醉茄素A对缺血性脑卒中大鼠的神经保护作用及其机制

doi:10.13481/j.1671-587X.20210414

Abstract

Abstract: Objective

To investigate the protective effect of Withaferin A on the neurological function of the rats with ischemic stroke， and to clarify its possible mechanism.

Methods

A total of 75 SD rats were randomly divided into sham operation group， model group， positive control group （nimodipine group，12 mg·kg^－1），low dose of Withaferin A group （25 mg·kg^－1） and high dose of Withaferin A group （100 mg·kg^－1），and there were 15 rats in each group. The rat models of ischemic stroke were established by improved Longa line embolization. After successful modeling， the drugs were administered by gavage once per day for 14 d. Subsequently， the neurological function， the content of water in brain and the volume of cerebral infarction were evaluated. The activities of superoxide dismutase （SOD）， glutathione peroxidase （GSH-PX）， catalase （CAT） and the levels of malondialdehyde （MDA） in brain tissue of the rats were determined by biochemical method. The apoptotic rates of cells of the rats were detected by TUNEL staining， and the mRNA and protein expression levels of nuclear E2-related factors 2 （Nrf2） and heme oxygenase-1 （HO-1） in brain tissue of the rats were respectively detected by RT-qPCR and Western blotting methods.

Results

Compared with sham operation group， the score of neurological function， the volume of cerebral infarction， the content of water in brain tissue， the apoptotic rate of cells and the level of MDA in brain tissue of the rats in model group were significantly increased （P<0.05）， while the activities of SOD， GSH-PX and CAT， as well as the mRNA and protein expression levels of Nrf2 and HO-1 in brain tissue of the rats were significantly decreased （P<0.05）. Compared with model group， the scores of neurological function， the volumes of cerebral infarction， the contents of water in brain tissue， the apoptotic rates of cells and the levels of MDA in brain tissue of the rats in high dose of Withaferin A group and nimodipine group were significantly reduced （P<0.05）， and the activities of SOD， GSH-PX and CAT as well as the mRNA and protein expression levels of Nrf2 and HO-1 in brain tissue of the rats were significantly increased （P<0.05）； However， there were no significant differences in the related indexes in low dose of Withaferin A group （P>0.05）.

Conclusion

Withaferin A can improve the level of antioxidant in the rats with ischemic stroke， reduce the oxidative damage of brain tissue， inhibit the apoptosis of nerve cells of brain tissue， and improve the neurological function of the rats； its mechanism may be related to the activation of Nrf2/HO-1 signaling pathway.

Key words: ischemic stroke, withaferin A, oxidative stress, nuclear factor erythroid-2 related factor 2/heme oxygenase-1 signaling pathway

CLC Number:

R285.5

Dimi ZHOU,Lu GAN,Lin CHEN,Chengfang ZHOU,Liang ZENG. Neuroprotective effect of Withaferin A on ischemic stroke rats and its mechanism[J].Journal of Jilin University(Medicine Edition), 2021, 47(4): 919-925.

Figures/Tables 6

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Fig. 2

Tab.4

References 20

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Gene	Primer sequence
Nrf2	F：5'-GGACCTAAAGCACAGCCAACACAT-3'
Nrf2	R：5'-TCGGCTTGAATGTTTGTCTTTTGTG-3'
HO-1	F：5'-CTTTTTTCACCTTCCCGAGCATC-3'
HO-1	R：5'-GGTCTTAGCCTCTTCTGTCACCCTGT-3'
GAPDH	F：5'-TGGAGTCTACTGGCGTCTT-3'
GAPDH	R：5'-TGTCATATTTCTCGTGGTTCA-3'

Group	Neurological score	Water content in brain tissue (η/%)	Volume of cerebral infarction（V/ mm³）
Sham operation	0.00±0.00	75.42±1.70	0.00±0.00
Model	2.89±0.53^*	83.87±2.00^*	37.55±3.60^*
Nimodipine	1.33±0.42^△	78.23±1.65^△	16.09±2.07^△
Withaferin A
Low dose	2.76±0.50	82.69±1.54	35.01±4.36
High dose	1.76±0.44^△	79.58±1.43^△	24.85±2.44^△

Group	SOD[λ_B/（kU·L^－1）]	GSH-Px[ρ_B/（mg·L^－1）]	CAT[λ_B/（kU·L^－1）]	MDA[c_B/（μmol·L^－1）]
Sham operation	29.58±1.49	18.48±1.22	2.39±0.23	33.50±2.27
Model	19.47±1.00^*	7.29±0.45^*	1.02±0.10^*	48.94±2.06^*
Nimodipine	33.56±1.23^△	22.46±1.10^△	2.51±0.32^△	39.55±1.96^△
Withaferin A
Low dose	20.30±1.13	8.60±0.60	1.13±0.11	47.54±2.06
High dose	30.22±1.54^△	20.77±1.59^△	2.46±0.23^△	42.88±2.45^△

Group	Nrf2		HO-1
Group	mRNA	Protein	mRNA	Protein
Sham operation	1.00±0.01	0.95±0.03	1.00±0.03	0.60±0.06
Model	0.18±0.06^*	0.12±0.03^*	0.27±0.04^*	0.05±0.02^*
Nimodipine	3.16±0.47^△	1.35±0.06^△	1.82±0.20^△	0.73±0.08^△
Withaferin A
Low dose	0.21±0.07	0.16±0.03	0.35±0.06	0.12±0.03
High dose	5.20±0.60^△	1.55±0.08^△	2.99±0.32^△	0.93±0.04^△

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Neuroprotective effect of Withaferin A on ischemic stroke rats and its mechanism

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