Study of cynaroside on apoptosis and expression of inflammatory factor in model cells of Alzheimer’s disease

doi:10.11958/20221922

Abstract

Abstract:

Objective To explore the effect and possible mechanism of cynaroside on apoptosis and expression of inflammatory factor in model cells of Alzheimer's disease (AD) by down-regulating mitogen-activated protein kinase kinase kinase 3 (MEKK3). Methods PC12 cells were cultured in vitro and incubated with different doses (6.25 mg/L, 12.5 mg/L, 25 mg/L) of cynaroside for 24 h to establish an AD cell model induced by Aβ_25-35. In addition, MEKK3 small interfering RNA or overexpression vector was transfected into PC12 cells, and Aβ_25-35 induced AD cell model was established after incubation with 25 mg/L luteolin for 24 h. CCK-8 was used to detect inhibition rate of cell proliferation, and flow cytometry was used to detect apoptosis rate. ELISA was used to detect inflammatory factors (TNF-α, IL-6, IL-1β). The protein expression of MEKK3 was detected by Western blot assay. Results Cynaroside could decrease the proliferation inhibition rate, apoptosis rate, inflammatory cytokines (TNF-α, IL-6, IL-1β) and MEKK3 protein expression in PC12 cells induced by Aβ_25-35 (P<0.05). Knockdown of MEKK3 could reduce the proliferation inhibition rate, apoptosis rate and inflammatory cytokines (TNF-α, IL-6, IL-1β) of Aβ_25-35-induced PC12 cells, while overexpression of MEKK3 showed the opposite effect. Upregulation of MEKK3 expression attenuated effects of cynaroside on the proliferation inhibition rate, apoptosis rate and inflammatory factors of Aβ_25-35-induced PC12 cells. Conclusion Cynaroside may inhibit apoptosis and expression of inflammatory factors of AD model cells by down-regulating MEKK3, which has potential value in the treatment of AD.

Key words: Galuteolin, Alzheimer disease, PC12 cells, MAP kinase kinase kinases, apoptosis, inflammatory factor

CLC Number:

R742

Figures/Tables 11

References 16

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组别	增殖抑制率	凋亡率
对照组	0.02±0.01	6.35±0.16
模型组	44.15±1.84^a	23.83±0.88^a
模型+木犀草苷低剂量组	32.36±1.84^b	20.37±0.89^b
模型+木犀草苷中剂量组	24.06±1.28^bc	16.29±1.12^bc
模型+木犀草苷高剂量组	12.45±0.71^bcd	11.93±0.56^bcd
F	1 482.537^＊＊	675.957^＊＊

组别	增殖抑制率	凋亡率
对照组	0.02±0.01	6.35±0.16
模型组	44.15±1.84^a	23.83±0.88^a
模型+木犀草苷低剂量组	32.36±1.84^b	20.37±0.89^b
模型+木犀草苷中剂量组	24.06±1.28^bc	16.29±1.12^bc
模型+木犀草苷高剂量组	12.45±0.71^bcd	11.93±0.56^bcd
F	1 482.537^＊＊	675.957^＊＊

组别	TNF-α/（ng/L）		IL-6/（ng/L）
对照组	100.06±5.79		64.78±4.99
模型组	400.08±14.63^a		442.40±32.12^a
模型+木犀草苷低剂量组	332.88±20.38^b		303.79±15.73^b
模型+木犀草苷中剂量组	244.86±12.78^bc		222.45±14.55^bc
模型+木犀草苷高剂量组	148.91±11.88^bcd		126.42±9.40^bcd
F	721.179^＊＊		621.541^＊＊
组别		IL-1β/（ng/L）		MEKK3
对照组		93.60±5.55		0.15±0.02
模型组		275.42±20.46^a		0.67±0.04^a
模型+木犀草苷低剂量组		235.50±13.86^b		0.51±0.03^b
模型+木犀草苷中剂量组		176.98±12.97^bc		0.39±0.04^bc
模型+木犀草苷高剂量组		125.28±8.96^bcd		0.22±0.02^bcd
F		286.230^＊＊		411.612^＊＊

组别	TNF-α/（ng/L）		IL-6/（ng/L）
对照组	100.06±5.79		64.78±4.99
模型组	400.08±14.63^a		442.40±32.12^a
模型+木犀草苷低剂量组	332.88±20.38^b		303.79±15.73^b
模型+木犀草苷中剂量组	244.86±12.78^bc		222.45±14.55^bc
模型+木犀草苷高剂量组	148.91±11.88^bcd		126.42±9.40^bcd
F	721.179^＊＊		621.541^＊＊
组别		IL-1β/（ng/L）		MEKK3
对照组		93.60±5.55		0.15±0.02
模型组		275.42±20.46^a		0.67±0.04^a
模型+木犀草苷低剂量组		235.50±13.86^b		0.51±0.03^b
模型+木犀草苷中剂量组		176.98±12.97^bc		0.39±0.04^bc
模型+木犀草苷高剂量组		125.28±8.96^bcd		0.22±0.02^bcd
F		286.230^＊＊		411.612^＊＊

组别	增殖抑制率/%	凋亡率/%	TNF-α/（ng/L）	IL-6/（ng/L）	IL-1β/（ng/L）
模型+si-NC组	44.19±1.96	23.65±1.58	399.83±23.91	448.76±20.57	272.70±35.62
模型+si-MEKK3组	14.56±0.83	13.35±0.97	198.54±19.89	144.50±22.08	133.54±10.81
t	41.762^＊＊	16.667^＊＊	19.416^＊＊	30.248^＊＊	11.215^＊＊