照组；B：模型组；C：低剂量Rg1组；D：中剂量Rg1组；E：高剂量Rg1组

Fig.3 Detection results of apoptosis related proteins in rat brain slices

表1 大鼠脑片中凋亡相关蛋白检测结果

Table 1 Detection results of apoptosis?related proteins in brain slices of rats

组别	Cleaved Caspase⁃3/GAPDH	Bax/Bcl⁃2
空白对照组（n=15）	0.35±0.09	0.19±0.06
模型组（n=15）	1.14±0.21^*	1.87±0.23^*
低剂量Rg1组（n=15）	0.95±0.13^#	1.42±0.17^#
中剂量Rg1组（n=15）	0.76±0.10^##′	1.15±0.19^##′
高剂量Rg1组（n=15）	0.56±0.12^##′#″	0.47±0.13^##′#″
F值	25.955	85.387
P值	<0.001	<0.001

表2 大鼠脑片中5?HT、Ach含量和TChE活力检测结果（x±s）

Table 2 Detection results of 5?HT, Ach levels and TChE activity in brain slices of rats (x±s)

组别	5⁃HT/（pg·g^-1）	Ach/（g·g^-1）	TChE/（U·mg^-1）
空白对照组（n=15）	158.35±19.25	2 254.10±180.28	0.31±0.04
模型组（n=15）	107.39±14.69^*	1 604.56±144.50^*	0.69±0.09^*
低剂量Rg1组（n=15）	119.51±15.07^#	1 803.68±150.14^#	0.51±0.07^#
中剂量Rg1组（n=15）	130.73±16.17^##′	1 997.89±141.87^##′	0.43±0.06^##′
高剂量Rg1组（n=15）	142.39±18.11^##′#″	2 098.90±150.58^##′#″	0.36±0.05^##′#″
F值	19.160	36.697	73.807
P值	<0.001	<0.001	<0.001

图4 大鼠脑片中BDNF?TrkB信号通路相关蛋白检测结果A：空白对照组；B：模型组；C：低剂量Rg1组；D：中剂量Rg1组；E：高剂量Rg1组

Fig. 4 Detection results of BDNF?TrkB signal pathway related proteins in rat brain slices

表3 大鼠脑片中BDNF?TrkB信号通路相关蛋白检测结果（x±s）

Table 3 Detection results of BDNF-TrkB signaling pathways related proteins in brain slices of rats (x±s)

组别	BDNF/GAPDH值	TrkB/GAPDH值
空白对照组（n=15）	1.23±0.13	1.53±0.18
模型组（n=15）	0.49±0.09^*	0.39±0.10^*
低剂量Rg1组（n=15）	0.64±0.10^#	0.75±0.16^#
中剂量Rg1组（n=15）	0.78±0.08^##′	1.11±0.12^##′
高剂量Rg1组（n=15）	0.93±0.11^##′#″	1.33±0.15^##′#″
F值	37.724	49.695
P值	<0.001	<0.001

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人参皂苷Rg1对阿尔茨海默症大鼠BDNF⁃TrkB信号通路的影响

Effects of Ginsenoside Rg1 on BDNF-TrkB Signaling Pathways in Rats with Alzheimer′s Disease

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