Magnolol attenuates the locomotor impairment, cognitive deficit, and neuroinflammation in Alzheimer’s disease mice with brain insulin resistance via up-regulating miR-200c

Figures & data

Table 1. All antibodies information and sources in Western blot in this study

ID	Catalog number	Company (country)	Molecular weight	Dilution ratio
TNF-α	ab215188	Abcam (Cambridge, UK)	26 kDa	1/1000
IL-6	ab229381	Abcam (Cambridge, UK)	24 kDa	1/1000
CRP	ab259862	Abcam (Cambridge, UK)	25 kDa	1/100
GAPDH	ab9485	Abcam (Cambridge, UK)	37 kDa	1/2500
Rabbit IgG	ab205718	Abcam (Cambridge, UK)		1/2000

Figure 1. Magnolol ameliorated the impairment of locomotor activity and cognitive function, and regulated the levels of oxidative-related factors in AD model mice with brain insulin resistance. (a-b) OFT was performed to measure the locomotor activity of the mice. (c) NOR evaluation was conducted to measure the discrimination ability of the mice. (d-f) MWM test was used to evaluate the spatial learning and memory ability of the mice. (g-i) The secretions of MDA (g), GSH (h), and SOD (i) in hippocampus tissues of the mice were detected by ELISA. (*P < 0.05, ***P < 0.001, vs. Control; ^{^}P < 0.05, ^{^^}P < 0.01, ^{^^^}P < 0.001, vs. Model). (AD: Alzheimer’s disease, OFT: open-field test, NOR: novel object recognition, MWM: morris water maze, MDA: malondialdehyde, GSH: glutathione, SOD: superoxide dismutase).

Figure 2. Magnolol down-regulated the levels of inflammation-related factors and up-regulated the level of miR-200c in AD model mice with brain insulin resistance. (a-c) The secretions of TNF-α (a), IL-6 (b), and CRP (c) in hippocampus tissues of the mouse were detected by ELISA. (d-e) The expressions of TNF-α, IL-6, and CRP in hippocampus tissues of the mice were detected by Western blot. GAPDH was used as an internal control. (f) The expression of miR-200c in hippocampus tissues of the mouse was detected by RT-qPCR, and U6 was used as an internal control. (***P < 0.001, vs. Control; ^{^}P < 0.05, ^{^^}P < 0.01, ^{^^^}P < 0.001, vs. Model). (AD: Alzheimer’s disease, TNF-α: tumor necrosis factor-α, IL-6: interleukin-6, CRP: C-reactive protein).

Figure 3. miR-200c antagomiR offset the effects of Magnolol on locomotor activity and cognitive function and levels of oxidative-related factors in AD model mice with brain insulin resistance. (a) The expressions of miR-200c in hippocampus tissues of the mice were quantified by RT-qPCR. U6 was used as an internal control. (b-c) OFT was performed to measure the locomotor ability of the mice. (d) NOR evaluation was employed to measure the discrimination ability of the mice. (e-g) MWM test was conducted to evaluate the spatial learning and memory ability of the mice. (h-j) The secretions of MDA (h), GSH (i), and SOD (j) in hippocampus tissues of the mice were detected by ELISA. (*P < 0.001, **P < 0.001, ***P < 0.001, vs. AntagomiR-NC; ^{^}P < 0.05, ^{^^^}P < 0.001, vs. AntagomiR; ^#P < 0.05, ^##P < 0.01, ^###P < 0.001, vs. AntagomiR-NC+ Magnolol 10; ^&P < 0.05, ^&&P < 0.01, ^&&&P < 0.001, vs. AntagomiR-NC+ Magnolol 15). (AD: Alzheimer’s disease, OFT: open-field test, NOR: novel object recognition, MWM: morris water maze, MDA: malondialdehyde, GSH: glutathione, SOD: superoxide dismutase, PCO: protein carbonylation).

Figure 4. miR-200c antagomiR offset the effect of Magnolol on the levels of inflammation-related factors in AD model mice with brain insulin resistance. (a-c) The secretions of TNF-α (a), IL-6 (b), and CRP (c) in hippocampus tissues of the mice were detected by ELISA. (d-e) The expressions of TNF-α, IL-6, and CRP in hippocampus tissues of the mice were detected by Western blot. GAPDH was used as an internal control. (*P < 0.001, ***P < 0.001, vs. AntagomiR-NC; ^{^^}P < 0.05, ^{^^^}P < 0.001, vs. AntagomiR; ^##P < 0.01, ^###P < 0.001, vs. AntagomiR-NC+ Magnolol 10; ^&&&P < 0.001, vs. AntagomiR-NC+ Magnolol 15). (AD: Alzheimer’s disease, TNF-α: tumor necrosis factor-α, IL-6: interleukin-6, CRP: C-reactive protein).