The Chinese herbal formula Fuzheng Quxie Decoction attenuates cognitive impairment and protects cerebrovascular function in SAMP8 mice

Figures & data

Figure 1 The position of platform and entry points in the MWM test.

Abbreviation: MWM, Morris water maze.

Table 1 Active compounds in FQD

	OB (%)	DL	BBBp	Source	Target number
R-canadine	55.36687	0.77465	0.57189	Screened	56
Alexandrine_qta	36.913906	0.75268	0.87952	Screened	28
Aposcopolamine	66.646907	0.21999	0.40163	Screened	8
Arachidonate	45.57325	0.20491	0.57974	Screened	56
Berberine	36.86125	0.77665	0.56718	Screened	52
Berberrubine	35.73551	0.7269	0.17408	Screened	40
Berlambine	36.6809	0.81596	0.17347	Screened	47
β-sitosterol	36.913906	0.75123	0.98588	Screened	72
Celabenzine	101.8826	0.48772	0.05084	Screened	11
Deoxyharringtonine	39.27444	0.8116	−0.24531	Screened	2
Diop	43.593325	0.39247	0.26014	Screened	8
Epiberberine	43.09233	0.7761	0.39823	Screened	35
Ferulic acid	39.56	0.06	−0.03	Literature^Citation77^–^Citation79	56
Frutinone A	65.903731	0.34184	0.4604	Screened	27
Fumarine	59.262505	0.82694	−0.13192	Screened	46
Ginsenoside Rb1	6.24	0.04	−5.6	Literature^Citation80^–^Citation84	7
Ginsenoside Re	4.27	0.12	−4.39	Literature^Citation85^–^Citation87	4
Ginsenoside Rg1	10.04	0.28	−3.5	Literature^Citation88^–^Citation90	5
Ginsenoside Rg5_qt	39.563071	0.78506	0.21243	Screened	0
Ginsenoside Rh4_qt	31.112148	0.77829	−0.18125	Screened	5
Girinimbine	61.215303	0.31484	1.22221	Screened	23
Gomisin B	31.990424	0.82858	0.17993	Screened	15
Inermin	65.830931	0.53754	0.36121	Screened	35
Ligustilide	23.5	0.07	1.2	Literature^Citation91^–^Citation93	6
Ligustrazine	20.01	0.03	1.05	Literature^Citation94^–^Citation96	6
Magnograndiolide	63.70888	0.18833	−0.24386	Screened	7
Malkangunin	57.713844	0.62642	−0.17138	Screened	5
Mandenol	41.9962	0.19321	1.13821	Screened	8
Myricanone	40.597566	0.51262	−0.07932	Screened	25
Palmatine	64.60111	0.64524	0.37072	Screened	45
Panaxadiol	33.087961	0.79404	0.22661	Screened	3
Perlolyrine	65.947753	0.2747	0.1464	Screened	12
Senkyunone	47.663947	0.24435	0.5013	Screened	6
Stigmasterol	43.829852	0.75665	1.00045	Screened	61
Suchilactone	57.518824	0.55573	0.27669	Screened	32
Wallichilide	42.310679	0.70639	0.73257	Screened	7

Note:

a Compounds with the suffix “_qt” refer to the deglycosylated form of their prototypes.

Abbreviations: BBBp, blood–brain barrier permeability; DL, drug likeness; FQD, Fuzheng Quxie Decoction; OB, oral bioavailability.

Table 2 Shared genes of putative FQD targets and AD-related genes (DRTs)

Target/protein name	UniProt ID	Gene
ATP-binding cassette subfamily A member 1	ABCA1_HUMAN	ABCA1
Acetylcholinesterase	ACES_HUMAN	ACHE
Apolipoprotein E	APOE_HUMAN	APOE
Cholinesterase	CHLE_HUMAN	BCHE
Brain-derived neurotrophic factor	BDNF_HUMAN	BDNF
Voltage-dependent L-type calcium channel subunit α1S	CAC1S_HUMAN	CACNA1S
Calmodulin 1	CALM_HUMAN	CALM1
Caspase 3	CASP3_HUMAN	CASP3
C-C chemokine receptor type 2	CCR2_HUMAN	CCR2
Choline O-acetyltransferase	CLAT_HUMAN	CHAT
Neuronal acetylcholine receptor protein, α7 chain	ACHA7_HUMAN	CHRNA7
Cytochrome C, somatic	CYC_HUMAN	CYCS
Cytochrome P450 2D6	CP2D6_HUMAN	CYP2D6
24-Dehydrocholesterol reductase	DHC24_HUMAN	DHCR24
Dual-specificity tyrosine-phosphorylation-regulated kinase 1A	DYR1A_HUMAN	DYRK1A
Glycogen synthase kinase 3β	GSK3B_HUMAN	GSK3B
Heme oxygenase 1	HMOX1_HUMAN	HMOX1
5-Hydroxytryptamine 2A receptor	5HT2A_HUMAN	HTR2A
Low-density-lipoprotein receptor	LDLR_HUMAN	LDLR
Low-density-lipoprotein receptor-related protein 8	LRP8_HUMAN	LRP8
Amine oxidase (flavin-containing) A	AOFB_HUMAN	MAOB
Microtubule-associated protein 2	MTAP2_HUMAN	MAP2
Mitogen-activated protein kinase 1	MK01_HUMAN	MAPK1
Mitogen-activated protein kinase 3	MK03_HUMAN	MAPK3
Microtubule-associated protein tau	TAU_HUMAN	MAPT
Nitric oxide synthase, inducible	NOS2_HUMAN	NOS2
Nitric oxide synthase, endothelial	NOS3_HUMAN	NOS3
Urokinase-type plasminogen activator	UROK_HUMAN	PLAU
Prostaglandin G/H synthase 2	PGH2_HUMAN	PTGS2
Sodium-dependent serotonin transporter	SC6A4_HUMAN	SLC6A4
Tumor necrosis factor	TNFA_HUMAN	TNF
Very-low-density-lipoprotein receptor	VLDLR_HUMAN	VLDLR

Abbreviations: FQD, Fuzheng Quxie Decoction; DRTs, direct regulatory targets.

Figure 2 Intersections of putative FQD targets and AD-related genes; 32 shared genes were obtained, which were defined as DRTs.

Abbreviations: AD, Alzheimer’s disease; DRTs, direct regulatory targets; FQD, Fuzheng Quxie Decoction.

Figure 3 PPI network of DRTs and PRTs.

Notes: (A) PPI network with 32 DRTs as seeds. Pink nodes represent the 32 DRTs, blue nodes represent the proteins that have been reported to have interactions with the DRTs, and yellow nodes represent 245 PRTs with a DC value greater than twice the average. (B) PPI network of the 245 chosen PRTs, with nine DRTs (pink nodes).
Abbreviations: DC, degree centrality; DRTs, direct regulatory targets; PPI, protein–protein interaction; PRTs, possible regulatory targets.

Figure 4 GO enrichment of 245 PRTs. Most significantly enriched terms are displayed.

Abbreviations: GO, gene ontology; PRTs, possible regulatory targets.

Figure 5 The top-ten enriched pathways of the 245 PRTs analyzed by FunRich 3.0.

Abbreviations: PRTs, possible regulatory targets; VEGF, vascular endothelial growth factor; VEGFR, VEGF-receptor.

Figure 6 Effect of Fuzheng Quxie Decoction (FQD) on spatial learning and memory ability of SAMP8 mice.

Notes: (A) Ratio of time in platform quadrant to total swimming time; (B) ratio of swimming distance in platform quadrant to total swimming distance. (^▲▲P<0.01 vs control group; *P<0.05, **P<0.01 vs model group; n=10).

Figure 7 Effect of Fuzheng Quxie Decoction (FQD) on memory-retention ability of SAMP8 mice.

Notes: (A) Avoidance latency that mice spent to come into dark compartment for the first time; (B) error times in the dark compartment. (^▲P<0.05, ^▲▲P<0.01 vs control group; *P<0.05, **P<0.01 vs model group; n=10).

Figure 8 Effect of Fuzheng Quxie Decoction (FQD) on cerebrovascular morphology in the cortex of SAMP8 mice.

Note: H&E staining, magnification 100× and 400×.

Figure 9 Effect of Fuzheng Quxie Decoction (FQD) on neuronal morphology in hippocampus CA1 region of SAMP8 mice.

Note: Nissl staining, magnification 200×.

Figure 10 Effect of Fuzheng Quxie Decoction (FQD) on VEGF expression in brains of SAMP8 mice.

Notes: (A) VEGF-positive cells in cortex by immunohistochemistry (magnification 400×); (B) quantification of VEGF-positive cells in cortex; (C) VEGF bands detected by Western blot analysis; (D) quantification of VEGF expression, normalized as ratio to β-actin. ^▲▲P<0.01 vs control group; *P<0.05, **P<0.01 vs model group (B and D).

Figure 11 Effect of Fuzheng Quxie Decoction (FQD) on hypoxia-induced factor 1α (HIF1α) expression in brains of SAMP8 mice.

Notes: (A) HIF1α bands detected by Western blot analysis; (B) shows the quantification of HIF1α expression normalized as the ratio to β-actin. (^▲P<0.05 vs control group; *P<0.05, **P<0.01 vs model group; n=5).

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