Jiaotaiwan improves brain glucose metabolism in a mouse model of Alzheimer's disease by activating the PI3K/AKT signaling pathway

doi:10.12122/j.issn.1673-4254.2024.05.11

Abstract

Abstract:

Objective To investigate the effect of Jiaotaiwan on brain insulin-PI3K/AKT pathway in a mouse model of Alzheimer's disease (AD). Methods Fifty 3-month-old male APP/PS1 double transgenic mice were randomized into AD model group, low-, medium- and high-dose Jiaotaiwan treatment groups, and donepezil treatment group. Cognitive functions of the mice were assessed using water maze and open field tests, and neuronal pathologies were observed with HE staining and Nissl staining; immunohistochemistry was used to detect amyloid Aβ deposition in the brain. Fasting serum insulin levels of the mice were measured, and the expressions of Aβ₄₂, insulin-PI3K/AKT pathway components and downstream glucose transporters in the brain tissue were detected with RT-qPCR and Western blotting. Results The AD mouse models exhibited obvious impairment of learning and memory abilities, significantly reduced hippocampal neurons, and obvious Aβ amyloid plaques in the brain tissue with increased Aβ₄₂ protein expression (P<0.05) and insulin resistance index, decreased hippocampal PI3K expressions, lowered expressions of AKT and InR, reduced expressions of GLUT1, GLUT3, and GLUT4, and increased expression of GSK3β in both the hippocampus and cortex. Treatment with Jiaotaiwan and donepezil both effectively improved memory ability of the mouse models, increased the number of hippocampal neurons, reduced Aβ amyloid plaques and increased the expressions of PI3K, AKT, InR, GLUT1, GLUT3 and GLUT4 in the hippocampus and cortex. Conclusion Jiaotaiwan improves learning and memory abilities of APP/PS1 double transgenic mice and delay the development of AD by activating the PI3K/AKT pathway and regulating the expression levels of its downstream GLUTs in the brain.

Key words: Alzheimer's disease, Jiaotaiwan, glucose metabolism, insulin PI3K/AKT pathway, glucose transporter proteins

Yan WANG, Yuqing RUAN, Can CUI, Xiu WANG. Jiaotaiwan improves brain glucose metabolism in a mouse model of Alzheimer's disease by activating the PI3K/AKT signaling pathway[J]. Journal of Southern Medical University, 2024, 44(5): 894-903.

Figures/Tables 7

Fig.1 Results of Morris water maze test and open field test. A: Comparison of escape latency of the mice among the groups (Mean±SD, n=8). B: Swimming trajectories during escape latency in each group. C: Open field behavior trajectories of the mice in each group. ***P<0.001 vs Control group; ##P<0.01, ###P<0.001 vs model group.

Tab.1 Results of water maze spatial exploration test and open field test in each group of mice (Mean±SD, n=8)

Group	Times of platform crossing	Time of first platform crossing (s)	Time in platform quadrant (s)	Number of central area entry	Time in the central zone (s)
Control	4.25±0.83	24.17±2.72	31.68±2.56	14.88±1.45	32.91±1.78
Model	1.63±0.70***	37.69±2.65***	20.11±1.99***	6.25±1.85***	16.21±1.28***
JTWL	3.13±1.05^#	32.99±2.25^##	23.21±2.81	9.00±1.41^#	18.10±2.56
JTWM	4.00±0.71^###	28.12±1.05^###	26.89±3.00^###	12.25±1.85^###	20.91±1.99^###
JTWH	3.25±0.83^#	31.27±2.02^###	25.52±2.25^##	10.63±1.58^###	19.77±2.02^#
DON	3.38±0.99^##	33.71±1.41^#	21.01±2.16	9.75±1.98^##	19.9±1.66^#

Fig.2 Effect of Jiaotaiwan on hippocampal neuron morphology in each group. A: Representative HE staining results of the hippocampus in each group. B: Nissl staining of the hippocampus in each group (scale bar=100 μm).

Fig.3 Impact of Jiaotaiwan on aggregation of Aβ plaques and Aβ42 level in the hippocampus and cortex of the mice (Mean±SD, n=4). A: Immunohistochemistry showing Aβ plaques in the hippocampus and cortex (scale bar=100 μm. B: Quantification of Aβ42 protein expressions in the hippocampus and cortex using Western blotting. *P<0.05,***P<0.001 vs Control group; ##P<0.01,###P<0.001 vs Model group.

Tab.2 Effects of Jiaotaiwan on insulin resistance of the mice in each group (Mean±SD)

Group	Fasting glucose (mmol/L)	Fasting insulin (mIU/L)	HOMA-IR
Control	5.76±0.57	5.54±0.90	1.40±0.17
Model	11.68±1.34***	16.41±1.43***	8.52±1.24***
JTWL	8.98±2.44	13.98±1.16	5.57±1.53^###
JTWM	7.51±1.41^##	12.00±1.61^###	3.97±0.72^###
JTWH	9.60±3.71	13.43±3.07^#	5.29±1.12^###
DON	7.40±1.34^##	12.85±1.25^###	4.19±0.69^###

Fig.4 Effects of Jiaotaiwan on insulin-PI3K/AKT pathway in the hippocampus and cortex of the mice in each group. A: Western blots of the proteins in the insulin-PI3K/AKT pathway in the hippocampus and cortex in each group. B: Comparison of PI3K, AKT, GSK3β, and InR protein and mRNA expressions in the hippocampus and cortex among the groups (Mean±SD).*P<0.05,**P<0.01,***P<0.001 vs Control group; #P<0.05,##P<0.01,###P<0.001 vs Model group.

Fig.5 Effects of Jiaotaiwan on glucose transport in the hippocampus and cortex of the mice in each group. A: Western blots of the proteins involved in glucose transport in the hippocampus and cortex in each group. B: Expression levels of GLUT1, GLUT3, and GLUT4 proteins and mRNAs in the hippocampus and cortex in each group (Mean±SD).*P<0.05, **P<0.01, ***P<0.001 vs Control group; #P<0.05, ##P<0.01, ###P<0.001 vs Model group.

References 36

1	Kellar D, Craft S. Brain insulin resistance in Alzheimer's disease and related disorders: mechanisms and therapeutic approaches[J]. Lancet Neurol, 2020, 19(9): 758-66. DOI: 10.1016/s1474-4422(20)30231-3
2	Ou ZH, Deng LL, Lu Z, et al. Protective effects of Akkermansia muciniphila on cognitive deficits and amyloid pathology in a mouse model of Alzheimer’s disease[J]. Nutr Diabetes, 2020, 10(1): 12. DOI: 10.1038/s41387-020-0115-8
3	Macklin L, Griffith CM, Cai Y, et al. Glucose tolerance and insulin sensitivity are impaired in APP/PS1 transgenic mice prior to amyloid plaque pathogenesis and cognitive decline[J]. Exp Gerontol, 2017, 88: 9-18. DOI: 10.1016/j.exger.2016.12.019
4	Teipel SJ, Drzezga A, Bartenstein P, et al. Effects of donepezil on cortical metabolic response to activation during (18)FDG-PET in Alzheimer's disease: a double-blind cross-over trial[J]. Psychopharmacology, 2006, 187(1): 86-94. DOI: 10.1007/s00213-006-0408-1
5	Chen F, He YK, Wang PW, et al. Banxia Xiexin Decoction ameliorated cognition via the regulation of insulin pathways and glucose transporters in the hippocampus of APPswe/PS1dE9 mice[J]. Int J Immunopathol Pharmacol, 2018, 32: 2058738418780066. DOI: 10.1177/2058738418780066
6	袁琳, 李慧姣, 胡娜, 等. 交泰丸不同配比组方降糖作用及相关机制探讨[J]. 中国实验方剂学杂志, 2017, 23(8): 130-7.
7	Boccardi V, Murasecco I, Mecocci P. Diabetes drugs in the fight against Alzheimer's disease[J]. Ageing Res Rev, 2019, 54: 100936. DOI: 10.1016/j.arr.2019.100936
8	Blázquez E, Hurtado-Carneiro V, LeBaut-Ayuso Y, et al. Significance of brain glucose hypometabolism, altered insulin signal transduction, and insulin resistance in several neurological diseases[J]. Front Endocrinol, 2022, 13: 873301. DOI: 10.3389/fendo.2022.873301
9	Yan F, Liu J, Chen MX, et al. Icariin ameliorates memory deficits through regulating brain insulin signaling and glucose transporters in 3 × Tg-AD mice[J]. Neural Regen Res, 2023, 18(1): 183-8. DOI: 10.4103/1673-5374.344840
10	Davidy T, Yore I, Cukierman-Yaffe T, et al. A feasibility study of the combination of intranasal insulin with dulaglutide for cognition in older adults with metabolic syndrome at high dementia risk‑Study rationale and design[J]. Mech Ageing Dev, 2023, 213: 111825. DOI: 10.1016/j.mad.2024.111937
11	张韶君, 高江琴, 邱贵娟, 等. 黄连素通过调节磷脂酰肌醇-3-激酶/丝氨酸/苏氨酸激酶/葡萄糖转运蛋白4信号途径改善胰岛素抵抗的机制研究[J]. 中国药物与临床, 2017, 17(11): 1569-71.
12	Gomaa AA, Makboul RM, El-Mokhtar MA, et al. Evaluation of the neuroprotective effect of donepezil in type 2 diabetic rats[J]. Fundam Clin Pharmacol, 2021, 35(1): 97-112. DOI: 10.1111/fcp.12585
13	Hugon G, Goutal S, Sarazin M, et al. Impact of donepezil on brain glucose metabolism assessed using[¹⁸F]₂-fluoro-2-deoxy-D-glucose positron emission tomography imaging in a mouse model of Alzheimer’s disease induced by intracerebroventricular injection of amyloid-beta peptide[J]. Front Neurosci, 2022, 16: 835577. DOI: 10.3389/fnins.2022.835577
14	Shimada A, Hashimoto H, Kawabe J, et al. Evaluation of therapeutic response to donepezil by positron emission tomography[J]. Osaka City Med J, 2011, 57(1): 11-9.
15	邓晓威, 谢宁. 黄连素治疗2型糖尿病研究进展[J]. 中国中药杂志, 2014, 39(8): 1374-8.
16	Huang WY, Dong H. Coptidis rhizoma-contained traditional formulae for insomnia: a potential to prevent diabetes?[J]. Chin J Integr Med, 2018, 24(10): 785-8. DOI: 10.1007/s11655-018-3012-4
17	宋宗辉, 张艺雯, 王玲洁, 等. 肉桂醛的药理活性及其研究进展[J]. 解放军药学学报, 2018, 34(6): 550-4.
18	卫克昭, 姚平安, 刘晓宁, 等. 肉桂对糖尿病性心肌病大鼠的心脏保护作用[J]. 上海中医药杂志, 2018, 52(7): 69-74.
19	廖作庄, 徐灵源, 王金妮, 等. 肉桂多酚对链脲佐菌素致糖尿病小鼠的保护作用[J]. 西安交通大学学报: 医学版, 2019, 40(1): 162-6.
20	王雪萍, 张皓月, 王婷, 等. 交泰丸中小檗碱联合cinnamtannin D1的降血糖作用[J]. 中成药, 2018, 40(12): 2613-8.
21	Nguyen TT, Ta QTH, Nguyen TKO, et al. Type 3 diabetes and its role implications in Alzheimer's disease[J]. Int J Mol Sci, 2020, 21(9): 3165. DOI: 10.3390/ijms21093165
22	Suresh J, Khor IW, Kaur P, et al. Shared signaling pathways in Alzheimer's and metabolic disease may point to new treatment approaches[J]. FEBS J, 2021, 288(12): 3855-73. DOI: 10.1111/febs.15540
23	Sun YN, Ma C, Sun H, et al. Metabolism: a novel shared link between diabetes mellitus and Alzheimer's disease[J]. J Diabetes Res, 2020, 2020: 4981814. DOI: 10.1155/2020/4981814
24	Alves SS, Servilha-Menezes G, Rossi L, et al. Evidence of disturbed insulin signaling in animal models of Alzheimer's disease[J]. Neurosci Biobehav Rev, 2023, 152: 105326. DOI: 10.1016/j.neubiorev.2023.105326
25	Kim B, Elzinga SE, Henn RE, et al. The effects of insulin and insulin-like growth factor I on amyloid precursor protein phosphorylation in in vitro and in vivo models of Alzheimer's disease[J]. Neurobiol Dis, 2019, 132: 104541. DOI: 10.1016/j.nbd.2019.104541
26	Zhang N, Liu XY, Zhuang LL, et al. Berberine decreases insulin resistance in a PCOS rats by improving GLUT4: dual regulation of the PI3K/AKT and MAPK pathways[J]. Regul Toxicol Pharmacol, 2020, 110: 104544. DOI: 10.1016/j.yrtph.2019.104544
27	Akhtar A, Sah SP. Insulin signaling pathway and related molecules: role in neurodegeneration and Alzheimer's disease[J]. Neurochem Int, 2020, 135: 104707. DOI: 10.1016/j.neuint.2020.104707
28	Wang L, Li JJ, Di LJ. Glycogen synthesis and beyond, a comprehensive review of GSK3 as a key regulator of metabolic pathways and a therapeutic target for treating metabolic diseases[J]. Med Res Rev, 2022, 42(2): 946-82. DOI: 10.1002/med.21867
29	Zhang F, Zhang ZL, Kong DS, et al. Tetramethylpyrazine reduces glucose and insulin-induced activation of hepatic stellate cells by inhibiting insulin receptor-mediated PI3K/AKT and ERK pathways[J]. Mol Cell Endocrinol, 2014, 382(1): 197-204. DOI: 10.1016/j.mce.2013.09.020
30	Wadhwa P, Jain P, Jadhav HR. Glycogen synthase kinase 3 (GSK3): its role and inhibitors[J]. Curr Top Med Chem, 2020, 20(17): 1522-34. DOI: 10.2174/1568026620666200516153136
31	Xu AP, Zeng QT, Tang YS, et al. Electroacupuncture protects cognition by regulating tau phosphorylation and glucose metabolism via the AKT/GSK3β signaling pathway in Alzheimer's disease model mice[J]. Front Neurosci, 2020, 14: 585476. DOI: 10.3389/fnins.2020.585476
32	Yang W, Liu Y, Xu QQ, et al. Sulforaphene ameliorates neuroinflammation and hyperphosphorylated tau protein via regulating the PI3K/akt/GSK-3β pathway in experimental models of Alzheimer's disease[J]. Oxid Med Cell Longev, 2020, 2020: 4754195. DOI: 10.1155/2020/4754195
33	Biasibetti R, Almeida Dos Santos JP, Rodrigues L, et al. Hippocampal changes in STZ-model of Alzheimer's disease are dependent on sex[J]. Behav Brain Res, 2017, 316: 205-14. DOI: 10.1016/j.bbr.2016.08.057
34	李娟娥, 姜小帆. 交泰丸对糖尿病小鼠认知功能障碍的影响及机制[J]. 中国实验方剂学杂志, 2019, 25(17): 23-7.
35	王帅, 金磊, 海春旭, 等. PI3K/Akt信号通路在胰岛素抵抗中作用的研究进展[J]. 毒理学杂志, 2015, 29(4): 313-6.
36	Agrawal R, Vieira-de-Abreu A, Durupt G, et al. Insulin regulates GLUT4 in the ventromedial hypothalamus to restore the sympathoadrenal response to hypoglycemia in diabetic rats[J]. Am J Physiol Endocrinol Metab, 2018, 315(6): E1286-95. DOI: 10.1152/ajpendo.00324.2018

[1]	Jingjing YANG, Lixia YIN, Ting DUAN, Minzhu NIU, Zhendong HE, Xinrui CHEN, Xiaofeng ZHANG, Jing LI, Zhijun GENG, Lugen ZUO. High expression of ATP5A1 in gastric carcinoma is correlated with a poor prognosis and enhanced glucose metabolism in tumor cells [J]. Journal of Southern Medical University, 2024, 44(5): 974-980.
[2]	. Research progress in the role and mechanism of polysaccharides in regulating glucose and lipid metabolism [J]. Journal of Southern Medical University, 2021, 41(3): 471-474.
[3]	. Correlation of type 2 diabetes and impaired glucose regulation with chronic kidney disease in middle-aged and elderly individuals [J]. Journal of Southern Medical University, 2020, 40(10): 1457-1464.
[4]	. Effects of chronic intermittent hypoxia on glucose transporter 4 expression in rat skeletal muscles [J]. Journal of Southern Medical University, 2014, 34(07): 1061-.
[5]	. Glucometabolic state in hypertensive and normotensive patients: identifying candidates for oral glucose tolerance tests [J]. Journal of Southern Medical University, 2013, 33(01): 108-.
[6]	CHEN Jing1,HUANG Yong1,WANG Sheng-xu1,LI Qiu-shi1,LIANG Yong-jiu2,GUO Yong-ning3 1Department of Traditional Chinese Medicine,Southern Medical University,Guangzhou 510515,China;2Medical Department,Millennium Rest home,Guangzhou 510407,China;3Medical Department,Xingguang Rest home,Guangzhou 510407,China. ～(18)FDG PET cerebral function imaging in 10 vascular dementia patients receiving needling at Baihui(DU20),Shuigou(DU26) and Shenmen(HT7) [J]. Journal of Southern Medical University, 2006, 26(05): 610-612.
[7]	HUANG Yong¹, CHEN Jing¹, LAI Xin-sheng², TANG An-wu³, LI Dong-jiang³. Effects of needling in Baihui (DU20),Shuigou (DU26) and Shenmen (HT7) on glucose metabolism in the lentiform nuclus in patients with vascular dementia [J]. Journal of Southern Medical University, 2005, 25(11): 1405-1407.