Study on edaravone pretreatment prolonging thrombolytic time window by acute ischemic stroke and mechanism of ROS/TXNIP/NLRP3 pathway in rat

doi:10.3877/cma.j.issn.2095-123X.2023.02.001

Abstract

Abstract:

Objective

To investigate whether edaravone pretreatment can prolong the thrombolytic time window of acute ischemic stroke (AIS) rat model and its mechanism.

Methods

Sixty-four healthy SD rats were randomly divided into sham operation group, AIS group, recombinant tissue plasminogen activator (rt-PA) treatment group and rt-PA+edaravone treatment group. The AIS model was constructed by autologous thrombosis. The neurological function of rats in each group was evaluated after AIS, and the volume of cerebral infarction was measured by TTC method; Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling was used to observe the changes of cell apoptosis in rats; The levels of ROS and superoxide dismutase (SOD) in brain were detected by enzyme-linked immunosorbent assay; The content of malondialdehyde (MDA) in brain was detected by thiobarbituric acid; The content of hemoglobin in brain was detected by colorimetry; The content of hemoglobin in brain was measured by hemoglobin assay; The protein expression level of ASC, TXNIP, Caspase-1, IL-18 and IL-1β related to ROS/TXNIP/NLRP3 pathway were detected by Western blot.

Results

The Longa score of the rt-PA+edaravone treatment group was lower than that in the rt-PA treatment group after intravenous thrombolysis at 6 and 7 h, and the improvement of cerebral edema degree and cerebral infarction volume at different intervention time points were better than those in the rt-PA treatment group (P<0.05). The apoptosis rate was significantly lower than that in the AIS group and rt-PA treatment group, with a statistically significant difference (P<0.05). Compared with the rt-PA treatment group, the levels of ROS protein, MDA and hemoglobin of the rt-PA+edaravone treatment group after intravenous thrombolysis at 7 h were significantly reduced, and the SOD level after intravenous thrombolysis at 7 and 8 h were higher (P<0.05). The protein expression level of NLRP3, ASC, IL-18 and IL-1β of the rt-PA treatment group and rt-PA+edaravone treatment group after intravenous thrombolysis at 4.5 h were lower than those of AIS group, and the protein expression level of ASC, TXNIP, Caspase-1, IL-1 of rt-PA+edaravone treatment group after intravenous thrombolysis at 6 and 7 h were decreased (P<0.05). Compared with the rt-PA treatment group, the protein expression level of NLRP3 after intravenous thrombolysis at 6 h, the protein expression levels of ASC and TXNIP after intravenous thrombolysis at 7 h, and the protein expression level of IL-18 after intravenous thrombolysis at 7 and 8 h of the rt-PA+edaravone treatment group were decreased, with statistical significance (P<0.05).

Conclusion

Edaravone pretreatment can appropriately prolong the thrombolytic time window of rt-PA treatment in AIS rats, and its mechanism may be related to the participation of ROS/TXNIP/NLRP3 pathway.

Key words: Edaravone, Acute ischemic stroke, Rats, Cerebral thromboembolism, Reactive oxygen radical

Yuzhou Long, Hua Liu, Yunqian Zhang, Xingtong Li, Yunhu Fan, Zhengliang Shang, Zhenyu Song, Lihua Luo. Study on edaravone pretreatment prolonging thrombolytic time window by acute ischemic stroke and mechanism of ROS/TXNIP/NLRP3 pathway in rat[J]. Chinese Journal of Brain Diseases and Rehabilitation(Electronic Edition), 2023, 13(02): 65-74.

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References 25

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组别	只数	治疗前	治疗12 h后
组别	只数	治疗前	4.5 h溶栓	6 h溶栓	7 h溶栓	8 h溶栓
AIS组	3	3.33±0.58	3.33±0.58	3.67±0.58	3.67±0.58	3.67±0.58
rt-PA治疗组	3	2.67±0.58	1.67±0.58^ab	2.00±0.00^ab	2.33±0.58^b	3.00±0.00
rt-PA+依达拉奉治疗组	3	2.33±0.58	1.67±0.58^ab	1.67±0.58^abc	2.00±0.00^abc	2.33±0.58^bc
F值		2.333	8.333	11.231	10.501	3.500
P值		0.178	0.019	0.007	0.011	0.098

组别	只数	治疗前	治疗12 h后
组别	只数	治疗前	4.5 h溶栓	6 h溶栓	7 h溶栓	8 h溶栓
AIS组	3	3.33±0.58	3.33±0.58	3.67±0.58	3.67±0.58	3.67±0.58
rt-PA治疗组	3	2.67±0.58	1.67±0.58^ab	2.00±0.00^ab	2.33±0.58^b	3.00±0.00
rt-PA+依达拉奉治疗组	3	2.33±0.58	1.67±0.58^ab	1.67±0.58^abc	2.00±0.00^abc	2.33±0.58^bc
F值		2.333	8.333	11.231	10.501	3.500
P值		0.178	0.019	0.007	0.011	0.098

溶栓时机	只数	假手术组	AIS组	rt-PA治疗组	rt-PA+依达拉奉治疗组	F值	P值
4.5 h	3	71.82±0.23	75.76±0.60^a	73.21±0.56^ab	73.04±0.29^ab	27.150	0.002
6 h	3	69.57±0.60	75.89±0.54^a	74.36±0.65^a	72.32±0.37^abc	49.751	<0.001
7 h	3	72.32±0.54	77.68±0.69^a	76.52±0.68^a	73.87±0.73^abc	24.923	<0.001
8 h	3	70.91±0.64	77.19±0.24^a	76.99±0.50^a	75.44±0.69^a	57.670	<0.001

溶栓时机	只数	假手术组	AIS组	rt-PA治疗组	rt-PA+依达拉奉治疗组	F值	P值
4.5 h	3	71.82±0.23	75.76±0.60^a	73.21±0.56^ab	73.04±0.29^ab	27.150	0.002
6 h	3	69.57±0.60	75.89±0.54^a	74.36±0.65^a	72.32±0.37^abc	49.751	<0.001
7 h	3	72.32±0.54	77.68±0.69^a	76.52±0.68^a	73.87±0.73^abc	24.923	<0.001
8 h	3	70.91±0.64	77.19±0.24^a	76.99±0.50^a	75.44±0.69^a	57.670	<0.001

溶栓时机	只数	假手术组	AIS组	rt-PA治疗组	rt-PA+依达拉奉治疗组	F值	P值
4.5 h	3	2.55±0.27	27.38±2.91^a	20.11±2.13^ab	15.81±1.68^ab	54.781	<0.001
6 h	3	6.22±0.76	27.52±2.92^a	20.36±1.50^ab	15.43±0.88^abc	49.680	<0.001
7 h	3	3.74±0.43	27.13±2.88^a	25.23±2.68^a	15.55±1.65^abc	50.131	<0.001
8 h	3	4.56±0.48	36.42±1.49^a	32.12±0.79^ab	26.86±2.85^ab	140.901	<0.001

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