基于斑马鱼模型研究木蝴蝶苷A的抗阿尔茨海默病活性和作用机制

doi:10.3976/j.issn.1002-4026.2023.06.005

Abstract

Abstract:

To investigate the ameliorative effects of oroxin A on Alzheimer's disease (AD) and the underlying mechanism of action, a zebrafish AD model induced by aluminum chloride hexahydrate (AlCl₃) was used. Wild-type zebrafish AB larvae at 3 dpf(days post fertilization) were divided into different groups, including negative control group, AlCl₃ (80 μmol/L) model control group, AlCl₃ (80 μmol/L) combined with donepezil (6 μmol/L) positive control group, and AlCl₃ (80 μmol/L) combined with different concentrations (5, 10, and 20 μmol/L) of oroxin A test group. At 6 dpf, zebrafish behavior was monitored and analyzed using zebrafish light-dark locomotion test. Aβ deposition in zebrafish heads was assayed by thioflavin S staining. Acetylcholine assay kit tested acetylcholinesterase (AchE) activity. In addition, the expression of autophagy-related genes(beclin1、ulk1b、ulk2 and atg7) was tested by real-time quantitative polymerase chain reaction. Molecular docking was performed to validate the interaction between oroxin A and autophagy-related protein(beclin1、ulk1b、ulk2 and atg7). The results indicated that oroxin A significantly relieved the dyskinesia and inhibited Aβ deposition and AchE activity of zebrafish induced by AlCl₃. The expression of autophagy-related genes tended to be normal after oroxin A treatment. This study preliminarily revealed that oroxin A alleviated AlCl₃-induced AD symptoms in zebrafish, where the underlying mechanism of action is possibly associated with activated autophagy, providing a theoretical basis for the clinical application of oroxin A and its related research in treating AD.

Key words: Alzheimer's disease, aluminum chloride hexahydrate, autophagy, zebrafish, oroxin A

CLC Number:

R965

SHI Ruidie, GAO Xin, WANG Baokun, GAO Daili, JIN Meng, ZHANG Xiujun. Anti-Alzheimer's disease activity of oroxin A and its mechanism of action based on zebrafish model[J].Shandong Science, 2023, 36(6): 28-37.

Figures/Tables 6

Table 1

Fig.1

Fig.2

Fig.3

Fig.4

Fig.5

References 22

[1]	TATULIAN S A. Challenges and hopes for Alzheimer's disease[J]. Drug Discovery Today, 2022, 27(4): 1027-1043. DOI: 10.1016/j.drudis.2022.01.016. pmid: 35121174
[2]	TAM K, JU Y J. Pathological mechanisms and therapeutic strategies for Alzheimer's disease[J]. Neural Regeneration Research, 2022, 17(3): 543. DOI: 10.4103/1673-5374.320970. pmid: 34380884
[3]	SRIRAM N, SAH M K. Regulatory insights into progression of cancer and Alzheimer's disorder from autophagy perspective[J]. Molecular Biology Reports, 2021, 48(12): 8227-8232. DOI: 10.1007/s11033-021-06838-4. pmid: 34665401
[4]	FANG E F, HOU Y J, PALIKARAS K, et al. Mitophagy inhibits amyloid-β and tau pathology and reverses cognitive deficits in models of Alzheimer's disease[J]. Nature Neuroscience, 2019, 22(3): 401-412. DOI: 10.1038/s41593-018-0332-9. pmid: 30742114
[5]	SUN C, ZHANG SS, BA S K, et al. Eucommia ulmoides olive male flower extracts Ameliorate alzheimer's disease-like pathology in zebrafish via regulating autophagy, acetylcholinesterase, and the dopamine transporter[J]. Frontiers in Molecular Neuroscience, 2022, 15: 901953. DOI: 10.3389/fnmol.2022.901953.
[6]	国家药典委员会. 中华人民共和国药典2020年版二部[M]. 北京: 中国医药科技出版社, 2020.
[7]	CHENG X D, HUANG T T, WANG C H, et al. Natural compound library screening identifies oroxin A for the treatment of myocardial ischemia/reperfusion injury[J]. Frontiers in Pharmacology, 2022, 13: 894899. DOI: 10.3389/fphar.2022.894899.
[8]	鲁冰冰. 木蝴蝶苷A对食管鳞癌增殖的抑制作用及其机制的研究[D]. 郑州: 郑州大学, 2020.
[9]	任擎宇, 靳梦, 商学良, 等. 基于网络药理学和斑马鱼模型的金盏菊缓解阿尔茨海默病症状及作用机制[J]. 中国药理学通报, 2022, 38(3): 429-436. DOI: 10.12360/CPB202108075.
[10]	BASNET R M, ZIZIOLI D, TAWEEDET S, et al. Zebrafish larvae as a behavioral model in neuropharmacology[J]. Biomedicines, 2019, 7(1): 23. DOI: 10.3390/biomedicines7010023.
[11]	SHENOY A, BANERJEE M, UPADHYA A, et al. The brilliance of the zebrafish model: perception on behavior and alzheimer's disease[J]. Frontiers in Behavioral Neuroscience, 2022, 16: 861155. DOI: 10.3389/fnbeh.2022.861155.
[12]	PATIL D N, PATIL S A, SISTLA S, et al. Comparative biophysical characterization: A screening tool for acetylcholinesterase inhibitors[J]. PLoS One, 2019, 14(5): e0215291. DOI: 10.1371/journal.pone.0215291.
[13]	SAXENA M, DUBEY R. Target enzyme in Alzheimer's disease: acetylcholinesterase inhibitors[J]. Current Topics in Medicinal Chemistry, 2019, 19(4): 264-275. DOI: 10.2174/1568026619666190128125912. pmid: 30706815
[14]	SENGER M R, SEIBT K J, GHISLENI G C, et al. Aluminum exposure alters behavioral parameters and increases acetylcholinesterase activity in zebrafish (Danio rerio) brain[J]. Cell Biology and Toxicology, 2011, 27(3): 199-205. DOI: 10.1007/s10565-011-9181-y. pmid: 21240652
[15]	WALSH D M, SELKOE D J. Amyloid β-protein and beyond: the path forward in Alzheimer's disease[J]. Current Opinion in Neurobiology, 2020, 61: 116-124. DOI: 10.1016/j.conb.2020.02.003. pmid: 32197217
[16]	BUSCHE M A, WEGMANN S, DUJARDIN S, et al. Tau impairs neural circuits, dominating amyloid-β effects, in Alzheimer models in vivo[J]. Nature Neuroscience, 2019, 22(1): 57-64. DOI: 10.1038/s41593-018-0289-8. pmid: 30559471
[17]	DENG Z Q, DONG Y, ZHOU X T, et al. Pharmacological modulation of autophagy for Alzheimer's disease therapy: Opportunities and obstacles[J]. Acta Pharmaceutica Sinica B, 2022, 12(4): 1688-1706. DOI: 10.1016/j.apsb.2021.12.009. pmid: 35847516
[18]	PICKFORD F, MASLIAH E, BRITSCHGI M, et al. The autophagy-related protein beclin1 shows reduced expression in early Alzheimer disease and regulates amyloid beta accumulation in mice[J]. The Journal of Clinical Investigation, 2008, 118(6): 2190-2199. DOI: 10.1172/JCI33585.
[19]	JAEGER P A, PICKFORD F, SUN C H, et al. Regulation of amyloid precursor protein processing by the beclin1 complex[J]. PLoS One, 2010, 5(6): e11102. DOI: 10.1371/journal.pone.0011102.
[20]	MCALPINE F, WILLIAMSON L E, TOOZE S A, et al. Regulation of nutrient-sensitive autophagy by uncoordinated 51-like kinases 1 and 2[J]. Autophagy, 2013, 9(3): 361-373. DOI: 10.4161/auto.23066. pmid: 23291478
[21]	任李梅, 杨松, 卢姝言, 等. 人参总皂苷在巨噬细胞RAW264.7中降低脂多糖诱导的炎症和氧化应激[J]. 中国医院药学杂志, 2022, 42(9): 896-901. DOI: 10.13286/j.1001-5213.2022.09.04.
[22]	CARVALHO C, SANTOS M S, OLIVEIRA C R, et al. Alzheimer's disease and type 2 diabetes-related alterations in brain mitochondria, autophagy and synaptic markers[J]. Biochimica et Biophysica Acta (BBA)-Molecular Basis of Disease, 2015, 1852(8): 1665-1675. DOI: 10.1016/j.bbadis.2015.05.001.

基因名称	引物序列
rpl13a	上游: TCTGGAGGACTGTAAGAGGTATGC
	下游: AGACGCACAATCTTGAGAGCAG
beclin1	上游: GTTCAGGTGGTCTGCGTTTT
	下游: GCAAACAGAAGCCAGTGTCA
ulk1b	上游: AGGCCGAAAGTCTCACTTCA
	下游: AGCCATGTACATCGGAGACC
ulk2	上游: ACCTCTGATTGGCTGACAAAAT
	下游: GAGATTGCAAGAGGCTTGAGTT
atg7	上游: AGAGTCCAGTCCGATGTC
	下游: A GAAGTAACAGCCGAGACG

Anti-Alzheimer's disease activity of oroxin A and its mechanism of action based on zebrafish model

RichHTML

PDF (PC)

Like

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 6

References 22

Related Articles 15

Metrics

Comments

Recommended 0

[1]	CHEN Shanjun, WANG Huan, HU Kaiqing, BI Wenjie, CHENG Guidong, WANG Songsong, HAN Liwen, WANG Xiaojing. Screening of potential antiepileptic active ingredients in Rhizoma Gastrodiae based on zebrafish model and metabolomics technology [J]. Shandong Science, 2024, 37(3): 1-9.
[2]	MA Shijing, HE Chunyan, GUAN Tianzhu, YAO Xueshuang, ZHANG Junpeng. Exploration of antihyperlipidemia mechanism of Monopterus albus peptides based on hyperlipidemic zebrafish model and network pharmacology [J]. Shandong Science, 2024, 37(3): 27-38.
[3]	LIU Kechun, WANG Yongcheng, ZANG Xiaohan, XIA Qing, ZHANG Yun, ZHANG Shanshan, SUN Chen. Advancements in network pharmacology and zebrafish modeling for studying traditional Chinese medicine’s effective substances and mechanisms of action [J]. Shandong Science, 2024, 37(2): 29-35.
[4]	XIA Qing, ZANG Xiaohan, WANG Yongcheng, ZHANG Yun, LI Peihai, ZHANG Xuanming, LIU Kechun. Progress and development trends in the use of zebrafish as a model organism for evaluating cosmetic efficacy [J]. Shandong Science, 2024, 37(2): 36-46.
[5]	WU Yonghao, XIE Hebing, HOU Hairong, WANG Rongchun, CHEN Xiqiang. Protective effect of Feirepuqing Powder on hypoxia-induced sports injury in zebrafish model by inhibiting Hif1α [J]. Shandong Science, 2024, 37(1): 32-38.
[6]	ZOU Hong-yuan, ZHU Cheng-yue, ZHANG Shan-shan, KONG Rui-qi, ZHANG Yun, LIU Ke-chun. Protective effect of peony seed oil microcapsule powder on isoniazid-induced developmental toxicity in zebrafish [J]. Shandong Science, 2021, 34(4): 45-51.
[7]	MO Cai-lian, LI Jie, WANG Jia-zhen, LIU Xin , LIN Sheng-hua, MOU Yan-ling, ZHANG Yun, LIU Ke-chun, HE Qiu-xia. Applicability of a zebrafish thrombosis model in screening active ingredients of traditional Chinese medicine [J]. Shandong Science, 2021, 34(4): 52-59.
[8]	LIU Ke-chun, GAO Yan, ZHANG Yun, HE Qiu-xia, HAN Li-wen, LI Ning, JI Xiu-na, SUN Chen. Zebrafish as a model organism for hearing research [J]. Shandong Science, 2019, 32(5): 46-53.
[9]	WANG Rong-chun，HE Qiu-xia，HAN Jian，CHEN Xi-qiang，SUN Chen，WANG Xi-min，HAN Li-wen，LIU Ke-chun. Research on the effects of chloramphenicol on developmental and immune toxicity of juvenile zebrafish [J]. SHANDONG SCIENCE, 2017, 30(6): 35-40.
[10]	LIU Fa-sheng, WANG Rong-chun, GUO Jing-lan, CHEN Yan-li, XIA Qing, KONG Hao-tian, FENG Run-liang, HAN Li-wen, ZHANG Ai-ping, LIU Ke-chun. The immune-regulatory effects of active ingredients of asparagus using zebrafish model [J]. SHANDONG SCIENCE, 2017, 30(6): 29-34.
[11]	PENG Wei-bing, ZHOU Ling-xiao, FU Xian-jun, HE Qiu-xia, WANG Zhen-guo, LIU Ke-chun. Anti-inflammatory research on component combination of Kunhai ginger decoction based on zebrafish model [J]. SHANDONG SCIENCE, 2017, 30(2): 37-42.
[12]	SUN Chen, HE Qiu-xia, HAN Li-wen, CHEN Xi-qiang, WANG Xi-min, CHU Jie, HAN Jian, WANG Rong-chun, HOU Hai-rong, LIU Ke-chun. Toxicity of aspirin to gastrointestinal tract of zebrafish [J]. SHANDONG SCIENCE, 2016, 29(6): 50-55.
[13]	HOU Hai-rong,ZHANG Shan-shan,SUN Chen, CHEN Yu-feng, HE Qiu-xia,LI Xiao-bin, YU Tao, PENG De-jie, HAN Li-wen, LIU Ke-chun. The effect of fructo oligosaccharides and isomaltooligosaccharide on the intestinal peristalsis of zebrafish [J]. SHANDONG SCIENCE, 2016, 29(6): 56-61.
[14]	ZOU Yu,ZHANG Yun,HAN Liwen,HE Qiuxia，LI Zhiping，HOU Hairong，HAN Jian，WANG Ximin，LI Juanjuan，CEN Juan，LIU Kechun. Impact of isoniazid on developmental toxicity and behavior of zebrafish larvae [J]. SHANDONG SCIENCE, 2016, 29(5): 47-53.
[15]	HAN Jian, PENG Weibing, HAN Liwen, HE Qiuxia, ZHANG Yun, SUN Chen, WANG Rongchun, CHU Jie, LIU Kechun. Impact of Cu,Hg and Pb on liver growth of zebrafish larvae [J]. SHANDONG SCIENCE, 2016, 29(5): 54-59.