β-谷甾醇对阿尔茨海默病模型小鼠认知功能的改善作用及其机制

doi:10.13481/j.1671-587X.20230307

Abstract

Abstract:

Objective： To discuss the improvement effect of β-sitosterol on the cognitive function of the Alzheimer’s disease （AD） model mice， and to elucidate its possible molecular mechanism. Methods Sixty mice were randomly divided into control group， sham operation group， AD group， donepezil hydrochloride （0.6 mg·kg^－1）group， low dose （1.0 mg·kg^－1） of β-sitosterol group， and high dose （4.0 mg·kg^－1）of β- sitosterol group； and there were 12 mice in each group. The mice were injected with β amyloid protein 1-42 （Aβ_1-42） to establish the AD models，and β- sitosterol interventional treatment was conducted. The free activity numbers of the mice in various groups were observed by autonomous activity experiment； the scores of nesting behavior of the mice in various groups were observed by nesting behavior experiment；the identification time of new objects and the discrimination ratios （DR） of new and old objects of the mice in various groups were observed by new object discrimination experiment； Morris water maze experiment was used to observe the escape latencyies and numbers of crossing platform of the mice in various groups； real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of interleukin-17A（IL-17A） and p53 mRNA in hippocampus tissue of the mice in various groups； Western blotting method was used to detect the expression levels of IL-17A， p53，Caspase-3， cleaved Caspase-3，B-cell lymphoma 2 （Bcl-2）， and Bcl-2 associated X protein （Bax） in hippocampus tissue of the mice in various groups；enzyme-linked immunosorbent assay （ELISA） was used to detect the activities of superoxide dismutase （SOD） and levels of glutathione （GSH），tumor necrosis factor α（TNF-α）， and IL-17A in hippocampus tissue of the mice in various groups. Results The nesting behavior experiment results showed that compared with control group， the score of nesting behavior of the mice in AD group was significantly decreased （P<0.01）； compared with AD group， the scores of nesting behavior of the mice in donepezil hydrochloride group and low dose of β-sitosterol group were increased （P<0.05）. The new object discrimination experiment results showed that compared with control group， the identification time of new objects of the mice in AD group was decreased （P<0.01），and DR was decreased（P<0.01）； compared with AD group，the identification time of new objects of the mice in donepezil hydrochloride group and high dose of β-sitosterol group were significantly increased （P<0.01）， and DR was increased （P<0.05）. The Morris water maze experiment results showed that on the 3rd and 5th days after training， compared with control group， the escape latency of the mice in AD group was significantly increased （P<0.01）； compared with AD group， the escape latencies of the mice in latendonepezil hydrochloride group and low and high doses of β-sitosterol groups were increased （P<0.05 or P<0.01）. Compared with control group， the number of crossing platforms of the mice in AD group was increased （P<0.01）； compared with AD group， the numbers of crossing platform of the mice in donepezil hydrochloride group and low and high doses of β-sitosterol groups were decreased （P<0.01）. The RT-qPCR results showed that compared with control group， the expression levels of IL-17A and p53 mRNA in hippocampus tissue of the mice in AD group were significantly increased （P<0.01）； compared with AD group，the expression levels of IL-17A mRNA in hippocampus tissue of the mice in low and high doses of β-sitosterol groups were decreased （P<0.05）， while the expression levels of p53 mRNA in hippocampus tissue of the mice in donepezil hydrochloride group and high dose of β-sitosterol group were significantly decreased （P<0.05）. The Western blotting results showed that compared with control group， the expression levels of IL-17A， p53， and cleaved Caspase-3 proteins and the Bax/Bcl-2 ratio in hippocampus tissue of the mice in AD group were significantly increased （P<0.01）； compared with AD group， the expression levels of IL-17A and p53 proteins and Bax/Bcl-2 ratio in hippocampus tissue of the mice in low dose of β-sitosterol group were significantly decreased （P<0.05 or P<0.01）， while the expression levels of IL-17A and p53 proteins and Bax/Bcl-2 ratio in hippocampus tissue of the mice in donepezil hydrochloride group and high dose of β- sitosterol group were decreased （P<0.01）. The ELISA results showed that compared with control group， the SOD activity and GSH level in hippocampus tissue of the mice in AD group were decreased （P<0.01）， while the levels of TNF-α， IL-17A and IL-17A in hippocampus tissue of the mice were increased （P<0.01）； compared with AD group， the SOD activity and GSH level in hippocampal tissue of the mice in high dose of β- sitosterol group were significantly increased （P<0.01），while the IL-17A level was decreased （P<0.05）， and the level of TNF-α in hippocampus tissue of the mice in low dose of β-sitosterol group was decreased （P<0.05）. Conclusion β-sitosterol can reduce the neuroinflammation and inhibit the apoptosis，thereby improving the cognitive function of the AD model mice；its mechanism may be related to inhibiting the IL-17-p53 signaling pathway.

Key words: Alzheimer’s disease, β-sitosterol, Neuroin flammation, Apoptosis, Oxidatve stress

CLC Number:

R742

Xingye WANG,Xiangri KONG,Mengli JIN,Bingmei WANG,Mingquan LI. Improvement effect of β-sitosterol on cognitive function in Alzheimer’s disease model mice and its mechanism[J].Journal of Jilin University(Medicine Edition), 2023, 49(3): 599-607.

Figures/Tables 8

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References 28

1	BRIGAS H C， RIBEIRO M， COELHO J E，et al.IL-17 triggers the onset of cognitive and synaptic deficits in early stages of Alzheimer’s disease［J］. Cell Rep， 2021， 36（9）： 109574.
2	张钰，马兰. β淀粉样蛋白寡聚体参与阿尔茨海默病发病机制的研究进展［J］.中华老年多器官疾病杂志， 2021， 20（8）： 628-631.
3	AMOR S， PEFEROEN L A， VOGEL D Y， et al. Inflammation in neurodegenerative diseases： an update［J］. Immunology， 2014， 142（2）： 151-166.
4	TANG Y， LE W D. Differential roles of M1 and M2 microglia in neurodegenerative diseases［J］. Mol Neurobiol， 2016， 53（2）： 1181-1194.
5	DUGGER B N， DICKSON D W. Pathology of neurodegenerative diseases［J］. Cold Spring Harb Perspect Biol， 2017， 9（7）： a028035.
6	KWEON J H， KIM S， LEE S B. The cellular basis of dendrite pathology in neurodegenerative diseases［J］. BMB Rep， 2017， 50（1）：5-11.
7	STEPHENSON J， NUTMA E， VAN DER VALK P， et al. Inflammation in CNS neurodegenerative diseases［J］. Immunology， 2018， 154（2）： 204-219.
8	薛愉，邹和建，郑颂国. IL-17与自身免疫性疾病相关研究进展［J］.中华微生物学和免疫学杂志，2007，27（9）： 866-870.
9	OGITA N， OKUSHIMA Y， TOKIZAWA M， et al. Identifying the target genes of suppressor of gamma response 1， a master transcription factor controlling DNA damage response in Arabidopsis［J］.Plant J， 2018， 94（3）： 439-453.
10	ABATE G， FRISONI G B， BOURDON J C， et al. The pleiotropic role of p53 in functional/dysfunctional neurons： focus on pathogenesis and diagnosis of Alzheimer’s disease［J］.Alzheimers Res Ther，2020，12（1）： 160.
11	陈元堃，曾奥，罗振辉，等. β-谷甾醇药理作用研究进展［J］. 广东药科大学学报， 2021， 37（1）： 148-153.
12	刘雅谦，李琳，孙万成，等. 植物甾醇的抗炎性研究进展［J］. 中国油脂， 2022， 47（5）： 93-99.
13	SHI C， WU F M， ZHU X C， et al. Incorporation of beta-sitosterol into the membrane increases resistance to oxidative stress and lipid peroxidation via estrogen receptor-mediated PI3K/GSK3beta signaling［J］. Biochim Biophys Acta， 2013， 1830（3）： 2538-2544.
14	赵帅，陈冬梅，虎娜，等. β-谷甾醇通过PI3K/AKT通路影响颗粒细胞增殖及凋亡［J］. 宁夏医科大学学报， 2021， 43（4）： 339-344.
15	SHI C， WU F， XU J. Incorporation of β-sitosterol into mitochondrial membrane enhances mitochondrial function by promoting inner mitochondrial membrane fluidity［J］. J Bioenerg Biomembr，2013，45（3）： 301-305.
16	ZHANG J， KE K F， LIU Z， et al. Th17 cell-mediated neuroinflammation is involved in neurodegeneration of aβ1-42-induced Alzheimer’s disease model rats［J］. PLoS One， 2013， 8（10）： e75786.
17	闵喆，王淑楠，吴军，等. Tg-SwDI小鼠的筑巢行为研究［J］. 神经损伤与功能重建， 2014， 9（1）： 1-5.
18	SIVAKUMARAN M H， MACKENZIE A K， CALLAN I R， et al. The discrimination ratio derived from novel object recognition tasks as a measure of recognition memory sensitivity， not bias［J］. Sci Rep， 2018， 8（1）： 11579.
19	白鑫宇，刘萍，杨帆，等. 补阴益智汤配方颗粒对东莨菪碱所致小鼠学习记忆损伤的保护作用［J］. 中国实验方剂学杂志， 2017， 23（12）： 138-144.
20	CHEN J M， JIANG G X， LI Q W， et al. Increased serum levels of interleukin-18， -23 and -17 in Chinese patients with Alzheimer’s disease［J］. Dement Geriatr Cogn Disord， 2014， 38（5/6）： 321-329.
21	CRISTIANO C， VOLPICELLI F， LIPPIELLO P，et al.Neutralization of IL-17 rescues amyloid-β-induced neuroinflammation and memory impairment［J］. Br J Pharmacol， 2019， 176（18）： 3544-3557.
22	JAZVINŠĆAK JEMBREK M， SLADE N， HOF P R， et al. The interactions of p53 with tau and Aß as potential therapeutic targets for Alzheimer’s disease［J］. Prog Neurobiol， 2018， 168（104-127.
23	邹瑜，钟纯正，郭春宣，等. miR-130b通过调控p53抑制局灶性脑缺血大鼠神经细胞凋亡［J］. 中国老年学杂志， 2020， 40（3）： 602-605.
24	DEZOR M， DORSZEWSKA J， FLORCZAK J， et al. Expression of 8-oxoguanine DNA glycosylase 1 （OGG1） and the level of p53 and TNF-αlpha proteins in peripheral lymphocytes of patients with Alzheimer’s disease［J］. Folia Neuropathol， 2011， 49（2）： 123-131.
25	陈小玉，罗连响，潘韵琪，等. 组织透明化技术在神经退行性疾病中的应用研究进展［J］. 解放军医学杂志， 2022， 47（3）： 305-313.
26	郝淼淼，田海燕，刘晗，等. 水通道蛋白4低表达通过抑制类淋巴系统功能对AD模型615-619小鼠脑组织磷酸化Tau蛋白表达的影响［J］. 郑州大学学报（医学版）， 2022， 57（5）.
27	NAKANISHI A， MINAMI A， KITAGISHI Y， et al. BRCA1 and p53 tumor suppressor molecules in Alzheimer’s disease［J］. Int J Mol Sci， 2015， 16（2）： 2879-2892.
28	杨俊峰，刘强. 免疫与神经系统遗传病［J］. 中国实用内科杂志， 2022（4）： 273-277.

Primer	Sequence (5'－3')
β-actin	F: GACTGTTACTGAGCTGCGTTT
	R: AGGGTGAGGGACTTCCTGTA
IL-17A	F:GCTGACCCCTAAGAAACCCC
	R:GAAGCAGTTTGGGACCCCTT
p53	F: TGTTCCGGGAGCTGAATGAG
	R:GAGTGGATCCTGGGGATTGTGTC

Group	Number of autonomic activity	Score of nesting behavior
Control	110.33±10.24	3.17±0.58
Sham operation	107.50±15.10	3.00±0.74
AD	110.50±14.49	1.67±0.78^*
Donepezil hydrochloride	109.83±13.81	2.75±0.87^△
Low dose of β-sitosterol	107.17±14.41	2.67±0.89^△
High dose of β-sitosterol	103.00±14.86	2.25±1.06
^*P<0.01 vs control group; ^△P<0.05 vs AD group.

Group	Idenfification time of new object (t/s)	DR
Control	57.56±11.78	0.65±0.14
Sham operation	54.30±10.96	0.64±0.15
AD	34.47±7.94^*	0.36±0.11^*
Donepezil hydrochloride	50.88±11.30^△△	0.48±0.15
Low dose of β-sitosterol	39.37±10.68	0.51±0.14^△
High dose of β-sitosterol	44.53±4.01^△△	0.45±0.10
^*P<0.01 vs control group; ^△P<0.05, ^△△P<0.01 vs AD group.

Group	n	Escape latency(t/s)			Number of crossing platform
Group	n	(t/d) 1	3	5	Number of crossing platform
Control	12	80.73±5.10	47.47±12.57	35.81±11.81	2.70±0.65
Sham operation	12	81.36±4.51	54.06±13.00	41.14±7.79	2.80±0.97
AD	11	81.33±4.93	75.14±8.35^*	67.80±5.60^*	5.90±1.60^*
Donepezil hydrochloride	12	80.23±3.75	62.12±11.56^△	47.81±7.65^△△	3.10±1.30^△△
Low dose of β-sitosterol	12	77.10±5.95	61.91±9.87^△	52.79±12.06^△△	3.40±1.80^△△
High dose of β-sitosterol	12	80.19±4.96	56.57±10.40^△△	55.62±7.39^△△	4.00±1.21^△△
^*P<0.01 vs control group; ^△P<0.05, ^△△P<0.01 vs AD group.

Group	SOD[λ_B/(U·mL^－1)]	GSH[c_B/(μmol·L^－1)]
Control	56.97±5.51	123.47±27.99
Sham operation	54.77±6.30	123.71±22.85
AD	37.41±9.94^*	70.59±9.64^*
Donepezil hydrochloride	44.18±14.76	88.30±10.95
Low dose of β-sitosterol	40.15±7.89	93.70±16.22
High dose of β-sitosterol	53.56±8.13^△	109.59±13.59^△
^*P<0.01 vs control group; ^△P<0.01 vs AD group.

Improvement effect of β-sitosterol on cognitive function in Alzheimer’s disease model mice and its mechanism

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Group	TNF-α	IL-17A
Control	467.63±36.50	192.98±57.22
Sham operation	495.21±54.06	218.30±62.85
AD	571.12±52.05^*	489.49±45.26^*
Donepezil hydrochloride	547.58±32.65	454.13±92.60
Low dose of β-sitosterol	507.49±41.57^△	415.89±64.78
High dose of β-sitosterol	525.28±39.31	381.97±94.82^△

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